水稻光温敏雄性不育基因连锁标记的分离与鉴定

两系法杂交稻是一种利用水稻杂种优势的重要途径，主要依据水稻光温敏雄性不育系在不同条件下的育性转换用于配制杂交种．以培矮64S(PTGMS)与明恢63杂种自交的F2代为供试材料，采用随机放大多态性DNA(RAPD)技术分离与培矮64S的PTGMS基因连锁的分子标记．在F2代2个分别代表可育和不育的群体以及亲本株系中，采用100个RAPD引物扩放基因组DNA筛选多态性DNA片段．RAPD引物S8产生的DNA片段中，除重复顺序外，有一个长度为0．85kb片段为单拷贝．分子杂交表明，这一单拷贝标记与培矮64S的PTGMS基因连锁．     

Mapping of the rice （Oryza sativa L.） thermo-sensitive genic male sterile gene tms5 with EST and SSR markers

VELOPING“TWO-LINE”HYBRID RICE.THE POLLEN FERTILITY OF TGMS IS REGULATED BY THE TEMPERATURE OF ENVIRONMENT.THE POLLENS OF TGMS LINES ARE STERILE WHEN THE ENVI-RONMENT TEMPERATURE IS ABOVE A CRITICAL POINT,BUT FERTILE BELOW THIS POINT.SO FAR,A NUMBER OF T…     

Construction and characterization of a bacterial artificial chromosome library of rice 5460F

Thermo-sensitive genie male sterile (TGMS) rice has a number of desirable characteristics for hybrid rice production. Many studies have demonstrated that the sterility of TGMS rice is controlled by a single recessive gene. It has been mapped for the first time on chromosome 8 and namedtms 1. Several AFLP markers which tightly linked to thetms 1 gene have been identified recently. In order to develop a detailed physical map of thetms1 gene-encompassing region and finally clone thetms1 gene, a bacterial artificial chromosome (BAC) library of rice 5460F (the fertile mutant line of TGMS rice 5460S) using a modified vector pECBAC1 has been constructed. The constructed 5460F BAC library consists of 16 896 clones with an average insert size of 119 kb, which represents about 4.7 times rice haploid genome equivalents. Neither chloroplast nor mitochondrial DNA was detected from the library. The library was screened with three single copy sequence amplified fragment length polymorphism (AFLP) markers which tightly linked totms1 gene as probes and eight positive clones were identified.     

Rapid genome evolution in Pms 1 region of rice revealed by comparative sequence analysis

Pms1, a locus for photoperiod sensitive genic male sterility in rice, was identified and mapped to chromosome 7 in previous studies. Here we report an effort to identify the candidate genes for Pms1 by comparative sequencing of BAC clones from two cultivars Minghui 63 and Nongken 58, the parents for the initial mapping population. Annotation and comparison of the sequences of the two clones resulted in a total of five potential candidates which should be functionally tested. We also conducted com-parative analysis of sequences of these two cultivars with two other cultivars, Nipponbare and 93-11, for which sequence data were available in public databases. The analysis revealed large differences in sequence composition among the four genotypes in the Pms1 region primarily due to retroelement activity leading to rapid recent growth and divergence of the genomes. High levels of polymorphism in the forms of indels and SNPs were found both in intra- and inter-subspecific comparisons. Dating analysis using LTRs of the retroelements in this region showed that the substitution rate of LTRs was much higher than reported in the literature. The results provided strong evidence for rapid genomic evolution of this region as a consequence of natural and artificial selection.     

水稻光温敏核不育系不育性表达的同步性研究

以农垦５８Ｓ、７００１Ｓ、５０４７Ｓ、Ｗ６１５４Ｓ、ＫＳ－９、培矮６４Ｓ、安农－１Ｓ和５４６０Ｓ８个不同类型的水稻光温敏核不育为材料，在广州盛夏７月－８月份对各不育系的不育性表达进行了系统的比较研究，并对生转换的同步性进行了观察。结果表明，不育性表达的同步性以安农－１Ｓ和７００１Ｓ较好，各体不育怀整齐，其他６个不育系的不育性表达的同步性较差，株间、株内穗间颖花间的花粉育性差异显著或极显著。连续单株     

太谷核不育小麦的研究与利用

本文综述太谷核不育小麦的遗传理论与育种实践的进展概况。经研究鉴定太谷小麦的不育性是受控于一个显性雄性核不育基因,并开展了基因作用及表达研究;太谷小麦是轮回育种的理想工具,已取得了明显效果。     

Gene expression profiling under different photoperiod/temperature conditions in a photoperiod-/thermo-sensitive genic male sterile line of rice （Oryza sativa L.）

A photoperiod-sensitive genic male-sterile (PSGMS) line was found in 1973 as a spontaneous mutant of the rice variety Nongken 58 (Oryza sativa ssp. japonica) in Hubei Province, China[1]. The fertility of this line was determined by day length and temperat…     

白菜型油菜核育性相关基因片段的克隆与序列分析

通过ＲＡＰＤ标记,所获得的与育性基因紧密连锁的基因片段进行克隆与序列分析,结果表明,该育性基因片段为与油菜小孢子发育早期ＢＰ４调控基因５８％同源,并含有一ＭＡＰＤＳ盒高度同源的保邓列。Ｓｏｕｔｈｅｒｎ杂交结果表明该基因为单拷贝基因。     

谷胱甘肽S转移酶P1基因多态性与慢性阻塞性肺疾病关系的研究

目的:了解湖北地区健康汉族人群和慢性阻塞性肺疾病(COPD)患者谷胱甘肽S转移酶P1(GSTP1)基因多态性,探讨汉族人GSTP1基因多态性与COPD的关系。方法:用聚合酶链式反应—限制性片段长度多态分析(PCR-RFLP)检测68例健康汉族人和50例COPD患者GSTP1基因型。结果:健康人群和COPD人群中有3种基因型,但以Ile105纯合子基因型为主,两组人群的基因型和基因频率比较均无显著性差异(P>0.05)。结论:GSTP1基因多态性可能与COPD的发生无相关性。     

Fine mapping of the rice bacterial blight resistance gene Xa-4 and its co-segregation marker

An F₂ population developed from theXa-4 near isogenic lines, IR24 and IRBB4, was used for fine mapping of the rice bacterial blight resistance gene,Xa-4. Some restriction fragment length polymorphism (RFLP) markers on the high-density map constructed by Harushima et al. and the amplified DNA fragments homologous to the conserved domains of plant disease resistance (R) genes were used to construct the genetic linkage map around the geneXa-4 by scoring susceptible individuals in the population.Xa-4 was mapped between the RFLP marker G181 and the polymerase chain reaction (PCR) marker M55. The R gene homologous fragment marker RS13 was found co-segregating withXa-4 by analyzing all the plants in the population. This result opened an approach to map-based cloning of this gene, and marker RS13 can be applied to molecular marker-assisted selection ofXa-4 in rice breeding programs.     

包台型水稻不育系花药发育过程中的Ca2+分布

用焦锑酸钾沉淀法研究了包台型不育水稻及其保持系花药发育早期细胞中的Ca²⁺分布变化.从花粉母细胞时期开始,不育系与保持系花药中的Ca²⁺分布出现差异,不育系药壁外层和药室内壁及药隔中均有Ca²⁺出现,而保持系花药中基本没有Ca²⁺分布,在减数分裂时期表现更加明显,不育系药壁和药隔中的Ca²⁺相对于花粉母细胞时期明显地有所增加,特别是绒毡层细胞和药隔中木质部细胞的次生加厚壁上Ca²⁺增加尤其明显,而保持系花药中的Ca²⁺较少.笔者从花药发育的更早期去寻找水稻雄性不育的机理.     

Primary analysis of QTG contribution to heterosis in upland cotton

In this paper, we analyzed the contribution of pure DNA factors to heterosis using quantitative trait genes (QTG) in two randomly selected strains from a recombinant inbred line of Gossypium hirsutum. According to a set of QTL mapping results, combined with analysis of DNA recombinant fragment sources in the two strains and QTL association analysis with their field traits, we hypothesize a view of “dominance + overdominance + epistasis”. That is, additive and additive epistasis may be the genetic basis of heterosis, and dominance, overdominance and epistasis may be the modes of heterosis action. Based on the heterosis results of this study, we also suggest a molecular mechanism for heterosis, and explain, in detail, with two randomly selected strains as examples. The male and female parent-derived additive epistatic QTLs of upper half mean length in LG01 and LG03 produced a trait variance of 2.99–3.52 compared with the female parent-derived loci. The trait of bolls per plant was controlled by two pairs of additive epistatic QTLs in LG02 and LG07, which were derived from both female and male parents. The QTLs were reciprocally interacted and produced a trait variance of 0.86. An initial concept of “super-hybrid cotton” was raised according to the nature of additive effect, that is genetic stability.     

Characterization and mapping of a lesion mimic mutant in rice （Oryza sativa L.）

A rice initiation-type lesion mimic mutant (lmi) was identified, which was isolated from an indica rice Zhongxian 3037 through γ radiation mutagenesis. Trypan blue staining and sterile culture revealed that the mutant spontaneously developed lesions on the leaves in a developmentally regulated and light-dependent manner. Genetic analysis indicated that the lesion mimic trait was controlled by a single resessive locus. Using public molecular markers and an F2 population derived from lmi and 93-11, we mapped the lmi locus to the short arm of chromosome 8, nearby the centromere, between two SSR markers RM547 and RM331. The genetic distance was 1.2 and 3.2 cM, respectively. Then according to the public rice genomic sequence between the two SSR markers, lmi was further finely tagged by three CAPS markers: C4135-8, C4135-9 and C4135-10. And lmi locus was a co-segregated with marker C4135-10, providing a starting point for lmi gene cloning.     

抗广谱除草剂pursuit水稻的抗性遗传分析和利用

以4个恢复系测64、明恢63、特青和02428为母本,分别与抗除草剂pursuit(普杀特)水稻品种PT2杂交,遗传分析表明,PT2的抗性遗传符合一对显性核基因的模式.通过杂交一代和回交二代已将抗性基因转移到恢复系,初步培育出4个抗性恢复系测64-P、明恢63-P、特青-P和02428-P;它们与不育系杂交,选育到4个杂交稻组合.考种结果发现,抗除草剂组合基本上能保持原组合的产量水平,苗期喷药可完全淘汰假杂种.利用这项技术可望解决三系、两系或化学杀雄杂交稻制种过程中出现的纯度问题,且有利于新杂交稻组合的选育.对该项技术在生产上的应用前景和存在问题进行探讨.     

Biochemical characterization of the protein encoded by rice osRACD gene

A recombinant plasmid pET-racd was first constructed by cloning osRACD,the development-regulating gene that controls photoperiod fertility transformation in the photoperiod sensitive genic male sterile rice Nongken 58S,into a prokaryote expression vector pET28a(+).It was then transformed into E.Coli BL-21.Cutting with thrombin of the fusion protein extracted from transformants and PAGE separation yielded pure osRACD protein,which was further concentrated using ultrafiltration and renatured using glutathione oxidation/reduction refolding system for later functional study.As demonstrated by in vitro functional assay,the osRACD protein expressed in E.Coli B-21 shows remarkable activity in binding GTP specifically and hydrolyzing it.     

野栽远缘杂交来源的反向温敏核雄性不育系不育性遗传研究

利用3个来源于野生稻与栽培稻杂交后代的反向温敏不育系R6S、N13S、Tb7S为材料,开展了反向温敏不育系不育性遗传研究,结果表明反向温敏不育系N13S的育性是细胞核内1对隐性基因控制的;Tb7S的育性是受隐性核基因控制的,F2代的育性分离比为9∶7,不育性状表现为2对基因的独立遗传;R6S的F2代育性分离比为37∶27,受细胞核内的3个隐性基因位点分别位于不同染色体上而通过互补作用的独立遗传.为进一步分离、定位及克隆有关这些反向温敏不育基因及发展分子标记辅助选择选育反向温敏不育系奠定了良好的遗传学基础.     

Mapping of a new gene for brown planthopper resistance in cultivated rice introgressed fromOryza eichingeri

Wild rice species is an important source of useful genes for cultivated rice improvement. Some accessions of Oryza eichingeri (2n = 24, CC) from Africa confer strong resistance to brown planthopper (BPH), whitebacked planthopper (WBPH) and bacterial blight (BB). In the present study, restriction fragments length polymorphism (RFLP) and simple sequence repeats (SSR) analysis were performed on disomic backcross plants between Oryza sativa (2n = 24, AA) and O. eichingeri in order to identify the presence of O. eichingeri segments and further to localize BPH-resistant gene. In the introgression lines, 1—6 O. eichingeri segments were detected on rice chromosomes 1, 2, 6, or/and 10. The dominant BPH resistant gene, tentatively named Bph13(t), was mapped to chromosome 2, being 6.1 and 5.5 cM away from two microsatellite markers RM240 and RM250, respectively. The transfer and localization of this gene from O. eichingeri will contribute to the improvement of BPH resistance in cultivated rice.     

Fertility analysis of the Arabidopsis transformed with antisense rice osRACD gene

The full length osRACD cDNA sequence was subcloned into the pBI121 plasmid in the antisense orientation under the control of the CaMV35S promoter to construct the expression vector pBID, and the constructs were introduced into Arabidopsis plants by using the vacuum infiltration method. The siliques of the transformants stopped growing after anthesis, and they turned yellow or died later; and the siliques from the control plants transformed by the pBI continued growing after anthesis and matured normally. In vitro pollen germination demonstrated that the growth and elongation process of the pollens of the transgenic plants was inhibited, the pollen tubes were shorter and slightly fatter than the tubes of the control plants, which grew normally with long cyclindrical tubes. The above results suggest the function of osRACD gene involved in regulation of the growth and elongation process of pollen tube, its encoding protein may be one of the important factors in regulation of fertility transition of the photoperiod-sensitive genic male-sterile rice Nongken 58S.