Novel microsatellite markers for Begonia maxwelliana and transferability to 23 Begonia species of Peninsular Malaysia

Begonias are hyper-diverse and important horticultural plants. Six polymorphic microsatellite markers were developed from CT- and GT-enriched libraries of Begonia maxwelliana. The number of alleles per locus ranged from 2 to 12 and the observed heterozygosity ranged from 0.036 to 0.813. Null alleles were detected in one locus (Bma161) after Bonferroni correction. All the six markers were amplifiable in 23 selected Begonia species with the success rates of 17–100%. On average, species of the same section as B. maxwelliana (i.e. sect. Platycentrum) yielded higher transferability (91%). These markers will be useful for population genetic studies of the genus Begonia.     

Isolation and characterization of microsatellite loci for Hymenaea courbaril and transferability to Hymenaea stigonocarpa, two tropical timber species

Hymenaea courbaril is a tropical timber species, intensely exploited and found in the Amazon, Atlantic Forest and Brazilian Cerrado biome. Nine highly polymorphic microsatellite loci were developed from a genomic library enriched for AG/TC repeats. In a total of 41 individuals, from two natural populations, seven to 13 alleles per locus were detected and expected heterozygosity ranged from 0.75 to 0.90. Seven loci were effectively transferred to Hymenaea stigonocarpa. High levels of polymorphism make the present primers useful for population genetic studies and are a powerful tool to investigate mating system, gene flow and spatial genetic structure.     

Cross transferability of SSR markers to endangered Cedrela species that grow in Argentinean subtropical forests,as a valuable tool for population genetic studies

Species of Cedrela with a high economic value from Northwest and Northeastern Argentina are severely exploited. This work evaluates whether 51 nuclear SSRs, developed to study phylogenetically close species in the Meliaceae family (Cedrela odorata, Cedrela fissilis, Swietenia humilis and Swietenia macrophylla), can be used to study C. fissilis, Cedrela balansae, Cedrela saltensis and Cedrela angustifolia. A 62.8% of the total of 194 SSRs/species combinations showed a successful, homologous and cross-species amplification. As expected, a great success in SSRs transferability among Cedrela species was observed. Twenty-one screened SSRs showed a successful amplification pattern in all target species and many of them were polymorphic (9, 13, 13 and 7 SSRs for C. fissilis, C. balansae, C. saltensis and C. angustifolia, respectively). The high number of evaluated SSRs from the Cedrela genus and Meliaceae family, allowed us to obtain a suitable set of validated markers that are highly variable and easily scored, and also identify those which were less sturdy. We were able to retain a useful set of markers for three of the target species, but not for C. angustifolia. This could be due to its greater phylogenetic and morphological distances to the other three species. The lack of SSRs developed for our target species, transforms the transferred SSRs reported here in a valuable tool to monitor the genetic consequences of forest overexploitation on Cedrela species.     

SSRs transferability and genetic diversity of Tunisian Festuca arundinacea and Lolium perenne

SSR primers specific to Lolium perenne generated a total of 96 alleles and 124 genotypes within Festuca arundinacea and Lolium perenne accessions. Their highly transferability (100 %) across genera was evidenced. Six alleles specific to loci H01F02, H02C11 and K01A03 and only 5/96 common alleles between both species (60, 140, 144, 190 and 192) expressed the differentiation between species. Besides, based on the Wrights fixation indices, the genetic variation within each species was attributable to differences within populations with a significant deficiency of heterozygous. The unweighted pair group method with arithmetic averaging dendrogram based on the Nei’s distances and the principal coordinate analysis based on Jaccard coefficient similarity distinguished each genus independently of the geographical origin. However, typically continuous genetic diversity and a low level of gene flow (N_m: 0.29–2.47) expressed the relatively closely relationships of both genera and suggest a possible hybridization in nature.     

近缘种扩增法对黑叶猴微卫星位点的筛选及特征分析(英文)

微卫星位点近缘种筛选法使得在探讨各种灵长类种群遗传结构和生殖策略上更加便捷。我们利用138条人类微卫星引物在黑叶猴中进行筛选,得到了23个具有多态性位点。在28个检测个体中,每个位点的等位基因数为3到9个,期望杂合度为0.62,观测杂合度为0.50,其中有7个位点偏离Hardy-Weinberg平衡,9个位点存在无效等位基因现象。但是各位点之间均未检测到连锁不平衡现象。这些位点将在黑叶猴种群遗传结构的研究中发挥重要作用。     

Development and characterization of novel microsatellite markers in Puccinia striiformis f.sp. tritici and their transferability in Puccinia species

Puccinia striiformis f.sp. tritici (Pst) and P. striiformis f.sp. hordei (Psh) causing stripe rust disease in wheat and barley, respectively, are two devastating phytopathogens. Microsatellite/simple sequence repeat (SSR) markers are increasingly being utilized for analysis of genetic diversity, diagnosis, population structure and possible migratory routes of plant pathogens. In the current study, novel polymorphic SSR markers were designed for Pst using the genomic sequences of PST-78 isolate. A total of 1,191 SSR motifs, comprising 30% each of di- and tri-nucleotide type of repeats, 17% of penta-nucleotide, 15% of tetra-nucleotide and 8% of hexa-nucleotide repeats, were detected through in silico scanning of PST-78 genomic sequences. Polymorphism was detected by nine of the 50 designed SSRs (PsSSRs) in seven stripe rust pathotypes of wheat and barley. The mean number of alleles per SSR locus, mean polymorphism information content (PIC), mean heterozygosity, mean major allele frequency (MAF) and mean gene diversity were 2.33, 0.34, 0.33, 0.71 and 0.40, respectively. The dendrogram analysis suggested that newly developed PsSSR markers could distinguish stripe rust pathotypes based on their virulence phenotype. Further, the cross-genera and cross-species amplification test of these markers in 14 different rust pathotypes revealed that 9 PsSSRs are capable of amplification in Pst species infecting wild grass, followed by 6 PsSSRs in Pt, 3 PsSSRs in Pgt, 1 PsSSRs in Puccinia species on barberry and Melampsora lini. Thus, the transferability of PsSSRs to other species reduced with increasing genetic distance of target species. These newly designed SSR markers expand the available Pst SSR marker resources and allow better genetic studies.     

Individual assignment tests proved genetic boundaries in a species complex of Pacific abalone (genus <Emphasis Type="Italic">Haliotis</Emphasis>)

We conducted this study to find genetic evidence to distinguish the members of Pacific abalone species complex (Haliotis discus hannai, H. discus discus, H. madaka, and H. gigantea) based on microsatellite DNA markers, illustrating the potential of microsatellites for species-assignment. First, we addressed the transferability of H. discus hannai microsatellites to the three other members of Pacific abalone and five additional species (H. diversicolor aquatilis, H. midae, H. corrugata, H. fulgens, and H. rubra). Second, using the microsatellites we applied two types of individual assignment testing (the distance-based assignment and Bayesian model-based clustering) to individuals from the Pacific abalone species. A total of 24 microsatellites were subjected to PCR trials for nine Haliotis species, and the cross-species amplification performance of these markers turned out to drop precipitously even for less divergent congeners. Within the Pacific abalone species complex, four of the 24 markers were not transferable to H. gigantea, suggesting a solid genetic boundary between H. gigantea and H. discus hannai, H. discus discus, and H. madaka. Among the three latter abalones, both assignment tests achieved approximately 90% or more success rate of assignment. The feasibility of the microsatellite markers to classify species sheds light on the genetic management of the Pacific abalone species complex. Electronic Supplementary Material  Supplementary material is available in the online version of this article at and is accessible for authorized users.

Cross-species amplification of soybean (Glycine max) simple sequence repeats (SSRs) within the genus and other legume genera: implications for the transferability of SSRs in plants

We investigated the transferability of 31 soybean (Glycine max) simple sequence repeat (SSR) loci to wild congeners and to other legume genera. Up to 65% of the soybean primer pairs amplified SSRs within Glycine, but frequently, the SSRs were short and interrupted compared with those of soybeans. Nevertheless, 85% of the loci were polymorphic within G. clandestina. Cross-species amplification outside of the genus was much lower (3%-13%), with polymorphism restricted to one primer pair, AG81. AG81 amplified loci in Glycine, Kennedia, and Vigna (Phaseoleae), Vicia (Vicieae), Trifolium (Trifolieae), and Lupinus (Genisteae) within the Papilionoideae, and in Albizia within the Mimosoideae. The primer conservation at AG81 may be explained by its apparent proximity to the seryl-tRNA synthetase gene. Interspecific differences in allele size at AG81 loci reflected repeat length variation within the SSR region and indels in the flanking region. Alleles of identical size with different underlying sequences (size homoplasy) were observed. Our findings and the emerging patterns in other plant studies suggest that in contrast to animals, successful cross-species amplification of SSRs in plants is largely restricted to congeners or closely related genera. Because mutations in both the SSR region and the flanking region contribute to variation in allele size among species, knowledge of DNA sequence is essential before SSR loci can be meaningfully used to address applied and evolutionary questions.      

Development, characterization and transferability of microsatellite markers for the plant genus Phlox (Polemoniaceae)

In order to study diversification and microevolution in Phlox, we developed nine polymorphic microsatellite loci. In 20 individuals of Phlox pilosa from a single population, the average number of alleles per locus was 10.0 ± 5.1, and average observed and expected heterozygosities were 0.611 ± 0.234 and 0.769 ± 0.170, respectively. Most of these markers amplified successfully in 11 additional species of Phlox, representing a broad diversity of the genus, and some also amplified in more distantly related members of the Polemoniaceae. These microsatellite markers will be valuable for investigation of evolutionary processes in this important study system.     

Reverse ascertainment bias in microsatellite allelic diversity in owls (Aves, Strigiformes)

A rich source of markers may be overlooked by screening for polymorphism in the source species only. We screened 129 microsatellite loci isolated from the powerful owl (Ninox strenua) against two closely related species; Ninox connivens and Ninox novaeseelandiae. From the screening effort 20 polymorphic markers were isolated, including six loci which were originally discarded as they were monomorphic in the source species. Further cross-species amplification of all 20 loci across species from two families, Strigidae and Tytonidae, revealed unusually high levels of polymorphism within closely related species, and limited success within phylogenetically distant species. Routine screening of multiple species during the marker development phase can yield a wider range of polymorphic markers which can subsequently enhance cross-species amplification attempts.     

Characteristics and transferability of new apple EST-derived SSRs to other Rosaceae species

Genic microsatellites or simple sequence repeat markers derived from expressed sequence tags (ESTs), referred to as EST–SSRs, are inexpensive to develop, represent transcribed genes, and often have assigned putative function. The large apple (Malus × domestica) EST database (over 300,000 sequences) provides a valuable resource for developing well-characterized DNA molecular markers. In this study, we have investigated the level of transferability of 68 apple EST–SSRs in 50 individual members of the Rosaceae family, representing three genera and 14 species. These representatives included pear (Pyrus communis), apricot (Prunus armeniaca), European plum (P. domestica), Japanese plum (P. salicina), almond (P. dulcis), peach (P. persica), sour cherry (P. cerasus), sweet cherry (P. avium), strawberry (Fragaria vesca, F. moschata, F. virginiana, F. nipponica, and F. pentaphylla), and rose (Rosa hybrida). All 68 primer pairs gave an amplification product when tested on eight apple cultivars, and for most, the genomic DNA-derived amplification product matched the expected size based on EST (in silico) data. When tested across members of the Rosaceae, 75% of these primer pairs produced amplification products. Transferability of apple EST–SSRs across the Rosaceae ranged from 25% in apricot to 59% in the closely related pear. Besides pear, the highest transferability of these apple EST–SSRs, at the genus level, was observed for strawberry and peach/almond, 49 and 38%, respectively. Three markers amplified in at least one genotype within all tested species, while eight additional markers amplified in all species, except for cherry. These 11 markers are deemed good candidates for a widely transferable Rosaceae marker set provided their level of polymorphism is adequate. Overall, these findings suggest that transferability of apple EST–SSRs across Rosaceae is varied, yet valuable, thereby providing additional markers for comparative mapping and for carrying out evolutionary studies.     

Novel microsatellite markers obtained from Gansu zokor (Eospalax cansus) and cross-species amplification in Plateau zokor (Eospalax baileyi)

Zokor (Mysopalacinae) is a group of subterranean rodents. Although they are regarded as important components in the ecosystems they belong to, due to their underground lifestyle, very little is known about their ecology and behavior. Development of microsatellite markers can potentially assist in addressing behavioral and ecological questions such as dispersal, mating and social systems. Here we report the development of 11 polymorphic microsatellite loci from the genome of Eospalax cansus using 454 shot-gun sequencing. Genetic diversity was assessed using DNA samples of 47 individuals of E. cansus from a single location. Cross-amplification in Eospalax baileyi was also tested, with five polymorphic markers being amplified successfully. These markers are the first published microsatellites loci from E. cansus and will be invaluable for studies addressing ecological and behavioral questions involving E. cansus and E. baileyi, and potentially other species in Mysopalacinae.     

Genetic diversity and population structure of the endemic Azorean juniper,Juniperus brevifolia (Seub.) Antoine,inferred from SSRs and ISSR markers

Juniperus brevifolia is an important woody species endemic of the Azores archipelago (Portugal), found from coastal to mountain environments. Due to colonization and grazing pressure this species has suffered fragmentation, leading to extinction in one island and being threatened in others. The genetic diversity and population structure of J. brevifolia populations was studied to provide guidelines for restoration and conservation programmes. Nuclear Single Sequence Repeats (nSSR) from Juniperus communis and Juniperus przewalskii and Inter Simple Sequence Repeats (ISSR) were tested and results compared to test the transferability of the microsatellites to J. brevifolia. Samples from ten populations over three islands, divided as coastal, mid-altitude and mountain were analysed. Both marker systems revealed results statistically and strongly correlated with each other, and not dependent on population sample size. We observed positive fixation indexes, moderate to high levels of genetic diversity (h = 0.415 for nSSR and h = 0.245 for ISSR), low to moderate φ_pt genetic differentiation (0.070 for nSSR and 0.129 for ISSR) and high gene flow (N_m > 2.432). Regarding Nei's genetic distance the coastal communities clustered together reflecting the phenotypic plasticity, but no specific clustering was observed regarding φ_pt values. Therefore no populations with substantial genetic differentiation were identified, once the diversity is mostly observed within populations. However it is advised the continuous monitoring of J. brevifolia.     

Isolation and characterization of polymorphic microsatellite loci from a dinucleotide-enriched genomic library of starry flounder (Platichthys stellatus) and cross-species amplification

Starry flounder (Platichthys stellatus) is a rare fish species in China. Here, we reported 12 polymorphic microsatellite loci isolated from a dinucleotide-enriched genomic library of starry flounder (P. stellatus). The number of alleles, observed and expected heterozygosity per locus in 30 individuals ranged from two to six, from 0.2500 to 1.0000 and from 0.4512 to 0.7667, respectively. One locus significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium between pairs of loci was found. Cross-species amplification of these microsatellite loci in additional three fish species was performed. These polymorphic microsatellite loci would be useful for investigating genetic population structure and construction of genetic linkage map in P. stellatus. Guidong Miao and Changwei Shao have contributed equally.     

鲤科鱼蓝黑鲮微卫星位点的种间扩增(英文)

本研究使用105对微卫星引物对7种鲤科鱼类进行跨越种间PCR扩增,共得到14个多态性微卫星位点.其中9个扩增效果较好的位点用于分析来自帕吉勒提河(Bhagirathi, n=20)和戈达瓦里河(Godavari, n=25)的蓝黑鲮(Labeo calbasu)样品的遗传多样性.结果显示,前者在每个位点的平均等位基因数为7.33,而后者为8 1,期望杂合度介于0.795(Bhagirathi)和0.801(Godavari)之间;4个位点MFW11* (Godavari)、R1*(Godavari)、R3* (Bhagirathi) 和 Lr38*(Bhagirathi和Godavari)都表现出明显的杂合子缺失和哈迪温伯格平衡偏离;而任意两位点间都未观测到连锁不平衡现象;位点R3*极可能存在无效等位基因.上述结果表明这些多态性微卫星位点作为共显性标记在蓝黑鲮群体遗传学研究中有着较好的应用前景.     

Assessing and improving the transferability of current global spatial prediction models

Aim

Global-scale maps of the environment are an important source of information for researchers and decision makers. Often, these maps are created by training machine learning algorithms on field-sampled reference data using remote sensing information as predictors. Since field samples are often sparse and clustered in geographic space, model prediction requires a transfer of the trained model to regions where no reference data are available. However, recent studies question the feasibility of predictions far beyond the location of training data.

Innovation

We propose a novel workflow for spatial predictive mapping that leverages recent developments in this field and combines them in innovative ways with the aim of improved model transferability and performance assessment. We demonstrate, evaluate and discuss the workflow with data from recently published global environmental maps.

Main conclusions

Reducing predictors to those relevant for spatial prediction leads to an increase of model transferability and map accuracy without a decrease of prediction quality in areas with high sampling density. Still, reliable gap-free global predictions were not possible, highlighting that global maps and their evaluation are hampered by limited availability of reference data.