中国花生小核心种质与ICRISAT微核心种质的SSR遗传多样性比较

明确花生种质资源的遗传多样性和分布规律,对于发掘优良种质资源,选配优良亲本,拓宽育成品种的遗传基础具有重要意义。核心种质为种质资源的研究、评价和鉴定带来了方便。本研究从206对SSR引物中筛选26对引物对我国花生小核心种质和ICRISAT微核心种质共466份资源进行了遗传多样性分析,相似系数为0.49～0.99,鉴定出遗传差异最大的种质L2刚果(中国花生资源)与ICG12625(ICRISAT资源),相似系数为0.49。分析结果表明,多粒型花生的多态性信息量(0.761)和遗传多样性指数(0.97～1.11)均最大(平均相似系数最小,0.73～0.76),其次是普通型花生。中国花生种质资源与ICRISAT资源存在较大差异,尤其是ICRISAT的赤道型材料ICG12625,与中国花生资源的差异最大。相似系数和遗传多样性指数的分析结果均表明,我国花生种质资源的遗传多样性比ICRISAT资源丰富。     

DNA profiling and genetic diversity of Korean soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill) landraces by SSR markers

Approximately 7,000 accessions of Korean soybean (Glycine max (L.) Merrill) landraces, largely composed of three collections, the Korea Atomic Energy Research Institute’s soybean (KAS), the Korean Crop Experiment Station’s soybean (KLS) and the Korean Agricultural Development and Technology Center’s soybean (KADTC) collections, have been conserved at the Rural Development Administration (RDA) genebank in Korea. The accessions within collections were classified based on their traditional uses such as sauce soybean (SA), sprouted soybean (SP), soybean for cooking with rice (SCR), and OTHERS. A total of 2,758 accessions of Korean soybean landraces were used to profile and to evaluate genetic structure using six SSR loci. A total of 110 alleles were revealed by at the six SSR loci. The number of alleles per SSR locus ranged from 9 to 39 in Satt187 and Satt_074, respectively. The number of alleles ranged from 87 in the KADTC collection to 96 in the KLS collection, and from 63 in the SCR group to 95 in the SP group. Nei’s average genetic diversity ranged from 0.68 to 0.70 across three collections, and 0.64 to 0.69 across the usage groups. The average between-group differentiation (G _st) was 0.9 among collections, and 4.1 among the usage groups. The similar average diversity among three collections implies that the genetic background of the three collections was quite similar or that there were a large number of duplicate accessions in three collections. The selection from the four groups classified based upon usage may be a useful way to select accessions for developing a Korean soybean landrace core collection at the RDA genebank. DNA profile information of accessions will provide indications of redundancies or omissions and aid in managing the soybean collection held at the RDA genebank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding programs and could be used to develop a core collection.     

Assessment of genetic diversity of cultivated chickpea using microsatellite-derived RFLP markers: Implications for origin

Restriction fragment length polymorphism (RFLP) analysis was performed on 30 accessions of cultivated chickpea (Cicer arietinum L.) collected from 11 different countries representing the Near East, Central Asian and Hindustani regions. A synthetic digoxygenated oligonucleotide (GATA)₄ complementary to a microsatellite DNA sequence was used as a probe. The results revealed that simple repetitive sequences are abundant and polymorphic in the chickpea genome. The fragments detected were used lo estimate the genetic diversity within accessions and a similarity index between the genotypes of the accessions. The genetic distance data were used to construct a dendrogram depicting genetic relationships among the different accessions. The results indicate that the greatest genetic diversity occurs in Pakistan, Iraq, Afghanistan, south-east Russia, Turkey and Lebanon. Lower genetic diversity was found in Iran, India, Syria, Jordan and Palestine, Based on DNA markers, it is concluded that there are three centres of diversity for chickpea: Pakistan-Afghanistan. Iraq Turkey and Lebanon. India, which was previously considered as a secondary center of diversity for chickpea, showed lower diversity than the above regions.     

四川豌豆种质资源白粉病抗性及分子鉴定

豌豆白粉病是由白粉菌（Erysiphe pisi D. C.）引起的豌豆最重要的病害之一,控制豌豆白粉病最经济有效且环保的方法是种植抗病品种。在温室条件下对400份豌豆种质资源进行白粉病抗性鉴定,同时利用7个与已知豌豆抗白粉病基因连锁的分子标记进行基因型鉴定。结果表明,在400份资源中,有8份表现免疫,3份表现抗病,5份表现抗感分离,其余384份均为感病;16份抗性资源中有10份来自于四川省中部不同纬度地区,其余6份为国外引进资源;7个分子标记将400份种质资源分为39个标记基因型,其中16份抗性资源分为7个标记基因型。上述抗性资源及其标记基因型可有效应用于豌豆白粉病抗性育种的研究中。     

Genetic diversity of flax accessions originating in the Alpine region: a case study for an ex situ germplasm evaluation based on molecular marker

Agro-biodiversity is currently experiencing severe genetic erosion due to mankind’s unsustainable activities. Because of initiatives following the goal of the conservation of biological diversity, so far seven million crop accessions are being conserved ex situ in gene banks worldwide. Many of these accessions are landraces being rich in gene diversity, silently awaiting their proper characterisation. This is a very critical part of any long-term strategy to enhance the productivity and resilience of crops and agricultural systems and—most importantly—to ensure the preservation of our cultural and biological heritage. In this study of an ex situ germplasm evaluation we analysed 27 flax (Linum usitatissimum L.) accessions originating in the Alpine region, provided by five local gene banks/providers. Based on genomic microsatellite markers (gSSRs), a varying extent of accession-specific gene diversity (expected heterozygosity, H_E) was revealed ranging from 0.05 to 0.51. Admixture of individuals between accessions was uncovered, pointing towards past processes related to gene bank management activities (e.g. intentional selection, unintentional cross-pollination during regeneration) or towards the evolution of the landrace itself (e.g. same regional origin, traditional naming), highlighting the co-existence of cultural and biological diversity. Such an genetic analysis of accessions stored ex situ not only produces valuable agronomic and breeding data, but also is useful for the clarification of past processes leading to duplicates within and between collections or mislabelling, contributing to the potential for rationalisation of collections, which in turn can help ensure that the limited resources available for regeneration are used most efficiently and effectively.     

Phenotypic and marker-assisted evaluation of spring and winter wheat germplasm for resistance to fusarium head blight

Fusarium head blight (FHB) caused by Fusarium species, is among the most devastating wheat diseases, causing losses in numerous sectors of the grain industry through yield and quality reduction, and the accumulation of poisonous mycotoxins. A germplasm collection of spring and winter wheat, including nine reference cultivars, was tested for Type II FHB resistance and deoxynivalenol (DON) content. Genetic diversity was evaluated on the basis of Simple Sequence Repeat (SSR) markers linked to FHB resistance quantitative trait loci (QTLs) and Diversity Arrays Technology (DArT) markers. The allele size of the SSR markers linked to FHB resistance QTLs from known resistance sources was compared to a germplasm collection to determine the presence of these QTLs and to identify potentially novel sources of resistance. Forty-two accessions were identified as resistant or moderately resistant to Fusarium spread, and two also had very low DON concentrations. Genetic relationships among wheat accessions were generally consistent with their geographic distribution and pedigree. SSR analysis revealed that several resistant accessions carried up to four of the tested QTLs. Resistant and moderately resistant lines without any known QTLs are considered to be novel sources of resistance that could be used for further genetic studies.     

28个小麦微核心种质抗叶锈性分析

选取在成株期表现高、中、低抗叶锈的28个小麦微核心种质,利用39个以Thatcher为背景的近等基因系(或单基因系)作为已知基因的鉴别寄主,接种8个小麦叶锈菌致病型进行苗期抗叶锈基因推导,结合成株期抗病鉴定,初步明确了这些品种(系)的抗性和可能携带的抗病基因。利用19个与Lr基因紧密连锁或共分离的分子标记,对28个微核心种质进行抗叶锈病基因的进一步鉴定,推测新克旱9号可能含有Lr17、Lr2b、Lr14a和Lr33;兴义4号可能含有Lr26、Lr36和Lr37;紫皮可能含有Lr2b和Lr34;大白皮含有Lr1;毕红穗含有Lr1、Lr10和Lr34;中优9507含有Lr10;小白麦、红粒当年老、老麦、蝉不吱、苏麦3号和车锏子含有Lr1和Lr34;红花早可能含有Lr1、Lr34、Lr14a和Lr2b;江西早、泡子麦、三月黄、有芒扫谷旦、阜阳红、成都光头和酱麦可能含有Lr34;敦化春麦和甘肃96可能含有Lr28;欧柔可能含有Lr34、Lr16、Lr11、Lr3bg和Lr33;此外,新克旱9号、兴义4号、红花早、红粒当年老、欧柔、有芒扫谷旦、成都光头、甘肃96、小红皮、定兴寨、中优9507和红冬麦中可能含有未知抗病基因;在这28份种质中,不含Lr9、Lr19、Lr20、Lr21、Lr24、Lr29、Lr35、Lr38和Lr47基因。研究结果表明,测试的微核心种质中含有比较丰富的抗叶锈病基因,可为育种提供丰富的抗源。     

Characterization of resistance response of pea (Pisum spp.) against rust (Uromyces pisi)

Components of resistance to the pea rust (Uromyces pisi) were studied at the histological level in seedlings of seven pea (Pisum spp.) accessions showing partial resistance under field conditions. Resistance was characterized macroscopically by an increased latent period and a decreased infection frequency in spite of a compatible interaction (high infection type). Histological investigations revealed little differences among accessions in spore germination and appressoria formation, differences being more evident once the stomata were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with significant levels of early aborted colonies in accessions IFPI3260, PI347321 and PI347347. Furthermore, a reduction in the number of haustoria per colony, hyphal tips per colony and smaller colony size were found in all genotypes studied. Resistance was not associated with early‐ or late‐acting hypersensitivity in any of the genotypes studied.     

枳壳EST-SSR标记的开发

GenBank上已公布的枳壳EST序列为开发新的SSR标记提供了宝贵的数据资源。本研究利用在线SSR鉴定软件SSRIT分析来自枳壳EST数据库的11029条Unigene序列。分析结果共发现327条EST序列含有348个SSR位点,占总数的2.96%。其中,三核苷酸重复的SSR类型最多,共有161个,占检索总数的46.26%。Primer 3.0设计合成58对EST-SSR引物,其中36对能扩增出产物,6对引物产生多态性分离,分别占所设计引物总数的62.07%和10.34%。本文研究成果为今后枳壳遗传多样性分析、遗传图谱构建及比较基因组等研究方面奠定了基础。     

Assessment of genetic diversity in sesame (Sesamum indicum L.) genotypes, using EST-derived SSR markers

A total of 16,619 ESTs sequences (SSRs) of sesame (Sesamum indicum L.) were mined from Genbank. From sequences, 156 primer pairs were designed and characterized to determine the diversity among 49 sesame accessions. Twenty SSRs were found to be polymorphic and the number of alleles ranged from two to five per locus. The allele size varied from 101 to 399 bp. The average PIC value of the 20 SSR loci was 0.72 ranging from 0.49 (SEM-12-68) to 0.90 (SEM-12-27). Dendrogram analysis grouped the 49 genotypes into five separate clusters exhibiting a genetic similarity coefficient from 0.59 to 1.0. Hence, these EST-derived SSRs markers could be useful in assessing the diversity of sesame accessions and could also help in identifying diverse parents for sesame improvement programs.     

Genetic diversity and relationships among Lablab purpureus genotypes evaluated using RAPD as markers

Summary Genetic variation in 40 accessions of Lablab purpureus was evaluated using random amplified polymorphic DNA as markers. A high level of genetic variation in this species was detected but this was mainly restricted to the difference between cultivated and wild forms. Of the cultivated genotypes, genetic variation among Asian collections was significantly higher than that among African collections. The three most divergent cultivated genotypes were all from Asia. Four of the five wild accessions, two from Zimbabwe and the other two from Zambia, were closely related. The other one, CPI 31113 collected from Uganda, was highly divergent. The two commercial forage varieties used in Australia, Rongai from Kenya and Highworth from India, were not very different.     

Genetic diversity of wild coffee (Coffea arabica L.) using molecular markers

Genetic diversity was studied using RAPD markers among119 coffee (Coffea arabica L.) individuals representing 88 accessions derived from spontaneous and subspontaneous trees in Ethiopia, the primary centre of species diversity, six cultivars grown locally in Ethiopia, and two accessions derived from the genetic populations Typica and Bourbon, spread in the 18^th century, which gave rise to the most currently grown cultivars. Twenty-nine polymorphic fragments were used to calculate a similarity index and construct dendrograms. The Ethiopian material was separated from the Typica- and Bourbon-derived accessions and classified in four groups: one with most of the collected material from southwestern Ethiopia and three from southern and southeastern Ethiopia. Almost all detected diversity was found in the southwestern group while the southern and southeastern groups presented only 59% of identified markers. The genetic distances were low between the southwestern group and the southern and southeastern groups, and between the southwestern group and the Typica- and Bourbon-derived accessions. The cultivated coffee derived from the genetic populations Typica and Bourbon appeared little differentiated from wild coffee growing in the southwest. The results supported the hypothesis that southwestern Ethiopian coffee trees could have been introduced recently in the south and southeast. A separate analysis of the 80accessions classified in the southwestern group allowed identifying particular spontaneous- and subspontaneous-derived accessions and redundancies in the collected material from southwestern Ethiopia. RAPD markers did not detect any within-collection polymorphism except for two trees that were identified as off-types in the CATIE field genebank. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and population structure of ‘Khao Kai Noi’, a Lao rice (Oryza sativa L.) landrace,revealed by microsatellite DNA markers

Rice (Oryza sativa L.) is the main food for people in Laos, where it has been grown and eaten since prehistory. Diverse landraces are grown in Laos. ‘Khao Kai Noi’, a landrace favored for its eating quality, is held in the nationwide collection of traditional landraces in the Lao national genebank. Genetic diversity is crucial for sustainable use of genetic resources and conservation. To investigate the genetic diversity of ‘Khao Kai Noi’ for conservation, we genotyped 70 accessions by using 23 polymorphic simple sequence repeat markers. The markers generated 2 to 17 alleles (132 in total), with an average of 5.7 per locus. The total expected heterozygosity over all ‘Khao Kai Noi’ accessions was 0.271. Genetic variation was largest among accessions and smallest within accessions. Khao Kai Noi accessions were classified into three different genetic backgrounds, but there was unclear association between the three inferred population and name subgroups and geographical distribution. Most of the accessions were clustered with temperate japonica and showed genetic relatedness to rice from neighboring provinces of Vietnam, suggesting a Vietnamese origin. The results of this study will contribute to the conservation, core collection and future breeding of the Khao Kai Noi population.     

四个小麦抗穗发芽分子抗性标记有效性的验证与评价

成熟期穗发芽是一种世界性灾害, 严重影响小麦品质和产量。本试验利用已报道的4个与穗发芽抗性相关的标记, 即STS标记MST101、STMS标记wmc104、QTL位点Xgwm155与Vp1B3, 结合穗发芽率分析,对95份中国小麦地方品种和历史品种的穗发芽抗性进行筛选, 旨在从中筛选出抗穗发芽品种, 并对这4个分子标记的有效性进行比较, 筛选出可用于种质资源筛选和分子标记辅助育种的高效分子标记。结果表明, Vp1B3和Xgwm155与穗发芽抗性相关, 而MST101和wmc104与穗发芽抗性无关。比较而言, Vp1B3更能有效地用于筛选穗发芽抗性品种, 但将Vp1B3和Xgwm155结合起来筛选抗穗发芽小麦品种, 会提高选择效率。     

豌豆抗白粉病资源筛选及分子鉴定

由豌豆白粉菌引起的白粉病是豌豆生产上的重要病害,利用抗病品种是防治该病害最经济有效的方法。本研究在控制条件下苗期接种鉴定了396份豌豆资源对2个不同地理来源的豌豆白粉病菌分离物EPBJ和EPYN的抗性,用4个与豌豆抗白粉病基因er1连锁的SCAR标记对66份免疫或抗病资源进行标记基因型鉴定。结果表明,在鉴定的396份资源中,有101份资源表现免疫或抗病,其中对分离物EPBJ和EPYN免疫的资源分别为59份(14.9%)和60份(15.2%),对2个分离物均免疫的资源有54份(13.6%);在鉴定的82份中国资源中,有8份对2个分离物均表现免疫。分子标记将66份免疫或抗病资源鉴定为13个标记基因型,同一地理来源的抗性资源分属不同的标记基因型,其中8份来自中国云南的抗性资源分属7个标记基因型。研究表明,中国存在有效的豌豆白粉病抗源,抗性资源具有丰富的遗传多样性。     

Analysis of population structure and genetic diversity reveals gene flow and geographic patterns in cultivated rice (<Emphasis Type="Italic">O. sativa</Emphasis> and <Emphasis Type="Italic">O. glaberrima</Emphasis>) in West Africa

To fully exploit the diversity in African rice germplasm and to broaden the gene pool reliable information on the population genetic diversity and phenotypic characteristics is a prerequisite. In this paper, the population structure and genetic diversity of 42 cultivated African rice (Oryza spp.) accessions originating from West Africa (Benin, Mali and Nigeria, Liberia etc.) were investigated using 20 simple sequence repeats (SSR) and 77 amplified fragment length polymorphisms (AFLP). Additionally, field trials were set up to gain insight into phenotypic characteristics that differentiate the genetic populations among rice accessions. The analysis revealed considerably high polymorphisms for SSR markers (PIC mean?=?0.78) in the germplasm studied. A significant association was found between AFLP markers and geographic origin of rice accessions (R?=?0.72). Germplasm structure showed that Oryza sativa accessions were not totally isolated from Oryza glaberrima accessions. The results allowed identification of five O. glaberrima accessions which grouped together with O. sativa accessions, sharing common alleles of 18 loci out of the 20 SSR markers analyzed. Population structure analysis revealed existence of a gene flow between O. sativa and O. glaberrima rice accessions which can be used to combine several interesting traits in breeding programs. Further studies are needed to clarify the contributions of this gene flow to valuable traits such as abiotic and biotic stresses including disease resistance.     

Genetic analysis of resistance to Phoma tracheiphila in three Citrus and Poncirus progenies

Mal secco is a serious tracheomycosis, caused by the fungus Phoma tracheiphila (Petri) Kanc, et Ghik., which attacks some species of the genus Citrus. The considerable amount of time and space required to evaluate host plant response has discouraged research on the mechanism of inheritance involved in the susceptibility to this disease. The aim of the present work was to study segregation of the resistance trait in three progenies obtained by crossing Citrus latipes (Swing.) Tan., as a female parent, with the male parents Poncirus trifoliata (L.) Raf., C.aurantium L. and C. volkameriana Pasq. Resistance determination was based on natural infections observed over 10 years. Results allowed us to hypothesize three alternative genes (A, B and C) which determine dominant resistance as opposed to susceptibility: the presence of a single allele can thus confer resistance. In addition, we suggest the existence of a fourth gene which, in conditions of dominance, is able to determine susceptibility by nullifying the action of the dominant B allele. Response to the pathogen was also associated with the production of chitinase a PR (pathogenesis-related) protein reported to be present in great quantities in mal secco-resistant Citrus genotypes. Segregation analysis of one F₂ progeny confirmed the hypothesis that the resistance trait is controlled by at least two dominant genes. Further evidence is needed to ascertain whether this hypothesis is applicable to other Citrus species, in which the possibility of other genetic mechanisms being implicated cannot be ruled out, thus making the genetic basis of mal secco resisance more complex.     

黄麻核心种质的遴选

遴选黄麻核心种质可为黄麻种质创新及新品种选育奠定基础。本研究以300份黄麻种质资源为基础,基于SSR分子标记和农艺性状考察,结合地理来源构建核心种质。结果表明,11个农艺性状变异系数变幅在13.06%～84.87%,表现出丰富的遗传多样性。按农艺性状聚类分析可划分为8个类群,按分子标记聚类可划分为10个类群。结合2个聚类分析、地理位置并按比例取样,建立一个由108份品种(系)组成的预选核心种质。采用44对SSR引物对其进行遗传差异分析,在遗传相似系数为0.65时,可把108份品种(系)分成圆果种和长果种两大类。根据遗传差异分析,剔除遗传相似系数大于或等于0.85的遗传冗余,获得84份品种(系)的核心种质,其中圆果种60份和长果种24份。比较84份核心种质与300份种质的农艺性状变异系数及Shannon-Wiener指数发现,两者之间相差不大,表明遴选的84份核心种质可以最大限度代表300份黄麻种质资源的遗传多样性加以利用和保存。     

Estimation of between and within accession variation in selected Spanish melon germplasm using RAPD and SSR markers to assess strategies for large collection evaluation

The population structure of 15 Spanish melon (C. melo L.)accessions, mostly of Group Inodorus, was assessed by the analysis of 16individuals of each accession using 100 random amplified polymorphic DNA (RAPD) bands produced by 36 primers, and allelic variation at 12microsatellite (SSR) loci (23 alleles). A relatively high level of polymorphism (25.6%) was detected using RAPD markers, and eight SSR loci (66.7%) were useful in discriminating accessions. Cluster analysis using RAPD- and SSR-based genetic distance estimates resulted in similar and consistent groupings of most of the accessions studied. The mean genetic distance and standard error among accessions estimated by RAPD variation was 0.421 ± 0.099, and mean SSR-based genetic distance estimate was 0.285 ± 0.141. Albeit many dominant markers examined were fitted to a 3:1 test ratio, deviation from this ratio and from Hardy-Weinberg expectations for many SSR loci suggests that some populations were in genotypic disequilibrium. Moreover, a higher level of genetic variation was observed between Cassaba market classes than within accessions, suggesting that, depending upon the accession, allelic fixation has occurred in these populations. The relatively high level of heterogeneity observed (different band morphotypes and cluster grouping within a particular market class), however, indicates that the Spanish melons examined possess a relatively broad genetic background. An appraisal of accession population structure such as the one reported herein indicates that bulk sampling techniques coupled with molecular analysis techniques that employ a unique array of discriminating markers can provide information leading to effective strategies for diversity analyses of large collections. This revised version was published online in August 2006 with corrections to the Cover Date.