Tauroragus derbianus derbianus) and non-native Least Concern Cape eland (Taurotragus oryx oryx) bred under different management strategies were studied in Senegal, Western Africa. The aims of the study were to compare the population genetic parameters of the two species and to test for the presence of interspecific hybrids. In total, blood and tissue samples from 76 Western Derby elands and 26 Cape elands were investigated, using 12 microsatellite markers. No hybrid individuals were detected in the sampled animals within the multispecies enclosure in Bandia Reserve, Senegal. The parameters of genetic polymorphism indicated much lower genetic diversity in Western Derby elands compared to Cape elands. On the other hand, the coefficient of inbreeding was low in both species. It is hypothesized that this could be a positive effect of strict population management of Western Derby elands, which, despite the loss of genetic diversity, minimizes inbreeding.Breeding management of small populations may have a critical influence on the development of population characteristics in terms of levels of genetic diversity and inbreeding. Two populations of antelope sister species – Critically Endangered Western Derby eland ( Problems connected with maintaining diversity are common in captive populations, whose sizes are limited by space and the individuals are scattered among institutions worldwide so the gene flow is restricted5, which can vary from intensive interventions7 to simple monitoring8. However, for the decision-making process, it is essential to know some basic information about the kinship and genetic variability of the individuals9 which is usually recorded in studbooks. Even if the studbooks for captive wildlife exist, for example, European studbooks10 or International studbooks11, knowledge of kinship across these populations is often limited and can be as low as only 50% or less in some ungulate species kept in zoos12.To eliminate the negative impact of the phenomena affecting small populations (e.g. loss of variability due to genetic drift and inbreeding), it is necessary to apply appropriate genetic managementex situ conservation programmes have been created, such as the European Endangered Species Programmes (EEPs) for species like the Cuvier’s gazelle 13, or the recently established EAZA Ex situ Programmes10.However, there are also species with better background information recorded, where a high proportion of their pedigree is known (% PK). These are mainly endangered species for which special 14 but even the studbooks do not guarantee reliable information concerning genetic parameters of polymorphism within the populations. The reason for the lack of reliable genetic data is mostly because the studbook analyses rely on the assumption that the founders are unrelated and non-inbred, which is not always the case15. Only genetic monitoring can reveal the true genetic polymorphism of populations16, including the ones without studbooks8.Pedigree data are especially valuable in the evaluation of ancestry and kinship17. Even if the hybridization occurs in nature, it can also be caused by human interference, usually by keeping related, but initially geographically isolated, taxa together; for example, in wildlife reserves or mixed-species exhibits in zoos, which is currently very popular20.Loss of genetic variability is not the only issue of genetics in captive breeding of wildlife. The populations can face problems with interspecific hybridization, which is considered as a very important factor that endangers biodiversity and the existence of many speciesSmall isolated populations of two eland species with different conservation statuses and different geographical origins are kept in Bandia Reserve in Senegal. Both populations had a similar number of founding animals, but they have been managed differently and thus may serve as an ideal model for evaluation of the influence of population management on the genetic parameters of the population.Taurotragus derbianus derbianus) are represented worldwide by less than 200 free-ranging individuals in Senegalese Niokolo Koba National Park (NKNP)21. A unique conservation programme for this antelope takes place in two Senegalese wildlife reserves, Bandia and Fathala, where about 100 of these antelopes live in semi-captive conditions22. When their founders were captured in NKNP in 2000, the total WDE population was already very limited and WDE was considered as Endangered. The population in NKNP was estimated to total only around 100 individuals at that time23 and all the founders of the captive programme were captured from just one herd24. The founders were transported to Bandia Reserve where the first captive breeding of WDE started. In 2006, the primary selected animals from Bandia Reserve were transported to Fathala Wildlife Reserve25.Western Derby elands  were introduced to Bandia Reserve in 1996 to increase its attractiveness for safaris for visitors. The only management applied in the population of CE in Bandia Reserve involves the culling of surplus males and older calf-less females for meat production. Since the initiation of both breeding programmes, no animals have been imported into the populations of WDE and CE. For a comparison of the background of both studied populations, see the overview in Table According to the WDE Conservation Strategy, the conservation programme’s “…aim is to manage the population to retain as high as possible genetic diversity…”26. The length of pregnancy in the WDE is approximately 9 months25, so all WDE offspring which have been born in the multispecies enclosure in Bandia Reserve since April 2013 should be considered as potential hybrids, in other words, 26 potential hybrids were born up from this period until June 201722.In Bandia Reserve, all WDE were kept in fenced areas, separated from other species, until July 2012 when reserve managers decided to remove some of the fences in the reserve, and thus two previously separated breeding herds of WDE were merged and mixed with other animal species. Since that time, this WDE herd has been in physical contact with CE32. Such hybridization would jeopardize the entire conservation programme and thus there is an urgent need for monitoring, as well as genetic variability assessment, and subsequent evaluation of the appropriateness of the applied population management.Even though a hybrid of the Derby eland has never been observed, there is a risk of its hybridization with CE in Bandia Reserve, considering the previous experience with other Tragelaphini antelopes that are characterized by “unusual readiness to hybridize in captivity”T. derbianus derbianus) and Cape elands (T. oryx oryx) living in a multispecies enclosure in Bandia Reserve, Senegal;Test for the presence of interspecific hybrids between semi-captive Western Derby elands  from Bandia and Fathala reserves in Senegal, and CE (n = 26) from Bandia Reserve, Senegal.22 with the exception of 25 animals, from which the samples were not available for various reasons . Samples from CE were obtained by random sampling of live calves at the age of 1–3 years, animals that died naturally, and surplus males and older calf-less females culled for meat production by the staff of Bandia Reserve.Blood and tissue samples of WDE were acquired systematically by continuous whole population sampling, and included all living individuals recorded in the studbook from 201725. Tissue samples were collected from both species either from dead individuals by scalpel or from living unanesthetized animals by Biopsy & DNA darts Pneu-Dart, Inc.All samples were collected in Senegal in the period 2005–2017 by the NGO Derbianus Conservation  in cooperation with the Directorate of National Parks of Senegal (DPN) and Society for the Protection of Environment and Fauna in Senegal (SPEFS), who are recognized on an international level . The samples were then imported to the Czech Republic  and provided by the NGO for research purposes. No animal was sacrificed for this study. All sampling procedures were performed by certified veterinarians, except the collection of tissues from dead animals. Blood and hair samples were collected only from anesthetized WDE individuals during transport between breeding herds and the reserves, which regularly occurs because of the genetic management of the populationImmediately after collection, the hair and tissue samples were fixed in 96% ethanol, and the blood samples were treated with anticoagulants. Then, all the samples were transported to the freezers in the shortest possible time to be stored at −20 °C until processing in a laboratory which has approval (No CZ 11712934) for storage and usage of animal material according to § 48(1)(i) of Act No 166/1999s concerning veterinary care and amending certain related laws, as amended, pursuant to Article 17(1) of Regulation of the European Parliament and the Council (EC) No 169/2009 and Article 27(1) of Commission Regulation (EU) No 142/2011.® DNeasy® Blood & Tissue Kits. In the case of blood samples, Geneaid™ Genomic DNA Mini Kits (Blood-Cultured Cell) were used. Twelve microsatellite loci were selected for the analyses because microsatellites are fast evolving markers suitable for analyses of recent population structure33.Genomic DNA from hair and tissue samples was extracted by Qiagen™ Thermal Cyclers from Bio-Rad using the Qiagen® Multiplex PCR kit according to the enclosed protocol34. During reactions, 12 microsatellite loci were amplified with already published fluorescently labelled primers that were chosen based on cross-species amplification, including 5 previously used for WDE6; for details see Table ™) with the GeneScan™ 500 LIZ™ dye Size Standard.The PCRs were carried out in T100® Version 2.2.0, SoftGenetics LLC®35 and then binned by AutoBin36. Large allelic dropout, presence of stutter bands, and null alleles were tested in Micro-Checker 2.237. Departures from the Hardy–Weinberg equilibrium and linkage equilibrium of all microsatellite loci were checked by Genepop 4.238.Data from the fragmentation analyses containing information about allele lengths of individual loci were manually scored using GeneMarker39 with 1,000,000 steps of Markov Chain Monte Carlo repetitions after 100,000 steps of the burn-in period. The analysis was run for 1–5 clusters (K) always with 5 repetitions for each K. The results were post-processed by Structure Harvester to evaluate the best supported K40.Population structure and the presence of hybrids were tested by the Bayesian clustering method in Structure 2.3.441. To compare the populations of WDE and CE, basic population structure characteristics were obtained using different programs intended for population genetics calculations. Values of observed (Ho) and expected (He) heterozygosity were calculated in GenAlEx 6.50242, inbreeding coefficient (FIS), fixation index (FST) and allelic richness (Ar) were obtained via FSTAT 2.9.3.243, which was also used for the counting of the confidence interval of FST. The confidence interval of FIS was evaluated using Genetix 4.05.Factorial correspondence analysis was done in Genetix 4.05In total, 76 samples of WDE and 26 samples of CE were genotyped. Analysis in Micro-Checker 2.2 did not detect any genotyping errors, including the presence of null alleles. Linkage disequilibrium was not detected. The populations were in Hardy–Weinberg equilibrium at all of the studied loci, except CE at ETH10 (p = 0.025) and WDE at CSRM42 (monomorphic locus in WDE).The highest likelihood was obtained for two clusters see Fig. .Figure 1Factorial correspondence analysis visualized the relationships between all tested individuals of the tested populations Fig. . WesternThe results of the selected population structure characteristics to compare the populations of WDE and CE are presented in Table Taurotragus derbianus) and common elands (Taurotragus oryx) are considered as sympatric only in South Sudan46. So far, there is no information about the occurrence of hybridization between them, but it cannot be excluded, due to the high relatedness of the taxa (separation estimated at 1.6 million years before present)47 and due to the presence of hybrids between other Tragelaphini species32. The present study did not detect any individual of hybrid origin. However, not all suspect individuals were sampled and thus some possible hybrids might have been overlooked. In the WDE population, 16 samples from totally 26 suspect animals were tested while three of them have died before being sampled and never reproduced. Moreover, samples were taken randomly in the CE population. It may be concluded that until June 2017, no hybrid was detected, and considering this, the probability of ongoing hybridization is low. However, the risk of hybridization between eland species in Bandia Reserve still exists, as the species remain in direct contact48. Also, one cannot exclude that post-zygotic reproduction isolating mechanisms may exist, and that the embryo may be lost during its development. Should this be the case, the fitness of the whole herd would decrease. Respecting the uniqueness and conservation status of the WDE, we propose continuous monitoring of hybridization between WDE and CE until they have separated breeding facilities. If the hybrids occur, it may have severe consequences for the genetic integrity of the species, as shown i.e. in the giant sable antelope (Hippotragus niger variani)49 and bontebok 50.Derby elands  are much higher in CE Table . Factori54, they have their limitations55, which must be considered regarding the results. They do not reflect the whole genome and therefore provides only partial information about the level of polymorphism, and often it is impossible to correlate specific traits with microsatellite parameters.Genetic management of the Critically Endangered WDE might positively affect the level of inbreeding of the whole population in semi-captivity. The values of the coefficient of inbreeding Table  are comp56. However, this is not always possible in the case of Critically Endangered species, in which animal numbers have decreased to such an extent that backup breeding programmes must be established with only a few remaining founders57. An example of an extreme situation is the attempt to save the subspecies of northern white rhinoceros (Ceratotherium simum cottoni) via hybridization with the conspecific southern white rhinoceros (Ceratotherium simum simum), despite their natural long period of geographic, and thus genetic, isolation58. Nevertheless, there should be always an effort to avoid the possibility of both inbreeding and outbreeding depression, if possible16.The genetic background of the population should always be considered from the onset of the establishment of a conservation programme. In optimal circumstances, there are more suitable candidate populations in the wild from which intended founders of the backup population can be selected. All founders should ideally be unrelated and their numbers sufficient to establish more breeding pairs to avoid kinship in the first generation8. Even though such actions can be risky either from the epidemiological or genetic point of view59, they may be crucial for species survival if managed properly60. In the case of the WDE semi-captive population, further importation of new individuals from NKNP to reduce inbreeding and increase genetic diversity has been already recommended25. The results of the present study regarding the genetic diversity of the WDE semi-captive population are in accordance with these previous conclusions and support the idea of introducing new founders from the wild. It also corresponds with the recommendations of Ochoa et al.61 to promote mutual and continuous gene flow via translocations between populations in the wild and captive populations which should function as a source of genetic variation for reintroduction programmes. The One Plan Approach as an integrated approach to species conservation consisting of management strategies and conservation actions by all responsible parties for all populations of a species, whether inside or outside their natural range, should be a priority63.Mitigating genetic threats of small isolated populations, such as inbreeding, and the negative consequences associated with the loss of genetic diversity can involve conservation actions such as translocations61. Although the WDE semi-captive population was considered as a whole in the present study, the individuals are kept in two separate reserves, and regular animal transport takes place between the breeding herds, mostly within each reserve. The transport of animals from Bandia Reserve to Fathala Wildlife Reserve took place in 2006, 2008, 2009 and 2011, with a total of 32 individuals  being translocated and organized into one bachelor herd at first, and later, two breeding herds25. Since only a limited number of females succeeded in reproducing in Fathala Wildlife Reserve, the population in Fathala suffered multiple founder effects, and pedigree analysis suggests considerably lower genetic diversity in Fathala Wildlife Reserve than in Bandia Reserve64.However, in the case of WDE, the promotion of gene flow is needed not only between the wild and captive populations, but also within the captive population. Otherwise, they could suffer from high differentiation as was described in the subpopulations of Arabian oryxet al.7 concluded that population management including interventions possibly influencing genetic diversity via regular changes of breeding males does not always offer distinct advantages when compared to unmanaged populations. However, their study did not reflect genetic parameters. In contrast, the results of the present study indicates that genetic management could have a positive effect on the genetic background of populations. Considering the results of this and a previous study by the present authors6, it may be concluded, that for highly inbred populations such as WDE, genetic management keeps inbreeding at a low level, despite low genetic polymorphism and high relatedness of the population.Wespi Supplementary info.


Freezing grapes is a winemaking technique known as cryoextraction that intends to modify the composition of the final wines. The changes that take place in the frozen grapes facilitate the transfer of certain compounds from the grape skins into the musts because of the grape’s unstructured tissues. For this study, the white grape variety Muscat of Alexandria was selected. Two different cryoextraction procedures have been analyzed as follows: (i) Ultrafast freezing, and (ii) liquid nitrogen freezing. The wines obtained using liquid nitrogen freezing exhibited higher levels of terpenoids, as well as higher levels of hydroxylic compounds and fatty acids than both the wines obtained through traditional methods and ultrafast freezing wines. In any case, both freezing techniques produced wines of a more intense aroma compared with those wines obtained by traditional methods. In fact, liquid nitrogen freezing produced the wines with the most intense aroma and were the best valued by the tasting panel. Numerous studies have been reported in the literature dealing with the phenomena involved in the extraction of aroma components from grape skins into grape juice during the preferential maceration of the juice and skin from white grapes ,2,3. In As a modification of the classical prefermentative maceration at low temperature, different techniques have been developed that involve freezing the grapes; a method known as cryoextraction . FreezinThe freezing speed of the grapes is a particularly important aspect to be taken into account, since it is directly related to the degree of disorganization of the berrys’ structure ,15,16. CThe effect of the different cryogenic techniques can be more easily evaluated in those grape varieties with specific compounds of interest in their skin. One such example is provided by the variety Muscat of Alexandria, where the level of the varietal characteristic aroma are particularly high in their berrys’ skin. In fact, this grape variety has been frequently used to evaluate the effect of different cultivation conditions as well as winemaking techniques on the resulting wines . Some suThe work described here intends to determine the influence that cryoextraction may have on the elaboration of white wine from Muscat of Alexandria grapes. Two freezing methods have been evaluated and these have been related to their specific freezing rates. The final objective is to determine the effect that each one of these two freezing procedures may have on the wines obtained and particularly with regard to their content in aroma related and phenolic components as well as on their actual sensory qualities.Muscat of Alexandria grapes grown in the area of Jerez de la Frontera  were used to produce white wine. Three different vinification procedures were implemented .Approximately 900 kg (100 kg × 3 replicates × 3 type of wines) of grapes from this variety were used to produce the wines. Three different tanks were used for each wine type. The first wine was the “reference” (R) sample, whose grapes had been crushed at room temperature (20 °C) and then pressed. The second wine was denoted as “ultra fast mechanical freezing” (UF). The grapes that were used to produce this second wine had been deep-frozen in a highly powerful freezing chamber that would bring the grapes down to −28 °C in 15 min. The third wine was labeled as “liquid nitrogen freezing” (LN) and the grapes used to produce this third wine type were frozen for less than 1 min by means of liquid nitrogen. The grapes frozen by either technique were stored in the same chamber at −18 °C for 2 h. The bunches were then allowed to thaw at room temperature for 3 h. The semi-thawed grapes were then crushed and pressed.−1 sulfur dioxide was also added as potassium metabisulfite. The yeast Saccharomycescerevisiae var. bayanus, commercially known as “viniferm PDM” commercialized by  was used for the fermentation process in 35 liter containers. The fermentation started 24 h after the inoculation of the yeast and carried on for 11 days, never exceeding 20 °C, until the end of the process when the reducing sugars reached below 5 g L−1. The wine from each winemaking process was cold stabilized in a refrigerator at 4 °C. Sulfur dioxide was added to 80 mg L−1 of total sulfur dioxide and it was filtered through 0.4–0.6 micron filtering plates.The must for each winemaking process was corrected to a pH of 3.3 using tartaric acid and 40 mg L−1 of tartaric acid.Both grape juices and wines were characterized by determining three routine parameters  as well as total polyphenols. All the determinations were carried out in triplicate. The density measurements were performed by means of an Anton Paar DMA 4500M electronic densimeter  in order to determine the sugar levels in the musts based on their direct correlation with density. It was also used to determine ethanol level in 200 mL wine distillate samples. The acidity was calculated by acid-base titration using a Crison-Hach model pHmatic 23  automatic titrator by adding NaOH 0.1 M until pH = 7.0. It was expressed as g L−1 gallic acid) were used to express the result as gallic acid equivalents. For most wines and must, the results from this method correlated with the results from the Folin-Ciocalteau method [The polyphenol content of the wines was determined according to the protocol by Mazza et al. after a minor modification . Brieflyu method .−1 in Milli-Q water containing 5 g L−1 of tartaric acid). The Erlenmeyer flask was placed on a 15-samples magnetic stirrer. The stir bar was stirred at 1250 rpm at 25 °C for 120 min. Then, the stir bar was removed from the wine sample and submerged for a few seconds into distilled water in order to remove any NaCl and subsequently gently wiped dry using a lint-free tissue. It was then transferred into a glass thermal desorption tube to be thermally desorbed by means of a commercial TDU thermal desorption unit (Gerstel) connected to a programmed-temperature vaporisation (PTV) injector CIS-4(Gerstel) through a heated transfer line. The PTV was installed on an Agilent 6890 GC-5973 MS system . An empty baffled liner was used with the PTV. The thermodesorption unit was equipped with a MultiPurpose Sampler (MPS) 2 L autosampler (Gerstel) with capacity for 98 coated stir bars. The desorption temperature was programmed from 40 to 300 °C (held for 10 min) at 60 °C min−1 under a helium flow (75 mL min−1) and the desorbed analytes were cryofocused using the PTV system with liquid nitrogen at −140 °C. Finally, the PTV system was programmed from −140 to 300 °C (held for 5 min) at 10 °C s−1 for analysis by GC–MS. The capillary GC–MS analyses in the electron impact mode were performed on an Agilent 6890 GC-5973N MS system , equipped with a DB-Wax capillary column , 60 m × 0.25 mm I.D., with a 0.25 micron coating. Helium was used as the carrier gas at a flow rate of 1.1 mL min−1. The detector temperature was 250 °C. The GC oven was programmed as follows: Held at 35 °C for 10 min, then ramped at 5 °C min−1 up to 100 °C. Then it was raised to 210 °C at 3 °C min−1 and held for 40 min. The peak identifications were carried out based on the Wiley library by analogy with mass spectra and this was confirmed by pattern retention indices, whenever possible, or according to the retention data reported in the literature. The quantitative data for the identified compounds were obtained by measuring the relative area of the molecular ion peak of each compound relative to that of 4-methyl-2-pentanol, the internal standard [The stir bar sorptive extraction (SBSE) technique was used in conjunction with GC-MS to determine volatile components content of each wine. The extractions were carried out using 10 mm × 0.5 mm (length × film thickness) PDMS commercial stir bars, supplied by Gerstel . Each sample was run in duplicate and the average values of the three types of wine corresponding to each winemaking processes were used for later discussions. A volume of 25 mL of each wine type was pipetted and placed into a 100-mL Erlenmeyer flask containing 5.85 g of NaCl and 50 L of a solution formed by 4-methyl-2-pentanol  so that any influences from external stimuli on the judgments would be minimized. A panel formed by twelve members who were considered as experts took part in the tasting sessions. The panel members were either winemakers or equally experienced laboratory staff members selected for their consistent assessments.The wine samples from the 3 replicates of each specific winemaking process were blended together before starting the sensory analysis. The sessions consisted of presenting the three types of wines to the judges for comparative purposes. Exactly 50 µL of wine was poured into a tasting glass (UNE-EN ISO 3591:1977) fitted with a lid to minimize aroma losses. Each glass was identified by a 3-digit code. The room temperature was set at 22 °C and the samples were presented randomly.The sensory analysis of the wines was carried out in two sessions. In the first session, the judges were requested to perform an ordering test (UNE-EN ISO 8587:2010) to determine their preferences. This type of test is classified as discriminative and it is especially applicable when there are several samples to be compared, since it minimizes the consumption of the samples and the sensory fatigue of the judges. After presenting the wine samples before the judges, they were asked to evaluate and order them according to their preferences (from lowest to highest preference). The wine ID codes were noted down on the corresponding forms. Each judge was encouraged to include additional comments at his/her discretion, since such comments could contribute to a deeper interpretation of the quantitative data.In the second session, the judges were asked to grant a score on the impact notes for each wine sample according to a 1 to 5 scale. The specific impact notes were selected from the conclusions of the first session.p-value was less than 0.05. The statistical analyses were carried out by means of an SPSS V27 .Wines were prepared in triplicate, i.e., three different fermentation tanks were used for each grape juice and analyses were done in triplicate. An analysis of variance (ANOVA) was performed on the continuous variables, i.e., sugar, total acidity, pH, total polyphenols and volatile compounds and the Xi-square test was applied to the discrete variable, i.e., the sensory panel results. The differences were considered as relevant when the The grapes were frozen, thawed and then pressed. The resulting musts were analyzed in order to ascertain whether there were differences in their properties that could be attributed to the different treatments applied to the grapes. The results for total sugars, pH and total acidity in the musts are shown in −1). This result is a consequence of the precipitation of potassium salts during the freezing process and the lower solubility of the acidic salts when the grapes were defrosted—both effects had already been reported in the literature [Regarding the total acidity of the musts, it should be mentioned that the reference must, which was obtained without submitting its grape to any specific treatment, had the highest value  showed the lowest sugar content (216 ± 5) when compared to the musts obtained after freezing the grapes (263 ± 4 (UF) and 235 ± 4 (LN)) . One of Different grape freezing techniques have different effects on the final wines. When the grapes are frozen by means of liquid nitrogen (LN) the wines that are obtained have a greater total sugar content than those obtained when the grapes are frozen by ultra fast mechanical freezing (UF). The difference in sugar content might be a result of a faster defrosting process in the case of ultra fast mechanical freezing, while when grapes have been frozen by means of liquid nitrogen the effect of cryoscopic decrease would be more evident and, consequently, the musts obtained should exhibit higher sugar contents. In an effort to avoid the enzymatic oxidation of the grapes during the defrosting process, the grinding process was started just three hours after the defrosting of the grapes had begun. At that point, the thawing of the grapes was more advanced in those grapes that had undergone ultra fast mechanical freezing when compared to those which had been frozen by Liquid Nitrogen.The alcoholic level of the R wine was 11.2 ± 0.21, 15.2 ± 0.36 for the LN wine, and 13.4 ± 0.41 for the UF wine. These values reflect the differences in must sugar content initial values and underline the first major effect caused by the two freezing techniques used.Cryoextraction should produce wine with higher levels of polyphenols as they are easily extracted from the grape skins . However−1 of gallic acid). No significant differences were found between the two wines obtained from frozen grapes (LN and UF). For this reason, it can be assumed that during the short freezing process, the freezing procedure does not matter, the grapes are degraded to a sufficient extent so that more polyphenols are extracted from their skin during the grinding and pressing process than in the case of berries that had not been subjected to freezing at all, even if both grinding and pressing are processes that are carried out at lower temperatures. This effect has been previously described in the literature although for longer freezing times [It was confirmed that the wines from frozen grapes presented higher phenolic contents with no significant differences between them  were found between the different winemaking techniques used in this study. Most of these compounds are present in grape skins, therefore, any differences between the wines from frozen grapes should be attributed to differences in the degradation level of the grapes skin over the freezing process. It has been previously described that freezing processes may affect some volatile compounds such as the thiols in Sauvignon blanc grapes [The aromas of Muscat of Alexandria wines were studied by analyzing the concentrations of terpenes, alcohols, aldehydes, fatty acids and ethyl esters and significant differences , which was given a value of 100%. The results are presented in It can be seen in β-myrcene content in the LN wine was almost 600% that in the reference wine (R), whereas nerol (301%), geraniol 332%) and eugenol (310%) exhibited three fold levels compared to the R wine. It is also worth highlighting other components that differed from those in the reference wine by a factor of two or more. For example, in the LN wine these components include terpineol (229%), although linalool was present at a lower level than in the R wine. Terpineol is related to mint aroma  and nero2% and euThe data obtained from the determination of alcohol contents confirm that the LN wines always presented higher concentrations than the UF wines followed by the R wine. It is worth mentioning that, as can be seen in Regarding the analysis of aldehydes, their behavior is more heterogeneous with respect to the different techniques used in the winemaking process. Nonanal is the component with the most noticeable concentration difference with respect to the R wine: The LN wine reached a value of 482% and the UF wine went as high as 390%. Nonanal has been associated to citrus fruits aromas . The LN The study of fatty acids produced similar results to those corresponding to terpene-like components. The LN wines exhibited higher values for fatty acids than the UF wines. Their content in octanoic acid (153%), nonanoic acid (208%), lauric acid (163%) and decanoic acid (207%) were over that in the R wine by even more than 50%. A similar trend was detected in the UF wines for palmitic acid, with 190% its content in the reference wine, which is even higher than the levels found in the LN wines . It is aFermentation conditions can affect the composition of the fatty acids in a particular wine. Under anaerobic conditions, yeast produces medium-chain fatty acids and when fermentation is carried out in aerobic or semi-aerobic conditions more unsaturated fatty acids are produced . HoweverEthyl esters are formed because of the esterification of fatty acids during the alcoholic fermentation of must. The acetates from higher alcohols and the ethyl esters from fatty acids are associated to floral and fruity aromas in young wines . TherefoOn tasting the wines, all the members in the panel considered the reference wine to be the least aromatic of all the three wine types. The LN and UF wines were given significantly higher scores for fruit and flower notes with respect to the R wine that had be produced by traditional methods . This reThe panelists were able to differentiate the LN from the UF wines by its remarkable menthol notes, which made the former wine unique and attractive. The compounds responsible for these notes seem to be monoterpenes and their derivatives ,42. β-MyRegarding the overall rating , the LN Freezing procedures reduced acidity, increased alcoholic strength and, above all, produced wines that were more aromatic, since they contain greater amounts of the components that are more closely associated to Muscat grape wine specific characteristics.The wines produced using liquid nitrogen to freeze the grapes presented higher concentrations of terpenes in all the cases, whereas the wines produced using the ultrafast freezing chamber exhibited similar although just slightly higher values when compared to the reference wine. β-citronerol, nerol and geraniol contents were particularly high in the wines from frozen grapes using liquid nitrogen, with more than double their concentration with respect to the control wine. These results were consistent with the score granted by the tasting panel who judged them as characteristic wines of special interest.With regard to the difference between wines produced by the two freezing techniques applied in this study, liquid nitrogen frozen grape wines, not only contained more of the compounds that provide Muscat wines with their characteristic fruity and mentholated notes, but they were also more appreciated by the judging panelist than those wines obtained from fast frozen grapes.


This highly educated and experienced health consumer cohort reported they were significantly more familiar with the healthcare system generally than genetic medicine specifically (p < 0.0001). Consumers perceived that genetic testing would be significantly more important in the future than it is currently (p < 0.00001). Consumers agreed that genetic testing should be promoted (91.4%), made available (100%), better funded (94.2%), and offered to all pregnant women (81.6%). The preferred learning modality about genetics was internet sites (62.7%) followed by talks/presentations (30.8%). Benefits of genetic testing, reported in qualitative responses, included the potential for additional information to promote personal control and improve healthcare. Perceived concerns included ethical implications (including privacy and discrimination), and current limitations of science, knowledge and/or practice. This study demonstrates that even knowledgeable consumers have little familiarity with genetic medicine but are optimistic about its potential benefits. Ethical concerns, particularly concerns regarding genetic discrimination should inform legislation and policy. Consumers are supportive of online resources in increasing genomic literacy.As genetic testing becomes increasingly utilized in health care, consumer awareness and understanding is critical. Both are reported to be low in Australia, though there are limited studies to date. A consumer survey assessed perceived knowledge, awareness and attitudes toward genetic medicine, prior to consumers’ genomics forums in Queensland in 2018 and 2019. Data was analyzed using  The demand and utilization of genetic and more recently genomic testing continues to increase , yet allPrevious research has focused on attitudes, awareness and knowledge of health consumers. Throughout various countries, reported genetic literacy is low and consumers are typically uncertain about how it applies to healthcare . ContrarAustralia has a publicly funded health system which supports access to genetic and genomic testing, when clinically indicated . Despite its availability there are limited studies about consumer knowledge, awareness, and attitudes toward genetic testing in Australia. Previous studies reveal health consumers in Australia are interested in genetic testing but fear genetic discrimination . They shThis study involved the administration of a survey assessing consumers’ awareness and attitudes toward genetics in healthcare prior to a genomics educational intervention in the form of a presentation during the health consumer forum in Queensland in both 2018 and 2019.This survey was initially administered as a marketing tool to shape the content of future educational sessions. An ethics application to report the findings in the literature was approved by the University of Queensland Human Research Ethics Committee (UQ #2019002633) with the stipulation that quotes be paraphrased.All delegates attending the Queensland Genomics panel presentation on genomics in medicine, at the Health Consumer Queensland annual forum in 2018 (Brisbane) and 2019 (Cairns) were invited to complete the survey. Any individuals present both years (2018 and 2019) noted this on their 2019 survey and only their first response was included in the analysis.Surveys were administered and collected prior to the genomics education session. In 2018 the 23-item survey included: 5-items assessing demographic information ; 5-items assessing familiarity with genetic diseases and testing (Yes/No) . Ranks on scales of “1–5” which were reported as >5 were adjusted to “5.” Participants choosing both “agree” and “disagree” were categorized as “neither agree nor disagree.”t-tests were performed to test to detect any differences between means in normally distributed data  and Mann-Whitney U tests were used to detect differences in means in datasets which were not normally distributed .Descriptive statistics were performed to summarize and describe the characteristics of the data. Student’s All quotes in open-ended fields were transcribed verbatim and analyzed thematically to identify perceived benefits and concerns regarding genetic testing. Codes were categorized to identify overarching themes. Codes were reconciled between two researchers (CW and AM). Themes were reported as well as the frequency with which they appeared in the text. Illustrative quotes were paraphrased to preserve consumer privacy.n = 51) and 2019 (n = 15). Seven individuals in the 2019 survey reported having attended the Queensland Genomics presentation for the Health Consumer Queensland annual forum in 2018, and were therefore removed from statistical analyses, resulting in 59 unique individuals.A total of 66 individuals participated in the survey in 2018 (n = 41 (69.5%)], 16 were males (27.1%), and 2 were not specified (3.4%). Ages of participants ranged from 26 to 80 years of age with the average age being 47.3 years. This cohort was highly educated with 50 participants (84.7%) having some form of tertiary education and 28 participants (47.5%) having undertaken graduate education.The majority of participants were female [Of the participants who completed each question, 40/56 (71.4%) reported knowing someone with a genetic disorder personally, 11/59 (18.6%) had never heard of genetic testing before attending the forum, and 10/58 (17.2%) reported personal experience with genetic testing. Finally, 34/58 (59.6%) did not know how to find genetic services in Queensland.T-tests found these scores to be significantly different . There were no correlations between these scores and age or education.The average self-reported familiarity with the healthcare system in Queensland was 6.64 (range 1–10), and the median was 7. The average genetic medicine familiarity score was 2.97 (range 1–8) with a median of 1. refer to  (t = 8.7z-score −4.634 p < 0.00001). There were no correlations between these scores and age or education for either time point.Participants perceived that genetic testing moderately affected healthcare in Queensland today with a mean of 4.97 (range 1–10) and median of 5 . ParticiOf the participants who provided a response, 48/51 participants (94.1%) agreed that the use of genetic testing should be promoted with 52/52 (100%) agreeing that genetic testing should be available for those who want to use them, 49/52 (94.2%) agreeing that there should be more funding and 40/49 (81.6%) agreeing that genetic tests should be offered to all pregnant women . There wU-tests (p < 0.00001), videos (p = 0.00001), discussions with health care providers (p = 0.001) and talks and presentations (p = 0.019). Talks and presentations were significantly preferable to printed material (p = 0.003) and videos (p = 0.003) but not discussions with healthcare providers (p = 0.265). Mann-Whitney U test found that individuals under 45 years of age (n = 25) also preferred internet sites above videos (p = 0.001), discussions with healthcare providers (p = 0.032), and printed material (p = 0.00005), but there was no differences between their preference for internet resources as compared to talks and presentations (p = 0.172). Those over the age 45 years (n = 29) preferred internet sites to videos (p = 0.031), discussions with physicians (p = 0.033) and printed materials (p = 0.028), but there were no other significant preferences  followed by talks/presentations 16/52 (30.8%). The average ranked score for each modality was calculated . InterneU-tests  found thferences .Forty-seven written responses identified two main categories of benefits . More thn = 32) and knowledge limitations inhibiting interpretation and utilization (n = 12).Forty-seven individuals provided responses describing their concerns with genetic testing which could be broadly categorized into two themes, namely ethical considerations . Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.AM-L, CW, DB, KC, and EE established the study and applied for ethics. AM-L, LF, SI, and RL developed the survey which was administered by KC. Presenters for the Queensland Genomics panel at the Health Consumer Queensland forum were KC, AM-L, NW, GH, DB, and EE (2018), and EE, SI, GH, and LF (2019). CW and AM-L co-wrote the manuscript. All authors critically reviewed the manuscript.NW was the co-founder, minor equity holder, and board member of genomiQa. The remaining 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.


Assessments of genetic diversity have been claimed to be significantly efficient in utilising and managing resources of genetic for breeding programme. In this study, variations in genetic were observed in 65 pineapple accessions gathered from germplasm available at Malaysian Agriculture Research and Development Institute (MARDI) located in Pontian, Johor via 15 markers of simple sequence repeat (SSR). The results showed that 59 alleles appeared to range from 2.0 to 6.0 alleles with a mean of 3.9 alleles per locus, thus displaying polymorphism for all samples at a moderate level. Furthermore, the values of polymorphic information content (PIC) had been found to range between 0.104 (TsuAC035) and 0.697 (Acom_9.9), thus averaging at the value of 0.433. In addition, the expected and the observed heterozygosity of each locus seemed to vary within the ranges of 0.033 to 0.712, and from 0.033 to 0.885, along with the average values of 0.437 and 0.511, respectively. The population structure analysis via method of delta K (ΔK), along with mean of L (K) method, revealed that individuals from the germplasm could be divided into two major clusters based on genetics (K = 2), namely Group 1 and Group 2. As such, five accessions  were clustered in Group 1, while the rest were clustered in Group 2. These outcomes were also supported by the dendrogram, which had been generated through the technique of unweighted pair group with arithmetic mean (UPGMA). These analyses appear to be helpful amongst breeders to maintain and to manage their collections of germplasm. Besides, the data gathered in this study can be useful for breeders to exploit the area of genetic diversity in estimating the level of heterosis. Genetic diversity of 65 pineapple accessions from MARDI’s germplasm had been assessed by using 15 polymorphic SSR markers where moderate level of variations was observed.Two major clusters that separated the 65 accessions into two major groups were identified.All accessions can be differentiated from one another except for SG Spinal local and LT_SG that had a very low genetic distance.Ananas comosus (L.) Merr.), primarily due to the variations of naming customs among breeders and cultivators from within or between nations  that amplified A. comosus genome when the SSR actually reflects species-specific that conserves sequences across various species of the same family  markers revealed three clusters that distinguished Spanish, Queen, and Cayenne groups . On the The description of plant accessions in a systematic manner may suggest sectors that are organised in an excellent shape, thus enhancing and facilitating plant germplasm usage. Hence, this study assessed the genetic diversity of 65 pineapple accessions gathered from germplasm available at Malaysian Agriculture Research and Development Institute (MARDI), which were collected from many locations, including all South East Asian nations, Australia, and Pakistan.A total of 10 plants from 65 accessions were collected from MARDI’s pineapple germplasm in Pontian, Johor, in order to gather their young leaves. The collected plants had been grown in peat soil, which were taken care of by employing standard cultural practices (no hormone is used to induce flowers), as recommended by Fruit Programme established at the Horticulture Research Centre of MARDI. In the beginning, 56 SSR markers that relatively yielded a high number of alleles and heterozygosity were opted for optimization  was added to the plate wells by using a multichannel pipette and mixed well before the samples were incubated for an hour in 60°C water bath . After the plate was cooled down for 5 min, a ratio of 350 μL chloroform: isoamyl alcohol (24:1) was included into each well, which was then mixed and centrifuged  for 15 min at 5500 rpm. In addition, an equal volume of cold isopropanol was applied for DNA precipitation and then, it was washed using 70% ethanol. Finally, the produced pellets of DNA were air-dried and re-suspended in 50 μL of Tris-EDTA (TE) buffer. The gel electrophoresis was carried out by using 0.8% ethidium bromide pre-stained agarose gel, and then, further visualised under ultraviolet light using Quantum ST4 3000 , in order to determine the quality of the extracted DNA. After that, the Thermo Labsystems Floroskan Ascent™  was used to determine the extracted DNA’s concentration, which was later normalised to 30 ng μL−1.A modified high-throughput plant DNA extraction technique was employed to extract total genomic DNA from individual plants . Firstly−1 of DNA template. The thermal conditions of the PCR were set to: initial denaturation for 5 min at 95°C, next followed by 30 cycles  by using a thermal cycler . The PCR mixture consisted of 10X PCR buffer , 50 mM MgCl2 , 2 mM dNTPs , 10 μM of forward primer anchored with M13 tail (Schuelke 2000), 10 μM of unlabelled reverse primer and 5 μM of M13 tail , 5U Taq Polymerase , and 30 ng μLpair see , then anhttp://taylor0.biology.ucla.edu/structureHarvester). Lastly, the UPGMA dendrogram was formulated via Power Marker software 3.5 by calculating the genetic distance (Nei 1983) and subsequently, by employing the MEGA 7.0.18, the outcome was visualised . This genotypic data were then converted into several different formats by using CONVERT software . ScoringInitially, 25 out of 56 SSRs displayed exceptional amplifications after optimisation. Meanwhile, the rest were discarded due to non-specific binding and poor amplification in 2% pre-stained (ethidium bromide) agarose gel electrophoresis. As a result, 15 polymorphic SSRs with good amplification’s quality had been selected and used for further analysis across the 65 accessions of pineapple germplasm, while the others were disregarded due to monomorphic alleles and poor amplification during fragment analysis. More information pertaining to each SSR marker is presented in Evidently, the scoring error, the large allele dropout, and null alleles were not recorded for all the 15 SSR markers after they were analysed via Micro- Checker 2.2 hence the population is probably in Hardy Weinberg equilibrium (HWB). Nevertheless, 59 alleles had been identified with a mean of 3.93 alleles per locus at the range of 2.0 to 6.0 alleles. In comparison, Meanwhile, the pairwise genetic distances appeared to range from 0.0014 until 0.4949 and displayed an average of 0.2284 with exclusion of data. MRTxSS194JK and Yankee had the highest genetic distance (0.4949), while SG Spinal Local and LT_SG had the lowest genetic distance (0.0014). However, the lowest genetic distance is suspected due to missing alleles therefore SG Spinal Local and LT_SG cannot be differentiated between one another or both could possibly be from the same accession. Hence, these data can also be used to identify redundant accessions or varieties as this is one of the major concerns in pineapple plantations. Vegetative plant materials can easily be exchanged from one region to another. Due to non-standard nomenclatures in pineapple, the accessions can easily mix up amongst one another and subsequently cause homonym and synonym. Accessions that have the value of genetic distances below 0.1000 are highly likely suspected to be of the same accessions. However, more markers will be tested on these accessions to verify it in the future. Morphological evaluations will also be conducted to certify the accession. Additionally, the values of PIC seemed to range from 0.1040 (TsuAC035) until 0.6970 (Acom_9.9) at the average of 0.4330. Referring to bracteatus and erestifolius were clustered together in one group while the other accessions from var. ananassoides was clustered in another group.The analysis exhibited that after a total of 10 runs for every K value (1–10), along with burn-in from 10,000 until 100,000 iterations; the optimal K value was determined as 2 (K = 2), thus implying that individuals from the germplasm can be segregated into two major clusters of genetic. Besides, the Table of Evanno displays the values of ΔK for every K (1–10), whereby the optimal number of K is represented by the highest yield of ΔK (K = 2), as given in These observations are in line with the results obtained from dendrogram via UPGMA analysis, where the two major clusters (Groups 1 and 2) had been noted see . In factThis study reveals that MARDI’s pineapple germplasm has a moderate range of diversity. In fact, the data retrieved appear useful for plant breeders, especially in estimating the level of heterosis, where the offspring of the two breed lines carried superior traits or characteristics, in comparison to both of their parents. Furthermore, it was noted that the higher the diversity between two breed lines, the higher the chances of heterosis. In addition, the outcome of this study can be applied as a significant tool for breeders to maintain and to manage their germplasm collections systematically and efficiently.


Mauritia flexuosa L.f. to understanding the relation between environment and phenotypic traits. Twenty-five fruits were collected from each of the 10 individuals sampled in each study site: Chapada dos Guimarães (CG), Vila Bela da Santíssima Trindade (VB), and Alta Floresta (AF). We analyzed the genetic divergence, using the standardized Euclidean distance, the sequential method of Tocher, unweighted pair group method with arithmetic mean (UPGMA), and the projection of the distances onto 2D plane, and calculated the relative importance of the traits evaluated. The analysis showed the partition of individuals into three main groups: Two groups comprising the majority of individuals. Fresh fruit weight, pulp rate, fresh pulp weight, and moisture rate were the traits that most helped explaining the difference between materials. The results shown in the current study evidenced the influence of these three different environments on the biometric traits of M. flexuosa. Such influence has led to the formation of Alta Floresta and Vila Bela da Santíssima Trindade individuals in different groups, whereas the Chapada dos Guimarães individuals were able to permeate the two other groups, although they showed stronger tendency to group with individuals from Vila Bela da Santíssima Trindade.The environmental heterogeneity may reflect the different morphological and phenotypic traits of individuals belonging to a single species. We used 14 morphological traits of  Mauritia flexuosa L.f.is distributed in these three different areas.Mato Grosso State presents wide environmental heterogeneity due to the rich and complex plant biodiversity found in its three different biomes, namely, Pantanal, Brazilian Savannah; the Amazon; and the ecological tension zones (ETZ), which is the transition site between the two largest Brazilian plant biomes, the Brazilian Savanna and the Amazonian Forest . This enBuriti is a dioecious palm tree, up to 40 m tall , which iAll parts of this palm tree can be used. Its leaves can be used in craftwork and its fruits are consumed fresh or processed as juice, porridge, soup, and ice cream . The fruThe phenotypic differences are set by environmental variations because even fruits belonging to a single species and distributed in different locations are subject to temperature, photoperiod, and rainfall rate variations, as well as to other variables that may be linked to certain aspects of the species’ genetic composition . The difMauritia flexuosa L. f. fruits and seeds grown in the Amazon and Brazilian Savannah biomes, as well as in the ecological tension zones.Morphometry studies allow understanding the different phenotypic traits of buriti fruits and seeds. Such studies are an important instrument for the detection of genetic variability within populations, as well as to find the relation between this variability and the environmental factors. They also provide important information that help featuring the ecological and genetic aspects concerning conservation and breeding programs. The aim of the current study was to analyze the influence of the environment on the morphometry of The quantitative data about the 14 morphometric traits of fruits and seeds were collected from buriti individuals. Individuals 1–10 represent Alta Floresta; 11–20, Vila Bela da Santíssima Trindade; and 21–30, Chapada dos Guimarães. The combination between individual 18 (VB) and 29 (CG) was the most dissimilar one, since it presented the highest standardized Euclidean distance estimate (dii’ 6.36); whereas the lowest divergence was observed between the pair 24 and 30, both collected in CG (dii’ 0.00).The analysis resulted in the formation of five different groups  clustering method according to the standardized Euclidean distance is shown in Subgroups 1a, 1b, and 2a comprised seven individuals each, and these individuals derived from different locations. Subgroup 1a comprised individuals from CG and VB. Subgroup 1b comprised individuals from VB, whereas subgroup 2a comprised individuals from AF. Subgroup 2b comprised five individuals, three from AF and two from CG. On the other hand, group 3 was represented by four (CG) individuals; three of them were allocated in subgroup 3a, whereas individual 26 was allocated alone in subgroup 3b.By analyzing the projection of the distances in the plane , it was The quantitative morphometric traits explaining the divergence among the 30 buriti individuals were Fresh fruit weight (37.50%), Pulp rate (17.86%), Fresh pulp weight (14.68%), and Moisture rate (11.91%) . The traThe scatter plot  was baseThe genetic structure of natural populations can be influenced by many ecological factors. Groups of individuals established in different regions holding specific environmental features tend to genetically differentiate themselves as populations. It happens due to the gene flow limitation and to the different selection pressures faced by each population .Attalea speciosa Mart.) populations and found strong correlation between the individual’s genetic distance and place of occurrence. The first 15 most divergent pairs derived from different locations. The CG site had the highest intra-population variation; this variation can be attributed to genetic differences among individual parent plants, even within a small geographic area [M. flexuosa fruits and seeds.The dissimilarity matrix generated through the standardized Euclidean distance showed strong correlation between the individual’s genetic distance and place of occurrence. The 10 most divergent pairs of genotypes came from distant populations and, when the first 20 pairs were taken into consideration, just one pair of individuals was in the same location. Similar results were found by Santos , who stuhic area . Matos ehic area  found inThe clustering methods used in the present study corroborate the aforementioned observation, as it demonstrated the tendency to group individuals coming from the same location. The VB and AF individuals were found in isolated groups, whereas the CG individuals were merged between the two locations, a fact that can be explained by the individual features of each area wherein the samples were collected. Some of these features are: Geographical isolation, different climate, phytophysiognomy, and matrix where these populations were found. The environmental conditions may influence the morphological and physiological traits of fruits and seeds ,15,16,17The AF and CG individuals came from areas called palm swamps; however, the AF palm swamp has perennial stream, whereas the CG has intermittent streams. On the other hand, the VB individuals came from a floodable forest called “buritizal”, which is located in the riverbanks of the Guaporé River. This river remains flooded for 6–8 months throughout the year and, in the other months, it keeps water availability because of the shallow water tables .Although the region where the fruits were collected in VB is an ecological tension zone (Amazon-Brazilian Savannah), its phytophysiognomy shows predominant features of Savannah regions. Mariotti  performeBased on this information, it was possible to see that most CG individuals were closer to the VB one, because of the phytophysiognomy similarities between these sites. However, the fact that some CG individuals were grouped with AF individuals may be linked to the availability of similar resources in these two areas.The scatter plot based on the two traits showing the greatest relative dissimilarity contribution allowed seeing that AF individuals presented mean FFW higher than that found in the other two locations, a fact that can be explained by the different soil classes found in the research sites. The presence of acrisols in AF may have favored the FFW because such soils present sandy textures in surface soil horizons, as well as more clayey textures in subsurface horizons, where most buriti roots are often found. Thus, they enable water storage and cation exchange capacities higher than those enabled by sandy soils in the entire soil profile, as it was found in the CG and VB soils. On the other hand, the CG individuals, overall, showed pulp rate higher than that of the other individuals because this area is located at higher altitudes; thus, it favors conditions conducive to higher annual fruit productivity.Swietenia macrophylla King.) in which the fresh fruit mass was the second characteristic, with 21.47% in explaining the genetic diversity. In addition, the characteristics cited above were linked to the species adaptability to the spatiotemporal variability of habitats [Fresh fruit weight, pulp rate, fresh pulp weight, and moisture rate were the traits that most helped to explain the difference between materials. Partially similar results were found in research carried out by Leão et al. , in the habitats .Mauritia flexuosa L.f.) monodominant formations located in Alta Floresta (AF), Chapada dos Guimarães (CG), and Vila Bela da Santíssima Trindade (VB) counties, Mato Grosso State, Brazil , is of the Aw type—tropical climate, with dry winter. It has a rainy season in summer, from November to April, and a clear dry season in winter, from May to October (July is the driest month). The average temperature of the coldest month is over 18 °C. Precipitation is greater than 750 mm per year, reaching 1800 mm. In Alta Floresta, the climate is of the tropical humid or subhumid (Am) type and presents four months of drought. It is also characterized by presenting an average temperature of the coldest month always above 18 °C, presenting a short dry season that is compensated by the high total precipitation .Neosol, which presents sandy or sandy loam texture. This soil class comprises nonhydromorphic, discolored quartz sands and presents yellow or red color [According to Ribeiro and Walter , the loced color .Vila Bela da Santíssima Trindade in Southwestern Mato Grosso State, Alto Rio Guaporé Valley, holds transitional features between two of the largest Brazilian biomes, namely, Brazilian Savannah and Amazon Forest . In addiTwenty-five fruits were collected from each of the 10 different buriti palm trees in each study site. The collected samples were at least 100 m away from each other. This condition was necessary to decrease the number of specimens derived from common parents because parental diversity increases genetic variability within the sampling groups . The indAfter the fruits were assessed, they were transferred to identified and sealed polyethylene bags and, subsequently, stored in a cooler until the end of the collection process. Next, they were kept under refrigeration in the laboratory of the Study Center for Limnology, Biodiversity, and Ethnobiology of Pantanal , Mato Grosso State University (UNEMAT) – Cáceres campus.The height  and diameter of each sampled fruit were measured in a digital caliper , whereas the total weight was measured using an analytical scale . The fruits were peeled and pulped after their morphometric parameters were taken. Then, the peel, pulp, seed, and other structures were separated. All the structures mentioned above were separately weighed and classified as fresh weight. Moisture was determined through the oven method, at 105 ± 3 °C for 24 h, according to the recommendations in the Rules for Seed Analysis . Next, tThe means of each assessed trait were obtained, namely, fresh fruit weight (FFW), fresh peel weight (FPW), fresh pulp weight (FPuW), fresh seed weight (FSW), other fresh structures (OFS), height (H), diameter (D), dry fruit weight (DFW), dry peel weight (DPW), dry pulp weight (DPuW), dry seed weight (DSW), other dry structures (ODS), and moisture content (MC).standardized Euclidean distance as dissimilarity measure [Mauritia flexuosa L. f. fruits and seeds from plants distributed in the Amazon and Brazilian Savannah biomes, as well in the ecological tension zones (ETZ):n is the number of studied traits.The multivariate analysis was used to estimate the genetic divergence through the  measure  and to eThe grouping methods were intended to separate an original group of observations into several subgroups, in order to obtain homogeneity within and heterogeneity among the subgroups. Among these methods, optimization and hierarchical methods were employed on a large scale . Subsequently, to obtain the dissimilarity matrix, we used methodologies that made it possible to visualize the separation of the individuals evaluated in groups of similarity, which promoted a clearer and more visual interpretation of the results.The sequential Tocher method, the hierarchical clustering method (UPGMA), and the projection of distances in 2D format were used as clustering techniques according to the standardized Euclidean distance . The cluMauritia flexuosa L. f. plants distributed in the Amazon (AF) and Brazilian Savannah (CG) biomes, as well as in the ecological tension zone (VB). Two large, distinct groups were formed: Alta Floresta (AF) and Vila Bela da Santíssima Trindade (VB) individuals, whereas individuals in Chapada dos Guimarães (CG) permeated these two groups, although they showed greater trend to group with VB individuals. The greatest intrapopulation variation was observed between CG individuals, whereas the AF group showed the smallest variation. The fresh fruit weight and the pulp rate were the main traits contributing to the clustering results.There was environmental influence on the morphometry of fruits and seeds from


Cucumis sativus L.). For the genetic diversity study, 280 cucumber accessions collected from four continents  by the National Agrobiodiversity Center of the Rural Development Administration in South Korea and 20 Korean commercial F1 hybrids were genotyped using 151 Fluidigm SNP assay sets. The heterozygosity of the SNP loci per accession ranged from 4.76 to 82.76%, with an average of 32.1%. Population genetics analysis was performed using population structure analysis and hierarchical clustering (HC), which indicated that these accessions were classified mainly into four subpopulations or clusters according to their geographical origins. The subpopulations for Asian and European accessions were clearly distinguished from each other , while the subpopulations for Korean F1 hybrids and Asian accessions were closely related (FST = 0.34). The highest differentiation was observed between American and European accessions (FST = 0.41). Nei’s genetic distance among the 280 accessions was 0.414 on average. In addition, 95 commercial F1 hybrids of three cultivar groups  were genotyped using 82 Fluidigm SNP assay sets for cultivar identification. These 82 SNPs differentiated all cultivars, except seven. The heterozygosity of the SNP loci per cultivar ranged from 12.20 to 69.14%, with an average of 34.2%. Principal component analysis and HC demonstrated that most cultivars were clustered based on their cultivar groups. The Baekdadagi- and Gasi-types were clearly distinguished, while the Nakhap-type was closely related to the Baekdadagi-type. Our results obtained using core Fluidigm SNP assay sets provide useful information for germplasm assessment and cultivar identification, which are essential for breeding and intellectual right protection in cucumber.Genetic diversity analysis and cultivar identification were performed using a core set of single nucleotide polymorphisms (SNPs) in cucumber ( Cucumis sativus L.) is a major fruit vegetable belonging to the family Cucurbitaceae. It has two primary varieties: cultivated cucumber (C. sativus var sativus) and its wild relative (C. sativus var. hardwickii) [Cucumber (dwickii) . Cucumbedwickii) , and it dwickii)  and divedwickii) . Cucumbedwickii) . Worldwidwickii) . In Koredwickii) .http://genebank.rda.go.kr accessed on 12 May 2020). According to the records of the Korea Seed & Variety Service (KSVS), approximately 50 commercial F1 hybrids have been registered from 2005 to 2020, and 547 cultivars have been reported for production and sale (http://seed.go.kr accessed on 12 May 2020). Among them, the Baekdadagi-type is approximately 25%, accounting for the largest percentage of annual production [After early domestication in India, China, and Western Asia, cucumber was introduced to Europe and the Americas . Becauseoduction .Evaluation of genetic diversity is critical for the management, conservation, and breeding processes of genetic resources . FurtherThe genetic diversity of cucumbers has been evaluated based on the following types of molecular markers: isozyme ,17,18, rRecently, a cucumber Fluidigm SNP assay was developed for 240 core SNPs (core SNP marker sets) selected from more than 10,000 SNPs discovered through GBS of 88 commercial F1 hybrids . To dateGenotyping results  of 280 aGenotyping of the 95 commercial F1 hybrids  using 82The population structure of 300 accessions was analyzed based on the genotyping results using 151 SNP markers. Estimation of the optimal number of subpopulations (subpops.) was found to be the most suitable when the subpopulations were classified into two (delta K = 2) and four (delta K = 4) A.Population classification revealed that Subpop.1 was primarily composed of Asian accessions (ASs) and Korean PVP F1 hybrids (KF1s), while Subpop.2 mainly comprised European accessions (EUs), American accessions (AMs), and African accessions (AFs) B. WithinFst values calculated to estimate the genetic distance among the four subpopulations was 0.35–0.51, with an average of 0.43 , representing the genetic diversity among individuals belonging to the subpopulations 1, 2, 3, and 4, were 0.3290, 0.2225, 0.3910, and 0.2206, respectively, while the observed heterozygosity (OH) was 0.2928, 0.2814, 0.2667, and 0.4839, respectively. The OH was higher than the EH for Subpop.2 and Subpop.4. The range of pairwise  of 0.43 . The lowThrough HC based on the genetic distance reported by Cavalli-Sforza and Edwards , it was PCA was performed based on the genotyping results of the 95 commercial F1 hybrids using 82 SNP assay sets. Consequently, three clusters were formed according to the cultivar group. Cluster I was primarily classified as Baekdadagi-type, Cluster II as Gasi-type, and Cluster III as Nakhap-type . HoweverBased on the results of the analysis of 95 commercial F1 hybrids using 82 SNP assays, Nei’s genetic distance was calculated and HC was performed . The anaTo date, genetic diversity evaluation and cultivar identification of cucumbers have been performed using various types of markers. In particular, the use of SSR markers for genetic resource evaluation and cultivar identification has been reported in several studies ,22,30,34In the present study, 151 SNPs of 240 core Fluidigm SNP assay sets, first developed for cucumber cultivars, were used to evaluate the genetic diversity and cultivar identification ability of cucumber accessions at the NAC of RDA and commercial F1 hybrids in South Korea. In the 280 collected accessions and 95 commercial F1 cultivars, the success rates of marker analysis were high , indicating that the core Fluidigm SNP assay sets can be used for genotyping cucumber germplasm with diverse genetic origins. For the collected accessions, the average proportion of the heterozygous marker genotype was high (32.1%), which was similar to that of 20 PVP F1 hybrids (29.80%) or 95 commercial F1 hybrids (38.54%). In contrast, the proportion of the heterozygous marker genotype was relatively low at 9.07% in seven breeding lines considered to have a high genetic fixation level. This implied that the collected accessions were mostly F1 cultivars or genetically heterogeneous heirlooms. Population structure analysis results revealed that accessions with similar geographic locations were classified as having similar population structures. Broadly, ASs, including KF1s and accessions derived from other continents , showed a tendency to be distinguished from each other, and even within the ASs, KF1s were clearly distinguished. However, high levels of genetic admixture were observed for several accessions in the subpopulation, which were presumed to be due to hybridization and migration among the continents. The EH, a general statistic for evaluating genetic variation within a population , was higFST analysis [FST values among Subpop.1, Subpop.2, and Subpop.4 were also relatively high . According to Wang et al. [Additionally, in the pairwise analysis , which ig et al. , 1234 cug et al. . The EUsIn the current study, 82 Fluidigm SNP assay sets were used to show that identifying the 95 commercial F1 hybrids is possible at a high level. These SNP markers were able to classify the 95 commercial F1 cultivars by cultivar group, and cultivar-specific SNP marker sets could be generated for 77 cultivars for identification. Considering that the cultivars used in this study were commercial registration F1 hybrids (not PVP F1 hybrids), it could not be ruled out that the indistinguishable cultivars were the same but with different names. Because the genetic diversity of cucumbers is narrow [It is vital to understand the genetic relationships among cucumber accession to use them as breeding materials. In addition, efficient cultivar identification technologies that can protect cultivars and breeders’ intellectual property rights that are essential for improving the breeding industry. To the best of our knowledge, the present study is the first to provide information for developing and protecting cucumber cultivars using Fluidigm SNP assay sets. However, because our results were interpreted based on the limited resource information obtained from the NAC, further analysis with phenotypic information may add more value to what we have currently established. Our results can be useful for understanding the genetic characteristics underlying cucumber germplasm in South Korea for identifying new cultivars.http://genebank.rda.go.kr accessed on 12 May 2020). From these, 275 accessions from 30 countries across four continents  were collected, excluding five accessions whose origin was not disclosed. The resource characteristics included 78 heirlooms, 63 cultivars, 7 breeding lines, 1 genetic material, and 131 accessions without classification information   were selected from the cultivar inventory of the KSVS. A total of 95 commercial F1 hybrids were selected: 49 Baekdadagi-type, 23 Gasi-type, 6 Nakhap-type, and 17 cultivars that were difficult to classify . The classification of origin and resource characteristics of the 280 accessions used in this study followed the information disclosed by the NAC, and the cultivar groups of the 115 commercial F1 hybrids were classified according to the published cultivar names and opinions of cucumber breeders . ⓇGeneExTMPlant kit  according to the manufacturer’s instructions. The extracted DNA sample was used after quantitative and qualitative analysis using a spectrophotometer  and then diluted to a concentration of 20 ng·µL–1.Genomic DNA extraction for the Fluidigm SNP assay was performed by collecting the first fully expanded leaf from three seedlings per cultivar and then using a GeneAllFor genotyping 300 accessions for genetic diversity analysis, 151 of the 240 patent-pending cucumber Fluidigm SNP assay sets , locus-specific primer, and specific amplification primer)  were useTM Juno System . The SNPs were then called using the Fluidigm SNP Genotyping Analysis software (version 4.5.1).For genotyping 95 commercial registration F1 hybrids for cultivar identification, 82 of the 240 Fluidigm SNP assay sets described above were used . A totalThe population structure of 300 accessions was analyzed using an admixture-based model (Markov chain Monte Carlo (MCMC)-based Bayesian model) and considering independent allele frequencies n STRUCTURE 2.3.4 . The ranFST values between subpopulations was calculated using STRUCTURE 2.3.4. The genetic distance between accessions was calculated as described by Cavalli-Sforza and Edwards [Genetic distance between individuals in each subpopulation (expected heterozygosity) and pairwise  Edwards  using Po Edwards  and grou Edwards .PCA of the 95 commercial F1 hybrids was conducted using the pcaMethods package in R . An HC t


Avicennia marina trees. This disjunct zonation may be maintained through different positions in the tidal frame, yielding different dispersal settings. The spatial configuration of the landscape and coastal processes such as tides and waves is expected to largely influence the extent of propagule transport and subsequent regeneration. We hypothesized that landward sites would keep a stronger genetic structure over short distances in comparison with enhanced gene flow among regularly flooded seaward fringes. We tested this hypothesis from densely vegetated A. marina transects of a well‐documented mangrove system  and estimated local gene flow and kinship‐based fine‐scale genetic structure. Ten polymorphic microsatellite markers in 457 A. marina trees revealed no overall significant difference in levels of allele or gene diversities between sites that differ in hydrological proximity. Genetic structure and connectivity of A. marina populations however indicated an overall effect of geographic distance and revealed a pronounced distinction between channels and topographic setting. Migration models allowed to infer gene flow directionality among channels, and indicated a bidirectional steppingstone between seaward and nearest located landward stands. Admixed gene pools without any fine‐scale structure were found within the wider and more exposed Kidogoweni channel, suggesting open systems. Elevated kinship values and structure over 5 to 20 m distance were only detected in two distant landward and seaward transects near the mouth of the Mkurumuji River, indicating local retention and establishment. Overall, our findings show that patterns of A. marina connectivity are explained by hydrological proximity, channel network structure, and hydrokinetic energy, rather than just their positioning as disjunct landward or seaward zones.Mangrove ecosystems along the East African coast are often characterized by a disjunct zonation pattern of seaward and landward  A. marina trees, we show that connectivity patterns of landward–seaward A. marina populations in Gazi Bay (Kenya) are explained by hydrological proximity, channel network structure, and hydrokinetic energy, rather than just their positioning as disjunct landward or seaward zones. Genetic structure and connectivity of A. marina populations indicated an overall effect of geographic distance and revealed a pronounced distinction between channels and topographic setting.Using ten polymorphic microsatellite markers in 457  Exposed to the dynamic conditions of these intertidal environments , mangrove trees and shrubs display a number of adaptive strategies such as salt‐secreting glands, aerial roots, and the production of hydrochorous propagules   to transoceanic distances. Dispersal potential and the patterns of gene flow depend on the cumulative effect of a wide range of factors, such as propagule buoyancy and viability period, species‐specific propagule morphological traits (size and shape), landscape complexity, and the position of the parent tree in the tidal frame  in the Yucatan Peninsula, and found comparable genetic differences within populations from the same site as between populations from different sites. The authors ascribed this pattern to high autogamy rates, asynchronous phenology between populations, as well as limited dispersal due to the interaction between large propagule size and the physical barrier presented by the species’ intricate root system. Similarly, Ngeve et al. (Rhizophora racemosa G. Mey. (Rhizophoraceae) in a Cameroonian estuary complex. However, fine‐scale spatial genetic structure was not observed in all of the estuaries studied, and where absent, was explained by the recent recolonization of areas that are cleared for coastal development  Vierh. (Acanthaceae) zones in a Kenyan mangrove forest, indicating that there might be less genetic interchange between these intertidal zones than within each zone. Recently, Chablé Iuit et al. (R. mangle in the southern part of Quintana Roo (Mexico) and reported that the fine‐scale spatial genetic structure reflects contemporary processes such as restricted propagule dispersal and local hydrology.Previous studies found correlations between genetic differentiation and geographic distance  analyze the genetic structure between seaward and landward mangrove patches, positioned along a same transect perpendicular to a channel; (b) estimate patterns of connectivity in the light of channel network structure; and (c) examine the fine‐scale spatial genetic structure of mangrove patches located along the same channel. The local‐ and fine‐scale spatial genetic structure of A. marina can be hypothesized to maintain higher levels of connectivity among the more regularly flooded seaward sites and higher kinship values (relatedness) with a stronger structure over short distances in higher intertidal (landward) sites. We test this hypothesis of confinement in landward sites versus open connectivity between seaward sites using densely vegetated transects in a well‐documented mangrove area . The study site and species present an ideal case to undertake this study, given the disjunct (landward–seaward) pattern of A. marina in the area and the regional setting that is characterized by a series of open‐water channels. Throughout this manuscript, we use the term “landward” to refer to “higher intertidal” and “seaward” for “lower intertidal.” It is important to note that since channels have different orientations relative to the coastline, seaward does not necessarily mean oriented toward the sea. Instead, the terms “landward” and “seaward” reflect different geographical proximities  to one of the three major water channels that cross the area's mangrove forest.The goal of this study is to characterize the genetic structure and diversity of the mangrove species 22.1Avicennia marina (Forsk.) Vierh. (Acanthaceae), Sonneratia alba J. Smith (Lythraceae), Rhizophora mucronata Lam. (Rhizophoraceae), Ceriops tagal (Perr.) C.B. Robinson (Rhizophoraceae), Bruguiera gymnorrhiza (L.) Lam. (Rhizophoraceae), and Xylocarpus granatum J. Koenig (Meliaceae) , about 45 km south–southwest of Mombasa Figure . Climate2.2Avicennia marina is the most widely distributed of all mangrove species, found across the Indo‐Pacific, between latitudes 25°N and 38°S. It has been shown that A. marina is able to grow and reproduce across a relatively broad range of climatic, saline, and tidal conditions  distribution pattern across the intertidal zone, typically separated by formations of C. tagal and R. mucronata, with the trees from both zones often showing apparent physiognomic differences: trees in the landward fringe can show a rather shrub‐like appearance with an average height of ca. 3 m, and taller and more robust trees with an average height of ca. 10 m in the seaward fringe . SSR_pipeline  with the following conditions: an initial denaturation of 95°C for 15 min followed by 35 cycles of: 30 s denaturation at 95°C, 90 s annealing at 57°C and 80 s elongation at 72°C followed by a final extension of 30 min at 60°C. PCR products were separated on an ABI3730XL sequencer , and allele sizes were determined with GeneMarker V2.60 .Genomic DNA was extracted from approximately 20 mg of dried leaf tissue using the E.Z.N.A. SP plant DNA Mini kit . A multiplex polymerase chain reaction (PCR) consisted of in total 10 microsatellite markers Appendix . Six of 2.5A. marina. A linkage test between all pairs of loci  gave no genotypic disequilibrium at the 0.05 level using FSTAT (v.2.9.3) , namely whether two individuals could share an identical multilocus genotype by chance using GenAlEx , mean number of alleles (AM), effective number of alleles (AE), allelic richness (AR) for 46 diploid samples, observed heterozygosity (HO), unbiased expected heterozygosity (HE), and population inbreeding coefficient (FIS) using FSTAT and GenAlEx. The genetic structure among sites (FST), inbreeding within sites (FIS), and overall inbreeding (FIT) was calculated via AMOVA–FST at 999 random permutations using GenAlEx v.6.5, thereby allowing to estimate overall connectivity levels as Nm = FST/(1‐4FST) under the assumption of an island migration model within the Gazi Bay. An additional hierarchical AMOVA was performed and F‐statistics were calculated, considering three channels  as regions, and using 999 random permutations. Pairwise genetic differentiation (FST) was used to produce a PCoA at population level and together with a pairwise geographic Euclidean distance to perform a Mantel test using 1,000 permutations in GenAlEx (v.6.5). Pairwise genotypic differentiation was used to produce a PCoA at individual level. The overall FIJ kinship coefficient  repeats. The optimal K value was inferred with the ΔK statistic , and the Puechmaille , we opted to calculate from superposition of basic data from different FST matrices at locus level. The thickness of barrier lines thus will be based on the additivity of matrices accounting for the variability of different markers that we consider as a preferred informative and valid method over bootstrapping a single mean FST matrix.A Bayesian clustering analysis at individual level was carried out in STRUCTURE version 2.3.4 . We used the Bezier thermodynamic integration  was calculated as [Theta × M]/4  analysis   between neighboring individuals in the first distance class (0–5 m). Under an assumption of isolation‐by‐distance  and two‐dimensional  space, the neighborhood size can be estimated as Nb = 1/Sp , with a mean number of alleles (AM) ranging between 3.5 and 4.2, an effective number of alleles (AE) between 2.0 and 2.5, and an adjusted allelic richness (AR) between 3.3 and 4.1 (Table HO = 0.480) was very similar to the expected heterozygosity (uHE = 0.500). The within‐population inbreeding (mean ISF = 0.059) ranged from −0.014 to 0.147 and was significant only for site 5S (Table p > .05) differences in the levels of AR, HO, HE, and FIS, whereas two young stands (2L and 7S) showed slightly higher allelic richness  and gene diversity  compared to eight older stands. Overall, we observed very similar amounts for basic population genetic variables for most A. marina sites within the bay.In 3.2Avicennia marina within Gazi Bay showed an overall AMOVA–FIT = 0.122, FST = 0.067, and FIS = 0.059, though with all these low values at p = .001  and FSR = 0.041 (p < .001) with as much variance among channels (4%) than among populations (4%). Pairwise differentiation ranged from 0.008 for transects in close vicinity (3S and 4L) to 0.126 for more distant landward sites (6L and 8L). Both PCoA at individual and population level showed a gradient along the first axis of the locations 8L, 7S, and 1S, although A. marina individuals clustered as a single cloud  over 5.4 km  decreased significantly over the full distance (slope b = −0.03 at p < .001), with significantly (p < .05) higher kinship values at 0.3 km, 0.6 km, and up to a maximum distance of 1.5 km  and reached a marginally higher value at K = 5 with Delta K = 18  = −8334 when compared to K = 2 (mean LnP(K) = −8578). K = 5 showed the highest mean similarity score (0.988) for CLUMPAK, and K = 5 was obtained with the Puechmaille  and their neighbors, thereby separating the Gazi Bay transects according to the three major water channels  and from 3S toward 5S (Nem = 0.64). Lowest gene flow estimates were found from landward toward seaward stands . Various source–sink models as well as panmixia appeared less likely than this bidirectional steppingstone model. The outcome of this MIGRATE analysis supports the idea of well‐connected landward and seaward Avicennia populations along the Kidogoweni River, though dominated by an upstream movement reflecting flood tide rather than a downstream movement during ebb tide.MIGRATE was used to estimate the mutation‐scaled population sizes and immigration rates using the Brownian model. The specific testing on gene flow directionality between disjunct seaward and landward sites along the Kidogoweni channel gave best support for a bidirectional steppingstone model Table . EstimatM = 9.2–9.4  to the Kidogoweni channel (5S) andThe connectivity among the mouth of three major water channels in Gazi Bay was best supported from a unidirectional steppingstone model Table . Both unNem = 0.91 from the Kidogoweni channel (5S) to the Kinondo channel (1S).3.3A. marina in Gazi Bay revealed an average intragroup kinship FIJ = 0.061, however with an overall stronger kinship FIJ = 0.072 at a mean distance of 5.6 m (p < .001) and FIJ = 0.066 at a mean distance of 16.2 m (p < .001), with a log‐slope b = −0.008 (p < .001). A detailed analysis of the fine‐scale spatial genetic structure of each A. marina transect revealed only a significant different kinship value and slope within shortest distance class for sites of the southernmost channel, 7S and 8L  and FIJ = 0.036 (p = .001) for distance classes of 5 and 10 m, respectively, in transect 7S  and FIJ = 0.012 (p = .018) for distance classes of 0–5 and 10–25 m, respectively, in transect 8L , similarly for both 7S and 8L. The overall Sp value for Gazi Bay was 0.009 but with a considerable range. The Sp‐statistic was low (Sp = 0.020–0.021) for sites 7S and 8L (Mkurumuji) that showed a fine‐scale spatial genetic structure and appeared even much lower (Sp = 0.0010–0.0080) for all other sites lacking a clear genetic structure at short distances. The estimated neighborhood size was limited to Nb = 47–49 in sites 7S and 8L, respectively, whereas elsewhere the estimates ranged from Nb = 125 to Nb = 982.The spatial autocorrelation of individuals within transects of L Figure . The kinS Figure  and FIJ44.1vice versa. This is consistent with findings from Dahdouh‐Guebas et al. . Hence, only during a limited time frame of the monthly tidal cycle are landward sites hydrologically connected to the more seaward stands, and can propagules be transported directly between mangrove patches at different elevations in the intertidal zone. Importantly, these differences in hydroperiod also present different windows of opportunity for propagules to strand and develop roots , leaving a large amount of litter in a relatively long inundation‐free time window.Landward and seaward zones are characterized by important differences in hydroperiod (duration of submergence). Differences in hydroperiod present different timeframes for dispersal Increased potential for establishment in landward patches is also favored by the lower hydrodynamic forces from waves and currents, compared to landward zones. In a study on the mangrove forest structure dynamics in Gazi Bay, Di Nitto et al.  noted thNeosarmatium africanum) were shown to clear nearly 100% of the propagules in landward stands  revealed significant genetic differentiation between sites that are situated along different water channels . High longitudinal  connectivity was found for the Kidogoweni channel but not for Mkurumuji . Overall, this is consistent with findings that stream channels may act as corridors for dispersal  was not a subset of the older mangrove along Kidogoweni (5S), supporting the abovementioned flood‐mediated directionality from south to north. Significant genetic differentiation among channels may result from the tidal asymmetry in the region, characterized by stronger ebb than flood tide and resulting in a net export  found for A. marina in Gazi Bay is among the ranges reported for outcrossing trees in general  come close to patterns for “gravity‐dispersal” , the abovementioned studies on A. germinans considered first distance classes of 0–50 m ; Data curation (lead); Formal analysis (lead); Funding acquisition ; Investigation (lead); Methodology (lead); Resources (lead); Visualization (supporting); Writing‐original draft (lead); Writing‐review & editing (supporting). Tom Van der Stocken: Conceptualization (lead); Investigation (lead); Methodology (lead); Visualization (lead); Writing‐original draft (lead); Writing‐review & editing (lead). Abbie Allela Akinyi: Data curation (supporting); Formal analysis (supporting). Tim Sierens: Formal analysis (supporting); Methodology (supporting). James Kairo: Data curation (supporting); Funding acquisition . Nico Koedam: Conceptualization (lead); Funding acquisition ; Investigation (supporting); Project administration (lead); Resources (lead); Writing‐review & editing (supporting).Appendi S1‐S4Click here for additional data file.