元阳3个长期连续栽培水稻地方品种内部遗传异质性分析

水稻地方品种是稻种资源的重要组成部分,遗传多样性十分丰富,具有推广改良品种所缺少或没有的优质种质,是水稻育种和稻种起源、进化研究不可缺少的过渡材料。目前,对水稻地方品种间遗传多样性研究较多,而对其内部异质性研究甚微。本研究用24对微卫星(SSR)引物对云南元阳梯田3个栽种历史悠久的水稻地方品种的内部遗传异质性进行了分析。共检测出117个等位基因,香农指数为红脚老粳居群(0.5911)>白脚老粳A居群(0.4875)>月亮谷居群(0.3070)。结果显示:3个地方品种的内部遗传异质性丰富,且遗传异质性主要得益于个体间,而非居群间。     

中国水稻地方品种与选育品种遗传多样性的比较分析

利用98对SSR标记对202份中国水稻地方品种和选育品种的遗传多样性进行比较分析.结果显示供试品种具有较丰富的遗传多样性,共检测到等位基因1350个,每个位点的等位基因数(Na)变化范围为3～ 39,平均14个;Nei基因多样性指数变化范围(He)为0.125 ～0.955,平均0.733;多态信息量(PIC)变化范围为0.122 ～0.953,平均0.680;稀有等位基因数(Nr)913个;等位基因丰度(Rs)8.33.栽培稻地方品种和选育品种遗传多样性差异明显,地方品种等位基因数、Nei基因多样性指数、多态信息量、稀有等位基因数和等位基因丰度(Na=1219,He=0.747,PIC=0.710,Nr=756,Rs =8.50)均高于选育品种(Na =919,He =0.704,PIC =0.650,Nr=529,Rs =7.01).各染色体组水平的遗传多样性分析表明,选育品种仅在1号染色体上的遗传多样性高于地方品种,进一步分析显示选育品种的遗传改良在基因组水平上具有区间特异性.     

SSR标记揭示的云南地方稻品种遗传多样性及其保育意义

为了探索水稻(Oryza sativa L.)地方品种的遗传多样性及其有效保育方法,对采自云南省17个村寨的82个水稻地方品种和3个国际常用的典型籼稻和粳稻品种进行了微卫星(SSR)分子标记的分析.利用19对SSR引物在85个水稻品种中共扩增出了83个基因型,其分子量变异在100～500 bp之间.基于各品种SSR基因型遗传相似系数聚类分析而获得的UPGMA树状图表明各水稻品种之间存在较大的遗传多样性,其相似系数变异在0.15～0.90之间.但这些地方品种的遗传多样性并非呈均等的地理分布.这85个水稻品种在相似系数为0.52之处分为二组,其中一组包括几乎所有的籼稻品种,而另一组包括全部的粳稻品种,表明SSR标记能很好揭示水稻籼-粳分化.同时,有些来自不同采集地的同名品种表现出一定的遗传差异,说明同名异物的现象存在.云南水稻地方品种具有丰富的遗传多样性,对其有效保育十分重要和迫切,但只有根据遗传多样性的水平和分布特点,采用正确的保育对策和取样方法才能确保对云南水稻地方品种的有效保育.结果进一步表明,选用适当的微卫星引物,可以为准确鉴定籼稻和粳稻品种及研究其进化规律提供有效的分子标记方法,并有利于有目标的水稻遗传资源保育和育种创新.     

重庆大豆地方资源多样性评价及群体表型特点研究

对收集于重庆市的103份大豆地方品种进行了表型遗传多样性分析,并利用主成分分析和聚类分析对其进行了群体表型特点的研究,研究结果表明,重庆地区大豆地方种质资源以灰色茸毛、黄粒、褐色脐居多,个体之间数量性状上存在较大变异。主成分分析以3个主成分反映了10个农艺性状的大部分信息,将主要农艺性状归纳成产量因子、生长势因子及籽粒因子。聚类分析将103份地方种质资源聚为6个类群, 同一地区的大部分地方品种表现较为相似,少部分地方品种差异大,品种类群间的表型分化与地理分布既有一定的联系又不绝对相关。利用这些种质选配育种亲本应关注各类群间性状差异作选择,而不能仅关注地理来源选择。     

SSR标记揭示的云南地方稻品种遗传多样性及其保育意义

为了探索水稻(Oryza　sativa　L.)地方品种的遗传多样性及其有效保育方法,对采自云南省17个村寨的82个水稻地方品种和3个国际常用的典型籼稻和粳稻品种进行了微卫星(SSR)分子标记的分析。利用19对SSR引物在85个水稻品种中共扩增出了83个基因型,其分子量变异在100～500　bp之间。基于各品种SSR基因型遗传相似系数聚类分析而获得的UPGMA树状图表明各水稻品种之间存在较大的遗传多样性,其相似系数变异在0.15～0.90之间。但这些地方品种的遗传多样性并非呈均等的地理分布。这85个水稻品种在相似系数为0.52之处分为二组,其中一组包括几乎所有的籼稻品种,而另一组包括全部的粳稻品种,表明SSR标记能很好揭示水稻籼-粳分化。同时,有些来自不同采集地的同名品种表现出一定的遗传差异,说明同名异物的现象存在。云南水稻地方品种具有丰富的遗传多样性,对其有效保育十分重要和迫切,　但只有根据遗传多样性的水平和分布特点,采用正确的保育对策和取样方法才能确保对云南水稻地方品种的有效保育。结果进一步表明,选用适当的微卫星引物,可以为准确鉴定籼稻和粳稻品种及研究其进化规律提供有效的分子标记方法,并有利于有目标的水稻遗传资源保育和育种创新。     

利用微卫星标记分析山东地方鸡品种的遗传多样性

微卫星是近几年来应用较多的一种分子标记,可有效地进行基因鉴定与系谱分析,并可估算群体间的遗传距离。通过选用5个微卫星标记,检测了山东省5个地方鸡种：日照麻鸡、寿光鸡、莱芜黑鸡、济宁百日鸡、鲁西斗鸡以及一个外来鸡种——安卡黄鸡和一个外省地方鸡种——广西黄鸡共7个鸡种的遗传多样性。根据测试结果计算了每个等位基因的频率,以基因频率为基础分析了品种内的遗传变异和品种间的DA遗传距离,并讨论了微卫星多态性在应用于群体遗传变异及亲缘关系等方面的意义。结果表明：共检测到40个等位基因,其中等位基因数最多的位点为。ADL0136(10个);等位基因数最低的位点为ADL0146(5个);而且每个位点的等位基因分布并不均匀,都有一种或几种优势基因存在。在7个品种中,杂合度最低的为寿光鸡,杂合度值为0．3327,因为此鸡种多年来一直由寿光市慈伦种鸡场进行纯繁保种,未与其他鸡种杂交,因此杂合度最小;其他鸡种杂合度也都低于0．4,据分析可能是由于日照麻鸡、济宁百日鸡群体较小;莱芜黑鸡是正在选育的一个品种,个体间遗传关系也不远;安卡黄鸡和广西黄鸡自从引人嘉明公司后,群体近交现象普遍,因此各鸡种杂合度都偏低。由此可见,通过对杂合度的计算,微卫星可以较好地反应群体内的变异。各品种PIC值的变动范围从0．6196(寿光鸡)到0．7027(莱芜黑鸡),PIC值的大小与杂合度的高低体现了较高的一致性。对DA遗传距离的计算表明：日照麻鸡与济宁百日鸡的距离最近,而鲁西斗鸡与其他鸡种距离均较远。用UPGMA法进行聚类分析,结果7个鸡种被聚为3类：山东的4个地方鸡种寿光鸡、日照麻鸡、莱芜黑鸡与济宁百日鸡聚为一类;安卡黄鸡和广西黄鸡聚在一起;鲁西斗鸡独自为一类。这与几个鸡种的分化与选育历史是一致的,因此聚类图能够比较正确地反映7个品种之间的亲缘关系。     

水稻品种多样性田间稻瘟病菌群体遗传结构分析

利用稻瘟病菌的一段倒位重复序列Pot2设计的一对引物,采用rep—PCR分子指纹技术对来自石屏县净种杂交稻田块、净种糯稻田块以及间种杂交稻糯稻田间的251个稻瘟病单孢分离菌株进行扩增．结果表明,所有供试菌株均分别扩增到9—17条DNA带,大小从400bp到23kb左右,但大多数带主要集中在5—10kb之间．所有菌株共扩增出的DNA指纹带中,约65％的为多态性DNA带,35％的为共同扩增带．将供试菌株扩增带诺进行聚类分析,比较间裁与净载田间病菌群体遗传结构的组成差异结果表明,在不同遗传相似水平,菌株遗传宗群复杂度与栽培方式有一定相关性。间栽田间病菌遗传宗群较净栽田问复杂,为3—5个,且优势宗群群不明显;而在净栽糯稻或净栽杂交稻田间遗传宗群较为简单,只有1—3个,且优势宗群明显．本试验结果证明水稻品种多样性有利于稻瘟病菌稳定化选择。     

中国小麦地方品种内和品种间醇溶蛋白遗传多样性分析

为了揭示中国小麦地方品种内遗传异质性和品种间的遗传多样性,采用A-PAGE方法,对72份来自不同生态区的地方品种进行醇溶蛋白构成分析。结果发现,全部供试地方品种共观察到101条迁移率不同的务带,构成229种醇溶蛋白构型,每个品种醇溶蛋白条带数目为14—24。63份（87．5％）地方品种在品种内具有2种以上醇溶蛋白变异类型,其中,变异类型最多的品种二红皮小麦（ZM004659）30个子粒中有14种之多,多数品种具有2—3种变异类型。品种内醇溶蛋白构型一致的品种共有9个,占12．5％。这表明供试的大多数小麦地方品种内个体间在醇溶蛋白构成上具有遗传异质性。聚类分析表明,相同生态区的地方品种没有整齐地聚为一类。     

不同国家水稻品种的遗传多样性分析

探讨世界不同国家水稻品种的遗传多样性,旨在为各国品种资源的有效利用提供理论依据。本研究利用63对引物对36份来源于不同国家的水稻品种进行遗传多样性分析。共检测到269个等位基因,每个位点的等位基因数(Na)平均为4.54个,有效等位基因数(Ne)平均为3.22,基因多样性指数(H)平均为0.64,Shannon’s信息指数(I)平均为1.21,引物RM206、RM257、RM410、RM235、RM266的等位基因数较多在7条以上。所处纬度相近的国家或地区的水稻品种之间的遗传距离较近,被聚为同一类群,而所处纬度较远的国家或地区的水稻品种被分到了不同类群。结果表明,水稻品种之间的遗传差异与纬度和地理距离有很大的关系。     

浙江省薄皮甜瓜地方品种的表型遗传多样性

利用表型性状探讨了浙江省沿海地区27份薄皮甜瓜地方品种资源的遗传多样性。结果表明48个质量性状遗传多样性指数在0.17–1.98之间;33个数量性状的变异系数在4.56%–83.50%,表明其丰富的遗传多样性。形态学聚类分析表明,在相似系数0.30处可以将27个资源分成两大类,这与按生育期长短分类结果完全一致;亚类的划分与果实形状、种子形状、叶片颜色、果皮颜色、覆纹颜色等质量性状具有一定的相关性,但其划分依据相对独立。本研究结果进一步丰富了甜瓜的评价体系,并为今后优异基因资源的挖掘与利用提供重要依据。     

Diversity of microsatellites derived from genomic libraries and GenBank sequences in rice (Oryza sativa L.)

The growing number of rice microsatellite markers warrants a comprehensive comparison of allelic variability between the markers developed using different methods, with various sequence repeat motifs, and from coding and non-coding portions of the genome. We have performed such a comparison over a set of 323 microsatellite markers; 194 were derived from genomic library screening and 129 were derived from the analysis of rice-expressed sequence tags (ESTs) available in public DNA databases. We have evaluated the frequency of polymorphism between parental pairs of six inter- subspecific crosses and one inter-specific cross widely used for mapping in rice. Microsatellites derived from genomic libraries detected a higher level of polymorphism than those derived from ESTs contained in the GenBank database (83.8% versus 54.0%). Similarly, the other measures of genetic variability [the number of alleles per locus, polymorphism information content (PIC), and allele size ranges] were all higher in genomic library-derived microsatellites than in their EST-database counterparts. The highest overall degree of genetic diversity was seen in GA-containing microsatellites of genomic library origin, while the most conserved markers contained CCG- or CAG-trinucleotide motifs and were developed from GenBank sequences. Preferential location of specific motifs in coding versus non-coding regions of known genes was related to observed levels of microsatellite diversity. A strong positive correlation was observed between the maximum length of a microsatellite motif and the standard deviation of the molecular-weight of amplified fragments. The reliability of molecular weight standard deviation (SDmw) as an indicator of genetic variability of microsatellite loci is discussed. Received: 5 May 1999 / Accepted: 16 August 1999     

Cloning and characterization of a cDNA encoding a rice 13 kDa prolamin

Summary A cDNA library constructed from mRNAs obtained from developing rice endosperm was screened with a cDNA clone (RM7) of highest frequency of occurrence (1.8%). The translati) product directed by the mRNA which was hybrid-released from RM7 cDNA in a wheat germ cell-free system showed a molecular size of 13 kDa when coexisting with the protein body fraction of developing maize endosperm. A polypeptide sequence composed of 156 amino acids was deduced from the nucleotide sequence. By comparison with the 19 N-terminal amino acids obtained from Edman degradation of the isolated rice 13 kDa prolamin fraction, the signal sequence was determined as consisting of 19 amino acids. The deduced polypeptide is rich in hydrophobic amino acids such as Leu and Val, and also in Gln, but lacks Lys. Hence, the amino acid composition is consistent with that of rice 13 kDa rolamin. By homology with previously reported cereal prolamins, only a single octapeptide sequence, Gln-Gln-Gln-CysCys-Gln-Gln-Leu, which was observed in 15 kDa and 27 kDa zein, B- and -hordein, /- and -gliadin, and -secalin was conserved in the rice 10 kDa and 13 kDa prolamin. No repetitive sequences and/or sequences homologous to other cereal prolamins, except the above octapeptide, were observed for the mature 13 kDa prolamin polypeptide. The signal sequence region of the 13 kDa prolamin, however, shows homology of more than 65% in both the nucleotide sequence and the amino acid sequence with rice 10 kDa prolamin and maize zein.     

Distribution and characterization of over 1000 T-DNA tags in rice genome

We generated T-DNA insertions throughout the rice genome for saturation mutagenesis. More than 1,000 flanking sequences were mapped on 12 rice chromosomes. Our results showed that T-DNA tags were not randomly spread on rice chromosomes and were preferentially inserted in gene-rich regions. Few insertions (2.4%) were found in repetitive regions. T-DNA insertions in genic (58.1%) and intergenic regions (41.9%) showed a good correlation with the predicted size distribution of these sequences in the rice genome. Whereas, obvious biases were found for the insertions in the 5'- and 3'-regulatory regions outside the coding regions both at 500-bp size and in introns rather than in exons. Such distribution patterns and biases for T-DNA integration in rice are similar to that of the previous report in Arabidopsis, which may result from T-DNA integration mechanism itself. Rice will require approximately the same number of T-DNA insertions for saturation mutagenesis as will Arabidopsis. A database of the T-DNA insertion sites in rice is publicly available at our web site (http://www.genomics.zju.edu.cn/ricetdna).     

Molecularmarker diversity and hybrid sterility in indica-japonica rice crosses

 The partial sterility of hybrids between the indica and japonica rice subspecies of Asian cultivated rice is a serious constraint for utilizing inter-subspecific heterosis in hybrid rice breeding. In this study, we have investigated the relationship between molecular-marker polymorphism and indica-japonica hybrid fertility using a diallel set involving 20 rice accessions including 9 indica and 11 japonica varieties. Spikelet fertility of the resulting 190 F₁s and their parents was examined in a replicated field trial. Intra-subspecific hybrids showed much higher spikelet fertility than inter-subspecific hybrids except in crosses involving wide-compatibility varieties. The parents were surveyed for DNA polymorphism using 96 RFLP and ten SSR markers, which revealed extensive genetic differentiation between indica and japonica varieties. A large number of markers detected highly significant effects on hybrid fertility. The chromosomal locations for many of the positive markers coincided well with previously identified loci for hybrid sterility. The correlation between hybrid fertility and parental distance was low in both intra- and inter-subspecific crosses. The results suggest that the genetic basis of indica-japonica hybrid sterility is complex. It is the qualitative, rather than the quantitative, difference between the parents that determines the fertility of hybrids. Received: 3 January 1997/Accepted: 17 January 1997     

Assessment of genetic diversity and chemical composition among seven black locust populations from Northern China

Black locust (Robinia pseudoacacia L.) is one of the important biomass sources used to produce bioenergy and bioethanol. In this study, we examined variations in chemical composition as well as genetic diversity and differentiation among 165 black locust plants from seven populations in five provinces (Beijing, Hebei, Gansu, Shanxi, Shandong) of Northern China using microsatellite markers(SSR). The contents of cellulose, hemicellulose, lignin varied widely among seven populations. Of the microsatellite markers analysed, 14 showed polymorphisms, and 45 alleles were identified. The polymorphism information content (PIC) ranged from 0.2885 (Rops4) to 0.6837 (Rp200), and most of the microsatellite loci had PIC values > 0.5. Expected heterozygosity (He), observed heterozygosity (Ho), and Shannon's information index (I) detected relatively high genetic variation among populations. The percentage of polymorphic loci in three populations was 100%, and the average among all populations were 95.92%. Analysis of molecular variance (AMOVA) indicated that the degree of genetic differentiation among the seven populations was low (G_ST = 0.058; N_m = 4.05), and chemical compositions had no relationship with genetic or geographic distance. This study demonstrates that microsatellite markers efficiently assess of genetic diversity and genetic differentiation in black locust populations, and all seven populations exhibit high genetic diversity. The population from Feixian has the potential to be lingo-cellulosic biomass for bioenergy and bioethanol.     

Localization of the rice stripe disease resistance gene, Stv-bi, by graphical genotyping and linkage analyses with molecular markers

 We used graphical genotyping and linkage analyses with molecular markers to determine the chromosomal location of the rice stripe disease resistance gene, Stv-b ⁱ . The stripe resistance gene from the indica rice (Oryza sativa) cv ‘Modan’ was introgressed into several Japanese rice varieties. We found 4 RFLP markers in ‘Modan’, five susceptible parental rice varieties (‘Norin No. 8’, ‘Sachihikari’, ‘Kanto No. 98’, ‘Hokuriku No.103’ and ‘Koganebare’) and four resistant progeny varieties (‘St. No. 1’, ‘Aichi No. 6’, ‘Aoisora’ and ‘Asanohikari’). Graphical genotyping of the resistant progeny revealed a chromosomal segment ascribable to ‘Modan’ and associated with stripe resistance. The chromosomal segment from ‘Modan’ was located at 35.85 cM on chromosome 11. Linkage analysis using 120 F₂ individuals from a cross between ‘Koshihikari’ (susceptible) and ‘Asanohikari’ (resistant) revealed another 8 RFLP markers in the same chromosome. We performed a bioassay for rice stripe resistance in F₃ lines of the F₂ individuals using infective small brown planthoppers and identified an 1.8-cM segment harboring the rice stripe disease resistance gene, Stv-b ⁱ , between XNpb220 and XNpb257/ XNpb254. Furthermore, Stv-b ⁱ was linked by 0.0 cM to a RFLP marker, ST10, which was developed on the basis of the results of RAPD analysis. These DNA markers near the Stv-b ⁱ locus may be useful in marker-assisted selection and map-based cloning of the Stv-b ⁱ gene. Received: 26 September 1997 / Accepted: 4 November 1997     

Natural and human‐mediated selection in a landrace of Thai rice (Oryza sativa)

Landrace rice in Thailand consists of managed populations grown under traditional and long‐standing agricultural practices. These populations evolve both in response to environmental conditions within the local agro‐ecosystem and in response to human activities. Single landraces are grown across varying environments and recently have experienced temporal changes in local environments due to climate change. Here we assess the interplay between natural selection in a changing climate and human‐mediated selection on the population genetic structure of Muey Nawng, a local landrace of Thai rice. Genetic diversity and population structure of landrace rice were assessed by a STRUCTURE analysis of 20 microsatellite loci. The first exon–intron junction of the waxy gene was sequenced to determine genotypes for glutinous or non‐glutinous grain starch. Muey Nawng rice is genetically variable and is structured based on starch grain types and the level of resistance to gall midge pest. A strong positive correlation was found between genetic diversity and the percentage of gall midge infestation. Variation in the waxy locus is correlated with starch quality; selection for non‐glutinous rice appears to involve additional genes. The dynamics of genetic diversity within Muey Nawng rice depends on three factors: (a) a genetic bottleneck caused by strong selection associated with gall midge infestation, (b) selection by local farmers for starch quality and (c) variation introduced by farmer practices for cultivation and seed exchange. These results, when taken in total, document the ability of landrace rice to quickly evolve in response to both natural and human‐mediated selection.     

Comparative analysis of microsatellite DNA polymorphism in landraces and cultivars of rice

Genetic polymorphisms of ten microsatellite DNA loci were examined among 238 accessions of landraces and cultivars that represent a significant portion of the distribution range for both indica and japonica groups of cultivated rice. In all, 93 alleles were identified with these ten markers. The number of alleles varied from a low of 3 or 4 at each of four loci, to an intermediate value of 9–14 at five loci, and to an extra-ordinarily high 25 at one locus. The numbers of alleles per locus are much larger than those detected using other types of markers. The number of alleles detected at a locus is significantly correlated with the number of simple sequence repeats in the targeted microsatellite DNA. Indica rice has about 14% more alleles than japonica rice, and such allele number differences are more pronounced in landraces than in cultivars. The indica-japonica differentiation component accounted for about 10% of the diversity in the total sample, and twice as much differentiation was detected in cultivars as in landraces. About two-thirds as many alleles were observed in cultivars as in landraces; another two-thirds of the alleles in the cultivar group were found in modern elite cultivars or parents of hybrid rice. The majority of the simple sequence repeat (SSR) alleles that were present in high or intermediate frequencies in landraces ultimately survived into modern elite cultivars and hybrids. The greater resolving power and the efficient production of massive amounts of SSR data may be particularly useful for germplasm assessment and evolutionary studies of crop plants.     

Abundance, polymorphism and genetic mapping of microsatellites in rice

Dinucleotide microsatellites have been characterized and used as genetic markers in rice. Screening of a rice genomic library with poly(dG-dA)·(dC-dT) and poly(dG-dT)·(dC-dA) probes indicated that (GA)_n repeats occurred, on average, once every 225 kb and (GT)_n repeats once every 480 kb. DNA sequencing of ten randomly selected microsatellites indicated that the numbers of repeats ranged from 12 to 34 and that the patterns of microsatellites in rice were similar to those of humans and other mammals. Primers to these microsatellite loci as well as to four published microsatellite-containing sequences have been designed and degrees of polymorphism has been examined with 20 rice accessions. Multiple alleles, ranging from 5 to 11, have been observed at all the microsatellite loci in 20 rice accessions. Alleles specific to two cultivated subspecies, indica and japonica, were found in some microsatellite loci. Heterozygosity values of all the microsatellite markers were significantly higher than those of RFLP markers, based upon a parallel comparison. Ten microsatellite loci have been genetically mapped to four rice chromosomes. The genomic distribution of microsatellites appears to be random in rice.