小麦地方品种繁殖更新过程中的遗传多样性变化分析

采用分别保存于长期库及中期库的3个小麦地方品种的6份材料,进行了9项农艺性状及35个与农艺性状相关的微卫星标记检测,每份材料选取30个单株进行遗传多样性与遗传结构分析。结果表明:(1)更新前,3个小麦地方品种均为遗传异质性群体,在SSR位点上的异质度分别为57.14%、48.57%和5.71%。(2)在农艺性状表现上,只有温泉小麦3在株高和穗粒数上更新后比更新前显著增加,其他材料无显著差异。(3)在SSR位点多态性表现上,3个品种在更新后均发生了遗传多样性变化,8个与粒重、产量、生育期性状相关位点存在等位位点丢失现象,其中2个与粒重、生育期相关位点频率变化显著。(4)综合农艺性状调查与SSR分子标记检测结果发现,3个品种更新前后在多样性指数上无显著差异,遗传分化系数Gst分别为0.0269、0.0324和0.0380,即更新前后遗传差异分别为2.69%、3.24%和3.80%。上述结果建议,经繁殖更新的小麦种质资源能够比较完好地保持其遗传多样性和遗传结构。对于遗传异质性小麦地方品种在繁殖更新后存在遗传多样性丢失的危险,为了保证更新前后的遗传完整性,建议在繁殖更新过程中每个品种至少应保持300个单株的群体。     

中国花生核心种质的建立

以中国花生种质资源数据库中记录的6390份花生资源为材料,以其基本数据、特征数据和评价数据为信息,采用分层、层内分组聚类以及随机取样与必选资源相结合的方法,构建了由576份资源组成的花生核心种质,占基础收集品的9.01%。对核心种质的植物学类型组成和遗传多样性指数的分析,以及对各性状特征值、符合率和包含的主要抗病资源抗性等级及重要农艺性状资源的检测结果表明,本研究建立的核心种质是有效的。基础收集品中各种性状的遗传变异在核心种质中均存在,所用15个性状的各种特征值符合率均在90%以上,其中绝大部分性状的符合率达96%以上。     

中国花生核心种质的建立

以中国花生种质资源数据库中记录的6390份花生资源为材料,以其基本数据、特征数据和评价数据为信息,采用分层、层内分组聚类以及随机取样与必选资源相结合的方法,构建了由576份资源组成的花生核心种质,占基础收集品的9.01%。对核心种质的植物学类型组成和遗传多样性指数的分析,以及对各性状特征值、符合率和包含的主要抗病资源抗性等级及重要农艺性状资源的检测结果表明,本研究建立的核心种质是有效的。基础收集品中各种性状的遗传变异在核心种质中均存在,所用15个性状的各种特征值符合率均在90%以上,其中绝大部分性状的符合率达96%以上。     

Genetic differentiation in relation to seed weights in wild soybean species (Glycine soja Sieb. & Zucc.)

Seed weight is one of the most important botanical and phylogenetic characteristics. The study objective was to understand whether there is genetic difference in different seed weights of wild soybean (Glycine soja Sieb. & Zucc.). A total of 563 wild soybean samples, which belonged separately to genebank germplasm accessions (220 samples), one regional population samples (293 plants) and one natural population (150 plants), were analyzed using microsatellite markers. Of four size classes, the smallest seed size type had the highest coefficient of variation in seed weight; small and large seed types had relatively great genetic differences. In the national genebank germplasm accessions, genetic diversity gradually decreased from quantitatively dominant small and middling seed types to less frequent large seed types. In the regional and natural populations, generally, small to middling seed sizes had higher genetic diversity than the smallest and larger seed sizes. Cluster analysis revealed genetic differences in seed size traits. The semi-wild type (Glycine gracilis Skvortzow) was the most genetically differentiated from other seed sizes. However, it was also clearly shown that the phylogenic genetic differentiation among seed sizes was less than the genetic differentiation among geographical habitat populations in the wild soybean species.     

A note on the measurement of genetic diversity within genebank accessions of lettuce (Lactuca sativa L.) using AFLP markers

This paper discusses a statistical approach for measuring genetic diversity within genebank accessions of a self-fertilising species. This approach is applied to lettuce (Lactuca sativa L.), using AFLP marker data on a set of 1,390 accessions, representing six different lettuce types. Knowledge of the within-accession genetic diversity is important for decisions about the way accessions have to be maintained by genebanks. It is argued that if the within-accession diversity is small, as can be expected for a self-fertilising species like L. sativa, the best approach is to sample as many accessions as possible with only two plants per accession and estimate the within-accession diversity by the proportion of accessions of which the individuals are different.     

Variation among and within Capsicum species revealed by RAPD markers

 Germplasm characterization is an important link between the conservation and utilization of plant genetic resources. A total of 134 accessions from six Capsicumspecies maintained at the Asian Vegetable Research and Development Center were characterized using 110 randomly amplified polymorphic DNA (RAPD) markers. Ten pairs of potentially duplicated accessions were identified. Multidimensional scaling analysis of the genetic distances among accessions resulted in clustering corresponding to a previous species assignment except for six accessions. Diagnostic RAPDs were identified which discriminate among the Capsicumspecies. The diagnostic markers were employed for improved taxonomic identification of accessions since many morphological traits used in the identification of Capsicumare difficult to score. Three Capsicumaccessions, misclassified based on morphological traits, were reassigned species status based on diagnostic RAPDs. Three accessions, not previously classified, were assigned to a species based on diagnostic RAPDs. Definitive conclusions about the species assignment of three other accessions were not possible. The level of diversity between Capsicum annuumaccessions from the genebank and the breeding program were compared and no differences were observed either for RAPD variation or diversity. The utilization of genetic resources as a source of variance for useful traits in the breeding program may be the reason for the similarity of these two groups. Received: 1 September 1998 / Accepted: 28 December 1998     

种子老化诱导小麦染色体畸变及大麦醇溶蛋白带型频率变化的研究

通过时20份小麦种质发芽率和根尖细胞染色体畸变的测定,结果表明同一品种贮藏于中期库的低发芽率种质,其染色体畸变率明显高于贮藏于长期库的高发芽率种质.从总体上看,种子根尖细胞染色体畸变率与发芽率呈负显著相关,而与贮藏年限无显著相关.但对长期库和中期库的种质分别进行统计分析,种子根尖染色体畸变率与发芽率和贮藏年限均无显著相关.对大麦地方品种"普乃干木"的醇溶蛋白分析表明,该品种有4种biotype,随着发芽率的下降,其群体中的4种bio-type的频率发生了变化,当发芽率从95%降到34%时,其中一种biotype出现了消失,而有一种biotype频率从83%上升至95%,此结果表明,种子生活力下降和不同biotype种子存活能力存在差异,可能导致了异质种质材料遗传选择和漂变的发生.     

Use of microsatellites to evaluate genetic diversity and species relationships in the genus Lycopersicon

In order to determine how informative a set of microsatellites from tomato is across the genus Lycopersicon, 17 microsatellite loci, derived from regions in and around genes, were tested on 31 accessions comprising the nine species of the genus. The microsatellite polymorphisms were used to estimate the distribution of diversity throughout the genus and to evaluate the efficacy of microsatellites for establishing species relationships in comparison with existing phylogeny reconstructions. Gene diversity and genetic distances were calculated. A high level of polymorphism was found, as well as a large number of alleles unique for species. The level of polymorphism detected with the microsatellite loci within and among species was highly correlated with the respective mating systems, cross-pollinating species having a significantly higher gene diversity compared to self-pollinating species. In general, microsatellite-based trees were consistent with a published RFLP-based dendrogram as well as with a published classification based on morphology and the mating system. A tree constructed with low-polymorphic loci (gene diversity <0.245) was shown to represent a more-reliable topology than a tree constructed with more-highly polymorphic loci. Received: 19 February 2001 / Accepted: 26 March 2001     

我国作物种质资源保存研究与展望

我国于１９８４年１９８６年在北京建成两座国家种质库,９０年代初又在青海建立一座复份库,形成了国家库中,长期配套和异地结合的种质资源保存体系。截止１９９３年底,国家库保存种质资源达２７万余份,到１９９５年保存总数将达到３０余万份。与种子入库时,在国家库的管理,种子质量检测,种子干燥以及低温保存技术研究等方面取得一定进展,今后将继续扩大种质贮存、加强贮存种子生活力跟踪监测,无破坏性检测技术,贮存生理和     

小麦种子在不同保存条件下的生活力丧失特性研究

近年来种子在低温种质库和室温保存条件下的生活力丧失特性研究日益受到关注,但是与其重要性相比,目前很少见有文献报道此方面进展。本文中我们监测了在中期库、室温和人工加速老化条件下小麦种子的生活力变化,以了解种子在这些保存条件下的生活力变化特性。结果表明,三种条件下所有种子的存活曲线均呈反S形,这种特点不受保存条件的影响。随着保存温度的降低,种子生活力丧失速度降低,反S形曲线的平台期和种子寿命延长。我们采用T检验来判断平台期和种子生活力快速下降期之间的转折点（Pt）,发现小麦种子在不同条件下保存时,其Pt均十分接近,在82.2%到83.6%之间。本文讨论了将Pt以及其他生理指标作为种子保存过程中生活力快速下降的预警指标的可能性。     

Preservation of plant genetic resources in the biotechnology era

Thousands of years ago humans began domesticating crops as a food source. Among the wild germplasm available, they selected those that were best adapted for cultivation and utilization. Although wild ancestors have continued to persist in regions where domestication took place, there is a permanent risk of loss of the genetic variability of cultivated plants and their wild relatives in response to changing environmental conditions and cultural practices. Recognizing this danger, plant ex situ genebank collections were created since the beginning of the last century. World-wide, more than 6 million accessions have been accumulated including the German ex situ genebank in Gatersleben, one of the four largest global collections, housing 150,000 accessions belonging to 890 genera and 3032 species. This review summarizes the ex situ plant genetic resources conservation behavior with a special emphasis on German activities. Strategies for maintenance and management of germplasm collections are reviewed, considering modern biotechnologies (in vitro and cryo preservation). General aspects on genetic diversity and integrity are discussed.     

Siberian wild rye (Elymus sibiricus L.): Genetic diversity of germplasm determined using DNA fingerprinting and SCoT markers

Elymus sibiricus is a perennial, self-pollinating, allotetraploid grass native to northern Asia. It is widely used in cultivated pastures and natural grassland due to excellent cold and drought tolerance, good forage quality, and adaptability to a variety of habitats. Information on the genetic diversity and variation among worldwide E. sibiricus germplasm is limited but necessary for germplasm collection, conservation and effective commercial use. In this study we ana lyzed genetic diversity and variation of 69 E. sibiricus accessions from the species range and constructed DNA fingerprinting profiles of 24 accessions using SCoT markers. A total of 173 bands were generated from 16 SCoT primers, 154 of which were polymorphic with 89.0% of polymorphic bands (PPB) occurring at the species level. The PPB within 8 geographical regions ranged from 2.3 to 54.3 %. Genetic variation was greater within geographical regions (57.9%) than between regions (42.1%). The 24 accessions from Qinghai-Tibet Plateau, Mongolia Plateau, Kazakhstan, and Russia were distinguished by their unique fingerprinting. This is the first report using SCoT markers for identifying cultivars and accessions of E. sibiricus. The DNA fingerprinting profiles of E. sibiricus were useful in germplasm collection and identification. The genetic diversity of worldwide E. sibiricus germplasm has been substantially affected by ecogeographical factors. Our results suggest that collecting and evaluating E. sibiricus germplasm from major geographic regions and unique environments broadens the available genetic base and illustrates the range of variation.     

Farmers without borders—genetic structuring in century old barley (Hordeum vulgare)

The geographic distribution of genetic diversity can reveal the evolutionary history of a species. For crop plants, phylogeographic patterns also indicate how seed has been exchanged and spread in agrarian communities. Such patterns are, however, easily blurred by the intense seed trade, plant improvement and even genebank conservation during the twentieth century, and discerning fine-scale phylogeographic patterns is thus particularly challenging. Using historical crop specimens, these problems are circumvented and we show here how high-throughput genotyping of historical nineteenth century crop specimens can reveal detailed geographic population structure. Thirty-one historical and nine extant accessions of North European landrace barley (Hordeum vulgare L.), in total 231 individuals, were genotyped on a 384 single nucleotide polymorphism assay. The historical material shows constant high levels of within-accession diversity, whereas the extant accessions show more varying levels of diversity and a higher degree of total genotype sharing. Structure, discriminant analysis of principal components and principal component analysis cluster the accessions in latitudinal groups across country borders in Finland, Norway and Sweden. F_ST statistics indicate strong differentiation between accessions from southern Fennoscandia and accessions from central or northern Fennoscandia, and less differentiation between central and northern accessions. These findings are discussed in the context of contrasting historical records on intense within-country south to north seed movement. Our results suggest that although seeds were traded long distances, long-term cultivation has instead been of locally available, possibly better adapted, genotypes.     

High SSR diversity but little differentiation between accessions of Nordic timothy (Phleum pratense L.)

A large collection of genebank accessions of the hexaploid outcrossing forage grass species timothy (Phleum pratense L.) was for the first time analysed for SSR diversity on individual, population and regional level. Timothy is the most important forage grass species in the Nordic countries. Eighty-eight timothy accessions from Nordic countries and eight accessions around Europe were analysed with recently developed simple sequence repeat (SSR) markers. Timothy proved to be very polymorphic: the 13 selected SSRs amplified a total of 499 polymorphic alleles, the number of alleles per SSR locus varying from 15 to 74. Taking all SSR alleles together, the observed number in each accession ranged from 95 to 203. Levels of diversity were found to be significantly different between countries, vegetation zones and different cultivar types. However, the differentiation between accessions was low: most of the variation (94%) in the studied timothy material was due to variation within accessions and only 5% was between accessions and 1% between countries. Lack of geographical differentiation may reflect the outcrossing and hexaploid nature of timothy. Our results showed that neutral SSR markers are suitable for demonstrating levels of diversity but not alone adequate to resolve population structure in timothy. Nordic timothy material seems to be diverse enough for breeding purposes and no decline in the level of diversity was observed in varieties compared to wild timothy populations. Challenges in analysing SSR marker data in a hexaploid outcrosser were discussed.     

冰草属(Agropyron Gaertn.)植物遗传多样性取样策略基于醇溶蛋白的研究

对冰草属(Agropyron Gaertn.)植物的5个种22个居群分8个取样梯度,分别随机取3、6、9、12、15、18、21、24粒种子的混合样进行醇溶蛋白的A-PAGE分析,结果发现:平均Simpson指数依取样量的增加表现增大,以3粒种子0.922表现最低,以21粒0.933表现最高,变异系数(CV%)为0.41.醇溶蛋白谱带数依所提取的混合样的籽粒数目发生显著变化,冰草属内5个种表现趋势相同,22个居群不同取样整体平均以3粒种子提取的醇溶蛋白谱带数18.38为最少,以18粒种子提取的醇溶蛋白谱带28.24为最多,CV%为13.21.取样量达到12粒以上醇溶蛋白谱带差异不明显(P=0.01),图谱表现基本一致.建议在利用生化指纹进行冰草属居群间及种间的遗传多样性研究中,混合取样量最低应保持在12个个体及以上方能代表居群整体,反映居群的整体遗传特性.     

Molecular characterization of an international cacao collection using microsatellite markers

Plant germplasm collections invariably contain varying levels of genetic redundancy, which hinders the efficient conservation and utilization of plant germplasm. Reduction of genetic redundancies is an essential step to improve the accuracy and efficiency of genebank management. The present study targeted the assessment of genetic redundancy and genetic structure in an international cacao (Theobroma cacao L.) collection maintained in Costa Rica. A total of 688 cacao accessions maintained in this collection were genotyped with 15 simple sequence repeat (SSR) loci, using a capillary electrophoresis genotyping system. The SSR markers provided a high resolution among the accessions. Thirty-six synonymously labeled sets, involving 135 accessions were identified based on the matching of multilocus SSR profiles. After the elimination of synonymous sets, the level of redundancy caused by closely related accessions in the collection was assessed using a simulated sampling scheme that compared allelic diversity in different sample sizes. The result of the simulation suggested that a random sample of 113 accessions could capture 90% of the total allelic diversity in this collection. Principal Coordinate Analysis revealed that the Trinitario hybrids from Costa Rica shared a high similarity among groups as well as among individual accessions. The analysis of the genetic structure illustrated that the within-country/within-region difference accounted for 84.6% of the total molecular variation whereas the among-country/among-region difference accounted for 15.4%. The Brazilian germplasm contributed most to this collection in terms of total alleles and private alleles. The intercountry/interregion relationship by cluster analysis largely agreed with the geographical origin of each germplasm group and supported the hypothesis that the Upper Amazon region is the center of diversity for cacao. The results of the present study indicated that the CATIE International Cacao Collection contains a high level of genetic redundancy. It should be possible to rationalize this collection by reducing redundancy and ensuring optimal representation of the genetic diversity from distinct germplasm groups. The results also demonstrated that SSR markers, together with the statistical tools for individual identification and redundancy assessment, are technically practical and sufficiently informative to assist the management of a tropical plant germplasm collection.     

Population structure affected by excess gene flow in self-pollinating Elymus nutans and E. burchan-buddae (Triticeae: Poaceae)

Seed-mediated gene flow can considerably affect population genetic structure of strictly self-pollinating species, but little is known on the extent and nature of such gene flow among pastoral plant populations. Molecular fingerprints provide a powerful tool to address the relevant issues. Genetic structure of 22 populations of two self-pollinating pasture species, Elymus nutans and E. burchan-buddae, collected from various altitudes of the Qinghai–Tibetan Plateau was studied using fluorescence-based amplified fragment length polymorphism technique. Analysis of molecular variance revealed 42.97% and 37.63% among-population variation for the two Elymus species, respectively, indicating that the majority of the total variation presented within populations. This result contradicts the common genetic variation pattern for a selfing plant species: lower genetic variation within populations. Further analysis suggested higher level of gene flow among populations within the same region than among different regions across the sampled area for the two Elymus species. STRUCTURE analyses of the Elymus populations indicated an evident admixture genetic structure, particularly among neighboring populations from the same region, supporting the hypothesis of considerable seed dispersal among populations. The excess within-region gene flow of E. nutans and E. burchan-buddae might be caused by grazing animals that promote seed dispersal when moved around the pastoral lands during foraging. The among-population gene flow promulgated by grazing animals may promote the maintenance of genetic diversity in the pasture species, particularly in small and fragmented populations within a given region.     

Diversity of North European oat analyzed by SSR, AFLP and DArT markers

Oat is an important crop in Nordic countries both for feed and human consumption. Maintaining a high level of genetic diversity is essential for both breeding and agronomy. A panel of 94 oat accessions was used in this study, including 24 museum accessions over 100- to 120-year old and 70 genebank accessions from mainly Nordic countries and Germany, covering different breeding periods. Sixty-one polymorphic SSR, 201 AFLP and 1056 DArT markers were used to evaluate the past and present genetic diversity of the Nordic gene pool. Norwegian accessions showed the highest diversity, followed by Swedish and Finnish, with German accessions the least diverse. In addition, the Nordic accessions appeared to be highly interrelated and distinct from the German, reflecting a frequent germplasm exchange and interbreeding among Nordic countries. A significant loss of diversity happened at the transition from landraces and old cultivars to modern cultivars. Modern oat originated from only a segment of the landraces and left the remainder, especially black oat, unused. However, no significant overall diversity reduction was found during modern breeding periods, although fluctuation of diversity indices was observed. The narrow genetic basis of the modern Nordic gene pool calls for increasing genetic diversity through cultivar introduction and prebreeding based on neglected sources like the Nordic black oat.     

Variation of isozyme patterns among Arachis species

The genus Arachis contains a large number of species and undescribed taxa with patterns of genetic variation that are little understood. The objectives of this investigation were to estimate genetic diversity among species of Arachis by utilizing electrophoretic techniques and to establish the potential for use of isozymes as markers for germplasm introgression. One-hundred-and-thirteen accessions representing six of the seven sections of the genus were analyzed for isozyme variation of 17 enzymes. Section Rhizomatosae species were not included because they produce very few seeds. Seeds were macerated and the crude extract was used for starch-gel electrophoretic analyses. Although the cultivated species has few polymorphic isozymes, the diploid species are highly variable and two-to-six bands were observed for each isozyme among accessions. Because of the large number of isozyme differences between A. hypogaea and A. batizocoi (the presumed donor of the B genome), this species can no longer be considered as a progenitor of the cultivated peanut. Seed-to-seed polymorphisms within many accessions were also observed which indicate that germplasm should be maintained as bulk seed lots, representative of many individuals, or as lines from individual plants from original field collections. The area of greatest interspecific genetic diversity was in Mato Grosso, Brazil; however, the probability of finding unique alleles from those observed in A. hypogaea was greatest in north, north-central, south and southeast Brazil. The large number of polymorphic loci should be useful as genetic markers for interspecific hybridization studies.     

白菜型油菜黄子资源的初步遗传研究

对随机选取的国内外22份黄子白菜型油菜和22份褐子白菜型油菜进行了种皮色泽的显隐性关系鉴定、黄子性状的等位性测验以及遗传多样性分析。结果表明,黄子白菜型油菜与褐子白菜型油菜配组的杂交组合中,部分组合的F1种皮色泽呈现父本的种皮色泽,表现出花粉直感现象;自然界中存在多种白菜型油菜黄子类型,鉴定出的3种黄子白菜型油菜与褐色白菜型油菜的F2种皮色泽均为褐色,表明黄子性状对于褐子性状为隐性;分子标记方差分析结果显示,白菜型油菜的生长习性所解释的遗传变异大于种皮颜色所解释的遗传变异,表明国外不同生长习性的黄子白菜型油菜资源可用于国内黄子白菜型油菜遗传基础的拓宽。