Pratylenchus thornei) causes extensive yield losses in wheat worldwide and thus pose serious threat to global food security. Reliance on fumigants (such as methyl bromide) and nematicides for crop protection has been discouraged due to environmental concerns. Hence, alternative environment friendly control measures like finding and deployment of resistance genes against Pratylenchus thornei are of significant importance. In the present study, genome-wide association study (GWAS) was performed using single-locus and multi-locus methods. In total, 143 wheat genotypes collected from pan-Indian wheat cultivation states were used for nematode screening. Genotypic data consisted of  > 7K SNPs with known genetic positions on the high-density consensus map was used for association analysis. Principal component analysis indicated the existence of sub-populations with no major structuring of populations due to the origin. Altogether, 25 significant marker trait associations were detected with − log10  > 4.0. Three large linkage disequilibrium blocks and the corresponding haplotypes were found to be associated with significant SNPs. In total, 37 candidate genes with nine genes having a putative role in disease resistance  were identified. Genomic selection was conducted to investigate how well one could predict the phenotype of the nematode count without performing the screening experiments. Prediction value of r = 0.40 to 0.44 was observed when 56 to 70% of the population was used as a training set. This is the first report where GWAS has been conducted to find resistance against root lesion nematode (P. thornei) in Indian wheat germplasm.Root lesion nematode (RLN;  Triticum aestivum L.) is one of the most important cereal crops cultivated globally and is a major source of calories for the growing world population1. Green revolutions of the 1960s and 1980s led to the significant improvement in wheat production particularly in South-East Asia2. Record global wheat production of 761.5 million tons was achieved in 2019 which is expected to further increase by 20203. However, the current trends in wheat production increase appear to be insufficient for feeding the population of nine billion people predicted for 20504. To meet the growing human needs without increasing the area of cultivated land, which is simply not available, wheat grain production must increase. Although production is increasing, yields are still vulnerable to a variety of biotic and abiotic factors. Among biotic factors, plant-parasitic nematodes (PPNs) are one such limiting factor that cause losses of ~ 12.6% among different crop plants, representing an annual monetary loss of 216 billion US$5. The most common damaging PPNs are the root-knot nematodes (RKNs), cereal cyst nematodes (CCNs) and root lesion nematodes (RLNs). RLNs of genus Pratylenchus are obligatory endoparasites. Pratylenchus spp. are polyphagous in nature and feed on a wide range of crops including cereals, vegetables, legumes, coffee, etc. These are migratory endoparasites which penetrate into the root cortex, using their stylet and release cell wall degrading enzymes and migrate inter-cellularly6. Due to damage in the root cells, plants are unable to uptake water and nutrients from soil properly, producing symptoms similar to nutrient and water deficiency8. Completion of their life cycle takes 3–8 weeks depending on the species and conditions7. After embryogenesis and development, the first stage juvenile (J1) molts to the second stage juvenile (J2) within the egg which further hatches from the egg9. All juveniles and adult stages are mobile and can enter and leave the roots. They are distributed worldwide and have been reported on wheat cultivation in Syria, Mexico, Canada, Israel, Yugoslavia, Morocco, Iran, India, Turkey, Pakistan, Algeria, Italy, Australia, and USA10. Several of these countries recorded significant wheat yield loss due to the P. thornei infection11. In terms of economic importance, yield loss up to 62% has been recorded, due to P. thornei, in countries like Australia12.Bread wheat  to identify another source of resistance against P. thornei35. Several quantitative trait loci (QTLs) linked to P. thornei resistance have also been identified in wheat, and these resistance sources could be incorporated into wheat breeding programs37. Three major QTLs for P. thornei resistance on chromosome 2BS, 6DS and 6DL were identified in the synthetic hexaploid wheat CPI13387237. Toktay et al.38 reported QTLs for P. thornei resistance on chromosome 1B, 2B, 3B, 4D and 6D. Eight QTLs for P. thornei resistance were mapped on wheat chromosome 2A, 2B, 2D, 5D and 6D36. Out of these, two major QTLs namely “QRlnt.sk-2B” on chromosome 2B and “QRlnt.sk-6D” on chromosome 6D were fine mapped39. Only the single gene Rlnn1 originating from Australian spring wheat cv. Excalibur for P. neglectus resistance was mapped on the chromosome 7AL in a doubled haploid population40.Application of nematicides is quite effective in controlling nematodes41. However, not many GWAS studies have been conducted in wheat to find resistance against P. thornei. Dababat et al.42 found nine significant MTAs using GWAS associated with P. thornei resistance on chromosomes 1D, 2A, 3B, 5B and 7A.Association mapping (AM) or linkage disequilibrium (LD) mapping is used to search genotypic-phenotypic correlations in unrelated individuals. It could be a promising strategy for the identification of significant marker-trait associations for nematodes resistance. GWAS uses all recombination events that have occurred historically within the crop, resulting in much higher mapping resolution by exploiting the historical linkage disequilibriumP. thornei resistance in wheat genotypes which have been predominantly collected from India. Mainly single-locus and multi-locus GWAS methods were compared in order to identify significant marker-trait associations (MTAs). We also explored the overlap of the identified MTAs and the known QTLs from previous studies to locate the novel genomic regions which could be targeted in plant breeding. Lastly, we have conducted the genomic-selection on the genotypes to investigate the efficiency of genomic-selection in predicting the complex trait such as RLN resistance. This is the first GWAS study from India to find resistance against P. thornei in wheat.The present study was conducted to find out the novel sources of resistance i.e., MTAs and candidate genes significantly associated with P. thornei, 143 wheat genotypes predominantly from India were screened for nematode resistance. Screening experiments were carried out over two years by growing wheat genotypes in five different batches with total 5–7 replications. A histogram depicting the distribution of nematodes among genotypes is shown in Supplementary Fig. 2 = 0.55)  lower than 5% and heterozygosity higher than 10%. Marker coverage of the SNPs over the chromosomes is shown in Fig. p value  < 0.05). Further, a significant correlation was obtained for phenotypes with PC4-axis  wherein genotypes clustered towards the lower axis were resistant compared to the upper axis (data not shown).To estimate population structure, a PCA was conducted with 7,491 polymorphic SNP markers and 143 bread wheat genotypes Fig. . PC1 expFaster LD-decay was observed for the A-genome (5–7 cM) compared to the B and D-genome (8–9 cM) of the panel. However, local spikes in the long-range LD regions were also observed that indicates the population structure of the panel as the LD-decay was computed without correcting for population structure. Despite a lower number of markers on the D-genome, LD patterns follow a similar trend of declining LD-values over the genetic distances. Genome-wide and chromosome-wide LD decay is given in Fig. p value) > 4.0 as shown in circular Manhattan plots  were close to the distribution of the expected associations . These four methods identified 25 significant MTAs with − log10  on chromosome 1D located at 221 cM was detected in all four methods. MTA (AX-94692978) located on chromosome 1B at 24 cM was detected through CMLM, BLINK and FarmCPU. Likewise, MTA (AX-94620141) on chromosome 2B at 27 cM was detected through BLINK, FarmCPU and MLMM, respectively. These three MTAs on chromosomes 1D, 1B and 2B identified consistently in either three or four methods were considered to be the most significant of the study and could be useful for further analysis. Allelic effects of highly significant SNPs obtained consistently in more than one method is shown in the Fig. 9 on chroWe also generated the haplotypic blocks across the genome by using the SNPs. Interestingly, three main LD blocks that spanned around the regions of significant markers were used to generate haplotypes. Three large LD blocks and the corresponding haplotypes  which were found to be associated with significant SNPs are given in Fig. LD block one was detected on the chromosome 1D at 28 cM (209–237 cM region). Three haplotypes were generated on the basis of genotyping results by using 132 genotypes  Fig. a. HaplotLD block two was also identified on chromosome 1D at 161 cM. Two haplotypes were generated using 140 genotypes . Two haplotypes were generated using 141 genotypes Fig. c. HaplotAs nematode counting, used for resistance screening, is not straight forward and time consuming, we explored whether the genomic selection would predict the nematode count using genome-wide markers. For this analysis the population was divided into smaller subgroups starting from 20 to 80% of the population used as a training set and then genomic prediction was performed on the remaining set. We observed a prediction value of r = 0.40 to 0.44 when 56 to 70% of the population was used as a training set which is moderate for a complex trait like nematode count of the present study  to find the candidate genes. We identified 37 putative candidate genes  were screened against P. thornei that showed a varied response when tested under controlled environmental conditions. No reference source of resistance in wheat against P. thornei is available in India. Kranti and Kanwar19 evaluated resistance and susceptibility in several wheat varieties (including Indian lines) for P. thornei resistance by the reproduction factor (Pf/Pi). The reproduction factor is commonly used in plant nematology as a quantitative value to assess resistance. In this study we found six resistant genotypes having reproduction factor (Pf/Pi) value equal or less than one, 24 moderately resistant genotypes (RF = 1–2), 25 moderately susceptible (RF = 2–3), 17 susceptible (RF = 3–4) and 71 highly susceptible (RF = more than 4) (Supplementary Table 2) was found to be 0.55, which showed moderate heritable value. This value indicates that despite the complexity of the trait which mainly owes to environmental conditions significant genetic effects were still observed and measured in the present study. Although in previous studies comparatively higher heritability for this nematode was also observed ranging from 0.83 to 0.9745. Dababat et al.42 detected very high heritability values, 0.95, 0.97 and 0.97 for Heterodera avenae, P. neglectus and P. thornei, respectively. Thus, in the future more experiments are needed to ascertain the phenotype and minimize the errors.The use of resistant cultivar is considered as the economically feasible and eco-friendly method to control P. thornei resistance by GWAS. With the rapid advancement in high throughput sequencing techniques, GWAS has been widely used to identify and dissect complex traits in wheat47. GWAS has been successfully used to study disease resistance in wheat51. GWAS was also performed in wheat for cereal cyst and root-lesion nematode resistance42. However, nothing from Indian wheat genotypes were screened and reported which links the phenotype to genotype. To avoid false association which is often present in the genotypes of local origin, effective corrections for population structure is needed56. Thus, we employed four methods of GWAS to locate the MTAs. In the present study, PCA detected two groups of the population indicating the existence of sub-populations. Interestingly we did not see major structuring of the populations due to the origin possibly suggesting despite a smaller group the genotypes were diverse.In the current study, our aim was to identify novel MTAs associated with 57. The magnitude of LD is influenced by many factors like recombination rate, allele frequency, mating system, genetic isolation, population structure, selection, marker type61, etc. Many studies suggest that LD is not constant across the whole genome or along a single chromosome. It can occur over large distances but also decreases very quickly for nearby loci62. In our study, LD decay so observed was similar to earlier reports64 where LD decay in common wheat ranged between 0.22 to 10 cM using SNP markers65, 2 cM (in A and B genome) to 5 cM (in D genome) using SNP markers61. LD ranged from 8 cM (D genome) to 10 cM (A genome) and average LD for whole genome 5 cM was also reported64. The D-genome has a higher LD decay, which is believed to be mainly due to limited populations of Aegilops tauschii which contributed to the present wheat genome in the evolutionary history of wheat67.Contrary to traditional linkage analysis, association mapping offers higher mapping resolution. In GWAS, diverse germplasm is used that contains a large number of recombination events as well as LD across a lineage that could help to detect mutations associated with phenotypic traits of interest72. However, mixed linear model-based methods are computationally challenging for large data sets. Hence, to overcome this problem, CMLM is implemented. CMLM method implemented in the GAPIT package decreases the effective sample size of such large and computationally challenging datasets by clustering individuals into groups74. CMLM improves statistical power by 5 to 15% and reduces computing time compared to regular MLM75. However, due to confounding between population structure, kinship and multiple testing corrections, CMLM based analysis can be insufficient, which could lead to a false association. Therefore, we also used FarmCPU analysis to mitigate this problem as the latter method has improved statistical power to detect causative genotype–phenotype associations and reduced false positives76. This method implements both the fixed-effect model (FEM) as covariates and a random effect model (REM) containing the kinship matrix. MLMM, which is a modified version of the mixed linear model, as it fits loci of large effect as covariates, detects more MTAs with smaller effects and has higher power and a lower FDR than single-locus approaches77. A recently developed BLINK method is also used in the present study to decrease the confounding problems that may lead to the spurious association78. BLINK implements a multiple loci test method by combining a fixed-effect model (FEM) with Bayesian information criteria (BIC). This method uses linkage disequilibrium (LD) information to replace the bin method.For a complex trait like nematode resistance, using more than one GWAS analysis methods make results statistically more stringent and acceptable with higher confidence which could be further used for pre-breeding purposes. CMLM, FarmCPU, MLMM and BLINK were used to identify significant associations between genotypic and phenotypic data. In the present study, several common significant MTAs were detected through four association analysis methods. The mixed linear model has been widely used in controlling population structure and relatedness within GWAS and reduces the spurious associationsP. thornei resistance. Among these, five MTAs on chromosome 1D (221 cM), 1B (24 cM), 3B (8 cM), 1B (119 cM) and 2B (27 cM) were obtained in more than one method. The SNP marker AX-94470359 on chromosome 1D detected in the present study was found common in all four methods. Among MTAs detected in the present study, seven were considered novel as detected for the first time whereas nine MTAs were found to be overlapped with or near to previously reported QTL/MTA regions  by MLMM, FarmCPU and BLINK methods. In a previous study by Zwart et al.80 also identified QTL for P. thornei resistance in this region on chromosome 3B. Dababat et al.42 identified markers associated with P. thornei and P. neglectus resistance on chromosome 3B at 14.2 cM and 64.7 cM, respectively. Although, in previous studies, QTL/ markers associated with H. avenae and P. neglectus resistance were mapped on chromosome 5A, 6B and 7A83. In this study, two markers (AX-94920631 on chromosome 5A and AX-94478352 on chromosome 6B) were identified; no QTL was reported on these chromosomes for P. thornei resistance in previous studies.We identified three markers  on chromosome 3B using CMLM, FarmCPU and BLINK methods. Schmidt et al.In order to find out the allelic effect, the SNP alleles that decreased the nematode count were defined as favourable allele and those that resulted in an increase in nematode count were defined as unfavourable alleles. The allelic contribution may be quite effective in improving the resistance in wheat by marker-assisted breeding. The average phenotypic value (nematode count) of each significant SNP allele for nematode resistance was selected to calculate the allelic effect. Highly favorable allele detected in the present study can be used for the development of nematode resistant wheat genotypes.Since the nematode count is a tedious trait that needs uprooting of the plants which is a cumbersome process specifically when dealing with large populations. That often is practically impossible when large populations are screened. Thus, we have conducted the genomic selection of the trait to investigate how well one could predict the phenotype of the nematode count without performing the phenotyping by using genomic-selection. We observed a prediction value of r = 0.40 to 0.44 when 56 to 70% of the population was used as a training set  Cytochrome P450 superfamily; Cytochrome P450s are heme-containing membrane-bound enzymes that involve in plant defense, and play an important role in disease response including fusarium head blight disease of wheat90, (iii) Vacuolar protein sorting-associated protein Vps 28 and Zinc finger protein; these are found to be drought-responsive genes and also play a role in other multiple stresses response in Arabidopsis thaliana91. (iv) Myb domain; Myb family is the largest family of a transcription factor in wheat. These are known to play a role in fungal infection caused by Rhizoctonia cerealis and also characterized as genes responsive to heat stress93.In the present study, 37 putative candidate genes were identified for significant MTAs  markers, which will be further useful in marker-assisted breeding for P. thornei resistance. Further, we showed the effectiveness of the genomic-selection in successfully predicting the phenotype of the genotypes. Identified candidate genes need further validation through wet lab experiments.The results obtained in the present study should be highly useful for tapping resistance against The germplasm collection consisted of 143 wheat genotypes, predominantly collected from the pan-Indian wheat cultivation states, and was used for nematode screening and association analysis (Supplementary Table P. thornei population obtained from Indian Institute of Pulses Research (IIPR), Kanpur, India. Nematode population was maintained on carrot callus using the carrot disc method in the incubator at 22 °C for about three months94. Nematodes extraction was done from chopped carrot callus following modified Baermann’s funnel technique95. Seeds of wheat genotypes were surface sterilized with 0.2% (w/v) mercuric chloride (HgCl2) and then soaked in water for germination overnight. Germinated seeds were transferred to polyvinyl chloride (PVC) tubes (16 cm in height and 3.5 cm in diameter) filled with steam-sterilized soil. This soil was collected from the field, sieved and then sterilized twice at 121 °C for 2 h. At the bottom of each tube, a 20 μm sieve was fixed to prevent both root outgrowth and the nematode’s escape out of the tube. The tubes were placed in special holders on an irrigation system as described previously99. All the experiments were performed in the growth room under controlled environmental conditions . Completely randomized design (CRD) with a minimum five replicates in five different batches (over two years) of each genotype was used. The plants were irrigated with Hoagland media at a fixed interval during the experiment.Screening experiments were carried out using P. thornei mixed stage juveniles and adult nematodes in solution by pipetting at a depth of 1.0 cm near to root. Plants were not watered for the next 3–4 days after inoculation of nematodes. Nematode extraction was done after 90 days of transplantation from soil and root by Cobb’s sieving and decanting method combined with modified Baermann’s funnel technique100. Nematode suspensions were collected and stored in bottles at 4 °C for counting. Three aliquots of 1.0 ml were taken from each bottle and nematodes were counted under a stereo microscope (Nikon SMZ645) at 40X-fold magnification. Reproduction factor (Pf/Pi) values  were calculated as the ratio between the number of nematodes counted at the end of the test and the number of nematodes used as inoculum on the basis of reproduction factor (Pf/Pi) values  were generated. To control the errors all data points were manually checked and visually inspected by conducting outlier z-tests. The analysis of unbalanced nematode counts data was carried out by the mixed model using REML (restricted maximum likelihood)104.From the seedling of the grown plants, leaf samples were collected for DNA extraction. All DNA samples were quantified on 0.8% agarose gel followed by quantification on a spectro-photometer-based Nanodropsystem. Affymetrix 35 K Axiom wheat Breeder chip was used for genotyping. We obtained 16,406 SNPs after quality control on this breeder chip. We further filtered the SNPs based on the minor allele frequency of 5%, heterozygosity (< 10%) and missing values that reduce the total SNP numbers to 7,491. Full set of 7,491 polymorphic SNPs that have genetic positions on the high-density consensus map was used for further analysis103. To compute pair-wise LD between the markers TASSEL version 5.2.54 was used. The pairwise LD was determined by using the full set of mapped 7,491 polymorphic and genetically anchored SNPs. To display whole genome and sub-genomic LD pairwise r2 versus distances in cM, sliding window of 50 SNPs was used and the graphics of the LD-decay were generated in R package GGPLOT2105.To understand the population structure, PCA analysis was performed from a set of polymorphic SNPs implemented in TASSEL version 5.0 and displayed using R106 was used. To compute the kinship matrix a set of thinned markers over the 21 wheat chromosomes were used to generate the IBS based kinship matrix. Four methods of GWAS analysis were used including compressed mixed linear model (CMLM)73, multi loci mixed linear model (MLMM)77, Fixed and random model Circulating Probability Unification (FarmCPU)76 and Bayesian-information and linkage-disequilibrium iteratively nested keyway (BLINK)78. In total, 7,491 SNPs obtained after filtering were used for genome-wide association analysis. To visualize the false positives of the implemented methods, QQ-plots were generated from each method in R (using the R-package R-CM plot). Manhattan plots were generated in R package qqman and CMplot. A robust threshold value of − log10  = 4.0 was set to reduce false positives and to ascertain the comparison across the implemented GWAS methods of this study. To compute the haplotype-based association analysis haplotypes were determined over the whole genome. PLINK version 1.9 was used to generate the LD-blocks which in turn is based on the default algorithms of finding haplotype blocks from Haploview software107.For the genome-wide association analysis of SNPs to nematode counts, GAPIT R package (3.0)108. The predicted regression coefficients obtained for these analyses were checked against the actual values and were reported in this study.A set of polymorphic, genetically mapped markers discussed above were used for the genomic selection. The analysis was conducted by a cross-validation approach wherein a set of training sets were drawn from the total population with a starting set of size 20 to 120 that corresponds to the 14 to 84% of the population. As there were only a few marker data sets missing which we imputed using mean values. A set of 500 cycles were run for this analysis as implemented in the software rrBLUP R-packageP. thornei resistance, were searched for IWGSC sequence information in Ensembl plants for T. aestivum (http://plants.ensembl.org/Triticum_aestivum) and the high confidence genes were selected to explore the function. The protein products of genes present in the flanking sequence available for the SNP markers with maximum bases  were taken as putative genes.To identify the potential candidate genes and their gene ontology, the sequences of SNPs defined significant association for Supplementary Information.Supplementary Tables.


Human personality is 30–60% heritable according to twin and adoption studies. Hundreds of genetic variants are expected to influence its complex development, but few have been identified. We used a machine learning method for genome-wide association studies (GWAS) to uncover complex genotypic–phenotypic networks and environmental interactions. The Temperament and Character Inventory (TCI) measured the self-regulatory components of personality critical for health . In a discovery sample of 2149 healthy Finns, we identified sets of single-nucleotide polymorphisms (SNPs) that cluster within particular individuals  regardless of phenotype. Second, we identified five clusters of people with distinct profiles of character traits regardless of genotype. Third, we found 42 SNP sets that identified 727 gene loci and were significantly associated with one or more of the character profiles. Each character profile was related to different SNP sets with distinct molecular processes and neuronal functions. Environmental influences measured in childhood and adulthood had small but significant effects. We confirmed the replicability of 95% of the 42 SNP sets in healthy Korean and German samples, as well as their associations with character. The identified SNPs explained nearly all the heritability expected for character in each sample (50 to 58%). We conclude that self-regulatory personality traits are strongly influenced by organized interactions among more than 700 genes despite variable cultures and environments. These gene sets modulate specific molecular processes in brain for intentional goal-setting, self-reflection, empathy, and episodic learning and memory. Strong evidence for substantial heritability of human personality comes from family, twin, and adoption studies . HoweverHuman personality is a striking example of the challenges involved in identifying the specific genes and molecular processes that influence complex traits. Twin studies indicate that between 30% and 60% of the phenotypic variance in personality, as assessed by a variety of instruments, is genetic in origin –14. HoweThe Temperament and Character Inventory (TCI) measures two domains of personality hypothesized to be related to different genetic and neuronal networks . ImagingWe have chosen to apply strictly data-driven machine learning methods in a person-centered approach to GWAS to uncover the complex genotypic and phenotypic architecture of personality , 30, 31 Our discovery sample was the Young Finns Study, an epidemiological study of 2149 healthy Finnish children followed regularly from 1980  to 2012 ages, 35–50 years) . Childho–50 yearsWe replicated the results in two independent samples of healthy adults from Germany , 44 and All subjects completed the TCI to assess seven heritable dimensions of personality , 47. ThePeople at risk of unhealthy personality were identified as the bottom decile of the sum of TCI self-directedness and cooperativeness . Prior wp < E-20, RMSE 0.03).We also identified an empirical index of character functioning by clustering the 13 character subscales of the TCI  arrays . The Korp-value <0.01 without Bonferroni correction) for possible association with character, taking gender and ethnicity into account as covariates of the individual SNPs. Preselecting SNPs identified SNPs that have weak associations with character that are not individually significant genome wide after Bonferroni correction, but provided presumptive candidates for epistatic interactions in a SNP set. The preselection also identified SNPs with a strong additive effect individually, thereby providing a manageably sized initial pool of SNPs as candidates for both the additive and non-additive components of the genetic architecture of character. We accounted for ethnicity in each sample by using the first three principal components for ancestral stratification of SNP genotypes  to uncover naturally occurring  associations between patterns across different types of data, including genetics –59 and nhttp://phop.ugr.es/fenogeno. The PGMRA method and algorithm are also summarized in Supplementary Sections Our web server application for phenotype–genotype many-to-many relations analysis (PGMRA) in GWAS is published  and onliReplicability of results was evaluated in the three independent samples for SNP sets, phenotypic sets, and genotypic–phenotypic relations using multi-objective optimization techniques , as detaWe exhaustively identified 902 non-identical but possibly overlapping SNP sets in the Finnish sample using PGMRA without knowledge of the phenotype. The SNP sets were comprised of different numbers of SNPs and/or subjects, regardless of their phenotypic status. The SNPs were mapped to diverse functional classes of genetic variants that may be located on different chromosomes, frequently even within a single SNP set Figs. , 2a–d. SWe identified 342 non-identical but possibly overlapping character sets using the 13 character subscales without knowledge of the genotype. Character sets were labeled by a phenotypic identification “C” to distinguish them from the SNP sets. These fine-grained character sets were nested within five character supersets that were identified by recurrently applying PGMRA to minimize the cophenetic correlation coefficient , conscientiously considerate of others (CO5), self-deprecating (SD4), and otherwise low in self-directedness and self-transcendence. The people with an "apathetic" character were low in all aspects of character development Table .p < 1E-05). For example, the SNP set G_11_4 has a p-value of 1.22 E-19, whereas the best and average SNPs within this set have 9.00 E-05 and 3.29 E-02 p-values, respectively , showing that SKAT did not inflate results.We tested the association of SNP sets with character. The empirical index of character, a single quantitative measure of character functioning, was more strongly associated with SNP sets than with the average effects of their constituent SNPs according to SKAT Table . Forty-tForty-two SNP sets significantly associated with character are described in Table p < 1E-03, Table p < 4.7 E-03).We found that 55 of the 342 character sets were significantly associated with particular SNP sets , lncRNAs (24%), other RNA genes (6%), and targets of microRNAs (27%), as identified in the TRANSFACp = 0.01), as expected because the Germans had been screened to exclude psychopathology, including personality disorders, in themselves or their first-degree relatives . In other words, the SNPs that comprise different SNP sets strongly distinguished the character values of the subjects in each set, indicating that each individual SNP set contributed significantly to explain the total distributed heritability .We also considered environmental influences in the Finnish sample. There were direct associations of sets of environmental influences in childhood and adulthood with character sets  in our initial pool of SNPs was more than six orders of magnitude below what is required for genome-wide significance. We sought to evaluate the cooperative effects of groups of SNPs with possible non-additive gene–gene interactions and those with strong additive effects individually . Therefore, we included SNPs that were either weakly or strongly associated with character singly, and then compared their significance as a group vs. that of the best SNP within the group. Consequently, these candidate SNPs may have no main (additive) effect on the phenotype at all, but when organized as SNP sets, they presented consistent evidence of epistasis . In addition, the SNPs we identified were sufficient to account for nearly all the heritability expected from twin studies (about 50%), which includes both additive and non-additive effects.Our initial pool of SNPs was preselected to be the best candidates to have additive and/or non-additive effects on character. The threshold for possible association . We conclude that self-regulatory personality traits are strongly influenced by organized interactions among more than 700 genes, despite variable cultures and environments. We recommend studies that dissect detailed phenomic and genomic data, including brain images and physiological measurements, and integrate these in a multi-faceted view of each person. We also recommend an extended replicability analysis, in which a marker can be replicated at different multi-omic levels, such as genes, family of proteins, or pathways. The precision of our person-centered approach now allows such in-depth analysis and replication, even for complex traits in moderate-sized samples.Supplementary InformationSupplementary Information (Marked)Figure S1AFigure S1B-CFigure S1D-FFigure S2Figure S3Figure S4Figure S5Figure S6Figure S7Table S1Table S2Table S3Table S4Table S5Table S6Table S7Table S8Table S9Table S11Table S10Table S12Table S13Table S14Table S15Table S16Table S17Table S18Table S19Table S20Table S21Table S22Table S23


Leishmania (Viannia) braziliensis is an important Leishmania species circulating in several Central and South American countries. Among Leishmania species circulating in Brazil, Argentina and Colombia, L. braziliensis has the highest genomic variability. However, genomic variability at the whole genome level has been only studied in Brazilian and Peruvian isolates; to date, no Colombian isolates have been studied. Considering that in Colombia, L. braziliensis is a species with great clinical and therapeutic relevance, as well as the role of genetic variability in the epidemiology of leishmaniasis, we analyzed and evaluated intraspecific genomic variability of L. braziliensis from Colombian and Bolivian isolates and compared them with Brazilian isolates. Twenty-one genomes were analyzed, six from Colombian patients, one from a Bolivian patient, and 14 Brazilian isolates downloaded from public databases. The results obtained of Phylogenomic analysis showed the existence of four well-supported clades, which evidenced intraspecific variability. The whole-genome analysis revealed structural variations in the somy, mainly in the Brazilian genomes (clade 1 and clade 3), low copy number variations, and a moderate number of single-nucleotide polymorphisms (SNPs) in all genomes analyzed. Interestingly, the genomes belonging to clades 2 and 3 from Colombia and Brazil, respectively, were characterized by low heterozygosity (~90% of SNP loci were homozygous) and regions suggestive of loss of heterozygosity (LOH). Additionally, we observed the drastic whole genome loss of heterozygosity and possible hybridization events in one genome belonging to clade 4. Unique/shared SNPs between and within the four clades were identified, revealing the importance of some of them in biological processes of L. braziliensis. Our analyses demonstrate high genomic variability of L. braziliensis in different regions of South America, mainly in Colombia and suggest that this species exhibits striking genomic diversity and a capacity of genomic hybridization; additionally, this is the first study to report whole-genome sequences of Colombian L. braziliensis isolates. Leishmania (Viannia) braziliensis is the most important Leishmania species associated with cases of cutaneous leishmaniasis and muco-cutaneous leishmaniasis in several Central and South American countries  and describes a possible meiotic recombination event between them, with uniparental inheritance of maxicircles but biparental inheritance of minicircles, which may be crucial for survival of the parasite in the wild. Considering the close relatedness between L. braziliensis and L. peruviana, some authors have conducted genomic comparative analysis, which have identified a great number of interspecific SNP/indel differences between them as well as the presence of different gene and chromosome copy number variations supporting the classification of both organisms as closely related but distinct species  to evaluate phylogenomic relationships among six Colombian clinical isolates and one Bolivian clinical isolate. We also determined phylogenomic relationships and genomic plasticity among these isolates and 14 publicly available L. braziliensis genomes from Brazil in terms of genome plasticity (chromosomes/genes) and single point mutations.Despite these findings, knowledge about the genomic variability of in rural  comparedin rural . In addiin rural . ConsideAll procedures were approved by the Ethics committee of the Universidad de Antioquia (number VRI3445/2010) in accordance with resolution number 36836. The patients included into the study read and signed the informed consent.L. braziliensis genomic sequences reads, from Brazilian isolates publicly available from the DDBJ/ENA/GenBank database (http://www.ebi.ac.uk/ena) under accession codes PRJNA292004 , Medellín-Colombia’. Also, 14 NA292004  and PRJNNA292004  were dow2 as describe previously  fetal bovine serum; the cultures were maintained at 26°C with 5% COeviously . Once thCytb) and heat shock protein (Hsp70) were the genes selected to species identification through Sanger sequencing, as has been described previously (Leishmania sequences deposited in GenBank (L. braziliensis were selected for whole-genome sequencing.Cytochrome b proteins (eviously . The amp GenBank . Those iHiSeq X-Ten system (Illumina) was used to sequence the whole-genome DNA, later mate-paired libraries were built and finally subjected to paired-end sequencing (2 × 150-bp read length producing a median coverage at least of 40x per sample). Reads with adapter contamination, >10% uncertain nucleotides, or >50% low-quality nucleotides  were discarded .L. braziliensis genome sequence assembly (http://tritrypdb.org) using the SMALT program (version 0.7.4) . Exhaustive search option (−x and −y of 0.8), a sliding step of 3 and a reference hash index of 13 bases were the parameters using during the mapping. To prevent the mapping of non-Leishmania reads to the reference sequences, we used an identity threshold of y = 0.8. SAMtools (version 0.1.18) and Picard (version 1.85) were used to no-mapping read exclusion, sorting, file merging and elimination of PCR duplicates (L. braziliensis M2904 (available in tritrypDB).The paired-end Illumina reads of six Colombian clinical isolates, one Bolivian clinical isolate and of the fourteen Brazilian genomes (data downloaded from European Nucleotide Archive) were mapped to the reference MHOMBR75/M2904_2019 plicates . The reaL. braziliensis isolates and between each sequence generated by each isolate during the phasing. FastTree double precision version 2.1.10. (http://itol.embl.de) (L. braziliensis as genome sequence (REF) and used as outgroup the L. guyanensis genome assembly from European Nucleotide Archive accession number SRR8179913 (Lguy_SRR8179913) (http://www.ebi.ac.uk/ena) and the L. panamensis genome assembly from TriTrypDB http://tritrypdb.org: LPPSC1 (LpW).SNPs alignments from whole nuclear and mitochondrial (maxicircle) genomes unphased and phased (see “Phasing of heterozygous SNP sites” section), were used to evaluate the phylogenomic relationship among  2.1.10.  was usedembl.de) . To deteembl.de)  using thThe chromosomal somy was estimated calculating initially the median read depth of each chromosome (di). Positions with read depth of >1 standard deviation were removed and the di recalculated. Later, we calculated the median depth (dm) of whole genome (35 chromosomes) and the somy  of each chromosome, for that we used the formula previously described S = 3 × di/dm . The ranHG) and the full cell depth with somy effect (dFG) using the formula: (FG = S × dHGd). The statistical significance used in this study was set at a z-score cutoff of >2 and an adjusted p-value (Student’s t-test) of <0.05. Heatmaps were created using the Heatmap3 package in R (http://tritrypdb.org) to evaluate the genes with CNVs. P-values were adjusted for multiple testing with Benjamini-Hochberg method with a false-discovery rate (FDR) of <0.05. The GO terms were submitted to REVIGO , we calculated and related the average haploid depth per gene without somy effect , different options of this program were used to search random mapping of multiple hit reads and obtained optimal alignments. The merging and sorting of bam files and marking duplicated reads were implemented with the Picard program (version 1.85) (http://broadinstitute.github.io/picard/) as described previously (https://software.broadinstitute.org/gatk/), where SNPs were called among all the samples simultaneously. Later, we realigned around indels to remove these and retrieved only the SNPs. GATK Variant Filtration was used to filter Low-quality SNPs, according to the following criteria: QD < 2.0 || MQ < 40 || FS > 60.0 || ReadPosRankSum < −8.0. The SNP quality cutoff was set at 3000. Later, the Integrative Genomic Viewer program (IGV_2_3_47) was used to visualize all SNPs identified and the SnpEff program (version v4.1) to classify the SNPs based on their functional impact (L. braziliensis genomes using snp-sites program (https://github.com/tseemann/snp-dists), we generated a pairwise distance matrix; the results obtained were graphically represented. Lastly, we selected the SNPs that have a potential effect on gene function (high and moderate impact); from this selection, we identified the unique/shared SNPs between and within the clades and between the Colombian genomes, the data were included in an Excel matrix, which was used to perform the comparative analysis. Finally, to measure nucleotide diversity (π), we used the DnaSP software v.5.0.Initially, the reads were mapped to the reference MHOMBR75/M2904_2019 eviously . The SNPl impact . Once th program . Based oFrom allele frequency estimation data, we determined the Homozygous and heterozygous variants. Allele shifts of < 0.33 or > 0.66 were considered as homozygous variants while allele shifts between 0.33 and 0.66 as heterozygous variants . Once hohttps://faculty.washington.edu/browning/beagle/beagle.html) over 30 iterations. The algorithm also imputes missing genotypes from identity-by-state segments found in the data (Heterozygous SNPs were phased using BEAGLE v5.1 (the data . A custoCytb and Hsp70 genes identified that the isolates herein included corresponded to L. braziliensis (The L. braziliensis genome as the reference and L. guyanensis (Lguy_SRR8179913) and L. panamensis LP-PSC1 (LpW) genomes as outgroups. The approximately-maximum-likelihood phylogenetic trees built in FastTree double precision version 2.1.10  from nuclear genome . The second corresponded to SNPs from mitochondrial genomes (167 nucleotide positions). For both cases MHOM/BR75/M2904_2019 n 2.1.10  allowed iliensis   with extra copies, while the Lb7293733 genome presented three chromosomes  with extra copies. Additionally, we observed that in one clade 4 genome (Lb8102), there was an extra copy of chromosome 2 compared with the other genomes of the same clade , and the genomes with the lowest numbers were Lb7293737 and Lb7293742   Table 2.The next step was to evaluate the shared SNPs  within and between each clade. The comparison of genomes within each clade revealed that the genomes of clade 2 had the highest number of shared SNPs  compared with clades 1, 3, and 4, where the number of SNPs shared was 1,173 and 9,703, and 5,520, respectively  between the Colombian genomes included in the study. To analyze the unique SNPs in each genome, we observed that Lb7864 and Lb7616 were the genomes with the lowest number of SNPs , which represent less than 1% of the total, compared with the other genomes analyzed where the number of unique SNPs ranged between 9 and 13% (L. braziliensis genome (Chromosome 35) and compared the heterozygosity/homozygosity  along this chromosome among seven genomes  observed in some genomes Figure 5In contrast, we identified one genome of clade 4 (Lb8025) to be highly heterozygous throughout the genome (99% heterozygous SNPs) Figure 1L. braziliensis, we used DNA-seq technology and bioinformatic analyses to investigate the genetic structure and the phylogenomic relationships among L. braziliensis genomes from Colombian/Bolivian clinical isolates and among genomes of Brazilian L. braziliensis isolates archived in public databases. The phylogenetic analysis of nuclear and mitochondrial genomes revealed the presence of two different L. braziliensis populations in Colombia (clade 2 and clade 4)  , butterfclade 4) , arachniclade 4) , Triatomclade 4) , Lutzomy species  and planmariurus . (ii): Cmariurus . This cohabitats . Finally vectors .Leishmania and other trypanosomatids, such as Trypanosoma cruzi (Another interesting finding was associated with the clear swapping event (incongruence in the tree topologies) identified in clade 4, when tree topology of nuclear genome was compared with the mitochondrial genome . This wama cruzi . We wereLeishmania adaptation; therefore, these results indicate that Colombian strains could be changing to adapt to the eco-epidemiological conditions of the country as the Brazilian strains and that the amplification of some chromosomes may provide fitness advantages during host adaptation, which is consistent with the observed in other Leishmania species   Figure 2onensis) . Additio species . Despite species .L. braziliensis, results previously described in other Leishmania species . The results did not reveal important intraspecies variations in the numbers of genes with CNV . These r species  and suggL. donovani, L. tropica, L. infantum, and L. panamensis)  .L. braziliensis genomes involved analysis of nucleotide-level variations (SNPs). Comparing the number of SNPs found in this study         species with the highest genetic variability circulating in some regions of South America . Additionally, to analyze the genetic variability of L. braziliensis between the Colombian and Brazilian isolates, we observed that Colombian genomes were much more diverse , than the Brazilian genomes   ; L. infa n = 12)   , L. amaz; n = 2) , L. pana n = 22) , L. amaz00 SNPs)  and L. p; n = 2) , we can To evaluate unique/shared SNPs among the four identified clades, we identified a high number of unique SNPs  as a mechanism for introducing diversity into a population and as a strategy to survive in stressful conditions   . Therefore, this is the first study using Colombian clinical isolates and next generation sequencing to identify patterns of homozygosity in the whole genome of one of the most important New World Leishmania species, L. braziliensis. This pattern of diversity could be explained by a substantial bottleneck of adaptation that occurred in an environment/niche-dependent manner, which favored and fixed certain genotypes in the population and promoted the stochastic loss of others, as has been described for T. brucei (T. terrestris (Leishmania species (L. donovani complex) (Theileria parva and Saccharomyces cerevisiae)  observed in some Brazilian/Colombian genomes  (passage) . In Leisa block)  or low-la block)  mainly i. brucei  and recerrestris . Anothercomplex) , as wellevisiae) . The strL. braziliensis in Colombia and the occurrence of distinct phylogenetic groups in the sampled regions. The genomic changes, until date unknown in L. braziliensis, such as moderate changes in somy, CNVs, the introgression event, the increase of the heterozygosity and the duplication of genetic content in various chromosomes, identified in the Lb8025 genome , as well as the high number of homozygous SNPs (most of them unique for each clade) and the whole genome LOH, identified in Colombian/Brazilian genomes, suggest a striking genomic plasticity of this species, which could be genomic strategies used by L. braziliensis to favor its survival and adaptation to different ecological niches. These genomic findings can influence L. braziliensis epidemiology and future clinical and therapeutic outcomes.In conclusion, the findings in this study demonstrate the high intra-species genetic diversity of L. braziliensis is endemic. This at the end will permit not only to make a broader comparative genomic analysis of our Colombian isolates with genomes from other regions but also to expand the knowledge about of genetic variability of this species in South America.Although our study was focused to analyze and compare the genomic structure of Colombian/Bolivian isolates with Brazilian isolates, we consider that there are necessary additional studies that include genomes from other regions of South America where https://www.ebi.ac.uk/ena, ERS4385933, ERS4385934, ERS4385935, ERS4385936, ERS4385937, ERS4385938 and ERS4385939.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: The studies involving human participants were reviewed and approved by: This study was approved by the Ethics Committee of the Universidad de Antioquia (number VRI3445/2010) in accordance with resolution number 36836. Written informed consent was obtained from the patients from which the strains were isolated. The patients/participants provided their written informed consent to participate in this study.LP conceived and designed the study, analyzed and interpreted the data, and prepared the manuscript. MM interpreted the data and critically revised the manuscript and made important suggestions. LC-S critically revised the manuscript and made important suggestions. CM critically revised the manuscript and made important suggestions. JR conceived and designed the study and revised the manuscript. All authors contributed to the article and approved the submitted version.This work was funded by DIRECCIÓN DE INVESTIGACIÓN E INNOVACIÓN from Universidad del Rosario. JR is a Latin American fellow in Biomedical Sciences, supported by The Pew Charitable Trusts.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The relevant nickel complexes [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] and the corresponding hydrolysed derivatives (BzNH3)2[Ni(dtp)2] and (PhBuNH3)2[Ni(dtp)2] were prepared and fully characterized. The antimicrobial activity of the aforementioned amidodithiophosphonates against a set of Gram-positive and Gram-negative pathogen bacteria was evaluated, and [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] showed antiproliferative activity towards Staphylococcus aureus and Staphylococcus haemolyticus strains. density functional theory (DFT) calculations were performed to shed some light on the activity of reported compounds related to their tendency towards P–N bond cleavage.The reactions of 2,4-bis(4-methoxyphenyl)-1,3-dithio-2,4-diphosphetane-2,4-disulfide  with benzylamine (BzNH I, II, III and IV, respectively, see 2− moiety can coordinate in monodentate, bidentate (with either symmetric or asymmetric bonding), and polydentate modes. A huge variety of both discrete and polymeric structures are prevalent in the literature  and [Ni(PhBuNH-adtp)2] and the corresponding hydrolytic products (BzNH3)2[Ni(dtp)2] and (PhBuNH3)2[Ni(dtp)2] 2] .iPrNH2, BzNH2, and PhBuNH2  in toluene gave rise to the corresponding amidodithiophosphonate ammonium salts (iPrNH3)(iPrNH-adtp), (BzNH3)(BzNH-adtp), and (PhBuNH3)(PhBuNH-adtp) . The FT-IR spectra of (BzNH3)(BzNH-adtp) and (PhBuNH3)(PhBuNH-adtp) (BzNH-adtp) and (PhBuNH3)(PhBuNH-adtp) in DMSO-d6 solutions are shown in 31P{1H} NMR spectra of (BzNH3)(BzNH-adtp) and (PhBuNH3)(PhBuNH-adtp) show a singlet at 79.7 and 93.1 ppm, respectively, in agreement with what it was previously reported for (iPrNH3)(iPrNH-adtp) and similar known compounds , [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] 2] and [Ni(PhBuNH-adtp)2] were recorded in CDCl3 solution. 1H-NMR data are consistent with the proposed structure (1H-NMR spectra of [Ni(BzNH-adtp)2] 2] (BzNH-adtp) and [Ni(BzNH-adtp)2]; the singlet at 3.86 ppm is attributed to the protons of the OCH3 group. The protons of the aromatic portion display signals at 7.90 and 6.97 ppm, assigned to the methoxyphenyl P-substituent, and signals between 7.3–7.1 ppm can be assigned to the aromatic protons of the amine. The 31P{1H} NMR spectra of [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] recorded in DMSO-d6 show only a singlet at 75.8 and 76.2 ppm, respectively, and are comparable with the CP-MAS 31P value of 71.7 ppm measured for of [Ni(iPrNH-adtp)2] and consistent with the presence of the P–N bond.The reaction of the amidodithiophosphonate salts with NiCl-adtp)2] , as purpompounds  show theectively . Due to tructure . In part-adtp)2]  show thr-adtp)2]  shows th2] and [Ni(PhBuNH-adtp)2] from either acetonitrile or chloroform solutions, revealed that hydrolysis occurred, resulting in the formation of compounds (BzNH3)2[Ni(dtp)2]·2H2O and (PhBuNH3)2[Ni(dtp)2], featuring the anionic dithiophosphonato Ni complex [Ni(dtp)2]2– counterbalanced by two BzNH3+ and PhBuNH3+ cations, respectively (2] and [Ni(PhBuNH-adtp)2] and the corresponding Ni-dithiophosphonate ammonium salts (BzNH3)2[Ni(dtp)2] and (PhBuNH3)2[Ni(dtp)2], mainly regarding the melting points and the IR and NMR signals relative to the ammonium -NH3+ groups ectively . The obt+ groups . 3)2[Ni(dtp)2]·2H2O comprises half a molecule, with a NiII ion lying about a crystallographic inversion center, one dithiophosphonato ligand (ArPOS2)−, one benzylammonium cation, and one water molecule. The metal center is tetracoordinated in a square-planar geometry by four sulphur atoms belonging to two isobidentate ligands with Ni–S bond lengths of 2.2255(4) and 2.2232(4) Å, respectively, and an S–Ni–S angle of 87.92(2)°. The P–S1 and P–S2 bond lengths show very similar values [2.0409(5) and 2.0387(5) Å respectively], indicating an electron delocalization over the whole PS2− fragment and an S–P–S angle of 98.39(2)°; the P–O bond exhibits a length of 1.5094(10) Å (The asymmetric unit of the (BzNH94(10) Å . 3)2[Ni(dtp)2] (3)2[Ni(dtp)2]·2H2O (see above) with similar bond lengths and angles (The structure of (PhBuNHi(dtp)2]  displaysi(dtp)2] . The nicd angles , and an 2 moiety are comparable with those found in similar neutral and anionic dithiophosphonato nickel complexes described as bearing either a P=O or P–OH bond [2]− fragments are often engaged in strong H-bonds with counterions, which affects the bond lengths between the atoms involved. A better understanding of the nature of the P–O bond (and its charge distribution) may be gleaned if the bond lengths involved in the –POS2 fragment are considered together. The correlation reported in 2 fragments not directly bearing a negative charge  [III in 3)2[Ni(dtp)2]·2H2O (black triangle in 3)2[Ni(dtp)2]  ,38,39,403)2[Ni(dtp)2]·2H2O and (PhBuNH3)2[Ni(dtp)2] are mainly governed by the strong H-bonds involving the anionic complexes, the ammonium BzNH3+ and PhBuNH3+ cations and the crystallized water molecules in (BzNH3)2[Ni(dtp)2]·2H2O 2]·2H2O . 3)(PhBuNH-adtp) in toluene yielded a few crystals of the salt (PhBuNH3)2[(ArPS2)2O] (2)2O]2− counterbalanced by two 4-phenylbutylammonium cations. The structure was solved in the space group P-1, and the main structural and refinement parameters are reported as An attempt at crystallizing (PhBuNHrPS2)2O]  containi2)2O]2− anion and one PhBuNH3+ cation; the complete content of the asymmetric unit with disorder shown is reported in 2P–O–PS2)2− fragment are similar to those previously reported for the thionatedbis(phosphonic) acid monoanhydride [nhydride . The ani3)2[(ArPS2)2O] confirms that the hydrolytic process involves the cleavage of P–N bond in amidodithiophosphonates, while retaining the P–S, as already observed in (BzNH3)2[Ni(dtp)2]·2H2O and (PhBuNH3)2[Ni(dtp)2]. The isolation of compound (PhBuNHiPrNH3)(iPrNH-adtp), (BzNH3)(BzNH-adtp), and (PhBuNH3)(PhBuNH-adtp) and the relevant nickel(II) complexes [Ni(iPrNH-adtp)2], [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] against different Gram-positive and Gram-negative bacterial species, both in planktonic and in sessile life. In this context, microbial species described as being commensals or pathogens in humans were assayed, namely Staphylococcus aureus, Staphylococcus haemolyticus, Escherichia coli, and two strains of Pseudomonas aeruginosa, PA-01 and PA-02, that showed a different susceptibility pattern to disinfectants  and [Ni(PhBuNH-adtp)2] are active against both Staphylococcus spp. (S. aureus and of 17 and 15 mm against S. haemolyticus by [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2], respectively (S. aureus and S. haemolyticus was observed for the complexes’ ligand precursors (BzNH3)(BzNH-adtp), (PhBuNH3)(PhBuNH-adtp), and NiCl2·6H2O, showing that the coordination compounds are responsible for the antimicrobial activity. In contrast, the inability of [Ni(iPrNH-adtp)2] to inhibit bacterial growth suggests that the tendency towards hydrolysis of the complexes [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] could play an important role in their antimicrobial activity. The tendency to hydrolysis could be tentatively ascribed to the different nature of the alkyl/aryl amine substituents, as evidenced by the slight elongation of the P–N bond on passing from [Ni(iPrNH-adtp)2] to [Ni(PhEtNH-adtp)2] (1.619(5) and 1.641(4) Å, respectively)  and [Ni(PhBuNH-adtp)2] were observed to inhibit the growth of S. aureus up to just a 2-fold dilution of the stock solution . S. haemolyticus showed a MIC up to a 32-fold dilution in the case of [Ni(BzNH-adtp)2], corresponding to 6.25 μg/mL (1.56·10−5 M). In contrast, [Ni(PhBuNH-adtp)2] lost the ability to inhibit the bacterial growth after dilution (MIC > 100 μg/mL). Moreover, the bactericidal activity was assessed by evaluating the minimum bactericidal concentration (MBC), i.e., the lowest concentration of an antimicrobial required to kill a particular bacterium life in suspension (planktonic status). This approach is established when the substance under investigation can inactivate bacterial contamination in a fluid, such as water, saliva and urine. Neither [Ni(BzNH-adtp)2] nor [Ni(PhBuNH-adtp)2] showed any bactericidal activity 2] on biofilm formation by evaluating the minimum concentration required to inhibit the formation of the biofilm in vitro, i.e., the minimum biofilm inhibitory concentration (MBIC). Both complexes showed the ability to inhibit the biofilm growth, however they required relatively high concentrations in the case of S. aureus (MBIC = 100 μg/mL). Notably, for both complexes, lower MBICs were observed for the biofilms of S. haemolyticus (MBIC = 50.0 and 12.5 μg/mL for [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2], respectively, corresponding to 1.49·10−4 and 3.12·10−5 M). For the sake of comparison, MIC, MBC, and MBIC measurements were carried out on NiCl2·6H2O under the same experimental conditions, on the same strains of microrganisms. As expected, nickel chloride did not show any antibacterial activity towards S. aureus and S. haemolyticus (Both  and [Ni(PhBuNH-adtp)2] undergo hydrolysis and try to explain the different antimicrobial activity determined between these complexes and the analogous [Ni(iPrNH-adtp)2] and the free amidodithiophosphonate salts. DFT calculations were carried out on the starting amines iPrNH2, BzNH2, PhBuNH2, the corresponding ammonium cations, the relevant amidodithiophosphonate anions (iPrNH-adtp)−, (BzNH-adtp)−, and (PhBuNH-adtp)−, and the complexes [Ni(iPrNH-adtp)2], [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2]. As expected, all the starting amines feature a Kohn–Sham (KS) HOMO corresponding to the lone pair (LP) of electrons localized on the nitrogen atoms . This LP is therefore available to react with Lawesson’s Reagent, which features a positive charge on the P atom (Q = 0.878 |e|) accompanied by a low energy (−2.075 eV) KS-LUMO, antibonding in nature with respect to the P–S bonds of the P2S2 1,3-dithia-2,4-diphosphetane ring. The (iPrNH-adtp)−, (BzNH-adtp)−, and (PhBuNH-adtp)− anions feature strongly polarized P–N bonds . This results in Wiberg bond indices (WBI) sensibly lower than unity , reflected in optimized P–N bond lengths (dPN) in the range between 1.741 and 1.749 Å. All the anions feature virtual MOs antibonding with respect to the P–N bonds. Moving from the anions to the corresponding NiII complexes, the P–N bonds are slightly strengthened . In analogy to the corresponding ligands, these complexes feature low-lying virtual MOs, antibonding with respect to the P–N bonds. In addition, the charge on the P atoms, and therefore the polarization of the P–N bonds, increases, thus indicating a large electrophilic character of the P atoms 2], [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2], respectively). As a result, the eigenvalues of the virtual antibonding MOs (with respect to the P–N bonds) are more stable than those of the corresponding free anionic ligands  and result in an increased electrophilic character of the P atom. Therefore, the susceptibility to the hydrolysis of related amidodithiophosphonato NiII complexes is expected to be larger than that of the starting ligands. Following recent studies on different complexes featuring chalcogen donors ,47,48,49ry (DFT)  in order2] complexes in turn leads to the formation of the complexes (BzNH3)2[Ni(dtp)2] and (PhBuNH3)2[Ni(dtp)2] that have been isolated and structurally characterized (iPrNH-adtp)2], which has proved experimentally to be less prone to hydrolysis, as confirmed by the stability in solution and isolation in the solid state [− anions or in the corresponding [Ni(R-adtp)2] complexes. This indicates that hydrolysis of the compounds should be expected to occur through P–N bond breaking and dithiophosphonate anion formation, causing some doubt regarding the previously hypothesized emission of dihydrogen sulfide as the first step of the hydrolysis [3)2[(ArPS2)2O] (3)(PhBuNH-adtp). A similar salt was hypothesized as the intermediate in the in situ formation of a mixed cymene-ferrocenylphosphonodithiolate ruthenium complex, obtained by the hydrolysis of 2,4-diferrocenyl-1,3-dithiadiphosphetane 2,4-disulfide in the presence of ammonium hydroxide [These results are in line with the hypothesis that the antibacterial activity could be related to the hydrolytic process. The subsequent cleavage of the P–N bond in the neutral [Ni(adtp)cterized . This hyexes BzNH2[Ni(dtp)drolysis . The hydrPS2)2O]  during aydroxide .1H and 31P NMR measurements were carried out at 25 °C using a Bruker Avance 300 MHz  spectrometer at operating frequencies of 300.13 and 121.49 MHz, respectively. Chemical shifts for 1H-NMR are reported in parts per million (ppm), calibrated to the residual solvent peak set, with coupling constants J reported in Hertz (Hz). Chemical shifts for 31P NMR are reported in parts per million (ppm), calibrated to the external reference TPP 48.5 mM in acetone-d6. Infrared (IR) spectra were recorded on a Thermo Nicolet 5700 FT-IR spectrophotometer  using KBr pellets and reported in wavenumbers (cm−1).Starting materials and solvents were purchased from commercial sources TCI  and Aldrich  and, when necessary, the solvents have been distilled and dried according to the standard literature techniques. Melting point measurements were determined in capillaries, using melting point apparatus BUCHI M-560 . Elemental analyses were performed with an EA1108 CHNS-O Fisons instrument . Single-crystal X-ray diffraction data were collected at 100 K on a Rigaku FRE+ equipped with VHF Varimax confocal mirrors and an AFC12 goniometer  and HyPix 6000 detector diffractometer  . The striPrNH2, BzNH2, PhBuNH2, the corresponding ammonium cations, the relevant amidodithiophosphonate anions (iPrNH-adtp)−, (BzNH-adtp)−, and (PhBuNH-adtp)−, and nickel complexes [Ni(iPrNH-adtp)2], [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2]. For all the investigated compounds the geometries were optimized, starting from structural data when available, and symmetrized to achieve the highest possible point group. For all the neutral, anionic, and cationic species tight convergence criteria were adopted . The nature of the energy minima at the optimized geometries were verified by a vibrational analysis, computed by determining the second derivatives of the energy with respect to the Cartesian atomic coordinates and subsequently transforming to mass-weighted coordinates. Natural bonding orbitals , [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] using: (a) the agar diffusion method, (b) minimum inhibitory concentration (MIC) and (c) minimum bactericidal concentration (MBC), determined in accordance with the Clinical Laboratory Standard Institute CLSI  = 8.33·10−4, [Ni(BzNH-adtp)2] = 5.95·10−4 and [Ni(PhBuNH-adtp)2] = 5.00·10−4 M in 1/10 v/v DMSO/water). The antimicrobial activity was expressed as mm of inhibition diameter around the cavity, after the microbial growth at 37 °C. All experiments were performed in triplicate and the values shown are reported as the average ± standard deviation of the inhibition diameter. MIC and MBC were determined only against the microbial strains susceptible to Kirby–Bauer assay, according to the micro-broth dilution method  were obtained during an attempt at crystallizing (PhBuNH3)(PhBuNH-adtp) by slow diffusion of ethyl ether in a toluene solution of the salt. The very exiguous amount of compound prevented us from further characterizations.Few crystals of bis(4-methoxyphenyl)-tetrathiodiphosphonate (PhBuNHiPrNH2), benzylamine (BzNH2), and 4-phenylbutylamine (PhBuNH2) in toluene gave rise to the corresponding amidodithiophosphonate ammonium salts (iPrNH3)(iPrNH-adtp), (BzNH3)(BzNH-adtp), and (PhBuNH3)(PhBuNH-adtp) that were reacted with nickel chloride hexahydrate, yielding the corresponding amidodithiophosphonato complexes [Ni(iPrNH-adtp)2], [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2]. All the compounds were tested against a library of bacteria and fungi of clinical importance belonging to the genera Staphylococcus, Escherichia, and Pseudomonas, and Candida, but only the complexes [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] demonstrated some antimicrobial activity that was tentatively ascribed to their tendency towards hydrolysis. Theoretical and experimental results evidenced that [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] undergo hydrolysis and that during the hydrolytic process a cleavage of the polarized P–N bond occurs with consequent formation of a P–O bond and retaining of the two P–S bonds in the amidodithiophosphonate moiety. Even if hydrolysis was proven to occur both in the amidodithiophosphonate salts and in the corresponding nickel complexes, an increased polarization of the P–N bond was calculated for the latter, suggesting a higher tendency to undergo hydrolysis. The antibacterial inactivity of the salts can be tentatively explained by taking into account their high hydrophilicity associated with their ionic nature, which circumvents the penetration of the cellular membrane. On the contrary, the neutral complexes [Ni(BzNH-adtp)2] and [Ni(PhBuNH-adtp)2] can pass the cellular membrane and thus exploit their activity. The inactivity of the analogous [Ni(iPrNH-adtp)2], can be explained, taking into account its higher resistance to hydrolysis, demonstrated by its higher stability both in solution and in the solid state, also confirmed by a P–N bond that is slightly shorter than those determined for analogous phenyl-alkyl-amidodithiophosphonato complexes. Further studies are ongoing in order to better understand the role of the alkyl-aryl substituents of the amines in the final amidodithiophosphonato complexes.The reaction between Lawesson’s Reagent (LR) and isopropylamine (


Bemisia tabaci, was initially discovered in the United States in 1894 but was not considered an economic insect pest on various agricultural crops across the southern and western states. After the introduction of B. tabaci Middle East-Asia Minor 1 (MEAM1) into the United States around 1985, the insect rapidly spread throughout the Southern United States to Texas, Arizona, and California. Extreme field outbreaks occurred on vegetable and other crops in those areas. The sweetpotato whitefly is now regarded as one of the most destructive insect pests in vegetable production systems in the Southern United States. The direct and indirect plant damage caused by B. tabaci has led to substantial economic losses in vegetable crops. Bemisia tabaci outbreaks on vegetables in Georgia resulted in significant economic losses of 132.3 and 161.2 million US dollars (USD) in 2016 and 2017, respectively. Therefore, integrated pest management (IPM) tactics are warranted, including cultural control by manipulation of production practices, resistant vegetable varieties, biological control using various natural enemies, and the judicious use of insecticides.The sweetpotato whitefly, Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) is among the most economically important insect pests of various vegetable crops in the Southern United States. This insect is considered a complex of at least 40 morphologically indistinguishable cryptic species. Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) was initially introduced in the United States around 1985 and has since rapidly spread across the Southern United States to Texas, Arizona, and California, where extreme field outbreaks have occurred on vegetable and other crops. This pest creates extensive plant damage through direct feeding on vegetables, secreting honeydew, causing plant physiological disorders, and vectoring plant viruses. The direct and indirect plant damage in vegetable crops has resulted in enormous economic losses in the Southern United States, especially in Florida, Georgia, and Texas. Effective management of B. tabaci on vegetables relies mainly on the utilization of chemical insecticides, particularly neonicotinoids. However, B. tabaci has developed considerable resistance to most insecticides. Therefore, alternative integrated pest management (IPM) strategies are required, such as cultural control by manipulation of production practices, resistant vegetable varieties, and biological control using a suite of natural enemies for the management of the pest. Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), and the greenhouse whitefly, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae)  [Bemisia tabaci is a primary insect pest of vegetable crops throughout the Southern United States [The three main species that cause problems in vegetables in North America and Hawaii are odidae)] . Bemisiad States ,8,9.Bemisia tabaci was initially documented by Gennadius as Aleurodes tabaci, a whitefly found in Greece , as well as ornamental plants, including poinsettia (Euphorbia pulcherrima Willd. ex Klotzsch), hibiscus (Hibiscus rosa-sinensis L.), and chrysanthemum (Chrysanthemum morifolium Ramat. Hemsl.). These agricultural crops and ornamental plants may serve as sources of B. tabaci for vegetable plantings ..Bemisia Bemisia tabaci is a relatively new pest to Kentucky and has been observed to attack squash and tomato [B. tabaci outbreaks in Kentucky had been relatively uncommon [Bemisia tabaci infests greenhouse-grown plants throughout the year in Kentucky but is unable to survive winters in the crop fields [d tomato ,105. Thouncommon . Howeveruncommon . Bemisiap fields .Trialeurodes vaporariorum is the most common species infesting tomato in Tennessee, while B. tabaci sometimes infests greenhouse-grown tomato. However, B. tabaci infestations in field-grown tomato have been sporadic in 2014 in Tennessee; less than 5% of fields have problems . Thus, minimal documentation is available.Compared to the aforementioned states, the impact of B. tabaci have resulted in the acceleration of research to provide potential management strategies [Extensive economic losses owing to the plant damage and control efforts of rategies ,106. ThiB. tabaci on vegetable crops, such as tomato [Cucumis melo L.) [Phaseolus vulgaris L.) [Capsicum annuum L.) [Efficient, accurate, and practical monitoring and sampling methods are crucial to the implementation of any management strategy ,108 becas tomato ,112,113,s tomato , watermes tomato ,116,117,melo L.) ,119, comaris L.) ,121, eggnuum L.) . Most ofB. tabaci incidence. However, due to the lack of correlation between trap catches and field populations, the yellow sticky trap may not be utilized when estimating B. tabaci density in either research or pest management application [The yellow sticky trap is among the most widely used methods, especially in larger-scale monitoring of lication ,124,125.T. vaporariorum and B. tabaci using digital image analysis [Capsicum annuum L.), cucumber, tomato, and eggplant) utilizing computer images with high accuracy [Progress has been made in automating insect counting that could reduce effort and increase accuracy in monitoring and sampling insect pests ,128,129.analysis . More reCultural control relies on the manipulation of production practices to render the environment unfavorable to pests ,130.B. tabaci control on vegetable plants [B. tabaci density and incidence of whitefly-transmitted viruses in cucumber, green bean, squash, and tomato [Coriandrum sativum L.) intercropping strategy, sprinkler irrigation reduced B. tabaci and associated viruses on organic tomato crops [B. tabaci populations in okra [B. tabaci populations [Water and fertility management plays a vital role as a cultural tactic in e plants ,136,137.d tomato . Howeverto crops . Moreove in okra  and hydrB. tabaci and associated viruses on tomato and other vegetable crops in Florida was greatly reduced by maintaining fields free of host crops for at least two months during the rainy summer period [B. tabaci abundance and associated virus incidence by planting cucumber, squash, and tomato one month earlier than standard planting dates in a study in Egypt [One important cultural control strategy is to increase host-free periods or reduce intercrop migrations ,139,140.in Egypt .B. tabaci problems on vegetables [B. tabaci and the incidence of associated viruses in tomato [Cucurbita pepo L.) [B. tabaci and related viruses in watermelon [B. tabaci within synthetic UV reflective mulches obtained significantly higher yields than those grown with living mulch of buckwheat (Fagopyrum esculentum Moench) in Florida [Living mulches (a cover crop interplanted or undersown with a main crop and intended to serve the purposes of a mulch) and synthetic mulches have been adopted for mitigating getables ,145,146.pepo L.) ,148. Syntermelon , zucchintermelon , snap betermelon , and tomtermelon ,150. Siltermelon . FurtherB. tabaci can be physically excluded from the vegetable crops when using fine mesh materials [B. tabaci. Thus, an electric field screen has proven successful in the “repel and/or capture” of B. tabaci on tomato crops in ventilated greenhouses [In protective structures (such as greenhouses and screenhouses), aterials ,153. Howenhouses ,158,159.B. tabaci populations on vegetables [B. tabaci populations and the incidence of whitefly-borne viruses on tomato crops [UV-absorbing materials have been applied in high tunnels and other forms of protected culture to reduce getables ,162,163.to crops ,161,162.B. tabaci management. Studies associated with trap crops and barrier crops have mixed results. Squash, cucumber, and eggplant have been shown to be promising trap crops for protecting tomato [B. tabaci. Maize (Zea mays L.) was demonstrated to be a good barrier crop of tomato [B. tabaci populations on tomato crops [B. tabaci egg nor nymphal densities on common bean leaves were reduced by the trap crop eggplant or the barrier crop corn in North Florida [Trap crops (preferred host plants which are used to draw a herbivore away from a less-preferred main crop)  and barrg tomato ,167,168 g tomato  against to crops . Moreove Florida .B. tabaci from locating host plants has shown promise. Lower B. tabaci densities and/or incidence of associated viruses were observed on tomato intercropped with coriander [Capsicum annuum L.) [Phaseolus vulgaris L.) [Allium cepa L.) [Allium sativum L.) [Zingiber officinale L.) has been reported as one alternative strategy to suppress B. tabaci populations in okra [B. tabaci and a lower severity of squash silverleaf disorder compared with non-intercropped zucchini [Lactuca sativa L.), the number of B. tabaci adults on cucumber leaves was reduced by 69.7% [Intercropping to prevent oriander ,145,171,oriander , maize [oriander , chili pnuum L.) , cucumbenuum L.) ,174, Frearis L.) , onion (cepa L.) , and garivum L.) . Additio in okra ,178. Zuczucchini . By inteby 69.7% .B. tabaci populations [Host-plant resistance is an essential strategy in successful IPM program for the suppression of ulations .B. tabaci have been extensively implemented. Bemisia tabaci exhibited reduced host preference (antixenosis) and reproduction (antibiosis) on tomato cultivars with Mi gene  [Mi-1.2 has been shown to confer resistance to B. tabaci by decreasing infestation, oviposition, and the number of fourth instar nymphs [B. tabaci [Solanum peruvianum (reported as Lycopersicon hirsutum), the elevated type IV glandular trichomes ) were found to be highly correlated with a reduction in B. tabaci infestation [B. tabaci [B. tabaci resistance in tomato has concentrated on screening existing tomato varieties [B. tabaci eggs and nymphs in Florida [In tomato, numerous efforts for breeding resistance to eceptor) ,182,183.r nymphs . The typ. tabaci . In the estation . Neverth. tabaci ,188. Morarieties  or theirarieties ,192,193. Florida .B. tabaci and associated viruses was also detected in many additional cultivated vegetable crops, such as watermelon [Vigna radiata L.) [Capsicum annuum L., C. chinense Jacq., and C. baccatum L.) [Brassica oleracea L. var. viridis) [Citrullus ecirrhosus was observed to exhibit resistance against B. tabaci, and thus became exploited as a source of B. tabaci resistance in breeding cultivars of watermelon [B. tabaci and Mungbean yellow mosaic virus (MYMV) [B. tabaci  [B. tabaci adults and nymphs [C. annuum), IAC-1545 (C. chinense), and IAC-1579 (C. annuum) could be classified as good sources of resistance against B. tabaci and associated virus Tomato severe rugose virus (ToSRV) for cultivated pepper [B. tabaci [Resistance against termelon ,194, muniata L.) ,196, chiiata L.) ,198,199,atum L.) , and colviridis) ,202. A pication) . Chili pd pepper . CollardB. tabaci and/or associated viruses have been developed, such as transgenic tomato [B. tabaci fed on genetically modified bean for 8 days, the BGMV DNA amount in B. tabaci was significantly reduced 84% [B. tabaci showed a mortality rate of up to 98.1% [Transgenic vegetable plants that provide resistance to c tomato ,209,210,c tomato ,212,213,c tomato . For exac tomato . After Bto 98.1% .B. tabaci. Heavy reliance on insecticides against B. tabaci is still widespread in open field vegetable crop systems [B. tabaci, such as the insect growth regulators (pyriproxyfen and buprofezen), ketoenols (spiromesifen and spirotetramat), and diamides  [B. tabaci control [Applications of chemical insecticides are among the main tools used commonly by vegetable growers to control  systems ,59. The liprole) ,59. Oils control ,59.B. tabaci [B. tabaci on vegetables in Florida [B. tabaci populations on tomato plants in Southwest Florida during the period 1994–1996 [B. tabaci on vegetables in Texas as well [B. tabaci on cabbage and cantaloupe in Texas [The neonicotinoids are among the most effective group of insecticides. Imidacloprid was the first commercial neonicotinoid, eventually becoming the most widely used insecticide worldwide . Imidacl. tabaci . The app. tabaci . Imidacl Florida ,218, and994–1996 . Imidacl as well ,220. It in Texas .B. tabaci in cotton [B. tabaci on cotton crop in the Southwestern United States [B. tabaci on melons, and pyriproxyfen has the potential to be labeled against B. tabaci on melons and vegetables [® and Naturalis-L®), plus one soil application of imidacloprid, significantly reduced B. tabaci populations on spring cantaloupe in Texas [Pyriproxyfen, a juvenile hormone analog, has been utilized to successfully control n cotton ,222,223.getables . It was in Texas .B. tabaci on vegetable crops [B. tabaci on melon and collard with almost 100% nymphal mortality in laboratory bioassays and exhibit excellent promise against B. tabaci populations on melon and collard in field trials [B. tabaci on bell pepper compared to the neonicotinoid insecticide of imidacloprid 200 SL [Ketoenols are a relatively new class of insecticides that are derivatives of tetronic acids (spiromesifen) and tetramic acids (spirotetramat) . These cle crops ,230,231.d trials . It has B. tabaci [B. tabaci in tomato [B. tabaci control, TYLCV suppression, as well as reduced oviposition and nymph survival on tomato [Diamide insecticides are the most recent chemistry class in the market  and act . tabaci . Cyantran tomato ,238,239.B. tabaci on plants and influence the transmission of plant viruses [B. tabaci populations on vegetables, such as tomato [B. tabaci nymphs in zucchini squash and tomato under greenhouse conditions; and Saf-T-Side  OilTM, Natur’lTM (vegetable) oil, New DayTM liquid detergent or M-PedeTM reduced the number of B. tabaci adults on heavily infested cucumber plants in field in Florida [Soaps and oils have been long used as insecticides . Both so viruses . Soaps as tomato ,246,247,s tomato , zucchins tomato , collards tomato , and mels tomato . For exa Florida .B. tabaci on vegetable crops [B. tabaci populations, including neem-based products [Calotropis procera (Aiton) W.T. Aiton) and garlic [Jatropha curcas L. [Tulbaghia violacea Harvey) [B. tabaci populations in tropical open field-grown and greenhouse-grown tomato [B. tabaci [Piper callosum, Adenocalymma alliaceum, Pelargonium graveolens, and Plectranthus neochilus deterred the settlement and oviposition of B. tabaci adults on tomato plants [Various plant-based products have been extensively investigated for their activity against le crops ,252,253.products ,257,258,d garlic , extracturcas L. , as well Harvey) . A drencn tomato . Moreove. tabaci ,268,269.o plants .B. tabaci summarized various resistance mechanisms, several bioassays used to evaluate resistance, studies for monitoring resistance, known cases of insecticide resistance, and some strategies that can be used to mitigate resistance [B. tabaci species but focusing mainly on research published from 2010 to 2020 [B. tabaci, namely, chemical control with selective insecticides, rotation of insecticides with different modes of action, insecticide mixtures, and nonchemical control methods  [A review of insecticide resistance in sistance . More re to 2020 . Their rmethods) .B. tabaci IPM systems. There are at least 48 species of predators, 62 species of parasitoids, and 9 species of pathogens reported as natural enemies of B. tabaci [B. tabaci include the predators in the following genera: Brumoides, Verania, Cheilomenes, Macrolophus, Nesidiocoris, Geocoris, Orius, Chrysopa, Coccinella, and Amblyseius; the parasitoids in the genera Encarsia and Eretmocerus; the nematodes in the genera Steinernema and Heterorhabditis; as well as the fungi in the genera Beauveria, Cordyceps (formerly known as Isaria or Paecilomyces), and Verticillium [Biological control plays an essential role in . tabaci ,271. Theicillium .B. tabaci management [B. tabaci on vegetable crops over the past two decades.Numerous studies have been conducted and reviewed on the distribution, life history, bionomics, and ecology of natural enemies for nagement ,275,276.B. tabaci, only a few have been studied thoroughly and many are still under limited laboratory observations or qualitative field records [B. tabaci are coccinellid beetles, predaceous bugs, lacewings, and phytoseiid mites.Although over 150 arthropod species have been described as predators of  records ,273. MosHarmonia axyridis Pallas (Coleoptera: Coccinellidae) was demonstrated to suppress B. tabaci in greenhouse-grown tomato when released with a parasitoid, Encarsia formosa Gahan or Encarsia sophia Girault and Dodd (Hymenoptera: Aphelinidae) [Delphastus catalinae Horn (Coleoptera: Coccinellidae) is the most commonly used predacious natural enemy and commercially available for B. tabaci control on various vegetable crops in greenhouses [B. tabaci populations on tomato plants were attributed to the release of D. catalinae [D. catalinae are not known to occur in Northern Florida, Georgia, or South Carolina [The predatory linidae) . Delphasenhouses . For exaatalinae . This beatalinae . Field pCarolina .B. tabaci using mirids includes the inoculative and augmentative release of Macrolophus pygmaeus Rambur (Hemiptera: Miridae) and Nesidiocoris tenuis Reuter (Heteroptera: Miridae). For instance, M. pygmaeus could control B. tabaci populations in melon when released at 6 adults/plant into the initial infestation of 10 B. tabaci adults/plant in greenhouses [N. tenuis  resulted in a >90% reduction in B. tabaci populations in tomato under greenhouse conditions [N. tenuis did not contribute to B. tabaci suppression in protected sweet pepper (Capsicum annuum L. (Solanaceae) cv. Spiro) [B. tabaci was compromised when M. pygmaeus and N. tenuis coexisted on tomato plants but was improved when the parasitoid Eretmocerus mundus Mercet (Hymenoptera: Aphelinidae) was added into the mix [Macrolophus melanotoma Costa (Hemiptera: Miridae), and the parasitoid Er. mundus supported a low number of B. tabaci in greenhouse-grown eggplant [In vegetable greenhouses, successful biological control of enhouses . Two relnditions . However. Spiro) . Predati the mix . Additioeggplant .Chrysoperla rufilabris Burmeister (Neuroptera: Chrysopidae) provided significant B. tabaci suppression on watermelon in the field, but no effect in a legume field [Chrysoperla carnea Stephens (Neuroptera: Chrysopidae) on B. tabaci in greenhouse-grown tomato was also investigated, and the lowest number of B. tabaci was observed at a release rate of 10 C. carnea larvae/plant [B. tabaci in squash was >90% after a third predator release of 10 C. carnea larvae/plant and reached 100% after six releases [Bemisia tabaci populations on sweet pepper plants were reduced about 97% after the second predator release of 10 C. carnea larvae/plant [C. carnea with the predatory bug Orius albidipennis Reuter (Hemiptera: Anthocoridae) and the predatory mite Phytoseiulus persimilis Athias-Henriot (Acarina: Phytoseiidae) decreased B. tabaci populations and increased cucumber yields in greenhouses [The release of the lacewing me field . The preae/plant . Furtherreleases . Bemisiaae/plant . Integraenhouses .Amblyseius swirskii Athias-Henriot (Acarina: Phytoseiidae) is considered among the most effective natural enemies and utilized extensively on vegetable crops [A. swirskii at two release rates of 25 and 100 mites/m2 suppressed B. tabaci almost totally when an initial infestation was 8 B. tabaci adults per sweet pepper plant [A. swirskii provided a significant reduction in the B. tabaci populations in sweet pepper in greenhouses [A. swirskii/m2 and 12 Er. mundus/m2 was the most efficient strategy in protected sweet pepper crops at an initial infestation of 50 adults of B. tabaci per plant [N. tenuis or M. pygmaeus with A. swirskii significantly suppressed B. tabaci populations on sweet pepper under greenhouse conditions [The predatory mite le crops . Under ser plant . Similarenhouses ,291. Morer plant . Integranditions .Amblyseius barkeri Hughes or Amblyseius cucumeris Oudemans (Acarina: Phytoseiidae) suppressed B. tabaci populations on tomato plants in greenhouses [Amblyseius tamatavensis Blommers (Acarina: Phytoseiidae), was first reported in 2018 in Southern Florida [B. tabaci populations on bell pepper plants under laboratory conditions [The predatory mite enhouses . Another Florida  and has nditions .B. tabaci. The majority parasitoids of B. tabaci are from the genera Encarsia and Eretmocerus (Hymenoptera: Aphelinidae) [B. tabaci in Florida [Encarsia formosa, Eretmocerus eremicus Rose and Zolnerowich (Hymenoptera: Aphelinidae), and Er. mundus have been extensively investigated for B. tabaci control [Parasitoids as biocontrol agents play a crucial role in IPM program of linidae) ,59,271,  Florida . Encarsi control ,293.Encarsia formosa is the most widely used parasitoid for the biological control of B. tabaci on greenhouse-grown vegetable crops [En. formosa and Er. mundus resulted in B. tabaci parasitism of 62.0% and 77.9%, respectively, in sweet potato (Ipomoea batatas L.) field [le crops . For ins.) field .Eretmocerus species  have recently gained important focus in the biological control of B. tabaci [Er. eremicus significantly suppressed B. tabaci eggs and nymphs in mint under greenhouse conditions [Er. mundus provided high incidence of parasitism and significant control of B. tabaci in sweet pepper and tomato in greenhouses [Er. mundus with A. swirskii or M. caliginosus greatly reduced B. tabaci populations on greenhouse-grown sweet pepper and tomato plants [. tabaci ,299,300.nditions ,302. Morenhouses ,304. Into plants ,303.Entomopathogens  as biological control agents are a major component of IPM programs in regulating populations of insect pests ,306,307.Steinernema and Heterorhabditis are important biological control agents that are employed to control various destructive insect pests [B. tabaci management on vegetable crops [Steinernema feltiae Filipjev (Rhabditida: Steinernematidae) alone or combined with the insecticide imidacloprid significantly reduced B. tabaci survival in tomato and verbena  [Steinernema carpocapsae Wesier (Rhabditida: Steinernematidae) with the insecticides thiacloprid or spiromesifen provided high B. tabaci mortality of 86.5% and 94.3%, respectively, on tomato plants [Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) Guatemala strain; H. bacteriophora Chiapas strain; as well as Heterorhabditis indica Poinar, Karunakar, and David (Rhabditida: Heterorhabditidae) native strain from Sinaloa, Mexico killed up to 95% of B. tabaci nymphs on tomato leaves under laboratory conditions [B. tabaci in pepper caused by entomopathogenic nematodes was reported to be very low: 17.44% for H. bacteriophora, 35.26% for S. carpocapsae, and 18.48% for S. feltiae [S. feltiae only resulted in 32% and 28% B. tabaci mortality on tomato and cucumber plants, respectively [Entomopathogenic nematodes in the genera ct pests ,311,312.ct pests . Entomople crops ,317,318.alis L.) ,316,317.nditions . However feltiae . The appectively .Beauveria bassiana  Vuillemin; Cordyceps fumosorosea (Wize) Kepler, B. Shrestha and Spatafora (formerly known as Isaria fumosorosea (Wize) or Paecilomyces fumosoroseus (Wize) A.H.S. Brown and G. Smith); and Lecanicillium lecanii (Zare and Gams) are the most widely investigated entomopathogenic fungi that infect B. tabaci on vegetable crops [Entomopathogenic fungi are also major components of plant protection programs for managing economically important insect pests ,324,325.le crops .B. bassiana, the mean mortality of the fourth instar B. tabaci nymphs on cucumber was 91.8% [B. bassiana, the survival rate of nymphal B. tabaci was 4.2%, 9.6%, 13.4%, and 24.3% on cucumber, eggplant, tomato, and cabbage, respectively [C. fumosorosea (reported as P. fumosoroseus) and B. bassiana provided >90% control of B. tabaci nymphs on cucumber, cantaloupe melon, and zucchini squash in small-scale field trials [B. bassiana and C. fumosorosea (reported as I. fumosorosea) also resulted in 71%–86% mortality of B. tabaci nymphs on bean leaves under laboratory conditions [B. bassiana and C. fumosorosea (reported as I. fumosorosea) against B. tabaci on common bean plants was improved significantly when mixed with the non-ionic surfactant Silwet® L-77 [At 8 days post-inoculation of as 91.8% . Similarectively . Furtherd trials . The isonditions . A folloet® L-77 .B. tabaci populations has also been evaluated. The individual application of the entomopathogenic fungus Lecanicillium muscarium (Petch) Zare and Gams or integrating with the insecticide imidacloprid resulted in high mortality of B. tabaci on tomato, verbena, and cucumber plants under both laboratory and glasshouse conditions [C. fumosorosea (reported as P. fumosoroseus) and the insecticide azadirachtin were combined, up to 90% B. tabaci nymphal mortality was obtained, although the combined effects were less than additive effects [Cordyceps javanica  Kepler, B. Shrestha and Spatafora  Samson and Hywel-Jones) isolate caused up to 62.4% nymphal mortality of B. tabaci on bean plants, and an additive interaction was found when C. javanica (reported as I. javanica) was combined with the insecticides buprofezin or spiromesifen [B. bassiana was integrated with imidacloprid against B. tabaci in cantaloupe melon [The effect of integrating entomopathogenic fungi with chemical insecticides in suppressing nditions ,334,335. effects . Anotheromesifen . Howeverpe melon .B. tabaci populations if applied independently, they could be more effective at reducing losses due to B. tabaci infestations when used in combination as part of an IPM program [B. tabaci [B. tabaci infestations [B. tabaci populations while preserving beneficial organisms [Although the aforementioned control measures can impact  program ,336,337.. tabaci ,153 and . tabaci ,287,292,. tabaci ,303,304,. tabaci ,332. Undstations ,148; 2) rganisms ; and 3) rganisms ,220.B. tabaci and associated viruses plays an important role in B. tabaci management [B. tabaci and associated viruses utilizing genetic engineering technology, the cultivation and acceptance of genetically engineered vegetable crops tend to be deferred for the future considering ethical and legislative issues.Vegetable breeding for resistance to both nagement . New andB. tabaci populations on vegetables [B. tabaci populations have developed considerable resistance to insecticides from different chemical classes [B. tabaci, secondary pest outbreaks, and environmental degradation [B. tabaci problems, as it was noted that B. tabaci survived long-term under high temperature stress [The application of chemical insecticides remains the principal approach to suppress getables ,59. Neveradation . Furtherradation . Additioe stress .B. tabaci populations. There is a general trend in using two or more natural enemies together [B. tabaci IPM systems.Successful biological control has been achieved in suppressing together ,283,284 together ,336,337.B. tabaci management. Hyperspectral imaging along with machine-learning-based assessments might allow the rapid and accurate detection of B. tabaci on vegetables [B. tabaci infestations, especially at low densities. Subsequently, precision application of therapeutic management tactics, including insecticides, may be automated and coupled to monitoring systems, which makes them more practical for large-scale farming systems [Novel technologies provide more opportunities for getables ,128,129. systems . The preB. tabaci management, the biotechnological tool RNA interference (RNAi) is initiated by the introduction of double-stranded RNA into a cell, which results in the generation of loss-of-function phenotypes via the depletion of the target gene messenger [B. tabaci through injection [B. tabaci through RNAi, and these studies demonstrated its potential to manage B. tabaci at the laboratory level [Among the potential novel technologies that can be used as tools for essenger ,344. RNAessenger ,346,347.njection ,349, oranjection , or by enjection . To datery level ,353,354.