基于SSR标记的寒地水稻品种骨干亲本分析

富士光、藤系138、上育397和五优稻1是20世纪90年代至今寒地水稻品种选育的骨干亲本。利用50对SSR引物对上述骨干亲本及其衍生品种进行聚类分析和主坐标分析(principal coordinate analysis, PCO),结果表明,50对SSR引物在51份供试材料中共检测到150个等位基因,变化范围为2~6个,平均为3个;引物PIC的变化范围为0.0725~ 0.6845,平均0.3655;聚类分析将51份材料分为4类,4个骨干亲本分别被聚到4类中;PCO分析显示,四个骨干亲本相距较远,呈独立的分支,衍生品种围绕着骨干亲本分布;在检测出的39个稀有等位基因中,仅有3个存在骨干亲本中。表明近年寒地水稻品种遗传改良是围绕少数骨干亲本进行的,骨干亲本将大部分优良基因传递到了衍生品种中,SSR分析和PCO分析与系谱分析得到了一致的结果。     

用SSR标记鉴定大豆杂交组合F1的方法研究

为建立鉴定大豆杂种的方法,采用亲本间有多态性的3对SSR引物,对148个耐盐与盐敏感大豆品种正反交F1植株进行分子鉴定,结果表明,有81．8％的F1为真杂种,且3对多态性SSR引物检测结果一致;在亲本基因型纯合的情况下,采用1对在亲本间有多态性的SSR引物即可对F1真伪进行准确判断;亲本间分子量差异大的SSR位点可用高浓度琼脂糖电泳进行快速鉴定。利用该方法对2007年参加国家大豆区试的大豆杂交种品系H02—286的50粒种子进行纯度鉴定,进一步验证了SSR标记检测杂种真伪的可行性。     

利用SSR标记鉴定西瓜杂交种纯度的研究

以2个西瓜杂交品种(系)的种子黑公子和04-17及其亲本为材料,用SSR标记技术研究杂种与其双亲之间的扩增谱带多态性,以甄别真假杂种.结果发现,所试验的52对SSR引物中有13对引物分别在2个西瓜杂交种和其双亲之间存在扩增条带的多态性,表现为:多数SSR引物对自交系的扩增只出现1条带,但部分引物在某些自交系中扩增出2条带,杂交种条带均为父母本的互补型,很适合做杂交种纯度鉴定.用引物CMCT134b对黑公子和引物CMGA165对04-17进行了各100粒单种子SSR鉴定,所测纯度分别为96%和100%,与田间纯度95.6%和99.7%非常接近,表明SSR标记技术在西瓜杂交种子纯度室内快速检测中的应用前景.     

甘薯杂交F_1群体材料的遗传多样性研究

甘薯(Ipomoea batatas(L.)Lam.)作为世界上一种重要的粮食、饲料、工业原料及新型能源作物,从有性生殖F1选择优良实生系进行选择育种一直是甘薯育种的重要方式。为了优化甘薯杂交育种方法,合理选配杂交组合,提高育种效率,本试验利用SSR标记研究了甘薯杂交群体基于SSR标记及13个农艺性状的遗传多样性,得到了群体内的聚类图,并且筛选出了甘薯的高产株系。群体SSR标记的聚类分析结果显示,群体材料与各亲本遗传距离比较远,被聚为3类,而亲本单独聚在另外一类。13个农艺性状的聚类将亲本与部分群体材料聚在了一起,且将群体材料和亲本材料作为一个整体时,其遗传距离的变异高达30%以上,远远高于SSR标记所获得的遗传变异系数。     

我国茶树主要骨干亲本及其衍生品种(系)的SSR分析

铁观音、黄棪和福鼎大白茶分别是我国乌龙茶和红绿茶育种中的骨干亲本,由他们衍生出了一系列的优良品种,研究他们的遗传多样性及构建指纹图谱将有助于今后茶树育种工作中骨干亲本的合理利用和品种权的保护。本研究利用40对SSR引物对我国乌龙茶骨干亲本铁观音、黄棪及其衍生品种(系)和红绿茶骨干亲本福鼎大白茶及衍生品种进行了研究。结果表明,34份供试品种(系)的基因多样性指数（H）为0.54,平均遗传距离0.58,表明我国茶树主要骨干亲本及其衍生品种(系)具有较高的遗传多样性水平和较大的遗传变异,且90%的遗传多样性来自品种之间的遗传差异。聚类结果表明两套品种(系)各自聚为一类,遗传结构分析也显示两套品种(系)之间存在明显的差异。利用其中5对引物组合构建了供试材料的数码指纹图谱。     

铝对玉米生长和硝酸还原酶活性的影响

测定了生长在Al2（SO4）100μmol／L氮素营养液中的两个玉米品种（SC704和VA35的根系和叶片）的NADH-硝酸还原酶（EC1．6．6．）和NAD（P）H-硝酸还原酶（EC1．6．6．2）活性。结果表明铝的存在阻碍了玉米根系和叶片的生长、降低了营养液的pH值,降低了叶片的NADH-及NAD（P）H-硝酸还原酶活性（酶活性降低的程度SC704低于VA35）,增加了根系的NADH-和NAD（P）H-硝酸还原酶活性（VA35根系的比活性除外）。铝胁迫下根系的NADH-和NAD（P）H-硝酸还原酶活性的增加SC704大于VA35。耐铝品种SC704的高NR活性以及在铝胁迫下能维持更高的NR活性的特点说明硝酸还原酶与植物组织的Al解毒机制有关。同时,在铝胁迫下的硝酸盐代谢中NAD（P）H-硝酸还原酶具有更重要的作用。     

我国西南地区玉米地方品种遗传多样性的SSR分子标记分析

利用微卫星(SSR)标记技术和DNA混合取样方法,选取均匀覆盖玉米染色体组的42对SSR引物,检测了来自我国西南地区54个玉米地方品种的遗传多样性。在54个玉米地方品种中检测到256个等位基因,每个SSR标记的等位基因数为2～9个,平均6.1个,说明我国西南地区玉米地方品种遗传多样性丰富。根据遗传相似系数矩阵做出的树状图,将54个玉米地方品种大致划分成4类,来源于同一地区的多数玉米地方品种划分在同一类中,表明西南地区玉米地方品种的地理分布与其遗传背景存在内在联系。从54个玉米地方品种中选出11个,每个品种选取15个单株,共165个DNA单株样品,分析玉米地方品种的遗传结构及其品种内的遗传多样性。对于检测玉米地方品种的遗传多样性,DNA单株样品分析优于DNA混合样品分析,42对相同的SSR引物在11个玉米地方品种中检测到330个等位基因,平均等位基因数A=7.86,有效等位基因数Ae=3.90,平均期望杂合度He=0.69,实际观察杂合度H0=0.37。据遗传结构分析结果,固定指数(F)为0.25～0.79,表明玉米地方品种是典型的混合繁育系统;由于杂合体不足,玉米地方品种群体间及群体内的遗传结构均偏离了Hardy-Weinberg平衡;杂合性基因多样度比率(Fst)平均为0.07,表明品种间和品种内的遗传变异分别占总遗传变异的7%和93%。玉米地方品种内遗传多样性及品种间遗传距离分析结果表明,在我国西南地区,分布在四川的玉米地方品种具有最丰富的遗传变异。经综合分析推测,我国西南地区玉米地方品种最早引进到四川种植,由此向毗邻地区传播扩散。     

大豆杂种第二代粒茎比遗传变异的初步研究

大豆粒茎比系指大豆茎秆的生物产量与其 籽粒产量的比值。它是关系到大豆育种与栽培 的主要的经济性状。一个大豆品种高产与否,取 决于其粒茎比的大小。因此,研究大豆品种间 杂交材料粒茎比的遗传变异规律,作为选育高 粒茎比大豆品种之参考,便显得尤为重要。近 年来,国外一些科研工作者曾试图通过栽培措 施来提高大豆粒茎比LI121。但是,关于大豆粒茎 比遗传变异的研究,尚未见有报道。笔者从 1976年起,就杂交种第二代粒茎比的表现、变异 情况、亲本粒茎比对F2代粒茎比的影响,以及 粒茎比遗传变异与其他性状的关系等方面进行 了初步研究,以作为在大豆杂交育种时选择亲 本及处理杂交材料的参考。     

盐胁迫对不同品种玉米苗期生长与叶片光合特性的影响

为研究盐胁迫对不同玉米品种玉米苗期生理性状的影响, 以玉米杂交种隆平206、秦龙14以及隆平206亲本(母本L239和父本L7221)为试验材料, 设置不同盐浓度处理, 分析玉米苗期生长与叶片光合特性的变化。结果表明, 盐胁迫下玉米植株高和地上部分鲜重显著下降, 随着盐浓度的增加, 其下降幅度增加; 两杂交种品种比较, 盐胁迫对秦龙14的影响较隆平206大; 隆平206两亲本比较, 盐胁迫对父本L7221较母本L239植株高与地上部分鲜重的影响更大。与对照相比, 盐胁迫显著增加玉米叶片色素含量, 且随着盐浓度的增加, 其增幅逐渐增加。盐胁迫显著增加叶片净光合速率, 但单叶光合速率逐渐降低, 两杂交种品种表现一致。可见虽然盐胁迫下单叶光合速率下降, 单位面积叶片的光合作用对玉米有一定的补偿作用。盐胁迫对秦龙14单叶光合速率的影响较隆平206大, 隆平206两亲本比较, 盐胁迫对父本L7221的影响较母本L239更大。     

SSR与SRAP标记在玉米品种鉴定中的比较研究

利用SSR标记和SRAP标记对19个玉米品种及8份莱农15样品进行了分析,比较了2种标记的分辨能力及在亲缘关系和杂交种纯度鉴定中的表现.与SSR标记相比,SRAP标记用于玉米品种鉴定扩增住点数量更多,PIC值更高,具有更高的分辨率;在亲缘关系分析方面,SSR检测的遗传距离变幅更大,2种标记计算的遗传距离呈极显著正相关,分类结果基本一致;在杂交种纯度检测中,SRAP标记的期望位点在杂交种群体中检测率高于SSR标记相应位点,检测杂交种纯度结果更接近田问种植鉴定,因而准确度更高.SRAP在玉米品种鉴定中具有一定的优势,可作为SSR标记技术的有益补充,特别是在杂交种纯度鉴定中应用.     

Genetic analysis of maize germplasm in the Korean Genebank and association with agronomic traits and simple sequence repeat markers

We assessed genetic and phenotypic variation in 105 maize germplasm accessions from RDA-Genebank of Korea and performed association analyses for 11 agronomical traits and 100 simple sequence repeats (SSR). Genetic diversity (GD) analysis revealed a total of 1104 alleles at the 100 SSR loci. The average number of alleles per locus was 11.0. The average GD and polymorphic information content values were 0.73 and 0.70, respectively. The average major allele frequency was 0.41. Population structure analysis indicated that these maize accessions comprised two major groups and one admixed group based on a membership probability threshold of 0.80. The two major groups contained 35 and 46 maize accessions. A mixed linear model of association analysis revealed five marker-trait associations with a significance level of P?≤?0.01 involving five SSR markers. A general linear model showed 72 marker-trait associations involving 42 SSR markers. We confirmed the presence in the general linear model associations of the five significant marker-trait associations (SMTAs) identified in the mixed linear model. For these SMTAs, two loci were associated with stem diameter and one locus each was associated with ear row number, leaf width, and leaf length. These results should prove useful for breeding new inbred lines by selecting parental lines using molecular markers and will help to preserve maize genetic resources in Korea.     

Genetic diversity of sea-island cotton (Gossypium barbadense) revealed by mapped SSRs

In order to evaluate the genetic diversity of sea-island cotton (Gossypium barbadense), 237 commonly mapped SSR markers covering the cotton genome were used to genotype 56 sea-island cotton accessions. A total of 218 polymorphic primer pairs (91.98%) amplified 361 loci, with a mean of 1.66 loci. Polymorphism information content values of the SSR primers ranged from 0.035 to 0.862, with a mean of 0.320. The highest mean polymorphism information content value for the SSR motifs was from a compound motif (0.402), and for the chromosomes it was Chr10 (0.589); the highest ratio of polymorphic primers in Xinjiang accessions was from Chr21 (83.33%). Genetic diversity was high in Xinjiang accessions. AMOVA showed that variation was 8 and 92% among populations and within populations, respectively. The 56 sea-island accessions were divided into three groups in the UPGMA dendrogram: Xinhai5 was in the first group; accessions from Xinjiang, except the five main ones, were in the second group, and the other 34 accessions were in the third group. Accessions from the former Soviet Union and Xinjiang main accessions were closely related. Both PCA and UPGMA confirmed that Xinhai5 was distinct from the other accessions, and accessions from Xinjiang were in an independent group. Given the differences between principal components analysis and UPGMA results, it is necessary to combine molecular markers and pedigree information so that genetic diversity can be objectively analyzed.     

Genetic diversity among Chinese Hami melon and its relationship with melon germplasm of diverse origins revealed by microsatellite markers

Thick-skinned melon called Hami melon is the most widely cultivated and exported type of melon in China, and mainly grown in Xinjiang province. Here the genetic variation of 64 melon genotypes including 43 Xinjiang Hami melon accessions was analyzed using 36 simple sequence repeat (SSR) markers yielding 145 alleles. The polymorphic information content of SSR markers ranged from 0.09 to 0.83 (average 0.45). Based on the SSR markers, the melon accessions were clustered into 2 major groups (thick and thin-skinned melons). In addition, the sub-cluster analysis based on SSR markers partitioned different botanical groups, even separating similar agronomic trait groups (Xinjiang landraces var. ameri and var. inodorus). SSR analysis showed that 4 SSR markers (CMBR150, CMCTT144, CMBR84 and CMBR12) produced polymorphic bands of different sizes between these two botanical groups. Those four molecular markers might be related to melon fruit maturing time. A considerably low level of genetic diversity was detected in Xinjiang melon accessions. Genetic distances indicated the relatively narrower genetic base but specific taxonomic status of Xinjiang landraces compared with foreign reference accessions.     

Analysis of phenotypic and microsatellite-based diversity of maize landraces in India,especially from the North East Himalayan region

The maize landraces in the North East Himalayan (NEH) region in India, especially in the Sikkim state, are morphologically highly diverse. The present study provides details of phenotypic and molecular characterization of a set of 48 selected maize landrace accessions, including the ‘Sikkim Primitives’ which have a unique habit of prolificacy (5–9 ears on a single stalk). Multi-location phenotypic evaluation of these 48 accessions revealed significant genetic variability for grain yield and its components, leading to identification of several promising accessions. Cluster analysis and PCA using nine morpho-agronomic characters clearly separated ‘Sikkim Primitives’ from the rest of the accessions. PCA revealed two principal components describing 90% of the total variation, with hundred kernel weight, ear length, ear diameter, number of kernels per ear and flowering behaviour forming the most discriminatory traits. The accessions were genotyped using 42 microsatellite or simple sequence repeat (SSR) markers using a ‘population bulk DNA fingerprinting strategy’, with allele resolution using an automated DNA Sequencer. The study revealed a high mean number of alleles per SSR locus (13.0) and high Polymorphism Information Content (PIC) value of 0.60. The analysis also led to identification of 163 private/unique alleles, differentiating 44 out of 48 accessions. Six highly frequent SSR alleles were detected at different loci (phi014, phi062, phi090, umc1266, umc1367 and umc2250) with individual frequencies ≥0.75. Some of these SSR loci were reported to tag specific genes/QTL for some important traits, indicating that chromosomal regions harboring these SSR alleles were not selectively neutral. Cluster analysis using Rogers’ genetic distance also revealed distinct genetic identity of the ‘Sikkim Primitives’ from the rest of the accessions in India, including Sikkim. Mantel’s test revealed significant and positive correlation between the phenotypic and molecular genetic dissimilarity matrices. The study was the first to portray the patterns of phenotypic and molecular diversity in the maize landraces from the NEH region in India.     

Variation of DNA fingerprints among accessions within maize inbred lines and implications for identification of essentially derived varieties.

Genetic distances (GDs) based on molecular markers are important parameters for identifying essentially derived varieties (EDVs). In this context information about the variability of molecular markers within maize inbred lines is essential. Our objectives were to (1) determine the variation in the size of simple sequence repeat (SSR) fragments among different accessions of maize inbreds and doubled haploid (DH) lines, (2) attribute the observed variation to genetic and marker system-specific sources, and (3) investigate the effect of SSR fragment size differences within maize lines on the GD between maize lines and their consequences for the identification of essentially derived varieties. Two to five accessions from nine inbred lines and five DH lines were taken from different sources or drawn as independent samples from the same seed lot. Each accession was genotyped with 100 SSR markers that evenly covered the whole maize genome. In total, 437 SSR fragments were identified, with a mean of 4.4 alleles per locus. The average polymorphic information content (PIC) was 0.58. GD estimates between two accessions of the same genotype ranged from 0.00 to 0.12 with an average of 0.029 for inbred lines and 0.001 for DH lines. An average of 11.1 SSRs was polymorphic between accessions of the same inbred line due to non-amplification (8.1 SSRs), heterogeneity (4.0 SSRs) or unknown alleles (2.6 SSRs). In contrast to lab errors, heterogeneity contributed considerably to the observed variation for GD. In order to decrease the probability to be suited for infringing an EDV threshold by chance, we recommend to increase the level of homogeneity of inbred lines before applying for plant variety protection.     

Analysis of the genetic diversity of garlic (Allium sativum L.) by simple sequence repeat and inter simple sequence repeat analysis and agro-morphological traits

The genetic diversity of 39 garlic accessions was investigated using eight simple sequence repeat (SSR) primer combinations and 17 inter-simple sequence repeat (ISSR) primer combinations. A total of 109 polymorphic loci were detected among these accessions, with an average of 4.63 polymorphic loci per SSR primer combination and 4.29 polymorphic loci per ISSR primer combination. The mean effective number of alleles, the mean Nei's genetic diversity, and the mean Shannon's information index for SSR were 1.4799, 0.2870, and 0.4378, respectively; and those for ISSR were 1.4847, 0.2898 and 0.4415, respectively. Cluster analysis, using the unweighted pair-group method with arithmetic averages (UPGMA) based on the allele frequency data, classified the accessions into three groups. The results of principal component analysis (PCA) were consistent with those of the cluster analysis. PCA showed that each of these three groups exhibited significant variation in agro-morphological traits. These findings suggest that the eight SSR and 17 ISSR primers identified could define valuable markers for genetic diversity for use by plant breeders.     

SSR markers development and their application in genetic diversity evaluation of garlic (Allium sativum) germplasm

Garlic (Allium sativum), an asexually propagated vegetable and medicinal crop, has abundant genetic variation. Genetic diversity evaluation based on molecular markers has apparent advantages since their genomic abundance, environment insensitivity, and non-tissue specific features. However, the limited number of available DNA markers, especially SSR markers, are insufficient to conduct related genetic diversity assessment studies in garlic. In this study, 4372 EST-SSR markers were newly developed, and 12 polymorphic markers together with other 17 garlic SSR markers were used to assess the genetic diversity and population structure of 127 garlic accessions. The averaged polymorphism information content (PIC) of these 29 SSR markers was 0.36, ranging from 0.22 to 0.49. Seventy-nine polymorphic loci were detected among these accessions, with an average of 3.48 polymorphic loci per SSR. Both the clustering analyses based on either the genotype data of SSR markers or the phenotypic data of morphological traits obtained genetic distance divided the 127 garlic accessions into three clusters. Moreover, the Mantel test showed that genetic distance had no significant correlations with geographic distance, and weak correlations were found between genetic distance and the phenotypic traits. AMOVA analysis showed that the main genetic variation of this garlic germplasm collection existed in the within-population or cluster. Results of this study will be of great value for the genetic/breeding studies in garlic and enhance the utilization of these garlic germplasms.     

Analysis of Genetic Diversity in the North Eastern Himalayan Maize Landraces using Microsatellite Markers

Population DNA fingerprinting of 48 selected North Eastern Himalayan (NEH) landrace accessions was undertaken using 41 polymorphic fluorescent dye-labelled microsatellite/Simple Sequence Repeat (SSR) markers, using a DNA Sequencer. The analysis revealed a large number of SSR alleles (576), with high mean number of alleles per locus (13.8), and Polymorphism Information Content (PIC) of 0.63, reflecting the level of diversity in the NEH accessions and the informativeness of the SSR markers. The study also led to identification of 135 unique alleles, differentiating 44 out of the 48 accessions. Five highly frequent (major) SSR alleles (umc1545 _80bp, phi062 _162bp, umc1367 _159bp, umc2250 _152bp and phi112 _152bp) were detected indicating that chromosomal regions harbouring these S SR alleles might not be selectively neutral. Analysis of population genetic parameters, including Wright’s F statistics, revealed high level of genetic differentiation, very low levels of inbreeding, and restricted gene flow between the NEH landraces. AMOVA (Analysis of Molecular Variance) showed that 67 per cent of the total variation in the accessions could be attributed to within-population diversity, and the rest between the accessions. Cluster analysis of SSR data using Rogers’ genetic distance and UPGMA, showed significant genetic diversity among the landraces from Sikkim. This is the first detailed study of SSR allele frequency-based analysis of genetic diversity in the NEH maize landraces of India.     

QTL mapping for test weight by using F<Subscript><Emphasis Type="Bold">2:3</Emphasis></Subscript> population in maize

Test weight is an important trait in maize breeding. Understanding the genetic mechanism of test weight is important for effective selection of maize test weight improvement. In this study, quantitative trait loci (QTL) for maize test weight were identified. In the years 2007 and 2008, a F_2:3 population along with the parents Chang7-2 and Zheng58 were planted in Zhengzhou, People’s Republic of China. Significant genotypic variation for maize test weight was observed in both years. Based on the genetic map containing 180 polymorphic SSR markers with an average linkage distance of 11.0 cM, QTL for maize test weight were analysed by mixed-model composite interval mapping. Five QTL, including four QTL with only additive effects, were identified on chromosomes 1, 2, 3, 4 and 5, and together explained 25.2% of the phenotypic variation. Seven pairs of epistatic interactions were also detected, involving 11 loci distributed on chromosomes 1, 2, 3, 4, 5 and 7, respectively, which totally contributed 18.2% of the phenotypic variation. However, no significant QTL × environment (Q×E) interaction and epistasis × environment interaction effects were detected. The results showed that besides the additive QTL, epistatic interactions also formed an important genetic basis for test weight in maize.