用微卫星DNA标记检测中国主要杂交水稻亲本的遗传差异

选用分布于水稻(OryzasativaL.)12条染色体上的20对SSR(Simplesequencerepeats)引物,分析了具有多种质源和较大应用面积的24个水稻胞质雄性不育系、1个光(温)敏核不育系、3个保持系和5个生产中应用较广的恢复系。在以上33份杂交水稻亲本材料间共检测出102个等位基因(alleles),平均每对SSR引物可检测到5.1个等位基因。PIC(polymorphicindexcontent)值的变动范围为0.274～0.773,平均PIC值为0.554。从56对SSR引物中筛选出5对引物,能够有效地区分所有供试的水稻雄性不育系和恢复系。杂交水稻亲本的聚类分析表明:(1)我国水稻雄性不育系遗传变异丰富,但生产中主要应用的水稻雄性不育系遗传背景比较单一。(2)生产中应用面积较大的水稻雄性不育系遗传变异较恢复系差。(3)生产中主要应用的水稻雄性不育系与恢复系分别聚类于不同的类群,且遗传关系较远。     

用微卫星DNA标记对我国杂交水稻主要恢复系遗传差异的检测分析

选用分布于水稻（Oryza sativaL.)12条染色体上的25对SSR（Simple sequence repeats)引物,分析了生产中广泛应用的35个杂交 水稻恢复系,在35个杂交水稻恢复系材料间共检测出65个等位基因（alleles),平均每对SSR引物可检测到2.6个等位基因,PIC（Polymorphism index content）值的变动范围为0.206-0.682,平均值为0.414。聚类分析表明,我国杂交 水稻恢复系资源比较丰富,但其遗传差异较小,遗传背景单一,从而在很大程度上限制了我国水稻杂种优势的利用。     

56个杂交水稻骨干亲本SSR指纹图谱的构建及遗传相似性分析

以中国56个杂交水稻骨干亲本为研究材料,包括水稻雄性不育系和恢复系。从国标中公布的48对水稻SSR引物中筛选出14对稳定性好、多态性高、杂带少且在染色体上分布均匀的引物作为核心引物,建立56个杂交水稻骨干亲本的SSR指纹图谱,结果表明：14对SSR引物在56份材料中共扩增出48个多态性片段,平均每对引物可以检测3．43个等位基因。聚类分析得出56个品种间的遗传相似性系数在0．63～0．98之间,基本上反映了不同品种间的亲缘关系。     

太湖稻区与国内部分香稻SSR指纹图谱构建及遗传多样性初析

以太湖稻区和国内其他地区的39个香稻品种,以及籼型恢复系2个对照品种为研究材料,利用SSR分子标记进行DNA指纹图谱构建和遗传多样性分析。从40对国标中公布的水稻SSR引物中筛选出13对核心引物,在41个水稻品种中共检测到36个多态性片段。据此建立41个水稻品种的DNA指纹图谱,进行聚类分析,发现供试41个品种的遗传相似系数在0.58以上,而供试香稻品种的遗传相似系数在0.64以上,基本反映了不同品种间的亲缘关系。     

籼型杂交稻光合特性的配合力分析

以6个籼型杂交稻三系不育系和5个恢复系按不完全双列杂交设计配制的30个杂交稻组合及其亲本品种为材料,对其光合性状进行了测定和分析,结果表明:(1)杂交稻组合的光合特性存在显著或极显著的组合间遗传差异,光合特性的遗传变异主要来自基因的非加性效应;(2)胞间CO2浓度、蒸腾速率、叶绿素a、叶绿素b和类胡萝卜素含量受不育系的影响大于恢复系,而气孔导度、叶绿素a+b含量受恢复系的影响大于不育系;(3)杂交稻光合性状的广义遗传力均大于狭义遗传力,各性状主要受基因互作及环境的影响。狭义遗传力的大小依次为叶绿素b、叶绿素a+b、叶绿素a、气孔导度、胞间CO2浓度、类胡萝卜素和蒸腾速率,这些性状具有中等遗传力;(4)9个光合性状杂交稻F1表型值与父母本一般配合力效应值之和的相关系数均达极显著水平。因此,可以根据父母本一般配合力效应值之和来预测杂交稻组合光合性状的表现,有利于高效选育高光效杂交稻组合。     

红麻UG93细胞质雄性不育系、保持系及恢复系间的遗传多样性分析

利用8条核基因组ISSR引物和7对叶绿体基因组SSR引物(cpSSR),对9对红麻UG93细胞质雄性不育系/保持系及5个恢复系的细胞核、细胞质遗传多样性进行分析.结果表明:各材料的核基因组遗传相似系数在0.333～1.000之间,其中保持系间、保持系和恢复系间、恢复系间的平均相似系数分别为0.583、0.689和0.8...     

红莲型杂交稻(红莲2号)及其骨干亲本的RAPD分析与鉴定

利用RAPD技术,从248个随机寡核苷酸引物（10－mer）中筛出18个引物对红莲型杂交稻组合红莲2号及其亲本（T－07A、T－07B、YD6－05）,另6个红莲型胞质不育系的骨干恢复和汕优63及其亲本共14份水稻材料进行分析。共检测到173个多态性标记。聚类分析结果表明：不育系与保持系间因核背景相似,遗传差异很小;杂种（F1）的基因型更倾向于恢复系;恢复系与保持系间遗传距离的相对较大,但各恢复系之间的遗传距离较小。利用这些标记能有效地地区交组合中不育系,保持系、恢复系和杂种（F1）。     

A genetic linkage map of microsatellite,gene-specific and morphological markers in diploid <Emphasis Type="Italic">Fragaria</Emphasis>

Knowledge of the genetic diversity of a species is important for the choice of crossing parents in line and hybrid breeding. Our objective was to investigate European winter triticale using simple sequence repeat (SSR) markers and the coancestry coefficient (f) with regard to genetic diversity and grouping of germplasm. Three to five primer pairs for each of the 42 chromosomes were selected to analyse 128 European winter triticale varieties and breeding lines. SSR analysis resulted in the identification of 657 alleles with an average of 6.8 alleles per primer pair. The average polymorphism information content (PIC) for polymorphic markers was 0.54. Correlation between f and genetic similarity (GS) estimates based on Rogers Distance was low (r_f×GS(ABDR)=0.33). The analysis of molecular variance (AMOVA) revealed that 84.7% of the total variation was found within breeding companies, and 15.3% among them. In conclusion, SSR markers from wheat and rye provide a powerful tool for assessing genetic diversity in triticale. Even though no distinct groups within the European winter triticale pool could be detected by principal co-ordinate analysis, this study provides basic information about the genetic relationships for breeding purposes.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by H.C. Becker     

Prediction of hybrid grain yield performances in Indica Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) with effect-increasing loci

One hundred and fifty-one rice hybrids produced in two sets of half-dialell crosses and their parents (13 cytoplasmic male sterile lines and 19 restorers) were used to predict the F₁ performances of seven yield traits through the parental genetic distances (GD) based on SSR markers. The positive loci (PL) and effect-increasing loci (IL), which were screened from SSR polymorphic loci by the F₁ traits of 32 parents, together with total loci (TL), were utilized to estimate parental GD and the models were found to predict the traits of hybrids derived from different parents, fixed parents, and different environments, respectively. The results were as follows: (1) 550 polymorphic loci were detected from 174 SSR markers: a dendrogram based on these loci could separate all the sterile and restorer lines used in the present study, which indicated that parental genetic diversity of F₁ was large; (2) the correlations between F₁ traits and parental GDs based on IL ranged from 0.61 to 0.87 with a mean of 0.76, and they were higher than those on TL or on PL; (3) predictions based on IL for F₁ traits (except grain weight per plant) derived from different environments were ideal, but worse for F₁ traits derived from different parents; and (4) IL was more effective than TL and PL in predicting traits of F₁ with fixed parents, and predictions for fixed restorer combinations were more effective than those for fixed sterile line combinations. These results should facilitate molecular prediction for hybrid yield and other traits by means of both elite sterile and restorer lines.     

基于陆地棉茎尖转录组序列的genic-SSR标记的开发与评价

利用软件MISA和SSR Locator 对陆地棉（Gossypium hirsutum L.）Coker312茎尖转录组测序得到的73515条Unigene序列进行分析,查找到4507个SSR位点,分布于4039条转录本序列中,SSR出现频率为5.5%,非编码区的SSR位点要显著高于编码区（2853/1654）。SSR重复基元中,三核苷酸重复基元占主导地位（51.03%）,且AAG/CTT （20.08%）类型最多;其次是二核苷酸重复基元（28.76%）,主要为AG/CT（55.47%）。利用引物批量设计软件共开发1569对SSR引物,在陆地棉TM-1、海岛棉3-79 (G. barbadense L.)、阿非利加棉(G. herbaceum L.)、雷蒙德氏棉(G. raimondii Ulbrich.)四个棉种中进行初步评价,其中1117对引物能在四个棉种间扩增出稳定的条带,扩增率为71.2%。选择650对能稳定扩增出条带的引物,在涉及5个棉种（增加了亚洲棉（G. arboreum L.））的13份材料中进一步筛选,83对引物能在13份材料中扩增出特异性条带, PIC变幅在0.121～0.648之间,平均值为0.422,其中80对引物能在7份陆地棉材料中扩增出特异性条带,平均PIC值为0.336;挑选5对在本实验室作图亲本鲁棉研22和鲁原343间有明显多态的引物进行连锁分析,其中4对成功连锁到本实验室构建的遗传图谱上。本研究丰富了棉属genic-SSR的数量,为棉属遗传作图、性状关联分析提供了更多引物选择。     

利用SSR标记定位明恢63的2对恢复基因

选取珍汕97A和明恢63杂交组合的F2高可育和高不育单株构建基因池,利用302对SSR引物对其进行了多态性分析。结果表明,位于第1染色体上的RM1和位于第10染色体上的RM258,RM304在亲本,基因池间表现多态性,用F2单株验证证明它们与野败型恢复基因连锁,完全不育株分析表明,与恢复基因间的遗传距离分别为1.9,2.9和0.0cM,野败型,红莲型,BT型3种不育胞质恢复基因在第10染色体上可能为同一基因或家族成员。     

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.     

Genetic Diversity Analysis of Elite Pearl Millet Inbred Lines using RAPD and SSR Markers

Pearl millet [Pennisetum glaucum (L) R Br] is one of the widely grown cereal crops in the arid and semi-arid regions of Africa and India. We undertook a study to ascertain the genetic diversity in 21 elite inbreds (parental lines of 13 pearl millet hybrids in India) using 20 Random Amplified Polymorphic DNA (RAPD) and 21 Simple Sequence Repeat (SSR) markers. Based on Polymorphism Information Content (PIC) and unique banding profiles, 6 RAPD primers OPD12, OPA16, OPB6, OPA19, OPB5 and OPB1, and 3 SSR markers Xpsmp2208, Xpsmp2223 and Xpsmp2220, were found to be highly discriminative. The PIC values ranged from 0.28 to 0.48 for the RAPD and from 0.24 to 0.60 for the SSR markers. Cluster analysis and principal component analysis of the combined dataset of RAPD and SSR markers indicated moderate genetic divergence among the elite pearl millet germplasm, besides unraveling the genetic relationships among the male sterile lines and the restorers.     

Determination of genetic relationships among elite thermosensitive genic male sterile lines (TGMS) of rice (Oryza sativa L.) employing morphological and simple sequence repeat (SSR) markers

A set of morphological traits and SSR markers were used to determine the genetic relationship among 12 elite thermosensitive genic male sterile (TGMS) lines developed at three different research institutions of India. Agro-morphological data recorded on 20 morphological traits revealed a wide base of genetic variation and a set of four morphological traits could distinguish most of the TGMS lines. Analysis with 30 SSR markers (20 EST-SSRs and 10 genomic SSRs) revealed 27 markers to be polymorphic, amplifying a total of 83 alleles. Each SSR marker amplified 2-6 alleles with an average of 2.76 alleles per marker and a PIC value varying from 0.54 to 0.96. Cluster analysis based on SSR and morphological data clearly differentiated the lines according to their source of origin. Correlation analysis between morphological and molecular data revealed a very poor association (r = 0.06), which could be attributed to selection pressure, genetic drift, sampling error and unknown relationship among related lines. The SSR markers discriminated the genotypes distinctly and quantified the genetic diversity precisely among the TGMS lines. Data on the yield per plant indicated that the genotypes grouping under a similar cluster showed same heterotic behaviour as compared to the genotypes from different clusters when crossed to similar pollinators.     

华南杂交稻亲本间产量配合力评价与分子标记鉴定

选用华南地区广泛应用的8个杂交稻不育系与10个恢复系,按NCII设计配制两套不完全双列杂交组合61个,通过在湛江的两年种植试验,结合62对与产量性状QTL连锁SSR标记,对18个亲本的产量配合力进行综合评价和标记位点鉴定。结果表明：供试亲本间一般配合力存在较大差异,其中不育系珍汕97A、万金A和恢复系直龙、广恢3550的一般配合力相对较高;两套组合共检测出36个与亲本产量配合力显著相关的标记位点,其中RM1、RM216、RM231和RM542为两套材料所共有;检测出的杂合标记位点中,23个为增效标记位点,13个为减效标记位点,其中杂合标记位点RM231、RM209可使F1的单株粒重增加13.50 %和16.43 %。该结果可为华南地区杂交稻育种的亲本改良及选配提供了理论指导。     

SSR分子标记在山茶属观赏资源遗传多样性研究中的应用

采用微卫星(SSR)分子标记的方法,利用20对SSR引物对33份山茶属资源(含种和品种)的DNA样品进行了PCR扩增,从中筛选出10对扩增效果较好的SSR引物,并对33份资源进行了遗传多样性分析。结果显示:10对引物在33个植物材料中共检测出123个等位基因,每个SSR位点的等位基因数为3~22个,SSR2引物检测到的等位基因数量最多,达22个,平均每对引物含有12.3个等位基因,平均多态性信息量为0.74,变化范围为0.06~0.96。利用遗传距离矩阵按UPGMA方法进行聚类,结果表明33份植物材料可分为9个分支,很好的区分了品种间的亲缘关系,可为杂交育种的组合选择提供理论基础。不同花型的山茶属植物在10个微卫星位点上共发现42个特异等位基因,可能与不同的花型性状有关。     

An SSR-based linkage map of Capsicum annuum

There are five cultivated species of pepper, of which Capsicum annuum is the most widely cultivated as a vegetable or spice and the main experimental material of most pepper breeding programs. However, the number of simple sequence-repeat (SSR) markers known for C. annuum is limited. To develop SSR markers for Capsicum species, we constructed four SSR-enriched libraries from the genomic DNA of C.␣annuum, sequenced 1873 clones, and isolated 626 unique SSR clones. A higher percentage of these SSR markers were taken from dinucleotide motif libraries than from trinucleotide motif libraries. Primer pairs for the 626 SSR clones were synthesized and tested for polymorphisms; 594 amplified products were detected with the expected size. However, only 153 products were polymorphic between the parents of our mapping population. Using 106 highly reproducible pairs from the primer pairs, we constructed a linkage map of C. annuum in an intraspecific doubled haploid population (n=117) that contains nine previously reported SSRs as well as AFLP, CAPS, and RAPD markers and the trait of fruit pungency. The map contains 374 markers, including 106 new SSR markers distributed across all 13 linkage groups, and covers 1042 cM. The polymorphism information content (PIC) of these new SSR markers was calculated using 14 lines of Capsicum species. The average number of alleles per locus was 2.9 and the average PIC value was 0.46, even within C. annuum. The SSR markers developed in this study will be useful for mapping and marker-assisted selection in pepper breeding, and the linkage map provides a reference genetic map for Capsicum species.     

Differentiation and classification of parental lines and favorable genic interactions affecting F1 fertility in distant crosses of rice (Oryza sativa L.)

 This study was intended to investigate the extent of genetic differentiation in parental lines of rice hybrids and to analyze the genetic basis underlying the fertility phenomenon in distant crosses. Two subsets of rice material (111 entries in total) were used, including 81 doubled-haploid (DH) lines and 30 Indica and Japonica rice varieties or lines (as a control). The DH lines was derived from a heterotic Indica/Japonica cross (Gui630/02428) by anther culture. The materials in the control represent a broad spectrum of the Asian cultivated rice gene pool including landraces, primitive cultivars, historically important cultivars, modern elite cultivars, super rice and parents of superior hybrids. In accordance with the NC II design, 57 out of the DH lines were test-crossed to two important wide compatibility lines: photoperiod-sensitive genetic male sterile (PGMS) line N422s and thermo-sensitive genetic male sterile (TGMS) line Peiai64s. The F₁s and their parents, 182 entries in total, were examined for the performance of seven traits in a replicated field trial. All the rice materials was surveyed for polymorphisms using 92 RFLP markers selected from two published molecular marker linkage maps. Genotypes of the F₁ hybrids at the molecular-marker loci were deduced from the parental genotypes. The analysis showed that there were two types of genetic differentiation in the two subsets of rice material; that is, qualitative differentiation in the control and quantitative differentiation in the DH lines. In addition, favorable genic interactions (both intra- or inter-locus) contributed to better increase the fertility in hybrids of distant crosses through incorporation of a wide-compatibility line as the female parent. Favorable genic interactions can be applied in hybrid rice breeding programs by selecting parents with an appropriate extent of genetic differentiation. Received: 5 June 1997 / Accepted: 10 September 1997     

利用分子标记预测杂交水稻产量及其构成因素

利用AFLP、RAPD、SSR技术分析了10个恢复系和5个不育系的931个基因座,利用15个亲本配制了50个杂交组合,在泸州和重庆2个环境下同时种植,考察了产量及其构成因素,从931个基因座中筛选出了与之相关的阳性座位、增效座位、减效座位、非环境型座位,并分析了它们与杂种产量及其构成因素间的关系。结果表明,利用所有座位计算的遗传差异与产量及其构成因素的相关性,绝大多数性状未达显著水平,不能直接用于预测产量及其构成因素。阳性座位在一定程度上可以提高相关系数,因性状不同而存在差异,在多数性状上预测产量及其构成因素还有一定难度;增效座位和减效座位可以大幅度提高相关系数,在不同的环境下也表现一致,可以用来预测产量及其构成因素;非环境型座位计算的相关系数也较高,但低于增效座位和减效座位,说明环境对产量及其构成因素有较大的影响。     

DNA methylation affected by male sterile cytoplasm in rice (Oryza sativa L.)

Male sterile cytoplasm plays an important role in hybrid rice, and cytoplasmic effects are sufficiently documented. However, no reports are available on DNA methylation affected by male sterile cytoplasm in hybrid rice. We used a methylation-sensitive amplified polymorphism technique to characterize DNA methylation in four male sterile cytoplasms that are widely commercialized in China. In total, 12 pairs of selective primers in combinations of EcoRI and MspI/HpaII amplified 350 bands among four male sterile (A) lines and the corresponding maintainer (B) lines. Sites b1 and b3 were fully methylated only in all the B lines, while b2 was fully methylated only in all the A lines. These results implied a relationship of DNA methylation at these sites specifically with male sterile cytoplasms, as well as male sterility, since the only difference between the A and B lines was the cytoplasm. The DNA methylation was markedly affected by male sterile cytoplasms. WA-type and Yinshui-type cytoplasms affected the methylation to a much greater degree than G-type and D-type cytoplasms, as indicated by the number and degree of methylated sites, ratio of methylated sites, number of fully methylated sites, ratio of fully methylated sites, and polymorphism between A and B lines for these cytoplasms. The genetic distance between the cytoplasm and nucleus for the WA-type is much greater than for G- and D-types because the former is between wild and cultivated species and the latter is within indica subspecies between African and Asian cultivars. This difference in genetic distance may be responsible for the variation in methylation which we observed.