Genetic variation in central and peripheral populations of Excoecaria agallocha from Indo-West Pacific

Genetic diversity and differentiation were studied in Excoecaria agallocha L., a mangrove species growing on the coastlines of the Indo-West Pacific region. Twenty natural populations of E. agallocha were sampled from the Malay Peninsula (central population), and in Southern China, North Australia, Sri Lanka and Southern Japan (peripheral populations). Our results showed that central populations from Malay Peninsula possessed significantly higher genetic diversity than the peripheral ones (P < 0.05). Analysis of molecular variation (AMOVA) revealed that genetic variability was partitioned at 22.9% among regions, 23.6% among populations within regions, and 53.5% within populations. Genetic differentiation (G_ST = 0.300) among the six central populations was stronger than those from peripheral regions. Populations from North Australia clustered closely together in the dendrogram and were distinct from the rest of the populations. Those from Southern Japan, Southern China and Sri Lanka also clustered closely together, respectively. However, populations from Malay Peninsula did not cluster by region. The east coast populations of Malay Peninsula (including Pasir population) were more genetically similar to the populations from Southern China than those from the west coastline of Malay Peninsula. Our study suggests that ocean currents, land barriers, limited dispersal ability of seeds, and founder effect may play important roles in the distribution of genetic diversity in E. agallocha.     

Clonality in South African isolates and evidence for a European origin of the root pathogen Thielaviopsis basicola

Thielaviopsis basicola is a soil-borne fungal pathogen with a wide host range and a cosmopolitan distribution. It causes disease on many agricultural crops, and in South Africa is the causal agent of black pod rot of groundnuts and black root rot on chicory. Knowledge of the population diversity of T. basicola could provide valuable information regarding management strategies, the possible movement, origin, and reproductive strategies of the fungus. The objective of this study was to determine the population diversity of T. basicola isolates from groundnuts and chicory in South Africa using co-dominant polymorphic markers. These markers were also used to compare isolates from South Africa with those from other hosts and geographic regions. Seven loci revealed nine alleles and two genotypes, one on groundnut and one on chicory, differing at only two loci. T. basicola isolates from eight different countries and ten different hosts revealed 17 genotypes across the seven loci with 39 different alleles. The lack of diversity for the two South African host-related populations of isolates suggests that T. basicola was introduced into South Africa. Some evidence is provided for a European origin of the pathogen, possibly linked to trade in root crops.     

Genetic variability and population structure of Bergenia ciliata (Saxifragaceae) in the Western Himalaya inferred from DAMD and ISSR markers

Genetic variability and population structure of Bergenia ciliata (Haw.) Sternb., commonly known as “Pashanbheda” (Stone-breaker), collected from the Western Himalayan region of India were estimated using two DNA fingerprinting methods viz., directed amplification of minisatellite DNA (DAMD) and inter simple sequence repeats (ISSR). The cumulative data analysis of DAMD and ISSR markers for 74 accessions from eight populations showed 86.1% polymorphism. Analysis of molecular variance (AMOVA) showed highest percentage of variation within individuals of populations (73.6%) and 21.7% among populations. STRUCTURE and PCoA analyses on the hierarchical partitioning of genetic diversity showed strong admixture of individuals among the eight assumed geographical populations of B. ciliata. The data suggests that high genetic flow is one of the major factors responsible for low genetic differentiation. Preservation of genetic diversity of B. ciliata is important, both to promote adaptability of the populations to changing environment as well as to preserve a large gene pool for future prospection. The present study using DAMD and ISSR markers, therefore, provide the means of rapid characterization of accessions within the populations, and thus enable the selection of appropriate accessions for further utilization in conservation and prospection programmes.     

Genetic variation within the sand-fixation species Caragana microphylla (Leguminosae) in Horqin sandy land detected by inter-simple sequence repeats analysis

Caragana microphylla is the most dominant and constructive shrub species in the Horqin sandy land in the northeast of China. We evaluated it's the level of genetic variation within and among populations sampled from two different populations types in Horqin sandy land by using inter-simple sequence repeat polymorphism (ISSR) molecular markers. The results showed that eight ISSR primers generated 106 bands, of which 87 were polymorphic. At the species level, genetic diversity was relatively high (P = 82.08%, h = 0.2831, I = 0.4233). Genetic variation in natural populations (h = 0.2152, I = 0.3169) was more than that in plantation populations (h = 0.2021, I = 0.3040). Based on Nei's G_ST value, more genetic differentiation among plantation populations was detected (G_ST = 0.7787). Six populations of C. microphylla clustered into two clades. These results have important implications for restoring and managing the degraded ecosystem in arid and semi-arid areas.     

Genetic variation within the endangered mangrove species Sonneratia paracaseolaris (Sonneratiaceae) in China detected by inter-simple sequence repeats analysis

Sonneratia paracaseolaris, is a critically endangered mangrove species in China. Using inter-simple sequence repeats (ISSR) markers, we compared the genetic variation of introduced populations with that of natural populations to check whether the genetic diversity has been conserved. At the species level, genetic diversity was relatively high (P = 81.37%, He = 0.2241, and SI = 0.3501). Genetic variation in introduced populations (P = 75.78%, He = 0.2291, and SI = 0.3500) was more than that in natural populations (P = 70.81%, He = 0.1903, and SI = 0.2980). Based on Nei's G_ST value, more genetic differentiation among natural populations was detected (G_ST = 0.3591). Our data show that the genetic diversity of S. paracaseolaris was conserved in introduced populations to some extent, however, owing to the small natural populations and the threats they encountered, more plants should be planted to enlarge and restore the populations.     

Genetic diversity within and among the wild populations of Murraya koenigii (L.) Spreng., as revealed by ISSR analysis

Murraya koenigii (L.) Spreng., commonly known as curry leaf plant, is found in the different hilly regions of India. In the present study, fifty-nine accessions representing eight wild populations of M. koenigii were analyzed using thirteen ISSR primers. A total of 152 bands were amplified, out of which, 136 were polymorphic corresponding to 89.47% polymorphism across the accessions. The pairwise population genetic distances were calculated for all the populations that varied from 0.05 to 0.13 between the populations of M. koenigii. AMOVA and Nei’s genetic diversity analysis revealed higher genetic variations within populations than among the populations. The clustering of populations in the dendrogram was not in congruence with geographical affiliations. The results indicate that the ISSR method is sufficiently informative and powerful to estimate the genetic diversity in M. koenigii populations. As M. koenigii is an important wild plant genetic resource, therefore, information on genetic variability might be a potential source as breeding material for development of commercially valuable traits in M. koenigii plants.     

Genetic diversity and population structure of Leptosphaeria maculans isolates in Western Canada

Leptosphaeria maculans is a serious concern for canola production worldwide. For effective disease management, knowledge of the pathogen's genetic variability and population structure is a prerequisite. In this study, whole-genome sequencing was performed for 162 of 1590 L. maculans isolates collected in the years 2007–2008 and 2012–2014 in Western Canada. DNA variants in genome-wide and specific regions including avirulence (Avr) genes were characterized. A total of 31,870 high-quality polymorphic DNA variants were used to study L. maculans genetic diversity and population structure. Cluster analysis showed that 150 isolates were clustered into 2 main groups and 4 subgroups by DNA variants located in either Avr or small secreted protein-encoding genes and into 2 main groups and 6 subgroups by genome-wide variants. The analysis of nucleotide diversity and differentiation also confirmed genetic variation within a population and among populations. Principal component analysis with genome-wide variants showed that the isolates collected in 2012–2014 were more genetically diverse than those collected in 2007–2008. Population structure analysis discovered three distinct sub-populations. Although isolates from Saskatchewan and Alberta were of similar genetic composition, Manitoba isolates were highly diverse. Genome-wide association study detected DNA variants in genes AvrLm4-7, Lema_T86300, and Lema_T86310 associated with the years of collection.     

Genetic variation in Lumnitzera racemosa, a mangrove species from the Indo-West Pacific

The genetic structure of 18 populations of Lumnitzera racemosa from the Indo-West Pacific, including South China, Malay Peninsula, Sri Lanka, and North Australia, was assessed by inter simple sequence repeat (ISSR) markers. Our results showed a relatively high level of genetic variation at the species level (P = 87.04%, H_e = 0.260). The value of G_st was 0.642, suggesting significant genetic differentiation among populations. At the population level, however, genetic diversity was low (P = 32.17%, H_e = 0.097). When populations were grouped according to geographic regions, i.e., South China Sea, the East Indian Ocean, and North Australia, it was inferred from AMOVA that more than half the total variation (55.37%) was accounted for by differentiation between regions. A UPGMA dendrogram based on genetic distance also revealed a deep split between populations from these regions, indicating that Malay Peninsula and the Indonesia archipelago may play an important part on the genetic differentiation in L. racemosa. The high degree of population differentiation between regions and low genetic variation within populations recorded here highlights the need for appropriate conservation measures for this species, both in terms of incorporating further populations into protected areas, and the restoration strategies for separate regions.     

Population structure and diversity in cultivated and wild Luffa species

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福建泰宁野生铁皮石斛种群的ISSR亲缘关系分析

为了解铁皮石斛(Dendrobium officinale)种质间的亲缘关系,利用ISSR技术对34份铁皮石斛种质资源进行亲缘关系和遗传多样性分析。结果表明,9条ISSR引物在34份种质中共扩增出78条带,多态位点百分率达100%。UPGMA聚类分析表明,种质的相似系数为0.61~0.92,在相似系数0.626处,福建省泰宁的野生铁皮石斛与栽培铁皮石斛分为两大类。泰宁野生铁皮石斛种群的Nei’s基因多样性(H)和遗传分化系数(Gst)分别为0.3111和0.4609,均高于栽培种群(0.3056和0.4204),表明泰宁野生铁皮石斛具有较丰富的多样性和较高的种群分化系数。AMOVA分析表明,铁皮石斛种群内变异指数为74%,种群间变异指数为26%,表明不同种群间可能存在基因交流。这些为不同地域的野生铁皮石斛资源的有效保护及利用提供理论依据及技术参考。     

An analysis of genetic diversity in Marphysa sanguinea from different geographic populations using ISSR polymorphisms

The genetic diversity of five wild populations of marine polychaete Marphysa sanguinea found in China was investigated using Inter-Simple Sequence Repeat-PCR (ISSR-PCR) polymorphisms. The results of the ISSR-PCR showed that 108 (90.8%) of the 119 ISSR loci tested were polymorphic. The Shannon's information index value was 0.4981, Nei's gene diversity was 0.3418, and the coefficient of gene differentiation (Gst) was 0.3671, which indicated that the among-population component accounted for 36.7% of the total variation, while the within-population component accounted for 63.3%. A UPGMA tree showed that the five populations clustered into two branches. Populations from Dalian, Xingcheng, and Rushan clustered together, while the two Guangxi populations, A and B, clustered into a unique group. The results indicated that the genetic diversity among the five populations of M. sanguinea is high, which will provide useful information for the protection of biodiversity among marine polychaetes.     

基于ISSR技术研究诺丽种质资源的亲缘关系

为探讨我国诺丽(Morinda citrifolia)种质资源的亲缘关系,采用ISSR技术对诺丽种质资源的遗传关系进行分析。结果表明,10条ISSR引物对13份诺丽种质资源共扩增出183条带,其中多态性条带有159条,占86.9%。13份诺丽种质的遗传相似系数为0.464~0.784。聚类分析将13份诺丽种质资源聚为两类,其中诺丽小黑种单独聚为一类,与其他12份诺丽种质资源的亲缘关系较远。虽然按照外部形态特征不能将所有诺丽种质完全区分,但具有相同特征的多数种质还是聚在同一类或亚类中。     

基于RAD高通量测序的蓝花楹群体遗传分析

为了解蓝花楹(Jacaranda mimosifolia)种质资源的遗传多样性和群体遗传结构,对168份种质材料进行RAD-seq测序,构建了系统进化树并进行主成分、群体结构和遗传多样性分析。结果表明,比对参考基因组平均比对率为81.02%,平均测序深度23.18×,最终获得45 552个高质量的SNPs。群体遗传结构分析表明,供试蓝花楹可划分为2个大的类群,来自川、渝地区的种质材料基本归为一类;其余地区归为另一类。19个地区的蓝花楹在SNP水平上的遗传多样性较高,云南昆明(YNKM)居群的核苷酸多样性(π)和期望杂合度(He)最大,表现出最高的遗传多样性。因此,来自川、渝地区的蓝花楹具有相对较近的亲缘关系,推断来自同一祖先,而其余地区的种质可能是随机引种栽培。     

Genetic diversity and population structure in Tunisian Lavandula stoechas L. and Lavandula multifida L. (Lamiaceae)

The genetic diversity and population structure of 20 Tunisian Lavandula stoechas L. and Lavandula multifida L. populations, from different bioclimates, were analysed by starch gel electrophoresis using seven isozymes. The genetic diversity within populations varied according to species. Variation in L. multifida was higher than that observed for L. stoechas, and exclusive alleles were detected for taxa.

A high differentiation among populations, for each species, estimated by Wright's F-statistics was revealed. The genetic structure of populations from the same bioclimate was substantial. Nei's, R. [1978. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89, 583–590] genetic distance among pairs of populations was low. The UPGMA cluster analysis of genetic distance values revealed that populations for each species were not strictly clustered together according to bioclimate or geographic proximity.

For each species, the low genetic divergence among populations and their substantial structure indicate their recent fragmentation due to anthropic pressures. The dendrogram generated from pairwise genetic distance among all populations showed two distinct clusters each corresponding to one species. The high genetic divergence between the two species, based on isozymes, corroborates their taxonomic status, as previously reported using morphological traits. The strategy for the management and conservation of populations should be made for each taxa according to its level of diversity and bioclimate.     

Genetic diversity and structure of the noxious alien grass Praxelis clematidea in southern China

Praxelis clematidea (Asteraceae), a plant species native to South America, is a noxious weed in southern China. We examined the genetic variation and population structure of 12 populations (76 individuals) of P. clematidea from Fujian, Guangdong, and Hainan Provinces in China using inter-simple sequence repeat (ISSR) analysis. From an initial set of 69 ISSR primers, 10 were selected which yielded 80 reproducible bands. Polymorphic bands (P) were 100%, Shannon's information index (I) was 0.4226, and Nei's gene diversity (H) was 0.2791. We infer that the high levels of genetic diversity exhibited by P. clematidea may have contributed to its invasiveness. Gene flow among populations was 2.4930, which has led to homogenization. The coefficient of population differentiation (Gst = 0.1671) indicated low levels of genetic variation among populations and high levels of genetic polymorphism within populations. There was a negative correlation between population elevation and genetic diversity, while there was a significant positive correlation between genetic distance and geographic distance based on a Mantel test (r = 0.5820, P < 0.01). Some populations from different provinces clustered together in principal coordinate and UPGMA analyses indicating that human-mediated events may have contributed to the dispersal of the species.     

Genetic diversity and relationship of endangered plant Magnolia officinalis (Magnoliaceae) assessed with ISSR polymorphisms

Twenty-eight populations of the rare medicinal plant Magnolia officinalis (Magnoliaceae) were sampled across its natural range, and inter-simple sequence repeats (ISSRs) markers were used to assess the genetic variation within and among populations. Twelve primer combinations produced a total of 137 unambiguous bands of which 114 (83.2%) were polymorphic. M. officinalis exhibited a relatively low genetic diversity at population level (the percentage of polymorphic loci PPB = 49.8%, Nei’s genetic diversity H = 0.194, Shannon’s information index I = 0.286). However, the genetic diversity at species level was relatively high (PPB = 83.2%; H = 0.342; I = 0.496). The coefficient of gene differentiation (G_ST, 42.8%) and the results of analysis of molecular variance (AMVOA) indicated that genetic differentiation occurred mainly within populations. The estimated gene flow (Nm) from G_ST was 0.669. It indicated that the fragmentation and isolation of populations might result from specific evolutionary history and anthropogenic activity. Genetic drift played a more important role than gene flow in the current population genetic structure of M. officinalis. Conservation strategies for this rare species are proposed based on the genetic data.     

Genetic diversity analysis of sedges (Carex spp.) in Shandong,China based on inter-simple sequence repeat

As the plants of turfgrass, forage and environment protecting plants, Carex L. has important economic value. The aims of the study were to construct ISSR-PCR amplification reaction system on Carex and to investigate the genetic diversity of 16 Carex populations belonging to 10 species using inter-simple sequence repeat (ISSR) makers. A total of 120 polymorphic amplified bands were obtained from 6 primers, and the percentage of polymorphisms was 100%. Genetic similarity between accessions ranged from 0.4250 to 0.8667 with an average of 0.6459, suggesting that the collected accessions are genetically diverse. All accessions were grouped into 3 clusters according to the UPGMA dendrogram. Most of the populations from the same regions can be basically clustered together and molecular grouping of Carex spp. correlates with geographical distribution and ecological environment. However, a few appeared to be divergent with the geographical distribution. The results showed that ISSR maker is an effective tool for the study of genetic diversity in Carex. As for the genus Carex, such information is needed for successful management and preservation of species to ensure the maintenance of genetic variation.