Genetic variability and association of ISSR markers with some biochemical traits in mulberry (Morus spp.) genetic resources available in India

Utilizing intersimple sequence repeat (ISSR) markers, 18 mulberry (Morus spp.) germplasm collections were studied for genetic variability, phylogenetic relationship, and association with protein and sugar content. The genetic polymorphism exhibited by ISSR primers was 100%, and the genetic diversity recorded among the mulberry accessions had an average of 0.263 ± 0.094. Dendrogram (unweighted pair group method analysis) clustered the mulberry accessions into two major groups, one comprised the accessions collected from north or northeast regions of India, and the other comprised three subclusters and one isolate, i.e., Assamjati, a collection from Assam. Another subcluster contained accessions collected from Kerala, which belong to Morus indica. These accessions of M. indica from Kerala were found to be genetically diverse from north and northeast India. Multidimensional scaling of the ISSR data clearly separated the mulberry accessions according to their genetic diversity and protein content. Mulberry accessions were arbitrarily grouped into three classes viz. very low, moderate, and high in terms of protein and sugar content using standard statistical programs. Stepwise multiple regression analysis identified four ISSR markers (835_1,600, 835_5,600, 822_2,500, and 807_2,500) associated with protein content with highly positive correlation (p < 0.001) with linear curves with high F values (18.055 to 48.674; p < 0.001). In case of sugar content, four ISSR markers viz. 812₉₀₀, 817_1,500, 826_1,500, and 810_8,000 showed negative correlation. Hence, DNA markers for proteins seem promising and may be used in marker-assisted breeding program.     

Fine‐scale Population Genetic Structure of Two Dioecious Indian Keystone Species,Ficus hispida and Ficus exasperata (Moraceae)

Although Ficus (Moraceae) is a keystone plant genus in the tropics, providing resources to many frugivorous vertebrates, its population genetic structure, which is an important determinant of its long‐term survival, has rarely been investigated. We examined the population genetic structure of two dioecious fig species (Ficus hispida and Ficus exasperata) in the Indian Western Ghats using co‐dominant nuclear microsatellite markers. We found high levels of microsatellite genetic diversity in both species. The regression slopes between genetic relationship coefficients (f_ij) and spatial distances were significantly negative in both species indicating that, on average, individuals in close spatial proximity were more likely to be related than individuals further apart. Mean parent–offspring distance (σ) calculated using these slopes was about 200 m in both species. This should be contrasted with the very long pollen dispersal distances documented for monoecious Ficus species. Nevertheless, overall population genetic diversity remained large suggesting immigrant gene flow. Further studies will be required to analyze broader scale patterns.     

Primers for the amplification of sequence‐characterized loci in Cryphonectria cubensis populations

We describe the development of DNA markers for the fungal pathogen of Eucalyptus, Cryphonectria cubensis. These markers originated from cloned intershort sequence repeat polymerase chain reactions, which enrich for medium to highly repetitive DNA sequences. In total, 10 markers were isolated, eight of which were polymorphic, and these can subsequently be applied to study populations of C. cubensis.     

Genetic structure and essential oil composition in wild populations of Salvia multicaulis Vahl.

The phytochemical study on ten populations of Salvia multicaulis Vahl. revealed that their essential oil qualitative profiles contained a significant amount of monoterpene hydrocarbons, which were the most abundant compounds. Besides, α-Pinene was the major constituent in all studied populations' essential oils. Significant correlations were observed between edaphic parameters and some major essential oil compounds. According to clustering analyses of the chemical data, the S. multicaulis populations were divided into three chemotypes: β-caryophyllene, camphene and camphor, and limonene. The population genetics study showed significant molecular differences among the populations. The Mantel test indicated a significant positive correlation between the geographical distances and genetic diversity, exhibiting a low amount of gene flow and a considerable genetic differentiation value. We also detected four genotypes based on the Nei's genetic distance and structure analysis. The identified chemical and genetic similarities/differences among these populations were correlated with edaphic parameters and geographic distances, suggesting that environmental factors are the primary drivers of the chemical polymorphism of essential oils in S. multicaulis populations.     

Genetic diversity and phylogenetic relationship of kenaf (Hibiscus cannabinus L.) accessions evaluated by SRAP and ISSR

Understanding genetic diversity and phylogenetic relationships is useful for plant breeding. In this study, we assessed the genetic diversity in a panel of 84 accessions of kenaf from 26 countries using SRAP and ISSR markers. The kenaf accessions could be divided into L1 (60 cultivated varieties) and L2 (24 wild accessions) at the level of 0.145 genetic dissimilarity coefficient by UPGMA. The L₂ group was further divided into two subgroups (16 relative-wide and 9 origin wide accessions) at the level of 0.207 genetic dissimilarity. Out of the 9 wild accessions in the L₂ group, 6 were from Tanzania and the remaining 3 lines were from Kenya. These results suggest that the center of origin for kenaf might be Tanzania and Kenya.     

Genetic and allelopathic differences between populations of daisy fleabane Erigeron annuus (L.) Pers. (Asteraceae) from disturbed and stable habitats

The interactions between invasive plants and their habitats may vary at different phases of the invasion process and depend on the phenotypic plasticity or local adaptations of each species. In this study, we investigated whether habitat changes during the invasion process are related to variations in the physiological traits (allelopathic properties) and genetic differentiation of daisy fleabane (Erigeron annuus (L.) Pers.). E. annuus is a winter annual invasive species that originated in North America and is now distributed throughout Europe. Genetic and genotypic diversity analyses were performed for 37 populations of E. annuus based on inter simple sequence repeat (ISSR) polymorphisms. In total, 684 plants were analyzed; 342 were from stable habitats and 342 were from disturbed habitats. The genetic differences among the populations from the different habitats were studied using a Bayesian cluster analysis and an analysis of molecular variance (AMOVA) and by calculating the genetic and genotypic diversity parameters. A germination test using the juglone index was employed to examine the potential allelopathic properties of the plants from the different habitats. Bayesian cluster analysis, AMOVA and allelopathic effects evaluation revealed differences in the allelopathic potential and genetic structure of the E. annuus populations from the disturbed and stable habitats. This differentiation of populations could be associated with founder effects or with different selection pressures among habitats.     

“Barcoding” and ISSR data illuminate a problematic infraspecific taxonomy for Chrysanthemoides monilifera (Calenduleae; Asteraceae): Lessons for biocontrol of a noxious weed

Chrysanthemoides monilifera Tourn. ex Medik is a noxious weed in Australia and New Zealand. It is a widespread species in southern Africa, where it shows considerable morphological variation that has resulted in a confusing infraspecific taxonomy. We use DNA sequence data from the nuclear Internal Transcribed Spacer (ITS) region from 78 samples of this species from its indigenous distribution range and 10 samples from Australia and New Zealand to determine the regions of origin of the invasive plants. These data are augmented by a smaller study using ISSR markers. Bayesian Inference analysis was somewhat resolved, with many weakly supported nodes. Clades with support tended to correspond to infraspecific taxonomic entities, and were geographically coherent. In contrast, a neighbour-net analysis was not as well resolved and indicated considerable reticulation. All analyses of ITS data retrieved two major clades corresponding to Western and Eastern distributions, with some overlap. Samples from New Zealand and Australia correspond to the taxon C. monilifera subsp. monilifera, and are resolved as most closely related to samples from the greater Cape Town area. Biological control agent populations for C. monilifera subsp. monilifera should be sourced from this region in order to avoid host plant incompatibility problems.     

黄淮海地区玉蜀黍黑粉菌群体遗传结构分析

利用17个多态性ISSR引物对2017年在我国黄淮海地区采集的41株玉蜀黍黑粉菌Ustilago maydis样本进行了种群遗传结构分析。基于UPGMA法和贝叶斯模型对玉蜀黍黑粉菌群体遗传结构分析显示,采集的样本可以分为两大群,且种群的划分表现出了与来源地理纬度的一致性。因此按照各样本所属地理纬度将样本分为北纬33°-34°、北纬35°-36°、北纬37°-38° 3大种群。结果表明,分离自北纬33°-34°地区的玉蜀黍黑粉菌遗传多样性最丰富,Nei’s基因多样性（H）为0.2216,Shannon指数（I）为0.3300。分子方差分析（AMOVA）显示,变异中来自种群内部的变异为86%,种群间变异14%（φpt=0.141）。     

Activation of transposable elements and insertional polymorphism in Opuntia offspring as assessed by inter-retrotransposon amplified polymorphism markers

Activation of retrotransposon is a pivotal factor in the genesis of genetic polymorphism. Retrotransposon-based molecular markers are excellent tools for detecting genetic diversity and genomic changes associated with their activity. The objective of this study was to use IRAP markers to detect integration events of retrotransposon in Opuntia, and to compare IRAP and ISSR polymorphisms. To achieve these aims, five IRAP and five ISSR markers were analyzed on three varieties and their progenies. All IRAP primers showed an increase in the percentage of polymorphism, number of total bands, and polymorphic bands in the seedlings compared to their mother plants; that is, the offspring showed 13, 24 and 27 more bands than the mother plants from Tobarito, Montesa and Copena varieties, respectively. Conversely, sexual reproduction did not proportionally affect the variation in the number of ISSR bands, and neither did polymorphisms between mother plants and their offspring. Cluster and multivariate analyses based on IRAP and ISSR data revealed a clear separation between varieties, and there was no overlapping between seedlings of different varieties. The activation of retrotransposons in seedlings of opuntia with variable frequencies was evidenced. The presence of insertion and active retrotransposons may help to undertake studies on genetic diversity and evolution of Opuntia.     

Speciation and periodic restricted environments. The case of genus Ononis L. (subsections Natrix and Viscosae)

The morpho–environmental similarity between subsections Natrix and Viscosae has been pointed out as the reason for the genetic complexity of these groups of taxa. Based on this characterization a question emerges: could a very recent ongoing evolutionary process explain that morpho–environmental similarity? ISSR and cpSSR amplifications for 45 specimens belonging to taxa of Natrix and Viscosae subsections were developed, along their biogeographic distribution areas. Twenty-nine haplotypes were detected in the biogeographic area of both subsections, 79% were exclusive haplotypes, but the rest is shared between subsections Natrix and Viscosae species. Could that haplotype sharing be the result of potential hybridization between these taxa? Do current environmental conditions restrict the gene flow among taxa? The combination of ancestral genetic polymorphism, introgression, coalescence processes and periodic restricted environments (PRE) by glacial–interglacial environmental dynamics were discussed to explain the relevant percentage of exclusive haplotypes detected, as well as the persistence of shared haplotypes. These results are in accordance with the morpho–environmental proximity previously described for both subsections.