The diversity and sustainable development of crop genetic resources in the Lancang River Valley

This paper reviews the status of crop genetic resources in Yunnan province of China from 1978 to 1999. Results are presented of same research in the field of the diversity of cultivated and wild crop. Yunnan is one of the centre of origin or genetic diversity of more than 200 cultivated and wild crops. There are over 500 cultivated plants which account for over 80% of the total in China and more than 650 species of wild crops. In addition, there are more than 440 species of main wild flowers. According to our recently researches there are abundant species, subspecies and varieties of crop genetic resources in Yunnan Province. The Lancang River Valley is the richest genetic diversity centre of rice, maize, wheat, barley, buckwheat, legumes, ramie, sugarcane, vegetable, tea, actinidia and so on. For example, there are 59 varieties (including all varieties of Oryza sativa L. in China) in 5933 accession of Yunnan indigenous rice. The Lancang River Valley is one of the centre for genetic diversity of rice resources and a rich region for elite and rare rice resources of Yunnan, too. In order to protect the highly endangered crop genetic resources in the Lancang River Valley, it is necessary and very important to set up a collection, conservation, utilization and research system, enhancing their protection and utilization, in situ- and ex situ-conservation, farm management and sustainable production.     

云南地方稻种资源核心种质的微卫星分析

摘要：以来自云南地方稻(Oryza sativa L.)种资源核心种质中的113份材料为研究对象，运用36对微卫星引物，研究了籼(indica )粳( japonica )两个亚种间和云南5个稻作生态区间的遗传多样性分布趋势，并筛选了籼粳亚种、水陆生态型和不同生态区的特异指纹标记。结果表明，粳稻的遗传多样性大于籼稻，遗传分化水平较低；而5个生态区中滇西南水陆稻区遗传多样性最大，遗传分化水平较低；滇东北高原粳稻区的遗传多样性最小。这种遗传多样性的分布趋势与前人在形态和同工酶水平上对云南稻种资源多样性的考察，以及云南地方稻种资源核心种质在形态和同工酶水平上的遗传多样性分布趋势基本一致。另外，在所出现的416个指纹标记中，分别发现籼粳特异指纹标记6个，水陆特异指纹标记15个，不同生态区特异指纹标记3个。初步认为，云南地方稻种资源核心种质代表了云南省地方稻种资源的遗传多样性；从DNA水平上看，云南地方稻种资源的遗传多样性中心在云南省的西南部，粳稻的分化水平低于籼稻。微卫星标记是种质资源遗传多样性检测、分类和生态型确认以及核心种质研究中有用的工具。     

Evaluation of Genetic Diversity of Wild Rice (Oryza rufipogon Griff.) in Myanmar using Simple Sequence Repeats (SSRs)

As a part of an in situ survey of wild rice (Oryza rufipogon Griff.) in Myanmar (Burma), 16 strains of wild rice were collected, and analyzed for allelic diversity over 74 loci with simple sequence repeat (SSR) markers to obtain a basic information for their conservation. Three each of indica and japonica cultivars were added for a comparison. In the six cultivars and 16 strains of wild rice, three to 15 alleles were detected per locus with an average of 7.9. The wild rice revealed a large number of unique alleles throughout their chromosomes with much wider ranges of variation than those detected in the six cultivars of O. sativa L.. The alleles found in the wild rice were classified into those specific to wild rice, common to wild rice and cultivars, and those similar to indica or japonica cultivars. According to the classification, the genotype of each of the 16 strains of wild rice was schematically depicted. The genetic variation among individual strains within a collection site was larger than the variation among the collection sites.     

Genetic diversity and phylogenetic relationship of <Emphasis Type="Italic">Tadehagi</Emphasis> in southwest China evaluated by inter-simple sequence repeat (ISSR)

There are many valuable Tadehagi accessions in southwest China, but it is unknown that the genetic diversity and phylogenetic relationship of these Tadehagi resources. This report is the first study in which 41 primers of inter-simple sequence repeat (ISSR) were used to assess the genetic diversity of 36 Tadehagi accessions from 3 provinces in the southwest of China. Totally, 30 usable ISSR primers detected 163 polymorphic bands among the 36 accessions, which suggested high utility of ISSR primers in the genetic analysis of Tadehagi accessions. Genetic similarity coefficients among all of the accessions ranged from 0.54 to 0.92 with the average of 0.79 based on the ISSR data, indicating high level of genetic variation in Tadehagi resources from the southwest of China. As for the 3 population, Hainan population had the maximum average genetic similarity coefficients of 0.81, while similarity coefficient of Guangxi and Yunnan population was 0.75 and 0.74, respectively. All the 36 Tadehagi accessions were divided into 4 groups in the UPGMA dendrogram constructed from genetic similarity coefficients. The Tadehagi accessions from Yunnan and Guangxi provinces showed more genetic variation and occupied the bottom of the dendrogram. On the contrary, those from Hainan Province had less genetic variation and clustered in the middle and top of the dendrogram. The information on the genetic diversity and phylogenetic relationship from this study is propitious to construct a core germplasm collection and develop novel Tadehagi cultivars with desired economic traits.     

Genetic Diversity among Syrian Cultivated and Landraces Wheat Revealed by AFLP Markers

Genetic diversity among some important Syrian wheat cultivars was estimated using Amplified Fragment Length Polymorphism (AFLP) markers. Five Triticum aestivum L. and 10 Triticum turgidum ssp. durum were analyzed with 11 EcoRI–MseI primer pair combinations. Of the approximately 525 detected AFLP markers, only 46.67% were polymorphic. Cluster analysis with the entire AFLP data divided all cultivars into two major groups reflecting their origins. The first one contained T. aestivum L. cultivars, and the T. turgidum ssp. durum cultivars and landraces were grouped in the second. Narrow genetic diversity among all cultivars was detected with an average genetic similarity of 0.884. The lowest similarity index (0.9) was found between Cham5 and Hamary (durum wheat), whereas this value was 0.93 between Salamony and Bouhouth 4 (T. aestivum L.). The narrow genetic diversity level indicates that these genotypes could be originated from the same source. AFLP analysis provides crucial information for studying genetic variation among wheat cultivars and provides important information for plant improvement.     

Assessment of genetic diversity in cultivars and wild accessions of Luohanguo (<Emphasis Type="Italic">Siraitia grosvenorii</Emphasis> [Swingle] A. M. Lu et Z. Y. Zhang), a species with edible and medicinal sweet fruits endemic to southern China,using RAPD and AFLP markers

Genetic variation of wild populations and cultivars of Luohanguo (Siraitia grosvenorii), a plant species endemic to southern China, was assessed using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers. Based on the results for 130 individuals from seven populations, a high level of genetic diversity of Luohanguo was observed at the species level. The percentage of polymorphic loci (P) was 89.4%, Nei’s gene diversity (H _e) was 0.239, and Shannon’s information index (H _o) was 0.373 based on the combined AFLP and RAPD data. There was a high degree of genetic differentiation, with 45.1% of the genetic variation attributed to differences between the populations. The genetic diversity of the Luohanguo cultivars is much lower than that of wild populations (P = 41.8%, H _e = 0.141, H _o = 0.211), and a distinct genetic differentiation is observed between the cultivars and wild accessions. The pool of genetic variation in the wild populations provides an excellent gene resource for Luohanguo breeding.     

Genetic diversity of <Emphasis Type="Italic">Nelumbo</Emphasis> accessions revealed by RAPD

Total 65 lotus accessions in genus Nelumbo mainly collected from China, were subjected to random amplified polymorphic DNA (RAPD) markers to estimate the genetic diversity and to test the genetic basis of the relationships between morphotypes and molecular markers. Seventeen primers generated a total of 195 highly reproducible and discernible loci, among which 173 were polymorphic. Percent polymorphism varied from 66.7 to 100 with an average of 88.72, and five primers out of them, OPC05, OPG10, OPN20, OPP09 and OPS17, showed 100% polymorphism. A relatively high genetic diversity was detected among all the samples with the similarity coefficient values ranging from 0.45 to 0.85, and Nei’s gene diversity (h) 0.30, and Shannon index (I) 0.46. The UPGMA dendrogram clustered 65 accessions in four clusters and the clustering pattern showed two groups, N. nucifera ssp. nucifera and those accessions related to the American lotus, and some special cultivars, landraces, hybrids and the American lotus. Principal Coordinate Analysis (PCA) further indicated that the genetic diversity of Nelumbo accessions was not evenly distributed, instead, was presented by a clustered distribution pattern. Similar to the results revealed by the dendrogram, two main groups representing the two subspecies of N. nucifera, as well as some special landraces, cultivars of Chinese lotus, the Japanese lotus and hybrids out of the two groups were obtained. Neither the UPGMA dendrogram nor the PCA analysis exhibited strict relationship with geographic distribution and morphotypes among the accessions.     

Studying the extent of genetic diversity among Gossypium arboreum L. genotypes/cultivars using DNA fingerprinting

Genetic diversity is an area of concern for sustaining crop yield. Information on genetic relatedness/diversity among Gossypium arboreum L. cultivars/genotypes is scanty. We have used random amplified polymorphic DNA (RAPD) analysis to assess the genetic divergence/relationship among 30 genotypes/cultivars of G. arboreum. Of 45 primers surveyed, 63% were polymorphic. Out of the total number of loci amplified, 36% were polymorphic. The calculated genetic similarity between the cultivars/genotypes was in the range of 47.05–98.73%. Two genotypes, HK-244 and Entry-17, were the most distantly related. The average genetic relatedness among all the genotypes was 80.46%. However, most of the cultivated varieties showed a close genetic relationship, indicating a narrow genetic base in comparison to the non-cultivated germplasm. The calculated coefficients were used to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA) algorithm, which grouped the genotypes/cultivars into two major and three smaller clusters. The study is the first comprehensive analysis of the genetic diversity of G. arboreum germplasm and identifies cultivars that will be useful in extending the genetic diversity of cultivated varieties and future genome mapping projects.     

稻蟹共作对稻田水体底栖动物多样性的影响

稻蟹共作是一种生态型种养新模式,而底栖动物是该复合生态系统的重要组成部分。为研究稻蟹共作对稻田底栖动物多样性的影响,在水稻分蘖期、拔节期、扬花期和成熟期,采集不同放养密度的稻蟹共作稻田(仔蟹放养密度10 ind.m 2,低密度,LD;仔蟹放养密度30 ind.m 2,高密度,HD)和常规稻田(不放蟹,常规栽培,CK)水体的底栖动物,分析不同稻田生态系统底栖动物的种类构成、密度和多样性。结果表明:水稻分蘖期,3种放蟹密度处理稻田底栖动物的种类数、密度、多样性指数和Pielou均匀度指数均无显著差异(P>0.05);在拔节期和扬花期,养蟹稻田底栖动物的种类数和密度均小于常规稻田,且与放蟹密度呈反比;常规稻田底栖动物的Shannon-Wiener多样性指数在拔节期和扬花期均显著高于高密度养蟹稻田(P<0.05),而只在拔节期显著高于低密度养蟹稻田(P<0.05),且Pielou均匀度指数也在拔节期显著高于低密度养蟹稻田(P<0.05);成熟期,3种处理底栖动物的种类数和密度均达到最低点,Shannon-Wiener多样性指数无显著差异(P>0.05),高密度养蟹稻田的Pielou均匀度指数显著高于常规稻田(P<0.05)。研究结果可为稻蟹共作稻田底栖动物的多样性保护和稻蟹共作技术的可持续发展提供理论指导,促进稻蟹共作技术的发展和推广。     

Genetic Variation of Glutelin Acidic Subunit Polypeptides in Bangladesh Rice Genetic Resources

Bangladesh rice genetic resources collected from six distinct regions of the country were examined to obtain the diversity in glutelin acidic subunit polypeptides. Seed glutelins from 576 Bangladesh rice cultivars representing seven ecotypes were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IEF) electrophoresis and two-dimensional electrophoresis (2-DE: SDS-PAGE/IEF) analyses. Glutelin acidic subunit was separated into four bands, α-1, α-2, α-3 and α-4, and the variation in each of bands was detected by SDS-PAGE analysis. A higher molecular size component of α-1(α-1′) was identified in the cultivars tested. In case of α-2, α-2H with high molecular mass, α-2L with low molecular mass and α-2H/L were detected. α-3 band variation showed α-3H, α-3L, and α-3H/L, while for α-4, α-4H, α-4L bands were identified. In IEF analysis, a total of 16 bands with independent pI ranging from pI 6.30 to 7.52 were identified for the glutelin acidic subunit among the cultivars. The maximum and minimum numbers of IEF bands found were 13 and 9, respectively. The α-2L less cultivars were also lacking in pI 6.80 polypeptide in IEF. Result of 2-DE showed that pI 6.80 polypeptide was the main component of α-2L band. Transplanted Aman ecotype was the most diverse with respect to glutelin variation. Geographical distribution of glutelin variation in the Transplanted Aman ecotype differed according to the regions. The above results indicate that Bangladesh rice cultivars possess great genetic diversity in glutelin acidic subunit polypeptides. This study of indigenous rice cultivars from Bangladesh provides useful information regarding their breeding potential.     

Genetic diversity in bambara groundnut (Vigna subterranea (L.) Verdc) landraces assessed by Random Amplified Polymorphic DNA (RAPD) markers

Genetic diversity in 12 landraces of bambara groundnut (Vigna subterranea), an indigenous African legume, was evaluated using Random Amplified Polymorphic DNA (RAPD) markers. DNA from individuals of each landrace was also analysed to determine the level of heterogeneity within landraces. RAPDs revealed high levels of polymorphism among landraces. The percentage polymorphism ranged from 63.2% to 88.2% with an average of 73.1% for the 16 RAPD primers evaluated. The construction of genetic relationships using cluster analysis groups the 12 landraces in two clusters. RAPDs are useful for the genetic diversity studies in V. subterranea and can identify variation within landraces.     

Genetic diversity and relationships of common apricot (Prunus armeniaca L.) in China based on simple sequence repeat (SSR) markers

The common apricot (Prunus armeniaca L.) is adaptable to the diverse geographical areas throughout northern China where the original center for common apricot is worldwide. The diversity of 94 apricot samples, including 66 typical Chinese local cultivars, four kernel-using apricots, thirteen cultivars of Xinjiang (belong to Central Asian groups), ten exotic cultivars and one black apricot (Prunus dasycarpa Ehrh.) accession, were investigated using 21 simple sequence repeat markers. Totally, 662 genotypes were identified in the set of tested 93 common apricot cultivars, with an average of 31.52 genotypes per locus. The allele diversity per locus, with an average allele number of 15.14, and the heterozygosity rates, with an average expected heterozygosity value for the sample set of 0.792, were found to be higher than those in previous studies, implying that Chinese apricot germplasms presented a high level of genetic diversity. The clustering analysis and principal coordinate analysis outlined the genetic relationships of apricot cultivars tested from different geographical distributions. The results supported the traditional classification of apricot germplasm based on the ecological types, and showed that the richest diversity existed in the northern China and northwestern China local cultivars among common apricot germplasm resources tested. This study will facilitate the understanding of the evolution relationships and the conservation strategies of the genetic diversity in apricot in China.     

Studies on sampling schemes for the establishment of corecollection of rice landraces in Yunnan, China

The sampling scheme for a core collection from the basecollection of Yunnan rice landraces was studied, using 6,121accessions catalogued in the national genebank and 31 charactersincluding taxonomic characters, morphological characters andagronomic characters. Sampling scheme was studied at three levels,i.e., grouping principle, sampling proportion within group andsampling method from each group. Grouping principles used wereDingying's taxonomic system, Cheng-Wang taxonomic system,Yunnan rice ecological zone, Yunnan administrative district, singlecharacter and completely random sampling without grouping. Samplingproportions in each of the groups were square root, logarithm, indexof genetic diversity and fixed proportion. Clustered and randomsampling were carried out in each group. Five parameters, (indexof genetic diversity, variance of phenotypic value, variance ofphenotypic frequency, coefficient of variation and ratio ofphenotypic retained), in primary core collection were used toevaluate the validity of sampling scheme. The better samplingstrategies were grouping principles based on the two taxonomicsystems, decisions on sampling proportion in each group based onsquare root or logarithm, sampling method within group clustered.16%, 10% and 5% overall sampling ratio were usedto establish primary core collections. The results showed that a510% sampling ratio was suitable for a largecollection.     

The Distribution of <Emphasis Type="Italic">Perilla</Emphasis> Species

Perilla (Lamiaceae) contains one tetraploid species, P. frutescens (L.) Britt. and three diploid species, P. citriodora (Makino) Nakai, P. hirtella Nakai and P. setoyensis G. Honda. Tetraploid species have been traditionally cultivated in Asia for their seed oil and for their fragrant leaves that are used as medicine or as a garnish for fish. The center of diversity is still obscure. To conserve the genetic resources, it is important to know the diversity of the tetraploid species. The three diploid species, which are possible parents of the tetraploid species, are all believed to be indigenous to Japan. Their distribution in China and Korea was clarified on the basis of herbarium and field surveys. The tetraploid species is assumed to have originated somewhere around the mid-to downstream area of the Changjiang River. Though Perilla is not cultivated as often in these areas as in northern China, Korea, the Himalayan region, or Myanmar, these areas should also be important for the conservation of genetic resources of tetraploid Perilla crops because of the expected high genetic diversity.     

Genetic Diversity in Accessions of Wild Rice Oryza granulata from South and Southeast Asia

Oryza granulata, an upland wild rice species, represents an unique germplasm for possessing abilities of tolerance to shade and drought, immune to bacterial blight and resistance to brown planthopper. Although low degree of genetic variability has been revealed within its populations, little genetic information at the species level is available in determining rational conservation strategies. Here we used dominant DNA marker random amplified polymorphism DNA (RAPD) to assess the genetic variability among 23 accessions of O. granulata that collected from main distribution areas worldwide. Twenty decamer primers generated a total of 243 bands, with 83.5% of them (203 bands) being polymorphic. Calculation of Shannon index of diversity revealed an average value of 0.42 ± 0.25, indicating that O. granulata maintains a relatively high degree of genetic diversity on the species level. Analysis of genetic dissimilarity (GD) showed that genetic differentiation occurred among studied accessions, which supports the feasibility of current ex situ conservation strategies. We also suggested that information based on population studies, which could be achieved by international co-operation, is needed to conserve this widespread germplasm more effectively.     

Differentiation and distribution of indica and japonica rice varieties along the altitude gradients in Yunnan Province of China as revealed by InDel molecular markers

Indica–japonica variation represents the most significant genetic differentiation in Asian cultivated rice (Oryza sativa L. subsp. indica Kato or subsp. japonica Kato). Understanding the differentiation and distribution patterns of cultivated rice along altitude gradients will facilitate strategic utilization and conservation of rice germplasm from mountainous regions. In this study, we analyzed 203 varieties and 14 wild accessions of rice collected from localities across an altitude gradient between 450 and 2,350 m above sea level in Yunnan, China, applying the “InDel molecular index” developed recently. Results from PCA of the InDel data demonstrated significant genetic differentiation of rice varieties from Yunnan into indica and japonica types. A few cultivars and nearly all wild rice accessions showed only moderate or no differentiation. Further analyses demonstrated a clear distribution pattern of the rice varieties in the mountainous region: indica varieties were grown across the entire altitude gradient in the sampling areas, but most japonica varieties were found above 1,400 m. These results clearly indicated that indica rice could be cultivated in areas at much higher altitudes than those categorized by the traditional methods. The knowledge opens a new dimension for introducing indica rice varieties to mountainous regions at higher altitudes and for selecting rice germplasm in these regions. In addition, the pattern of significant indica–japonica differentiation in rice varieties from Yunnan suggested the exotic origin of cultivated rice, which did not support the hypothesis that Yunnan is a part of the center of origin of rice, although it is certainly one of the centers of genetic diversity for rice.     

广西地方食用木薯种质资源遗传多样性分析

为研究广西地方食用木薯材料的遗传多样性,以48份广西地方食用木薯为试验材料,分析其变异系数、遗传多样指数、聚类分析、多态性和遗传相似性系数。结果表明,收集的木薯材料的平均表型变异系数为37.9%,平均遗传多样性指数为0.807,其中主茎分叉角度的变异系数最大,为86.7%,块根内皮颜色的遗传多样性指数最大,为1.842。13对SSR引物共扩增出118个条带,其中多态性条带为106个,多态性比率为86.23%;分子标记聚类结果发现,遗传相似性系数在0.415~1.000之间;通过对比发现表型聚类和分子聚类结果不一致,其中表型聚类发现类群划分与地理来源之间没有关联,但与株型性状有一定的关联;在遗传相似系数为0.62时,SSR分子标记聚类为两类材料,第一类材料地理分布无规律,第二类材料大多分布在桂南。本研究结果表明,广西地方食用木薯资源遗传多样性具有一定的丰富度,可为创制优异食用木薯种质资源和新品种选育奠定基础。     

Molecular and morphological data reveals new insights into genetic diversity and population structure of Chinese cherry (<Emphasis Type="Italic">Prunus pseudocerasus</Emphasis> Lindl.) landraces

Chinese cherry (Prunus pseudocerasus Lindl.) is an indigenous fruit crop with high nutritional and ornamental value. It is widely cultivated in China, and many landraces have accumulated in large distribution regions. Genetic diversity analysis of Chinese cherry landraces is useful for breeding programs, as it helps to select genetic material to be used for further crossing. In this study, 110 Chinese cherry accessions from nine populations were assessed using fifteen morphological traits and seventeen ISSR markers. High average coefficient of variation (CV?=?17.43%) was detected based on morphological analysis. For molecular characterization, the global gene diversity (h?=?0.2816) and Shannon information index (I?=?0.4253) suggested a moderate high level of genetic diversity. Model-based STRUCTURE and principal coordinate analysis revealed three and two major gene pools based on morphologic and molecular data, respectively. Distinct distribution patterns of genetic variation between samples from southwest China and north China indicated two potential original domestication locations. We inferred that botanical features such as predominantly selfing characteristics, long lifespan, and insects-pollinated trait lead to the survival of abundant genetic variation and special genetic structure. This study provided morphological and molecular evidences for understanding genetic diversity and new insights into genetic consequence of populations for Chinese cherry landraces. Important implications for breeding programs and resources conservation were also provided.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

中国西南水稻白叶枯病菌遗传多样性垂直分布格局

针对我国西南稻区特有的立体气候和农业生态系统,利用rep-PCR和IS-PCR技术构建不同海拔稻区白叶枯病菌的DNA指纹图谱,聚类分析病菌的遗传多样性。结果表明:1西南稻区白叶枯病菌的遗传多样性十分丰富,共包含56种分子谱型,不同海拔稻区病菌的分子谱型种类明显不同;2白叶枯病菌的遗传多样性指数和丰富度指数在不同海拔稻区存在显著差异,其以中海拔稻区最高,低海拔稻区次之,高海拔稻区最低;3不同气候带白叶枯病菌的遗传多样性呈现出随着气候带温度降低而逐步减小的趋势;4聚类分析结果显示,56种分子谱型可分为9个遗传家系,低海拔稻区的菌株主要集中在lineage1和lineage2,中海拔稻区的菌株主要集中在lineage3~lineage6,而高海拔的主要集中在lineage3~lineage5。不同海拔稻区的菌株在变异程度上存在明显不同,因此,根据病菌致病型布局抗性基因时,应充分考虑菌株间遗传多样性的差异以及地区间病菌的变异强度,以减缓病菌在寄主选择压力下的变异速度,避免品种抗性丧失的周期过短。