Sorghum bicolor accessions. This genetic diversity resource is untapped, under-utilized, and has not been systematically incorporated into sorghum breeding programs. In this study, we characterized the germplasm collection using whole-genome SNP markers (DArTseq). Discriminant analysis of principal components (DAPC) was implemented to study the racial ancestry of the accessions in comparison to a global sorghum diversity set and characterize the sub-groups present in the Ugandan (UG) germplasm.The Plant Genetic Resources Centre at the Uganda National Gene Bank houses has over 3000 genetically diverse landraces and wild relatives of p > 3) to survival under cold stress under both field and climate chamber conditions, located on 3 chromosomes . To our best knowledge, the QTL on Sb09 with the strongest association was discovered for the first time.Population structure and phylogenetic analysis revealed the presence of five subgroups among the Ugandan accessions. The samples from the highlands of the southwestern region were genetically distinct as compared to the rest of the population. This subset was predominated by the caudatum race and unique in comparison to the other sub-populations. In this study, we detected QTL for juvenile cold tolerance by genome-wide association studies (GWAS) resulting in the identification of 4 markers associated (−log10This study demonstrates how genebank genomics can potentially facilitate effective and efficient usage of valuable, untapped germplasm collections for agronomic trait evaluation and subsequent allele mining. In face of adverse climate change, identification of genomic regions potentially involved in the adaptation of Ugandan sorghum accessions to cooler climatic conditions would be of interest for the expansion of sorghum production into temperate latitudes.The online version contains supplementary material available at 10.1186/s12870-022-03770-y. Sorghum bicolor [L.] Moench (sorghum) is the fifth most important cereal crop globally and shows remarkable diversity, including five different races, their intermediates, and several crop forms classified as grain, forage, sweet and broomcorn types [rn types . Sorghumrn types . Becausern types .S. bicolor are native [In Eastern Africa sorghum is traditionally grown as a food-fodder crop by smallholder farmers in low-input agricultural systems spanning highlands, lowlands, and semi-arid cropping regions. Uganda is located in eastern Africa, south of South Sudan. Archaeological evidence suggests that sorghum’s center of domestication is likely the Ethiopia-Sudan region in the north-east . For agee native , making S. bicolor accessions, including a vast range of landraces whose diversity has yet to be capitalized for use in breeding. This germplasm has not yet been fully characterised and evaluated, limiting its utilisation to date in sorghum improvement programs in Uganda and elsewhere. The considerable geographical and topological diversity of Uganda makes this genetic resource a potentially interesting reservoir for genetic analysis and diversity for adaptive traits of interest for sorghum breeding. For example, cool highland areas in the southwestern Uganda are potential sources of diversity for cold-tolerance traits that could help improve sorghum adaptation in temperate cropping regions.The country’s broad sorghum diversity reflects the variety of environments where the crop is grown, mainly on marginal agricultural lands ranging from extremely arid and semi-arid zones in eastern and northern Uganda to cool highlands in south-western regions. The Uganda National Genebank houses a large collection of Effective and efficient management of germplasm from a genebank collection is an essential prerequisite for farmers and breeders to identify, extract and exploit the extensive diversity. Genome-wide characterization of untapped genetic resources using genome sequencing technologies provides new opportunities for sustainable breeding and efficient usage of material.S. bicolor collection, representing different agro-ecological zones of the country, was investigated using genome-wide SNP markers and population genetic analysis. The primary objective was to genetically characterize Ugandan (UG) sorghum germplasm in the context of global sorghum diversity, in the absence of morphological data from flowering and maturity for classification into racial groups. As a case study for the value of this resource in adaptive breeding, we furthermore used the available genome data, in association with phenotypic data for juvenile cold tolerance traits, to identify genomic regions enriched with genetic variants associated with low temperature adaptation. The results demonstrate how genebank genomics can help facilitate discovery of economically or biologically important plant diversity and genes as a prerequisite for crop genetic improvement and climatic adaptation.In this present study, the diverse Uganda National Genebank In order to efficiently conserve and utilize the novel UG germplasm, we studied the underlying genetic variation and diversity. The population structure for the UG set was studied using all the 3333 samples and a subset of 12,742 markers from the complete dataset.Five sub-populations within the germplasm were inferred by the DAPC analysis based on Bayesian Inference Criterion (BIC) Fig. a. Four hWith the aim of genetically characterizing the novel Ugandan germplasm and racially comparing it to a well-established global diversity set of geo-referenced sorghum accessions a DAPC co-analysis was performed between the two. As Ugandan accessions outnumbered the global (UQ) set by almost three to one ratio, a random set of UG samples (10 samples from 10 random clusters) were selected for the comparison. All the underlying racial groups from the global collection were identified in the Ugandan germplasm Fig. . RelativTo confirm the racial distribution and diversity in the UG germplasm the DAPC co-analysis with the global set was replicated three times with three random unrelated UG samples. The racial clustering present among the UG set was found to be well dispersed Fig. .Due to significant adaptability differences between the lowland and highland races from the Ugandan diversity set, genetic diversity among the sub groups within the population were tested. Fst between each UG subpopulation from the various geographical regions compared to the entire population revealed genetic variance ranging from − 0.608 to 0.525 .A genome-wide association study (GWAS) was conducted for identification of regions of interest associated to juvenile survival under cold stress in two temperate-climate field environments  and one controlled-environment climate chamber experiment (CC). Highly significant differences for cold tolerance among genotypes -like superfamily protein coding gene and as a cleavage stimulation factor subunit 3 (Cstf3). This gene family has been shown to have anti-cold response functions in tomato [Arabidopsis thaliana [Comparing protein sequence of orthologous gene segments of sorghum, maize and rice, six genes involved in various cold acclimatization and tolerance were identified Table S. For exathaliana  .S. bicolor germplasm reflects the highly diverse environments where sorghum grows in Uganda. Accessions belonging to the arid regions of northern and eastern Uganda are well adapted to extreme drought and heat stress, whereas the ones from cold highlands of the southwestern region  genetic characterization of an underutilized novel germplasm and (ii) dissection of cold tolerance trait within this dataset. The population structure results indicate immense genetic and racial diversity within the germplasm predominated by admixed accessions. Contrast to other geographical regions in Uganda the accessions from the south-western highlands displayed a unique pattern, composed mainly of the caudatum race and genetically isolated from the other subpopulations. This manuscript can be used a base for precisely characterizing novel germplasm based on genetic data. The genomic and phenotypic data collected in this study provide an objective criterion for the selection of accessions for genetic diversity preservation and management, utilization in breeding programs and genetic relationship analysis with other germplasm collections.A comprehensive investigation involving survival traits, identified multiple key associations and genes underlying the response of juvenile sorghum seedlings to cold stress. The identification of multiple QTL associated to juvenile cold stress reported here can be used by breeders to enhance early-stage chilling tolerance in sorghum. The results provide important new insight for adaptive crops breeding to support the expansion and stability of sorghum production in the face of increasing abiotic stress constraints and climatic change.S. bicolor germplasm accessions (UG set) collected by the Plant Genetic Resources Centre at the Uganda National Genebank were used in this study  for DNA extraction. The DNA samples were then genotyped using DArTseq, an efficient genotyping-by-sequencing (GBS) platform which enables discovery of genome-wide markers through genome complexity reduction using restriction enzymes. Genotyping was performed fundamentally as described in references [Diversity Arrays Technology Pty Ltd.  and Discriminant Analysis of Principal Components (DAPC) were implemented using the R package Adegenet (2.1.3) . To avoihttps://github.com/knausb/vcfR). After an initial transformation using the PCA analysis, racial composition was subsequently identified for the clusters using discriminant analysis (DA).To select the UG subset representing the meta-population, we initially clustered the entire set into 10 groups followed by randomly sampling 10 accessions from each group. These randomly-selected samples were combined with the global dataset for DAPC co-analysis. To validate the racial assignment of the UG set groups, the DAPC co-analysis was repeated three times, each time using a different set of 10 random genotypes from each of the 10 clusters. The SNP data was converted to the genlight object bit-level genotype coding scheme using the function ‘vcfR2genlight’ of the vcfR tool . To elaborate the genetic relationship among the accessions a pairwise distance matrix was established using the tool VCF2Dis (https://github.com/BGI-shenzhen/VCF2Dis), which was then converted to a neighbour-joining phylogenetic tree using the R package ape (5.5) [To describe the population structure of the UG germplasm and evaluate the racial ancestry of each group in relation to geographical origin of the accessions, the filtered marker set  was pruned to exclude SNPs which were in strong LD using PLINK software . Pruningpe (5.5) . To studpe (5.5) .Juvenile survival under cold stress was evaluated through two field trials at Gross Gerau (GG), Germany (spring 2019 and 2020) and one climate chamber experiment. A UG subset of 444  and 255 (climate chamber) accessions representing all agro-ecological zones were used for the association study. For the field experiments Table S, all genp value) ≥ 3.0 was defined [r2) between each pair of SNPs. Haplotype blocks were calculated using an LD threshold of r2 > 0.7 implemented in the tool LDBlockShow [After eliminating markers and genotypes with more than 25% missing data points, the dataset was imputed using Beagle 5.1 . It was  defined . LinkagelockShow , as desclockShow .Sorghum bicolor reference genome v3.1.1 hosted by Phytozome 12 . Orthologous genes within selected haploblocks were identified by homology comparisons of the genomic sequences in maize and rice. Protein sequence alignments were conducted by using blastp option of DIAMOND [Candidate genes were selected based on the  DIAMOND .Additional file 1: Supplementary Figure 1. Scatter plot representing DA loading of Ugandan accessions based on races. (a) and (b) are replicate 2 and 3 respectively. Each dot represents an individual and the colour code is displayed in the index.Additional file 2: Supplementary Figure 2. Topographic map of Uganda showing the elevation.Additional file 3: Table S1. DAPC analysis of Ugandan lines based on races and geographical origin. Table S2. Population genetics indices for the sorghum accessions from different districts of geographical regions in the Uganda National GeneBank. Table S3. Statistical data of the field experiments and the climate chamber experiment. Table S4. Genome-wide association study reveals the genetic basis of juvenile cold tolerance trait in sorghum. Table S4. Summary of different cold tolerance traits QTL overlapping the haploblock regions. Table S6. Summary table of genes identified in the haploblock regions involved in cold stress tolerance. Table S7. Conditions of the field experiments conducted in Gross-Gerau, Germany  in 2019 and 2020. Table S8. Conditions of the climate chamber experiment.


Maintaining a genetic diversity in populations of endangered species is an important part of conservation biology and its programs focused on maintaining biodiversity. Gene banks are used to safeguard the genetic variability of populations. The management of genetic resources deposited in gene banks requires knowledge of the genetic profiles of the gamete donors and bioinformatics tools to process this information. In this work, we show how to use Genassemblage 2.2 software in managing the genetic variation deposited in a bank of cryopreserved semen. Our results showed that if we combine genetic profiles of gamete donors and bioinformatic techniques, we can identify a small group of semen samples that are enough to transfer all alleles detected across an entire set of samples. Consequently, we recommend the Genassemblage 2.2 software as a convenient tool in the management of genetic variation deposited in gamete banks.Eupallasella percnurus, which is designated as endangered in Poland. The semen samples (n = 192) were taken from four Polish lake minnow populations and frozen in the gene bank. Fin clips were taken and DNA extracted. Across 13 investigated microsatellite loci, 21–53 alleles were identified in each population and 66 in the entire group of samples. The module “Management of genetic variation in gamete bank” of Genassemblage 2.2 software was used to find the set of samples that will preserve 100% of the detected allelic diversity in the next generation. Our results showed that a small group of 4–19 semen samples was enough to transfer all alleles detected across this set of samples. We, therefore, recommend Genassemblage 2.2 as a convenient tool for the detection of genetic differences between donors, the construction of optimal sets of samples for conservation of genetic variation, and for managing genetic variation deposited in gamete banks. Consequently, it can be used in breeding human-dependent populations and to optimize the use of genetic diversity in samples in the gamete banks. It can be especially useful for conserving populations of species characterized by low genetic variation, such as the lake minnow.The management of genetic resources deposited in gene banks requires knowledge of the genetic profiles of the gamete donors and bioinformatics tools to process this information. In this work, we show how to use Genassemblage 2.2 software in managing the genetic variation deposited in a bank of cryopreserved semen. Our demonstration was based on the leuciscid fish species, lake minnow  Maintaining a genetic diversity in populations of endangered species is an important part of conservation biology and its programs focused on maintaining biodiversity. It can be performed using techniques of gene banking, which are used in conservation of populations endangered because of anthropogenic pressure or the deterioration of environmental conditions ,2,3. TheThe potential usefulness of gene banks in the conservation of genetic diversity depends on the genetic differences between the individuals from which the gametes are collected  and on tEupallasella percnurus  a member of Leuciscidae family, has been used as an example for the use of Genassemblage 2.2 (Poland) software in management of gamete banks. In Poland, this inhabitant of vanishing, small and shallow habitats is considered to be endangered facing extinction, and therefore, is strictly protected by law with the formal requirement of active protection [The freshwater native fish species, lake minnow otection . Active otection ,14. The otection . Management of genetic variation in gene banks can be performed if the genetic characteristics of the gamete donors are known and bioinformatic tools are used. Based on molecular markers, e.g., microsatellite DNA, single nucleotide polymorphisms (SNPs), etc., it should be possible to identify genetic differences among males and females used for breeding . ConsideTo the authors’ knowledge, no bioinformatic tools that can be used to manage genetic variation deposited in gene banks have been developed so far. Because of this, we developed such a tool and added it to the Genassemblage 2.2 software as a new module, “Management of genetic variation in gamete bank”.http://pracownicy.uwm.edu.pl/d.kaczmarczyk/main_page.htm (accessed on 4 November 2022). The new module analyzes allelic diversity detected across all loci included in the genetic profiles of males whose sperm samples are stored in a gamete bank. It calculates the overall number of alleles across all loci and indicates which samples should be used to obtain a target percentage of allelic diversity while using as few samples as possible.Genassemblage 2.2 is Windows-based software that constitutes a further development of Genassemblage 1.0 , includiHere, we present an example of how Genassemblage 2.2 can be used to manage genetic variation resources deposited in a bank of cryopreserved semen. As an example we used lake minnow males from four Polish populations that had donated semen to that bank.The investigated lake minnow populations were the following: Mikołajki Pomorskie (MP) , Drozdowo (DR) , Zielonka (ZI)  and Bełcząc (BE) . The semen was taken in 2015 from 192 males (n = 48 fish per population). The fish were captured using baited traps  and tranCa3, Ca4 and Ca12 [Z9878, Z10362 and Z13419 [https://www.ncbi.nlm.nih.gov (access data 4 November 2022); and Eupe1, Eupe2, Eupe4, Eupe5, Eupe6, Eupe7 and Eupe9, which were amplified using primer sequences described by Kaczmarczyk and Gadomski [DNA was extracted from all fin samples using a Genomic Mini AX Tissue SPIN DNA Extraction and Purification Kit . The extraction procedure was performed following the manufacturer’s recommendations. The integrity of the DNA samples was visually inspected after electrophoresis in 1.5% agarose gel stained with ethidium bromide. DNA yields were quantified by spectrophotometric analysis. The 13 microsatellite markers known to be polymorphic in this species  were useand Ca12 ; Z9878, d Z13419  . The lengths of the amplified DNA fragments were determined using an Applied Biosystems 3130 Genetic Analyzer against GS400LIZ size standards. Allele determination was performed using GeneMapper 3.0 software  according to the manufacturer’s recommendations.http://pracownicy.uwm.edu.pl/d.kaczmarczyk/genasemblage2_2/genassemblage_software2_2.htm (access data 4 November 2022) was used with the module “Management of genetic variation in gamete bank”. The example of input file  within each population, (2) in a group containing all four populations and individuals investigated in this study. As a measure of the allelic diversity, a number of alleles detected across all samples and loci were chosen with the software. To obtain the highest possible transfer of allelic diversity to the progeny, the value for the minimum level of genetic variation was set at 100%. The software identified in the input file the samples that contribute most to allelic diversity and enable transfer of a target percentage to progeny by using as few samples as possible. Consequently, all alleles identified in this sample group should be included in the genotypes of sperm donors chosen by Genassemblage 2.2. The details of how to perform this analysis are given in the user manual that can be downloaded from the author’s website, provided above.Results of calculations in the output file can be opened by an MS Excel  spreadsheet. This file contains such data as: (1) genotypes of males from the input file; (2) variation across all samples included in the input file ; (3) list of samples chosen by the software as optimal for transfer of a target percentage of genetic variation detected across all samples and their genetic profiles; (4) list of alleles at investigated loci that will be transferred by set of samples chosen by Genassemblage 2.2; (5) summary analyses, including number of samples in the input file, number of samples chosen by the software and actual percentage of allelic variation of the entire set that will be transferred. The allelic diversity described in number of alleles is given both in the Genassemblage interface and in the output file. Moreover, the alleles at each locus and across all samples and loci are shown in the output file . An examThe Genassemblage 2.2 identified 21, 27, 29, 53 alleles across all loci (nA) in the samples from Mikołajki Pomorskie (MP), Drozdowo (DR), Zielonka (ZI) and Bełcząc (BE), respectively . In the The example of the result file calculated for the Zielonka (ZI) population  shows thIn the case of other populations, a transfer of 100% of the allelic diversity (N’) is possible if we use 4, 5 and 13 samples from DR, MP and BE populations, respectively. The names of the selected samples from BE, MP and DR populations and the additional information described above are given in the The Genassemblage 2.2 software was designed to help in identification of a group of samples deposited in a gamete bank that are most valuable for conservation of the genetic variation of a species. The software can be used for: (1) detecting genetic differences between gamete donors, (2) assembling the samples from the gamete bank in sets that are optimal for transferring allelic diversity to the progeny, (3) management of resources deposited in the gamete bank by maximizing the effect of using a limited number of samples, (4) reduction of the likelihood of using sperm samples from closely related individuals and inbreeding in the next generations. In this paper, we have evidenced that it is not necessary to use all samples from the gamete bank to transfer the entire allelic diversity. In all investigated populations, as well as in the dataset containing all investigated samples, the same effect can be obtained if a small group of highly diverse samples were used. The Genassemblage 2.2 can detect and pinpoint them. This tool can be used in this way to obtain the most diverse progeny and to optimize the use of the available genetic diversity. The software is particularly useful in the case of populations of fish species of low genetic variation, as in our example, lake minnow, when only a small number of individuals differ from the others to a relatively large extent.Our results give evidence to the fact that Genassemblage 2.2 can serve as a convenient tool for detecting genetic differences between gamete donors and assembling the samples deposited in a gamete bank in optimal sets for conservation of the genetic variation. The Polish lake minnow populations, including those investigated in this study, demonstrate a considerable decrease in the genetic variation resulting from both founder and bottleneck effects and strong genetic drift and inbreeding. Likewise, their low allelic diversity is a well-known feature of this species in its entire range of the occurrence in Poland . The small genetic differences recorded in the Polish lake minnow populations indicate close relationships between the fish belonging to them, so it is particularly important to select samples that genetically differ to the largest extent . The lakIt should be noted that in this study, lake minnow populations and the number of analyzed markers were chosen for illustrative purposes. It is obvious that for better detection of the genetic differences between lake minnow individuals, more markers should be applied than we used in our work . The useCa4). Moreover, the application of Genassemblage 2.2 is not limited to microsatellite markers and can be used with a dataset including more than 100 samples. Other markers that inherit according to Mendelian Laws (such as SNP markers) can be used if numbers are used to name their alleles. This feature makes the software appropriate for conservation of disomic, partially tetrasomic and fully tetrasomic organisms and enables the use of various kinds of genetic data. It is important to note that the set of individuals or their gametes that are analyzed with the module “Management of genetic variation in gamete bank” can also be used to assemble breeding pairs or sets of pairs using other modules included in Genassemblage 2.2, i.e., “Select the best breeding pair and Select individuals for group spawning” [In this work, we have demonstrated that Genassemblage 2.2 software can operate with both disomic and tetrasomic loci such as  as a source of semen for breeding. In most cases, the neomales lack spermatic ducts, and therefore, fish must be killed in orFder to obtain semen . The potIn this work, we have demonstrated that Genassemblage 2.2 can optimize the use of the genetic diversity deposited in gamete bank. Consequently, this software can be an efficient tool for conservation of genetic diversity in human-dependent populations.


Objectives: As genetic testing is increasingly used in non-medical fields, the judgment of people’s potential conditions based on predictive genetic information inevitably causes genetic discrimination (henceforth GD). This article aimed to systematically investigate the disparity in attitudes and worrying scenarios concerning GD in China. Methods: A questionnaire survey of 555 respondents was conducted. Statistical tests were used to examine disparity in attitudes between gender, age, and education. A descriptive analysis was also conducted to explore other worrying scenarios. Results: It shows that (1) men are more tolerant of GD compared to women, and (2) participants aged between 18 and 30 years old possess the highest objection to GD. However, (3) no indication can attest to the relationship between educational level and perspective on GD. In addition, (4) the acceptance of gene testing in the three most common scenarios is ranked in descending order as follows: partner choice, insurance services, and recruitment. Moreover, (5) worrying scenarios relating to GD include: education, social occasions, medical services, fertility, shopping, and so on. Conclusions: Based on the results, suggestions proposed include developing a blacklist mechanism in the field of genetic data application and strengthening the security regulations for the commercial use of genetic data. In line with the development of gene technology since the 1980s , the usaRecruitment , insuranMeasures and regulations have been used by some countries to address the issues associated with GD. Research shows that North America and Europe are the regions with extensive policy-making activities, while Asia has moderate policy-making activities related to GD . In 2008What are the differences in the attitude towards GD among diverse types of people in China? Which scenarios of GD are of concern to Chinese people? In an attempt to answer these questions, we conducted an online questionnaire survey to explore the disparities between attitudes and worrying scenarios of GD and to propose some suggestions for balancing concerns about GD with technology development.Previous research shows that Chinese people were willing to explore using genetic information for disease prevention, while there was less awareness of the side effects  . NowadayThe paper is structured as follows: https://jinshuju.net (accessed on 1 May 2022), a professional survey company), and was distributed through social media platforms QQ and WeChat developed by Tencent. Tencent’s 2020 financial results show that QQ has 693.5 million monthly active accounts on smart terminals, and WeChat has 1.225 billion monthly active accounts on smart terminals.To systematically study GD concerns in China, an anonymous questionnaire survey was conducted from 27 May to 29 September in 2019. The questionnaire is hosted online by the “Jinshuju” website , consisting of four single-choice questions and one open question: (1) participants’ perspectives on GD; (2) participants’ willingness about participating in gene testing for job application related situations; (3) participants’ attitudes about attending gene testing in insurance services; (4) participants’ willingness about conducting gene testing in partner choice; and (5) participants’ understanding of other circumstances that may occur with regards to GD. In addition, three extra questions were used for the collection of demographic information  about the respondents. The overall setting of the questionnaire is represented in There are 14 questions in the questionnaire. In this paper, part of the questionnaire findings are reported  and the groups with a lower educational background did not fully complete or return the questionnaire, perhaps because they are not familiar with gene testing and GD and lack the knowledge to finish the survey. China’s illiterate population accounted for 4.59% of the population aged 15 and above in 2019. Gene testing and GD are unfamiliar to these sections of the population in China. Genetic technologies have increasingly developed only in the last 30 years [0 and theTo statistically investigate respondents’ views on gene testing and GD, a two-step difference analysis was conducted. The specific analyses involved the following three aspects:The disparities in attitude towards GD among different demographic groups, which contain factors of gender, age, and educational level, were investigated. Since the information collected was discrete random values and was not normally distributed, the chi-square test was selected, which is widely applied in clinical statistics  and suitA significant difference in willingness among participants to conduct gene testing in three real-life scenarios was identified: recruitment, partner choice, and life insurance. When there are two datasets being contrasted, the marginal homogeneity test, which is an extension of the McNemar test for two dependent sample studies, will be used to identify the differences. The Friedman test is used to detect the differences among the above three scenarios.A descriptive analysis contrasts the replies to the open-ended question, which was conducted to further explore the respondents’ concerns about other scenarios where GD might occur, the results of which are shown in There are 48.6% of the respondents who disapprove of GD and think it is unreasonable. Only 9.4% agree with GD and think it is reasonable. The rest of the respondents (42.0%) hold the opinion that the existence of GD depends on the situation, and their attitude varies depending on the application. To further compare the disparities in people’s views on GD, the specific investigation was undertaken in different demographical groups.p-value of 0.000 (<0.05), where men are more tolerant of GD than women. As shown in The attitude difference on GD between genders is statistically significant with a p = 0.000). Compared to the respondents aged between 18 and 30 years old, the ones aged 31–45 tend to accept GD in real life. The disparity between the two groups is most conspicuous with a p-value of 0.000. The approval rate of GD in the 18–30 year old group is 2.2%, while the one in the 31–45 year old group is 13.9% (The results show that there is a significant difference among different age groups (is 13.9% . Moreovep = 0.802 > 0.05), and there is little evidence to prove the correlation between an individual’s educational level and their perspective on GD according to the test.The differences between the five educational groups were not distinct. People with no formal high school qualifications have a slightly higher rate; with 16.7% thinking that GD is reasonable see . Most ofp-values of 0.000 and 0.000, respectively, which indicate that the respondents show significantly higher acceptance to attend gene testing in partner choice when compared with the contexts of recruitment and life insurance. Therefore, it can be concluded that partner choice is the area where the respondents are most inclined to adopt gene testing out of these three key scenarios.Respondents’ concerns about GD are not stereotypical in different contexts. Analysis shows that the acceptance of gene testing in partner choice is highest in the areas of recruitment and insurance services. As shown in Respondents’ understanding of other circumstances where GD might occur includes education, social, medical, and so on. Respondents were required to write down other scenarios in which they thought GD might occur. Some of the respondents left the answer blank or repeated one of the three scenarios studied above. Only 121 replies were valid in answering the question, among which two respondents gave two answers; thus, there are 123 valid answers in total for this question. The composition of the answers collected is shown in Based on the above statistics on the disparity between respondents’ views on GD and gene testing, several analyses of the reasons for these disparities were conducted. Then, we discuss the contributions and limitations of this paper.Women have more reservations about discrimination based on genes. On the one hand, it is probably because women are the ones who get pregnant and give birth, which to some extent influences their willingness to undergo gene testing. On the other hand, men, who tend to focus more on promotion and earning money for the family, might be more concerned about the health indications derived from gene testing to help them better prevent potential diseases. It might also be attributed to the willingness to accept innovation, where men show more interest in new technologies , while wThe knowledge of genes is updated very quickly, and many of its contents have not been studied systematically by previous students. Thus, people in the older group might have been less well educated about the knowledge of genes and GD when they were students. Moreover, people aged 31–45 years are more mature and likely more sophisticated on such discriminations. As a result, the younger age group is inclined to show more sensitivity and opposition toward GD.Among respondents with different educational backgrounds, those with a doctoral degree have the highest disapproval rate for GD. This might be attributed to the abundant educational experiences providing them with more opportunities to access relevant research and experiments, which may emphasize the negative effects of GD.Through the respondents’ acceptance of genetic testing in different scenarios, concerns about GD are reflected. In China, prenatal genetic diagnosis by gene sequencing  and geneAccording to the statistical results, discrimination in education is of the greatest concern to the respondents. The concerns in this area mainly focus on the school admissions-related process. Competition for admission to basic education and higher education in China is fierce. The prevalence of genetic testing poses a high risk of unfairly depriving predisposed students of their rights and opportunities to receive basic or higher education. Such discrimination based on gene testing information may result in obvious inequities in education , which hGD on social occasions is of great concern to the respondents. The discrimination based on genetic information may lead to vicious isolation in daily meetings. Persons with abnormal or criminal genetic composition are at risk of being ostracized by society or being exposed to violent treatments . The strMedical treatment is the third controversial scenario where GD occurs frequently. Since the 1990s, insurance has been the primary concern for the general public due to adverse selection based on genetic test results . Some meWorries about GD in other real-life scenarios such as fertility, leisure activities, and the sports industry have also been put forward by the respondents. The conduction of genetic testing gives rise to inequality by depriving certain groups of people of their pregnancy and fertility rights. In addition, artificial interference in genetic information such as genetic editing in embryos is of great concern to the public . Some reShopping is another field where genetic data is widely used. Businesses analyze consumers’ genetic data to price differently and customize discriminatory services based on their preferences. In the sports industry, the prevalence of gene testing poses a negative impact on justice . The selSome respondents also state the influence of GD in personal career promotions. The advancement of their careers might be negatively influenced by the consequences of gene testing. Since employers might seriously consider the talent development cost as well as the potential risk based on the predictive health condition of their staff, they are more inclined to select the individual who has proven higher talent and “safety”. In addition, genetic testing might be used by employers to select employees who will always be at work every day and potentially always be available to work. Many people may lose job opportunities .As for their concerns about the effect of GD on politics, people worry that the division of parties would be determined by personal genetics. People with similar genetic compositions would aggregate together, which is bad for national and ethnic solidarity.According to all the concerns stated above, the wider application of gene testing deserves wider attention and careful consideration. Genetic technology can benefit our lives only if the data has been applied appropriately and scientifically. Furthermore, consistent control of the occurrence of GD, as well as the prevention of privacy infringement, is of great necessity to establish a better environment for the developments of genetic technology in society.By answering the research question, “What are the differences in the attitudes of GD between different crowds of people in China? What are the scenarios of GD concern by the Chinese?” we claim as theoretical contributions to the wider discussion of GD literature. Our major contribution to GD literature includes: (1) having analyzed the attitudes of different groups in China towards GD; (2) having analyzed attitude disparities between three key scenarios, namely, insurance, partner choice, and recruitment; and (3) having explored other scenarios where Chinese people worry that GD might exist and impact their lives. This is a significant basis for follow-up related research, such as genetic technology acceptance, genetic technology diffusion, and gene industry research, especially as China is a major player and has the largest market in this field. Existing studies have put forward the idea that different groups have different concepts of GD, but few studies have been verified from an empirical perspective. This paper investigated it in the form of a questionnaire survey.The observations generated in our analysis also have practical implications for genetic technology and industry development. This study provides policy enlightenment not only for China but also for other countries. China has always upheld a relatively open attitude towards genetic technology. To some extent, the case of He Jiankui, a Chinese biophysics researcher who edited the genes of twin babies and was jailed for 3 years , caused Excessive faith in the predictions of genetic information leads to discrimination, bringing social and psychological risks  to indivNowadays, using genetic data for differential pricing in the insurance field in China has been banned. Similar systems should be set up in other fields as well. For instance, in order to protect labor rights, the application of genetic data should be prohibited in the hiring process; in order to maintain education fairness, the application of genetic data should be prohibited in the educational field; in order to avoid social isolation, private genetic data should not be open to the public; in order to protect personal medical data from being accessed and to avoid unfair treatment of patients, the management of medical gene data should be stricter; and in order to guarantee fair competition, sports competitions should not use genetic data to select players. To safeguard the diversity of the human gene bank, gene screening at the reproductive stage should be regulated, lest the excessive use of genetic data bring devastating disaster to human beings.Enacting regulations and laws to restrict the inappropriate use of genetic information and safeguard personal privacy is imperative. In order to promote industrial self-discipline and integrity, first, ethical guidelines on the application of gene testing are urged by Chinese geneticists for the improvement of genetics services in China, as well as filling the cultural gap between China and Western countries to reach an agreement on the ethical, legal, and social issues of genetics in the future ,52.Initiating genetic non-discrimination laws would protect the public from social discrimination and ease their concerns about GD, thereby enabling them to benefit from genetic technologies . In the Inevitably, there are some deficiencies in this research. Firstly, the representativeness of the sample is not guaranteed. The format of an online questionnaire survey excludes some groups of peoples, such as the elderly and those who are not able to conveniently access the Internet. This affects the representativeness of sampling. China has a large population that does not use social media. Secondly, the questionnaire design was not sufficiently refined to allow for explanatory research. This study mainly describes and analyzes the differences in attitudes towards GD among people of different genders, ages, and educational levels. It has not yet analyzed in depth the reasons for the differences. Additionally, the income data was not included in the design of the questionnaire since it is so sensitive that it may cause some respondents to be unwilling or unable to complete the questionnaire. What’s more, open-ended questions were posed for the study of other scenarios. This makes variance analysis impossible, which is significant for determining the different views of different groups.Through the form of a questionnaire survey, this study has analyzed different attitudes of various groups towards GD in the scenarios of partner choice, recruitment, and insurance purchase, as well as other real-life scenarios where Chinese people worry that GD might exist. To facilitate the benign development of industrial ecology, some pieces of policy advice are also suggested. With regard to future research, some in-depth research can be conducted. For example, insurance can be divided into health and life insurance, and thus a differentiation study can be conducted. Regarding partner choice, an in-depth interview can be carried out to study the causes of differences in views among different groups. What’s more, when it comes to specific scenarios, fairness in education, which is of great concern to the Chinese people, needs a special study in the future.


Migratory hoverflies are long-range migrants that, in the Northern Hemisphere, move seasonally to higher latitudes in the spring and lower latitudes in the autumn. The preferred migratory direction of hoverflies in the autumn has been the subject of radar and flight simulator studies, while spring migration has proved to be more difficult to characterize owing to a lack of ground observations. Consequently, the preferred migratory direction during spring has only been inferred from entomological radar studies and patterns of local abundance, and currently lacks ground confirmation. Here, during a springtime arrival of migratory insects onto the Isles of Scilly and mainland Cornwall, UK, we provide ground proof that spring hoverfly migrants have an innate northward preference. Captured migratory hoverflies displayed northward vanishing bearings when released under sunny conditions under both favourable wind and zero-wind conditions. In addition, and unlike autumn migrants, spring individuals were also able to orientate when the sun was obscured. Analysis of winds suggests an origin for insects arriving on the Isles of Scilly as being in western France. These findings of spring migration routes and preferred migration directions are likely to extend to the diverse set of insects found within the western European migratory assemblage. InsecMigratory hoverflies can be hugely abundant and are particularly valued as pollinators and predators of crops pests such as aphids ,6,13,14.To investigate if spring migratory hoverflies have an innate direction preference, and to see if this direction is influenced by the visibility of the sun or wind direction, we undertook a series of experiments to measure the vanishing bearings of captured, pre-reproductive, migratory hoverflies following their arrival into Cornwall and the Isles of Scilly, UK, in mid-June 2022. We predicted that: (i) hoverflies will continue to orientate northwards in a seasonally favourable direction when given a view of the sky with the sun; (ii) they will lose this ability when the sun is obscured; and (iii) they will use favourable winds to aid their flight north.. 2 (a)Syrphus vitripennis) and Lepidoptera, appeared on the Isles of Scilly and mainland Cornwall between 16 and 21 June. We captured 66 migratory hoverflies and performed vanishing bearing experiments on 16 (n = 4) and 17 (n = 25) June in the Isles of Scilly. Following these island experiments, we conducted further vanishing bearing experiments in Cornwall on 20 (n = 20) and 21 (n = 17) June. Five migratory species were caught: Syrphus vitripennis (n = 45), Episyrphus balteatus (n = 13), Scaeva pyrastri (n = 4), Eristalis tenax (n = 2) and Syrphus ribesii (n = 2) (a–c). Captured hoverflies were placed into a mesh insect cage and experimented upon within 15 min of capture.In 2022, a large influx of migratory insects, consisting of Diptera (the majority being  (n = 2) a–c. Capt (b)d); and (iii) during a thick sea mist that obscured the sun and with wind (e).Two researchers sat east and west, respectively, of the cage of captured hoverflies. Hoverflies were removed from the cage by hand, before being held in the air and released above the researchers' heads. In general, the hoverflies waited longer than 2 s before leaving, while on three occasions, the hoverflies left instantly. On departure, both researchers recorded the heading until the hoverfly was no longer visible (approx. 20 m), and this angle was recorded as the vanishing bearing. Headings were determined using a compass and/or the compass function on a Garmin Instinct Solar watch. Hoverflies were released during three distinct meteorological conditions: (i) sunny with no wind; (ii) sunny with wind d; and (iith wind e. (c)d). On the Isles of Scilly, the experiments were performed between the hours of 16.00 and 17.00 during a thick sea mist (e). At all locations, experiments were performed on a large open area and the migratory insects (released at least 40 m away from any landscape features above 5 m in height) always had a clear, unobstructed view of the sky.Experiments took place in two locations: the Isles of Scilly  and around the Exeter University campus in Penryn, Cornwall . At the sites around the Penryn campus, the experiments were performed in full sunlight between the hours of 11.00 and 16.00 d. On thesea mist e. At all (d)S. vitripennis females was examined under a dissection microscope.Hoverflies were examined in the field by eye to estimate reproductive state based on the fullness of the abdomen. In addition, the reproductive state of two  (e)Meteorological conditions were recorded on location at the same time of the experiments. The on-location measurements were cloud cover , sun visibility, windspeed  and wind direction. (f)www.ventusky.com) to visualize winds at 500 m altitude above sea level on the days leading up to the largest influxes.To estimate the potential flight origin, we used Ventusky ( (g)S. vitripennis females and all other hoverfly migrants. All data are provided in electronic supplementary material, file S1.Statistical analysis and graphing were carried out in R v. 3.5  using R . 3θ = 2.3°, Rayleigh test: r = 0.59, p < 0.0005, n = 41, 95% CI 343.5°–21.1°, figure 1f) after an average orientation period of 2.2 s (n = 30). Windspeed during recordings when the sun was visible ranged from 0 to 6 m s−1 (mean = 1.5 m s−1). Under zero-wind conditions, hoverflies headed in a north-northeasterly direction . There was no significant difference between the distribution of these vanishing bearings . When wind and sun were present simultaneously, the wind direction was either from the west , the south-southeast  or the east . Under these mixed windy conditions, hoverflies headed on average to the north-northwest . Owing to low replicates under westerly and easterly wind conditions, we were unable to test for an effect of wind direction on hoverfly vanishing bearings during sunny conditions. However, there was no significant difference between the distribution of south-southeasterly wind headings and vanishing bearings under these conditions . Released when the sun was obscured by a thick sea mist, and with a southwesterly wind (238°) blowing at 1.9 m s−1, hoverflies headed in a northeasterly direction . There was no significant difference between the distribution of wind headings and vanishing bearings under these conditions .Released when the sun was visible, hoverflies headed almost due north , two were S. ribesii , 13 were E. balteatus , four were S. pyrastri  and two were E. tenax . A Mardia–Watson–Wheeler test indicated a lack of significant directional bias by species and sex when comparing the distribution of S. vitripennis females with the remainder of the individuals . To estimate the potential origin of the migratory hoverflies, we visualized the wind conditions at 500 m above sea level at 10.00, 13.00 and 16.00 on the day preceding the largest influx. These wind conditions suggest an origin for these individuals in France (Mixed sexes and species of migratory hoverflies were analysed in this study. All females examined showed abdomens without significant egg development, and dissections of two female n France .Figure . 4The utilization of favourable winds has been investigated using radar studies in migratory hoverflies . This stFlight simulator experiments in autumn migrant hoverflies show that they fail to orientate south when the sun is obscured, indicating that other available cues are not used, at least in this experimental set-up . Surpris−1 [Our analysis of winds suggests that migrants arriving on the Isles of Scilly began their journey in western France, representing a minimum sea crossing of nearly 200 km . Radar s−1 . We noteThe insect migration assemblage is highly diverse, and the findings presented here on orientation behaviour and migratory routes undoubtedly extend to other members of the Syrphidae and perhaps other co-migratory Diptera and Lepidoptera that arrived together in the southwest of the UK. Many of these migratory insects play important ecological roles ,6,13; th


Genome-wide association studies (GWAS) of complex traits, that is, phenotypes that are influenced by many genetic variants, have discovered thousands of genetic loci  underlyin1s = n10 + n11 individuals with OSA in the population, n11 individuals are indeed observed to have OSA, and n10 individuals appear to have no OSA, despite having OSA. Define the misclassification rate as n10/n1s, the proportion of individuals with OSA who are erroneously classified. Using the same notation, the prevalence of OSA in the healthcare system or study is n1s/(n1s + n0s) = n1s/n. I performed a simulation study to demonstrate how misclassification of OSA may bias genetic effect estimates and reduce power (see https://github.com/tamartsi/OSA_misclassification for code). Using a simple logistic regression model, I assumed that OSA probability depends on a population-based constant, the intercept Misclassification of OSA reduces the power to discover genetic associations and biases effect-size estimates, in a manner depending on the OSA prevalence and on the misclassification rate. n = 20 000 in each simulation iteration.The simulations had To address the reduced power caused by OSA misclassification, Campos et al.  performeThe principle of leveraging genetic associations with OSA-related traits to discover OSA-specific genetic associations is useful. It could be extended to excessive daytime sleepiness (EDS), the most common presenting symptom of OSA , to insoAs shown in Knowledge about the specific OSA consequences associated with the variant can be leveraged, with the development of an appropriate statistical method, to compute unbiased effect size estimates for the variant-OSA association.


Solanum villosum (Sv) and Solanum scabrum (Ss). Spontaneously fermented (SF) and controlled fermented (CF) conditions using Lactobacillus plantarum LP90 and Leuconostoc mesenteroides LM58 were employed for 15 days and 120 h. From the fermented pickles, relish products were prepared using cooking oil and a variety of spices. The relish products were subjected to a consumer acceptability test. Results show a significant drop in pH to <3.5, increasing titratable acidity (TTA) to around 0.6 after 120 h and 15 days of CF and SF, respectively. LAF resulted in a 2.6–5 and 1.6–4.8‐fold significant rise in β‐carotene in pickles and their relish products. All pickles and relish products exhibited a significant decrease (p < .05) in vitamin C by 88.33%–95.90%. LAF significantly reduced total phenolic (26%– 43%) and Chlorophyll (16.45%–39.25%). On the other hand, LAF showed improvement in minerals content  and reduction of tannin (76.27%–92.88%) and oxalate (77.33%–90%) levels. LAF relish products were highly preferred by the consumers, with S. villosum controlled fermented relish (SvCFR) leading. All fermented relishes were stable at ambient (27°C) and refrigeration (4°C) temperatures after 6 months of storage. Generally, LAF is an effective method for ANS preservation, with improved nutritional quality and safety. LAF can therefore be recommended to small‐scale farmers, processors, and households for ANS preservation. Ultimately, this method enhances the nutrition and sensory quality, safety, and livelihood.African nightshade (ANS) is among many underexploited and neglected indigenous vegetables. This study assessed the effect of lactic acid fermentation (LAF) on nutritional and sensory quality in  Lactic acid fermentation (LAF) improved β‐carotene and minerals contents but reduced vitamin C, total phenols, and Chlorophyll levels. Also, it reduces antinutrients, tannin, and oxalates. Relish products were stable at ambient and refrigeration temperatures after six months of storage and consumers preferred all relish products. African nightshades (ANSs) are a good source of essential minerals , vitamins C, and β‐carotene. Also, they contain bioactive compounds such as lutein, zeaxanthin, polyphenol, flavonoids and have medicinal properties  are among widely distributed and consumed African indigenous vegetables (AIVs) in Tanzania Keller, . SolanumLeuconostoc mesenteroides, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus acidophilus, and Lactococcus lactis in ANS fermentation, has been reported  were collected in Arumeru District, Arusha, Tanzania. The leaves were transported under cold storage to the Food Processing Unit at the Nelson Mandela African Institution of Science and Technology (NM‐AIST) for further processing. Other ingredients, such as cooking oil, onions, and a variety of spices , were purchased in Arusha, Tanzania. Freeze‐dried probiotic Lactobacillus plantarum LP90 (100 billion CFU (colony‐forming units)/g) and Leuconostoc mesenteroides LM58 (100 billion CFU/g) were obtained from the Wuhan Healthdream Biological Technology Co., Ltd. The starter culture was stored in the freezer at −24°C.Africa nightshades (ANSs) (2.2S. scabrum (Ss) and S. villosum (Sv) leaves were sorted, washed, and then placed in a perforated vessel for water drainage before weighing. For controlled fermentation (CF), the leaves were blanched at 90°C for 5 min and immediately cooled in ice water  were inoculated separately according to Wafula (S. scabrum controlled fermented (SsCFP)) and Sv (S. villosum controlled fermented (SvCFP)) pickles were obtained. In SF, unblanched Ss and Sv leaves were likewise submerged into a brine solution  and incubated at 25°C for 15 days  and S. villosum spontaneous fermented pickle (SvSFP). The pH and titratable acidity (TTA) were recorded daily for SF and CF.In CF, blanched Ss and Sv leaves were submerged into a brine solution , and o Wafula  with sli Wafula,  with somS. scabrum controlled fermented relish pickle (SsCFRP), S. villosum controlled fermented relish pickle (SvCFRP), S. scabrum spontaneous fermented relish pickle (SsSFRP), and S. villosum spontaneous fermented relish pickle (SvSFRP), respectively. The relishes were then cooled, packaged in PET bottles, and stored at refrigeration and ambient temperatures for shelflife study.Pickle products SsSFP and SvSFP from SF and (SsCFP and SvCFP) from CF were cooked for 10 min mixing with cooking oil, onions, and different spices  employed in relish‐making. Four relish products of 2.4Titratable acidity (TTA) was determined, as described by Rajković et al. . About 12.5The total vitamin C content of the fresh leaves, pickle, and relish products was determined  were analyzed according to the method of , yeast and mold (potato dextrose agar), Lactobacillus  agar), and coliform (violet red bile agar) . The incubation conditions were: bacteria , molds and yeasts , coliforms , and Lactobacillus  at anaerobic conditions.Total bacteria, fungal counts, coliform, and i et al. , Rahimi i et al. , and Sini et al.  with sli2.12The shelflife study of relishes was conducted at refrigeration (4°C) and ambient temperatures according to Pala and Agnihotria , with so2.13S. scabrum spontaneous fermented relish (SsSFR), S. scabrum controlled fermented relish (SsCFR), S. villosum spontaneous fermented relish (SvSFR), and S. villosum controlled fermented relish (SvCFR), were tasted for consumer acceptability. A 9‐scale Hedonic test was used  in Kilimanjaro and Morogoro District Council  in Morogoro regions with high and low ANS production. About 370 untrained panelists, mainly ANS consumers, participated in the sensory evaluation of the relish products. They were selected based on their history of consumption of ANS. Four relish products, i.e., 2.14SD. SPSS software version 25 was used for data processing. The one‐way analysis of variance (ANOVA) and post hoc test (least significant difference) were used to compare the mean between the species of Solanum for the fermented pickles and relish products, and the differences were considered significant when p < .05.Experiments were conducted in triplicate, and data were expressed as mean values ± 33.1S. scabrum and S. villosum) at an initial pH of 7.4 were fermented spontaneously and under controlled conditions for 15 days and 120 h, respectively. A sharp drop in pH to 4.7 and pH to 3.5 was observed within 24 h for both spontaneous fermentation (SF) and controlled fermentation (CF), respectively , Weissella cibaria (21), Leuconostoc pseudomesenteroides (56), W. cibaria (64), or L. plantarum (75), in ANS fermentation had been reported   β‐carotene values than S. scabrum as opposed to their relish products. Likewise, for relish products, S. villosum had significantly high β‐carotene, unlike S. scabrum , oxygen, alkaline pH, and high temperature  in SsSFP and Chlorophyll b in SsSFP, SsCFP, SvSFP, and SvCFP, respectively , or phenolic compounds’ chemical oxidation masking the original green color of the leaves and reducing the Chlorophyll content , W. cibaria (64), or L. plantarum (75). Furthermore, after fermentation, a reduction of total polyphenol content to 3822.5–5681.5 mg/kg L. pseudomesenteroides (56) and Weissella cibaria (21) was noticed. The decrease in total phenolic compounds in fermented nightshade leaves can be due to detoxification and utilization of phenolic acids as a carbon source  ion power to bind with divalent minerals such as Ca2+ ion to form insoluble oxalates  substantially reduced tannins (76.27%–92.88%) and oxalate (77.33%–90%) levels of pickles and relish products, respectively Figures  and 5. A3.5Solanum retroflexum leaves through the degradation of Chlorophyll a and b. Furthermore, low pH is associated with color destruction in fermented vegetables due to the production of acids, concurrently with findings  and refrigeration (4°C) temperatures are presented in Tables On the other hand, β‐carotene losses amount to 46%–62% and 20%–29% for ambient and refrigeration storage for the relishes Tables  and 6. D4SsCFRP product was the most preferred over other relish products. All relish products were stable at ambient and refrigeration temperatures after six months of storage, hence contributing to product safety. Therefore, LAF can be recommended as the best preservation method for African nightshades (ANSs). From the study, LAF has been shown to retain nutrients, improve safety and shelf life, offer diverse products, subsequently contributing to minimize postharvest losses.Lactic acid fermentation (LAF) enhanced ANS pickle and relish products’ nutritional, sensory, and safety quality. Specifically, LAF improved β‐carotene and minerals contents but reduced vitamin C, total phenols, and Chlorophyll levels. LAF also reduced tannins and oxalate. The The authors declare no conflict of interest.Frank Sangija: Conceptualization (lead); Data curation (lead); Formal analysis (lead); Methodology (lead); Resources (supporting); Software (lead); Validation ; Writing – original draft (lead); Writing – review & editing . Haikael Martin: Conceptualization (supporting); Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (supporting); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (supporting); Validation (supporting); Writing – original draft (supporting); Writing – review & editing (supporting). Athanasia Matemu: Conceptualization ; Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (lead); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (lead); Validation ; Writing – original draft (supporting); Writing – review & editing (lead).


Genetic diversity among and within populations of all species is necessary for people and nature to survive and thrive in a changing world. Over the past three years, commitments for conserving genetic diversity have become more ambitious and specific under the Convention on Biological Diversity’s (CBD) draft post-2020 global biodiversity framework (GBF). This Perspective article comments on how goals and targets of the GBF have evolved, the improvements that are still needed, lessons learned from this process, and connections between goals and targets and the actions and reporting that will be needed to maintain, protect, manage and monitor genetic diversity. It is possible and necessary that the GBF strives to maintain genetic diversity within and among populations of all species, to restore genetic connectivity, and to develop national genetic conservation strategies, and to report on these using proposed, feasible indicators. Destruction of habitat, overharvest, and other societal activities are leading to widespread and precipitous declines in genetic diversity , an international treaty among nearly all countries  to conserve, sustainably use, and share benefits arising from biodiversity. The CBD had multiple frameworks since it came into force in 1993, including the Strategic Plan from 2002 to 2010, with four strategic goals for biodiversity and underlying targets for each CBD ; the StrTo navigate developments in the post-2020 GBF text with respect to genetic diversity, over the past three years, we provide a scientific synthesis of its past, present and possible future status. We aim to identify specific and science-informed improvements that could strengthen the GBF. We describe: the progression of wording around genetic diversity in GBF goals and targets, up to October 2022; highlight issues to be resolved in the final GBF draft, with suggestions for resolving them; share lessons from participating in this process; and reiterate connections between GBF wording and indicators to measure progress under the monitoring framework of the GBF.Box 1 Reflections on pre-2020 commitmentshttps://www.cbd.int/convention/text/) outlines many commitments—in situ and ex situ conservation, sustainable use, protected areas, research, public education—needed for the “conservation of biological diversity… [which] includes diversity within species, between species and of ecosystems.” The 2002–2010 commitments and the Strategic Plan for 2011–2020 contained a high-level goal and a target for genetic diversity  is needed to maintain unique adaptations to different local environments, while genetic diversity within populations is needed to allow each population to avoid inbreeding and quickly respond to changing conditions .Vital elements of the current text are worth highlighting. Specifically, genetic diversity Improvements in the post-2020 GBF draft are laudable, and we believe reflect an increasing acceptance among CBD policy makers of the importance of genetic diversity and of the demonstrated feasibility of measuring it with simple indicators. However, the current wording misses some elements, and so refining the text is critical to retain key principles and ensure robust monitoring and reporting, while avoiding perverse incentives and potential loopholes.Goal A currently includes maintenance of genetic diversity “within all species” or “within wild and domesticated species.” Either phrase is suitable, as both emphasize that genetic diversity within all species matters .The ambition remains insufficient at 95%, as highlighted by Frankham Frankham , who shoTherefore, a more scientifically justified Goal would be “maintain at least 99% of within population genetic diversity and maintain all distinct populations,” or “maintain sufficient genetic diversity for adaptive potential within populations and maintain all distinct populations.” For most populations, ‘maintain sufficient genetic diversity for adaptive potential’ implies near zero loss of current genetic diversity  which can be reported on using indicators for effective population size of 500 within each population to mitigate loss from genetic drift (see Implementation and Reporting below). No loss is consistent with CBD’s Mission (“To take urgent action across society… to put biodiversity on a path to recovery by 2030 for the benefit of planet and people” and Vision (“living in harmony with nature”), especially as many species already suffered high genetic diversity loss .); or targets on connectivity could explicitly add "genetic’  is also not mentioned as yet. In many situations, e.g., recent population fragmentation, maintaining genetic diversity will require appropriate levels of gene flow (which may necessitate restoring habitat connectivity or implementing periodic translocations). Therefore, it is recommended to add gene flow explicitly to the current Goal A: “ensure appropriate genetic connectivity/ gene flow” Frankham ; or targHoban and colleagues . However, it is sometimes understood as also preventing hybridization, maintenance of allele frequencies, and sustainable levels of offspring production, as mentioned by Hollingsworth and colleagues , and calculations are feasible , though they may be better suited as indicators for Targets 7 and 10. Lastly, we propose a complementary indicator: the number of populations or species being monitored with DNA-based methods . Decision makers helped us evaluate the feasibility of proposals, understand perspectives of other decision makers, and present proposed wording.It is vital to build relationships with decision makers- persons present in the rooms where policy is drafted and negotiated. For example, it is necessary to correspond frequently with the national representatives attending CBD meetings .https://www.youtube.com/watch?v=Oku8eTqH_hE), dozens of email chains to various CBD stakeholders, a side event for COP 15 which was co-sponsored by 50 + institutions and NGOs, an information booth at COP 15, and ad hoc responses to many inquiries several multi-page comment submissions to CBD, and ad hoc responses to many inquiries.A diversity of engagement modes is critical: webinars, mass emails to policy makers, direct and frequent personalized emails, numerous digital meetings, journal articles, navigating bureaucracies for institutional approval, and frequent submission of document comments—often with deadlines of days to several weeks. We created three policy briefs, 13 journal articles, four Statements sent to 500 + recipients, a SBSTTA  information document, 12 + webinars ; increasing this engagement requires both greater efforts by scientists to understand and join in policy discussions and more frequent and accessible opportunities for scientists to enter the policy realm Laikre . Bridge Progress depends on intensive and steadfast commitment from several ‘champions’ in research and policy, far outside their normal job obligations, to create outputs, bring people and groups together, and constantly track many moving parts.Our involvement with the post-2020 GBF led to several lessons about conservation policy, some of which have been highlighted previously . Communication between scientific groups, the CBD Secretariat, Parties, the IUCN, IPBES , GEO BON, and other stakeholders during implementation of the GBF is vital for capacity development and shared learning, which will take significant time and effort. We close by emphasizing that although “Ne > 500”, “maintain populations” and other feasible indicators can be implemented at national scales, Parties are urged to remember the spirit of the goal: little to no genetic diversity loss so that populations, species and nature retain adaptive potential.


Lolium perenne L.) and annual one (Lolium multiflorum Lam.). Development and implementation of the modern methods of plant varietal and species identification are of great importance. In recent years, molecular markers have been successfully used for these purposes, which increase the accuracy of the breeding material evaluation at a significant reduction of time and labor costs. The aim of this study was to assess the discriminatory potential of the new SCoT marking technique for the identification of Russian perennial (Lolium perenne L.) and annual (Lolium multiflorum Lam.) ryegrass varieties.Ryegrass is a promising crop for the development of meadow farming in the world. More than 1000 cultivated varieties widely used in feed production have been developed, based on the main species — perennial ryegrass . The polymorphic ratio of the tested markers was 30.8%. The marker SCoT-06 was characterized by the maximum number of PCR products and the highest level of polymorphism (50%). The effective number of alleles (ne) ranged from 1.35 to 1.58 with a mean of 1.48 per locus. The average value of the PIC and Shannon index (I) were 0.35 and 0.46, respectively. The unique PCR fragments were revealed for the identification of tested varieties. Analysis of molecular variance (AMOVA) showed that the level of genetic diversity between ryegrass species (59%) was more than between varieties within a species (41%). Based on binary matrix data, clustering and PCoA analysis  of the samples were carried out that divided them into two groups according to species.We found a set of markers that can be useful tools for ryegrass varieties identification. The level of intravarietal polymorphism turned out to be higher than the differences between varieties because of the possible significant heterogeneity of the varietal material. The information obtained can be used in breeding programs to create improved ryegrass varieties adapted to Russian climatic conditions. Perennial grasses are the main plants in natural meadow phytocenoses of the middle, north, and steppes of Russia, and are also used to create seeded meadows and pastures in modern agriculture. The widespread distribution of cereal grasses is due to their valuable biological properties — ecological plasticity, high productivity, longevity, winter hardiness, and vegetative regeneration. When grown in single-species crops and in grass mixtures with a legume component, they provide low-cost and high-quality forage (green and conserved in the form of silage and hay) and are used for landscaping, recultivation, and soil protection from water and wind erosion .Lolium perenne L.) and annual (Lolium multiflorum Lam.) ryegrass. On the basis of these species, more than 1000 cultivars have been created that are of great importance for fodder production and the development of animal husbandry in many countries [Two of the most promising crops in the worldwide practice of grassland culture are perennial , however, improved tetraploid cultivars are common . The croLolium L. In particular, Bolaric et al. [With the introduction of modern DNA technologies, new opportunities have emerged to improve the accuracy of source evaluation and selection efficiency. Various types of PCR-based molecular markers are widely used to study representatives of the genus c et al.  analyzedc et al. . ISSR mac et al. . Using Sc et al. .Dactylis glomerata L.) [Bromus L. [In recent years, the SCoT (start codon targeted polymorphism) technology has been successfully used to differentiate and identify species and cultivars of cereal grasses. The high variability of markers provides a special structure of 18-nucleotide primers designed to amplify sequences in the genome flanking the ATG start codon . The mairata L.) , as wellromus L. . Also, tromus L. .In Russia, at the moment, forage grasses remain a poorly studied area of molecular genetic research. The breeding of this group of plants is based, in most cases, on the phenotypic labeling of natural genetic variations between and within ecotypes. This approach causes certain difficulties due to the great variability of traits and properties, the complexity of the genetic system, and a high degree of plasticity in the interaction of the genotype and environmental conditions. Until now, Russian ryegrass cultivars are distinguished exclusively by morphological characteristics, which are often similar, despite the high genetic heterogeneity of culture. As a result, the accuracy of assessing the purity of varietal material is reduced, and the breeding process is laborious and time-consuming. The widespread introduction of molecular labeling methods will make it possible to choose the most suitable breeding strategy for the development of cultivars for various purposes, adapted to certain environmental conditions.(Lolium perenne L.) and annual (Lolium multiflorum Lam.) ryegrass.The aim of this study was to assess the potential of the new SCoT marking technique for the identification of Russian cultivars of perennial The object of the study was 8 Russian ryegrass cultivars created by breeders of the Federal Williams Research Center of Forage Production & Agroecology (FWRC FPA) . To isolate DNA, we used “bulk samples” formed from 30 seedlings of each cultivar grown in Petri dishes for 7 days Table .Table 1TGenomic DNA was isolated by the SDS method  with somThe studied ryegrass cultivars were genotyped using 25 SCoT markers , 0.5U Taq DNA polymerase, 0.8 μM primer, and 30 ng DNA. The reagents that are used are produced by the Russian company Evrogen. Amplification was carried out on a T100 Thermal Cycler device  at the following mode: 3 min – 94°C; then 35 cycles of 1 min – 94°C, 1 min – 50°C, 1 min – 72°C; and 5 min – 72°C. A 1.6% agarose gel was used to separate the amplification fragments.The size of the obtained PCR products was determined using the Image Lab version 6.0.1 software package on a GelDoc XR Plus device  in comparison with a 1kb DNA Ladder molecular weight marker . Based on the data obtained, a binary matrix was formed where the presence of the product was designated as “1,” and the absence – “0”. In the analysis only distinct and reproducible bands were considered.The POPGENE program of version 1.32 was used to calculate the effective number of alleles (ne), the Shannon index (I), and the genetic similarity and Nei distances . AllelicThe set of 25 SCoT primers was used for genotyping of ryegrass samples, consisting of 30 genotypes per cultivar. More informative of them, generating distinct and reproducible amplification fragments, were selected for DNA polymorphism estimation Fig. .Fig. 1ElA total of 107 PCR products were obtained with an average of 13 amplicons per one marker. The amplified DNA fragments ranged from 349 to 2718 b.p.  and NTSYSpc v2.02 software Table . The minWe used the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) to investigate the relationships between the different samples. The cluster analysis and UPGMA dendrogram arranged the samples in the two groups Fig. . The firFigure In addition, analysis of molecular variance (AMOVA) was conducted to study cultivars’ genetic variations based on 25 SCoT markers. AMOVA showed significant interspecies differentiation (59%), while between cultivars within groups, this indicator was 41%.Cross-pollinated species, which include ryegrass, are characterized by a high degree of genetic variability between and within populations; therefore, an increase in the number of individuals included in the general sample increases the reliability of the obtained results. For ryegrass, the sample from a cultivar or population should include at least 24 genotypes to provide the coverage of the most common alleles [Lolium perenne L. equaled 46.88% while in Lolium multiflorum Lam. − 34.88%.The total number of obtained PCR products and the level of polymorphism were compatible with the results, obtained in the research studies of other authors. So, Tabaripour and Keshavarzi  at genotLolium L. using SRAP markers . Other indicators of genetic diversity, determined according to the results of our studies, such as PIC values (0.35) and Shannon’s index (0.46 versus 0.24) were also somewhat higher.The average number of alleles per locus (13) and effective number of alleles (averaged 1.48) in our study was a significant indicator of the genetic diversity of investigated samples and it was higher than the values obtained by Cheng et al.  when anaAnalysis of molecular variance (AMOVA) was used to test genetic variation among the studied ryegrass cultivars. We expected the high level of differentiations because of the cross-pollination of this specie and the nature of the cultivars: most of them were developed as complex-hybrid variety populations. The obtained data on the ratio of genetic variability degree corresponds to the results, represented by Tabaripour and Keshavarzi , althougThe clustering results divided ryegrass cultivars into two groups. In the first cluster tetraploid accessions, mostly obtained by polyploidization based on wild or old local diploid varieties, were located. For example, VIK 66 and Duet were derived using the original wild-growing forms from Karelia, Tambov, and Leningrad regions. The closeness of these cultivars on the dendrogram was probably due to the presence of common sources of initial material during the breeding process. Annual ryegrass Rapid was also obtained by polyploidization with colchicine treatment based on the Moskovsky 74 cultivar. These two cultivars were located in the second cluster of the dendrogram at a close genetic distance. Despite the similarity in genetic status, they differ significantly in morphological characteristics and productivity, which is 25-30% higher in the Rapid cultivar.When developing new cultivars of ryegrass, Russian breeders often use the method of creating synthetic hybrid populations based on crossing forms that are similar in morphological characteristics, but originating from different geographic zones. The number of initial forms during free hybridization can exceed tens. A characteristic feature of populations created in this way is the maintaining of heterosis effect for 4–5 generations and a high degree of intrapopulation variability. The cultivars Karat and Fenix were obtained by the method of complex-hybrid populations and located on the common branch of the UPGMA dendrogram.The performed PCoA analysis clearly demonstrates the division of the studied cultivars by species differences, as was observed during the cluster analysis, except that the PCoA allocated separately VIC 22 accession in the group of perennial ryegrass. This cultivar was obtained from the hybridization of breeding samples from the central part of Russia (Tambov region) and wild-growing forms from the Baltic. It is characterized by high adaptive plasticity, resistance to frequent and low mowing, and is suitable for lawn use. The morphological and biological characteristics of cereal grasses for different purposes are often accompanied by varying degrees of genetic variability, which are evident in the works of other researchers. For example, Bolaric et al.  reportedPairwise comparative analysis of genetic relationship indexes revealed the greatest genetic similarity between perennial ryegrass cultivars VIK 66 and Duet (0.86). On the contrary, the most distant were VIK 22 and Agat, with a value of the genetic distance of 0.46. According to the results of the clustering and indicators of Nei’s genetic distances (0.32), a close relationship between Agat and VIK 66 cultivars was not found, despite the common origin. Agat cultivar was obtained from VIK 66 on the experimental plot with an artificial infection loading by selecting the forms with high disease resistance. One of the reasons for the genetic remoteness may be explained by the high level of intravarietal polymorphism revealed during the genotyping of individual plants within Agat cultivar (37%). Besides, it is possible that some of the common rare alleles, presented in both cultivars, were dropped out of analysis and calculation due to the use of a total DNA probe for each sample, consisting of different genotypes  , 28.Molecular and genetic analysis of the major species of ryegrass using SCoT markers was carried out for the first time in Russia. In studying the genetic variability of varietal material, we tested the potential of the new technology.The research results indicate the high efficiency of SCoT markers for the analysis of complex highly heterogeneous ryegrass cultivar populations. Using markers of this type, it was possible to assess the level of polymorphism between cultivars of perennial and annual ryegrass, which amounted to 38.1% and 42.8%, respectively, with an average value of 30.8%. The indicators of the primer informativeness included in the analysis varied from 0.29 (SCoT 32) to 0.37 (SCoT 02, SCoT 23, SCoT 32, SCoT 35. Markers have been identified that can serve to identify cultivars within the tested sample. Thus, using the SCoT 13 marker, it was possible to obtain unique DNA profiles for 3 cultivars at once: VIK 66, Karat, and Duet. The SCoT 02 marker turned out to be suitable for differentiating the cultivars of the one-year-old ryegrass Rapid and Moskovsky 74, and the SCoT 06, SCoT 22, and SCoT 35 markers — for the Fenix, Agat and VIK 22 cultivars, respectively.The level of intravarietal polymorphism revealed in our study turned out to be higher than the differences between cultivars, which indicates a significant heterogeneity of the varietal material and corresponds to the data on the sources and methods of its production. At the same time, a high degree of genetic diversity, due to the heterogeneous nature of the culture, can serve as an important factor in the success of breeding programs.Thus, it was found that SCoT markers are highly polymorphic and have the necessary discriminatory potential for distinguishing between species and cultivars of Russian ryegrass breeding. The research results are of practical importance for use in varietal identification and inbreeding cultivars for different purposes adapted to the various environmental conditions of the Russian Federation.