一个水稻苗期白化致死突变体asl6的鉴定及其基因定位

经60Coγ诱变处理粳稻"嘉花1号"得到一个稳定遗传苗期白化致死突变体asl6(albino seedling lethality 6).与野生型(WT)相比,该突变体从发芽出苗起一直表现白化,四叶期逐渐死亡,叶合色素含量几乎没有且没有完整的叶绿体结构.通过qRT-PCR分析发现,与叶绿体发育、叶绿素合成及光合作用相关的基因表达量明显下调.对利用asl6突变体与"培矮64S"杂交获得的F2代分离群体进行遗传分析,发现该突变表型受单个隐性核基因控制.利用图位克隆技术将该asl6基因定位于第2号染色体的InDel分子标记ID31982与SSR分子标记MM5712之间约293 kb的区域内.目前,该范围内没有叶色相关基因的报道,可能为一新的调控水稻叶绿体发育的基因.     

一个新水稻低温敏感叶色突变体tcm11的鉴定及基因定位

对粳稻"嘉花1号"经~(60)Coγ诱变处理获得的稳定遗传低温敏感叶色突变体tcm11(thermo-sensitive chloroplast mutant 11)进行了表型鉴定与遗传分析.在20℃条件下,该突变体三叶期之前幼苗均表现为黄色,光合色素含量明显下降,叶绿体发育不完整,从第4叶开始逐渐转为浅黄绿色直至最后死亡.而在32℃条件下,其表型与野生型相比没有明显差异,具有低温敏感属性.通过对培矮64S与tcm11杂交的F_2代分离群体进行遗传分析,发现该低温敏感突变体性状是受单个隐性核基因(tcm11)控制,利用图位克隆技术对tcm11进行定位,将其定位在第11号染色体的InDel分子标记ID13252与SSR分子标记MM1361之间一个约1 566 kb的区域内.这也为后续的研究奠定了基础.     

水稻高温白化复绿突变体tcd52的遗传分析和分子定位

粳稻品种“嘉花1号”经甲基磺酸乙酯（EMS）化学诱变处理,获得一个稳定遗传水稻幼苗高温白化复绿突变体tcd52.该突变体在高温（>24℃）条件下,二叶期叶色呈白色失绿,三叶期开始复绿,四叶期后与野生型没有明显差异;而在低温（20℃）条件下,tcd52突变体苗期叶色与野生型一致呈绿色,无白化现象.利用该突变体tcd52与“培矮64S”杂交构建F₂遗传群体,发现苗期的高温白化复绿叶色性状受到一对隐性核基因控制,并将该突变基因（tcd52）定位在水稻第5染色体上的ID05M16025和ID05M16113分子标记之间的127 kb区间内,经测序推定突变基因是编码PPR蛋白的基因LOC＿Os05g49920.结果表明：tcd52是一个受高温响应且影响水稻早期叶绿体发育的关键基因.今后将进一步对tcd52基因进行研究,以加深了解温度对水稻叶绿体分子发育机理.     

水稻低温敏感叶色突变体tcd32鉴定与遗传分析

对粳稻"嘉花1号"经60Coγ诱变处理获得的稳定遗传低温敏感叶色突变体tcd32进行了表型鉴定与遗传分析.在20℃条件下,该突变体表现为白色,光合色素含量明显下降,叶绿体发育不完整,植株最终枯萎死亡;在25℃条件下,三叶期之前幼苗表现为黄色,从第四叶开始逐渐转为黄绿色,光合色素含量也明显下降;而在30℃条件下,其表型与野生型(WT)相比没有明显差异.通过对培矮64S与tcd32杂交的F2代分离群体进行遗传分析,结果表明:该低温敏感叶色突变体性状是受一对隐性核基因(tcd32)控制,利用图位克隆技术将tcd32基因定位在水稻第3染色体上顶端537 kb区域内,发现TCD32是一个新的水稻早期叶绿体发育相关基因.     

水稻白化突变体alb21生理特性和基因定位

高等植物叶绿体的正常发育需要叶绿体基因和核基因相互协调,这些基因的突变将导致叶绿体发育的缺陷．通过同位素诱变,获得了1例水稻的白化突变体alb21,其在幼苗时期就表现出白化性状,生长1个月左右逐渐死亡．遗传学分析表明,该突变属于单基因隐性突变;电镜观察表明,突变体细胞内完全丧失了叶绿体结构,只有一些空泡状的结构．突变体中既没有检测出叶绿素a或b,也没有检测出叶绿素合成的前体——原脱植基叶绿素a,说明此突变体的叶绿素合成途径受阻．因此,推测Alb21基因的突变,导致叶绿体发育受阻,叶绿素a或b以及叶绿素合成的前体——原脱植基叶绿素a不能合成．利用本实验室开发的水稻InDel分子标记,将该突变基因定位在第3条染色体上分子标记R3M51—2与R3M52—5之间约1520kb范围内．这些结果为该基因的克隆及叶绿体发育过程中的功能研究奠定了基础．     

一个新的水稻幼苗黄叶突变体的遗传分析及其基因的初步定位

在60Coγ射线辐照的水稻突变体库中,发现了一个以粳稻品种日本晴为遗传背景的幼苗叶色黄化突变体syl11(seedling yellow leaf 11).与野生型相比,突变体幼苗第二和第三叶表现黄色,在其完全展开之前叶片自其顶端开始转绿,长到四叶期其叶色恢复正常;并且该突变体syl11幼苗黄色叶片光合色素含量明显下降.遗传分析表明,该突变体的遗传性状由1对隐性核基因控制.本研究以培矮64S/syl11的F2代突变型植株作为定位群体,应用微卫星(SSR)分子标记以及新发展的InDel分子标记,将基因syl11定位在水稻第11号染色体长臂上的RM26652和处于着丝粒附近的ID11974分子标记之间,其遗传距离分别为0.5 cM和0.7 cM.     

水稻黄绿叶基因YGLOSH的定位克隆

本文利用~(60)Co γ射线诱变光温敏感型核不育系广占63S(GZ63S),在后代中获得一个稳定遗传的黄绿化叶色突变体黄广占63S,突变体从苗期至成熟期均显示叶片黄化特征.遗传分析表明该性状受一对隐性核基因控制,命名为yglosh.利用BSA方法分析突变体黄广占63S与正常绿叶对照蜀恢881构建的F2群体,将该黄化基因定位于水稻2号染色体分子标记RM279和Pm6之间,物理距离约68kb.定位区间的cDNA测序分析发现,突变体中LOC_Os02g05890基因发生单碱基突变,形成终止子提前终止该基因的翻译.转基因互补实验确定LOC_Os02g05890为目标基因,可能参与叶绿体发育或者叶绿素生物合成途径.     

一个新的水稻矮秆突变体sd-sl的遗传与基因定位研究

新的矮秆基因的发掘、研究和利用对水稻育种和植物生长发育机制研究有重要的作用.用60Coγ射线辐照粳稻9522,获得一个能稳定遗传的突变体.该突变体表型为株高较野生型矮,叶片短而微卷.将该突变体与籼稻广陆矮杂交,F2代呈3∶1分离,说明该突变体受隐性单基因控制.通过InDel分子标记对F2代分离群体进行遗传定位,将该基因定位于第6染色体InDel标记OS604附近.随后又发展了多对有多态性的InDel分子标记,将该基因座位精细定位在InDel标记XL6-6和XL6-1之间,AP003490和AP005619上,两个引物之间的物理距离为118 kb.本研究为该克隆基因及其作用机理的探究奠定了基础.     

拟南芥叶色突变体ems3的基因定位

ems3是经甲基磺酸乙酯(EMS)诱变筛选得到的一拟南芥叶色突变体.通过背景纯化与遗传分析,发现ems3突变体是单基因隐性控制.利用图位克隆的方法对叶色基因EMS3进行了定位,结果表明:EMS3位于第5条染色体分子标记MHF15和MHF152之间55kb的区间内.生物信息预测,该区间包括有16个基因.这些结果为该基因的克隆及叶绿体发育过程中的功能研究奠定了基础.     

水稻矮秆基因d-ss的遗传分析与克隆

通过γ射线诱变,从粳稻品种9522的M2代中筛选出一株矮秆水稻(Oryza sativa L.)突变体,定名d-ss.d-ss突变体表现为叶色深绿、短宽的叶片、以及小而圆的籽粒.以d-ss突变体与籼稻品种龙特普杂交的F2代群体为基因定位群体,利用InDel分子标记将d-ss突变位点定位在5号染色体上的InDel标记ZZ5-6和ZZ1343之间,物理距离为412kb.最终通过图位克隆的方法获得了此基因,测序结果表明此基因在编码区发生了两处缺失突变.     

利用分子标记辅助选育香软型保持系“SH101B”、不育系“SH101A”及配组分析

为了给三系杂交稻新品种选育提供优良亲本,以"嘉花1号"水稻为转育亲本,与香型软米水稻杂交,再与"嘉花1号"水稻多代回交及自交,结合分子标记辅助选育香软型水稻新品系"SH101B"."嘉花1号"与"SH101B"水稻主要农艺性状和产量性状差异都不显著(p0.05)."SH101B"稻米直链淀粉含量(质量分数)为10.0%,明显低于"嘉花1号"水稻(15.2%).又将"SH101B"与三系不育系水稻杂交和回交,转育获得香软型三系不育系"SH101A",再与籼型恢复系"T201"进行配组,并对后代农艺性状、产量和稻米品质进行分析.结果显示,"SH101A×T201"组合稻米直链淀粉含量为12.8%,每亩产量达到848.1 kg(12 726.4 kg·hm~(-2)).     

Genetic analysis and gene mapping of a dominant presenescing leaf gene <Emphasis Type="Italic">PSL3</Emphasis> in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Leaf senescence as an active process is essential for plant survival and reproduction. However, premature senility is harmful to agricultural production. In this study, a rice mutant, named as psl3 (presescing leaf 3) isolated from EMS-treated Jinhui 10, displays obvious premature senility features both in morphological and physiological level. Genetic analysis showed that mutant trait was controlled by a single dominant gene (PSL3), which was located on rice chromosome 7 between SSR marker c7sr1 and InDel marker ID10 with an interval of 53.5 kb. The result may be useful for the isolation of the PSL3 gene.     

Genetic analysis and molecular mapping of a presenescing leaf gene psll in rice （Oryza sativa L.）

A rice psl1 （presenescing leaf） mutant was obtained from a japonica variety Zhonghua 11 via radiation of ^60Co-γ in M2 generation. Every leaf of the mutant began to wither after it reached the biggest length, while the leaves of the wild variety could keep green for 25--35 d. In this study, genetic analysis and gene mapping were carried out for the mutant identified. The SSR marker analysis showed that the mutant was controlled by a single recessive gene （psl1） located on chromosome 2. Fine mapping of the psl1 locus was conducted with 34 new STS markers developed around psl1 anchored region based on the sequence diversity between Nipponbare and 93-11. The psl1 was further mapped between two STS markers, STS2-19 and STS2-26, with genetic distances of 0.43 and 0.11 cM, respectively, while cosegregated with STS2-25. A BAC contig was found to span the psl1 locus, the region being delimited to 48 kb. This result was very useful for cloning of the psl1 gene.     

Genetic analysis and fine mapping of a lax mutant in rice

We have analyzed a lax mutant that exhibits altered panicle architecture in rice.The primary and secondary rachis-branches are normally initiated and each branch ends in a terminal spikelet,but all the lateral spikelets are absent and the terminal spikelet displays variegated structures in the mutant.An F2 population from the cross between the lax mutant and a japonica variety,W11,was constructed and analyzed.Using microsatellite and CAPS markers,the lax locus was mapped on the long arm of chromosome 1,co-segregated with a CAPS marker,LZ1,within an interval of 0.28 cM between a CAPS marker,HB2,and a microsatellite marker,MRG4389.RT-PCR analysis revealed that the expressions of the rice B-function MADS-box genes OsMADS2,OsMADS4,OsMADS16 and OsMADS3 were significantly reduced,whereas the expression of the rice A-function gene RAPIA was not altered.     

Genetic analysis and fine mapping of an enclosed panicle mutant esp2 in rice （Oryza sativa L.）

The phenomenon of panicle enclosure in rice is mainly caused by the shortening of uppermost internode.Elucidating the molecular mechanism of panicle enclosure will be helpful for solving the problem of panicle enclosure in male sterile lines and creating new germplasms in rice.We acquired a monogenic recessive enclosed panicle mutant,named as esp2 (enclosed shorter panicle 2),from the tissue culture progeny of indica rice cultivar Minghui-86.In the mutant,panicles were entirely enclosed by flag leaf sheaths and the uppermost internode was almost completely degenerated,but the other internodes did not have obvious changes in length.Genetic analysis indicated that the mutant phenotype was controlled by a recessive gene,which could be steadily inherited and was not affected by genetic background.Apparently,ESP2 is a key gene for the development of uppermost internode in rice.Using an F 2 population of a cross between esp2 and a japonica rice cultivar Xiushui-13 as well as SSR and InDel markers,we fine mapped ESP2 to a 14-kb region on the end of the short arm of chromosome 1.According to the rice genome sequence annotation,only one intact gene exists in this region,namely,a putative phosphatidylserine synthase gene.Sequencing analysis on the mutant and the wild type indicated that this gene was inserted by a 5287-bp retrotransposon sequence.Hence,we took this gene as a candidate of ESP2.The results of this study will facilitate the cloning and functional analysis of ESP2 gene.     

水稻苹果酸脱氢酶基因的克隆及其在E.coli中的表达

以玉米苹果酸脱氢酶的基因为探针，从水稻幼苗cDNA文库筛选得到一条水稻新基因，它与玉米ZHO2序列的一致性高达87％，被命名为RcMDH(rice cytoplasmic malic dehydrogenase gene)，分析发现RcMDH具有完整的读码框，编码332个氨基酸，预计分子质量为36．5ku,导入大肠杆菌中表达，经检测证实该产物具苹果酸脱氢酶酶活性。     

RAPD tagging of a salt tolerant gene in rice

A salt tolerant rice mutant was obtained through tissue culture. The inheritance of salt tolerance in F2 population of mutant crossed with its original variety under salt stress was studied. According to the standard, the ratio of salt tolerant: sensitive was about 3:1 suggesting that there was a major gene related to salt tolerance in the mutant. By RAPD analysis with 220 10-mer primers, the gene was tagged by a 1.0 kb DNA fragment named OPS12-10 which was located on chromosome 7. The genetic distance between the major salt tolerant gene was 16.4 cm.     

Mapping and characterization of a tiller-spreading mutant<Emphasis Type="Italic">lazy-2</Emphasis> in rice

Tiller angle of rice is an important agronomic trait that contributes to breed new varieties with ideal architecture. In this study, we report mapping and characterization of a rice mutant defective in tiller angle. At the seedling stage, the newly developed tillers of the mutant plants grow with a large angle that leads to a “lazy“ phenotype at the mature stage. Genetic analysis indicates that this tillerspreading phenotype is controlled by one recessive gene that is allelic to a reported mutant la. Therefore, the mutant was named la-2 and la renamed la-1. To map and clone LA, we constructed a large mapping population. Genetic linkage analysis showed that the LA gene is located between 2 SSR markers RM202 and RM229. By using the 6 newly-developed molecular markers, the LA gene was placed within a 0.4 cM interval on chromosome 11, allowing us to clone LA and study the mechanism that controls rice tiller angle at the molecular level.     

Genetic analysis and gene mapping of leafy head （lhd）, a mutant blocking the differen-tiation of rachis branches in rice （Oryza sativa L.）

Flowers, fruits and seeds are products of plant re- productive development and provide the important sources of foods for humans. Therefore, the moleculargenetic mechanisms of floral development have been ahotspot of research of plant developmental biology[1]. Rice is one of the most important staple food crops. Theoutcome of its reproductive development would determine the yield and quality of grains. Rice is also a model plantof cereals. Hence, the study of rice reproductivedevelopment, esp…