Identification, genetic characterization, GA response and molecular mapping of Sdt97: a dominant mutant gene conferring semi-dwarfism in rice (Oryza sativa L.)

Semi-dwarfism is an important agronomic trait in rice breeding programmes. sd-1, termed the 'Green Revolution gene', confers semi-dwarf stature, increases harvest index, improves lodging resistance, and is associated with increased responsiveness to nitrogen fertilizer. It has contributed substantially to the significant increase in rice production. In this paper, a novel semi-dwarf mutant in rice is reported. Genetic analysis revealed that only a single dominant gene locus non-allelic to sd-1, temporarily designated Sdt97, is involved in the control of semi-dwarfism of the mutant. The semi-dwarfism of the mutant could be partly restored to the tall wild-type by application of exogenous GA3, suggesting that the mutant gene Sdt97 may be involved in the gibberellin (GA) synthesis pathway and not the GA response pathway in rice. A residual heterozygous line (RHL) population derived from a recombinant inbred line (RIL) was developed. Simple sequence repeat (SSR) and bulked segregation analysis (BSA) combined with recessive class analysis (RCA) techniques were used to map Sdt97 to the long arm of chromosome 6 at the interval between two STS markers, N6 and TX5, with a genetic distance of 0.2 cM and 0.8 cM, respectively. A contig map was constructed based on the reference sequence aligned by the Sdt97 linked markers. The physical map of the Sdt97 locus was defined to a 118 kb interval, and 19 candidate genes were detected in the target region. This is the first time that a dominant semi-dwarf gene has been reported in rice. Cloning and functional analysis of gene Sdt97 will help us to learn more about molecular mechanism of rice semi-dwarfism.     

一个水稻矮秆突变体的遗传分析及基因定位

水稻（Oryza sativa）是我国重要的粮食作物之一。水稻矮秆材料的引入掀起了第1次＂绿色革命＂。但近年来,在水稻育种中矮生基因遗传单一的问题越来越突出,已经严重影响到水稻产量的持续提高。利用60Co-γ射线辐照籼稻亲本材料M804获得了一个性状能够稳定遗传的矮秆突变体MU101。对该矮秆突变体和台粳16号杂交获得的F2代的遗传分析表明,该矮秆性状受1对隐性单基因控制,并暂命名为ds1。利用已有的SSR分子标记将DS1基因定位在水稻第5号染色体上,通过扩大群体和开发新的Indel标记,进一步将DS1基因定位在2个Indel标记之间,两者间的物理距离大约为384kb。该研究为DS1基因的克隆及其在生产中的应用奠定了基础。     

Genetic Analysis and Molecular Mapping of a Rolling Leaf Mutation Gene in Rice

A rice mutant with rolling leaf, namely γ-rl, was obtained from M2 progenies of a native indica rice stable strain Qinghuazhan （QHZ） from mutagenesis of dry seeds by γ-rays. Genetic analysis using the F2 population from a cross between this mutant and QHZ indicated the mutation was controlled by a single recessive gene. In order to map the locus for this mutation, another F2 population with 601 rolling leaf plants was constructed from a cross between y-rl and a japonica cultivar 02428. After primary mapping with SSR （simple sequence repeats） markers, the mutated locus was located at the short arm of chromosome 3, flanked by RM6829 and RM3126. A number of SSR, InDel （insertion/deletion） and SNP （single nucleotide polymorphism） markers within this region were further developed for fine mapping. Finally, two markers, SNP121679 and InDe1422395, were identified to be flanked to this locus with genetic distances of 0.08 cM and 0.17 cM respectively, and two SNP markers, SNP75346 and SNPl10263, were found to be co-segregated with this locus. These results suggested that this locus was distinguished from all loci for the rolling leaf mutation in rice reported so far, and thus renamed rl10（t）. By searching the rice genome database with closely linked markers using BLAST programs, an e-physical map covering rl10（t） locus spanning about a 50 kb region was constructed. Expression analysis of the genes predicted in this region showed that a gene encoding putative flavin-containing monooxygenase （FMO） was silenced in γ-rl, thus this is the most likely candidate responsible for the rolling leaf mutation.     

Genetic analysis and gene mapping of a rice few-tillering mutant in early backcross populations (Oryza sativa L.)

A rice mutant, G069, characteristic of few tiller numbers, was found in anther culture progeny from the F1 hybrid between an indica-japonica cross, Gui630×02428. The mutant has another two major features: delayed tillering development and yellowing apex and margin on the mature leaves. As a donor parent, G069 was further backcrossed with the recurrent parent, 02428, for two turns to develop a BC2F2 population. Genetic analysis in the BC2F2 population showed that the traits of few-tillering and yellowing apex and margin on the mature leaves were controlled by one recessive gene. A pool of equally mixed genomic DNA, from few-tillering individual plants in BC2F2, was constructed to screen polymorphism with simple sequence repeat (SSR) markers in comparison with the 02428 genome. One SSR marker and three restriction fragment length polymorphism (RFLP) markers were found possibly linked with the recessive gene. By using these markers, the gene of few-tillering was mapped on chromosome 2 between RFLP marker C     

一个新水稻温敏感叶色突变体的遗传分析及其基因分子定位

在粳稻品种嘉花1号(Oryza sativa L. ssp. japonica ‘Jiahua No.1’)种子经⁶⁰Co γ射线辐照处理的后代中, 发现了1个低温敏感叶色突变体mr21。在较低温度(<25.0°C)条件下, 该突变体幼苗叶色呈黄色; 随着温度逐渐升高, 叶色由黄转绿,其临界温度约为27.5°C; 在低温条件下, 突变体幼苗总叶绿素含量以及叶绿素a、b的含量均较野生型嘉花1号明显下降, 表明该突变体的叶色性状具有明显的温敏感性。遗传分析表明, 该突变体叶色性状受1对隐性核基因控制, 暂将该突变基因命名为thermo-sensitive leaf-color 1(tsl-1)。以该突变体与籼稻9311(Oryza sativa L. ssp. indica ‘9311’)杂交的F₂代分离群体作为定位群体, 利用SSR分子标记将tsl-1基因初步定位在水稻(Oryza sativa)第1号染色体短臂上的MM1799与RM8132分子标记之间, 其遗传距离分别为2.4 cM和3.0 cM; 然后, 进一步利用扩大F₂代群体及新发展的分子标记将tsl-1基因定位在分子标记InDel2与InDel4之间的198 kb内。研究结果为今后对该基因的克隆和功能分析奠定了基础。     

一个水稻内颖退化突变体的形态特征及基因的精确定位

水稻产量和品质受花器官发育的直接影响,因此对水稻颖花发育机理的研究将有助于水稻产量提高和品质的改良。文章利用60Coγ射线辐照亲本8PW33(籼稻背景)获得一个性状能稳定遗传的内颖退化突变体(编号:MU102),并对其农艺性状和花器官进行了观察和分析。结果显示,相对于野生型,该突变体的株高、每穗总粒数及剑叶宽均显著增加,而结实率则显著降低,差异均达显著水平。解剖镜下观察表明,该突变体内颖退化,外颖弯曲呈现镰刀状,其余器官与野生型表型基本一致。扫描电镜观察显示,突变体与野生型叶片维管束的结构组成以及外颖表皮细胞组成、排列均正常,没有明显差异;与野生型相比,突变体内颖表皮细胞排列较为紧密,推测可能是内颖收缩退化导致的。遗传分析显示该突变性状是由隐性单基因控制,并命名为pd2。利用实验室现有的SSR分子标记将PD2基因定位于水稻第9号染色体上,通过进一步扩大群体和开发新的Indel标记,将PD2基因定位在2个Indel标记之间,两者间的物理距离大约是82 kb。在该物理区间内有一个已经克隆的内颖发育基因REP1,经过测序和比对分析,推测REP1与PD2为等位基因。     

一个新水稻温敏感叶色突变体的遗传分析及其基因分子定位

在粳稻品种嘉花1号（Oryza sativa L.ssp.japonica＇ Jiahua No.1＇）种子经60Coγ射线辐照处理的后代中,发现了1个低温敏感叶色突变体mr21。在较低温度（〈25.0°C）条件下,该突变体幼苗叶色呈黄色;随着温度逐渐升高,叶色由黄转绿,其临界温度约为27.5°C;在低温条件下,突变体幼苗总叶绿素含量以及叶绿素a、b的含量均较野生型嘉花1号明显下降,表明该突变体的叶色性状具有明显的温敏感性。遗传分析表明,该突变体叶色性状受1对隐性核基因控制,暂将该突变基因命名为thermo-sensitive leaf-color1（tsl-1）。以该突变体与籼稻9311（Oryza sativa L.ssp.indica＇ 9311＇）杂交的F2代分离群体作为定位群体,利用SSR分子标记将tsl-1基因初步定位在水稻（Oryza sativa）第1号染色体短臂上的MM1799与RM8132分子标记之间,其遗传距离分别为2.4cM和3.0cM;然后,进一步利用扩大F2代群体及新发展的分子标记将tsl-1基因定位在分子标记InDel2与InDel4之间的198kb内。研究结果为今后对该基因的克隆和功能分析奠定了基础。     

一个新的水稻白化转绿突变体的生理特性和基因定位

秋丰M来源于粳稻秋丰的自然白化转绿突变株。其主要特征为前三叶白化带绿,第四叶及以后叶片均为淡绿色,抽穗时,秋丰M的颖壳和前三叶一样仍出现带绿的白化现象。不同生长时期对野生型和突变型水稻叶片色素含量测定的结果与田间观察结果一致,秋丰M确实存在着一个叶色显著变化的过程。主要农艺性状的比较结果表明,秋丰与秋丰M除穗颈长和千粒重达到极显著差异外,其他农艺性状均无明显差异。遗传分析发现该突变性状受一对隐性核基因控制。以209株培矮64S×秋丰M F_2的隐性突变个体为定位群体,将突变基因定位在水稻第2染色体长臂上,位于 SSR 标记RM475和RM2-22之间,其遗传距离分别为17.3 cM和2.9 cM,并将该基因命名为gra_(t)。     

一个新的水稻叶形突变体(tll1)的遗传分析与精细定位

利用甲基磺酸乙酯(ethylmethane sulphonate, EMS)诱变粳稻品种日本晴获得了一个遗传稳定的叶形突变体 thread-like leaf 1 (tll1)。该突变体在杭州表现为矮化、窄叶, 极端时仅剩主脉, 呈细丝状。将该突变体分别与籼稻品种南京6号、浙辐802和9311进行正反交配组, 遗传分析表明该突变体性状由1对隐性单基因控制。通过SSR和STS分子标记对F₂代分离群体进行遗传定位, 将该基因初步定位在第12染色体SSR标记RM247和RM101之间。随后利用已公布的粳稻品种日本晴和籼稻品种9311的基因组序列, 发展了7对有多态的STS标记, 最终将该基因定位在FL13和FL14之间约94.3 kb的区间内, 为进一步克隆TLL1基因奠定了基础。     

一个水稻显性高秆突变体的遗传分析和基因定位

从水稻(Oryza sativa L.)的两个半矮秆籼稻品种6442S-7和蜀恢881杂交F2代群体中发现一个高秆突变体D111,其株高和秆长分别比亲本蜀恢881增加63.0%和87.0%。用205个微卫星标记分析D111及其原始亲本6442S-7和蜀恢881之间的基因组DNA多态性,结果未发现D111具有2个原始亲本都没有的新带型,证明D111的确是6442S-7和蜀恢881的杂交后代发生基因突变产生的。将D111分别与蜀恢881、蜀恢527、明恢63、9311、IR68、G46B等6个半矮秆品种和高秆对照品种南京6号杂交,分析F1和F2代株高的遗传行为,结果表明D111的高秆性状由一对显性基因控制,且该基因与南京6号的高秆基因紧密连锁或等位。以蜀恢527/D111 F2群体为定位群体,运用微卫星标记将D111显性高秆突变基因定位于水稻第一染色体长臂,与RM212、RM302和RM472的遗传距离分别是27.7 cM、25.5 cM和6.0 cM,该基因暂命名为LC(t)。认为D111是首例从半矮秆品种自然突变产生的水稻显性高秆突变体,LC(t)为首次定位的水稻显性高秆突变基因。此外,将上述基因定位结果与Causse等(1994)和Temnykh等(2000; 2001)发表的水稻分子连锁图谱进行比较,发现LC(t)基因恰巧位于与水稻“绿色革命基因”sd1相同或十分相近的染色体区域,因此,还就LC(t)基因与sd1基因之间的可能关系进行了讨论。     

Characterization and mapping of a novel mutant sms1 （senescence and male sterility 1） in rice

Plant senescence plays diverse important roles in development and environmental responses.However,the molecular basis of plant senescence is remained largely unknown.A rice spontaneous mutant with the character of early senescence and male sterility (sms) was found in the breeding line NT10-748.In order to identify the gene SMS1 and the underlying mechanism,we preliminarily analyzed physiological and biochemical phenotypes of the mutant.The mutant contained lower chlorophyll content compared with the wild type control and was severe male sterile with lower pollen viability.Genetic analysis showed that the mutant was controlled by a single recessive gene.By the map-based cloning approach,we fine-mapped SMS1 to a 67 kb region between the markers Z3-4 and Z1-1 on chromosome 8 using 1,074 F2 recessive plants derived from the cross between the mutant sms1 (japonica) × Zhenshan 97 (indica),where no known gene involved in senescence or male sterility has been identified.Therefore the SMS1 gene will be a novel gene that regulates the two developmental processes.The further cloning and functional analysis of the SMS1 gene is under way.     

水稻苗期耐盐突变体的遗传分析及基因定位

在籼稻品种R401辐射诱变的M2群体中筛选到一个苗期耐盐突变体, 在150 mmol/L的NaCl溶液处理下对照植株枯萎死亡, 而突变体植株依然存活。以粳稻品种Nipponbare(不耐盐)和耐盐突变体作亲本, 构建了一个F2群体, 调查该群体在150 mmol/L的NaCl溶液胁迫下的表现, 发现Nipponbare和耐盐突变体苗期耐盐性的差异受单个主基因控制, 耐盐为隐性, 将该基因暂时命名为SST(t)。利用该F2群体, 采用集团分离分析(Bulked segregant analysis, BSA)法将SST(t)定位在第6染色体上, 进一步对F2群体中137个典型的耐盐单株的分子标记进行分析, 将该基因定位在InDel标记ID26847和ID27253之间, 约2.3 cM (或406 kb)的区间内, 与两标记分别相距1.2 cM和1.1 cM。     

一个水稻显性高秆突变体的遗传分析和基因定位

从水稻(Oryza sativa L.)的两个半矮秆籼稻品种6442S-7和蜀恢881杂交F2代群体中发现一个高秆突变体D111,其株高和秆长分别比亲本蜀恢881增加63.0%和87.0%.用205个微卫星标记分析D¨1及其原始亲本6442S-7和蜀恢881之间的基因组DNA多态性,结果未发现D111具有2个原始亲本都没有的新带型,证明D1¨的确是6442S-7和蜀恢881的杂交后代发生基因突变产生的.将D111分别与蜀恢881、蜀恢527、明恢63、9311、IR68、G46B等6个半矮秆品种和高秆对照品种南京6号杂交,分析F1和F2代株高的遗传行为,结果表明D1¨的高秆性状由一对显性基因控制,且该基因与南京6号的高秆基因紧密连锁或等位.以蜀恢527/D111 F2群体为定位群体,运用微卫星标记将D111显性高秆突变基因定位于水稻第一染色体长臂,与RM212、RM302和RM472的遗传距离分别是27.7 cM、25.5 cM和6.0 cM,该基因暂命名为LC(t).认为D111是首例从半矮秆品种自然突变产生的水稻显性高秆突变体,LC(t)为首次定位的水稻显性高秆突变基因.此外,将上述基因定位结果与Causse等(1994)和Temnykh等(2000,2001)发表的水稻分子连锁图谱进行比较,发现LC(t)基因恰巧位于与水稻"绿色革命基因"sd1相同或十分相近的染色体区域,因此,还就LC(t)基因与sd1基因之间的可能关系进行了讨论.     

Genetic analysis and mapping of gene fzp(t) controlling spikelet differentiation in rice

Monocots and dicots have diverged for 120 million years. The floral morpha of cereals isunique and much different from that of dicot plants. Nevertheless, it has been found that most genes controlling flower development share a conserved sequence called MADS-box[1]. Therefore,it is likely that monocots and dicots could have similar basic characteristics of flower developmentbut the mechanisms of genetic regulation for flowering induction and floral differentiation might be different[2,3]. Du…     

Genetic analysis and mapping of gene fzp(t) controlling spikelet differentiation in rice

A mutant of spikelet differentiation in rice called frizzle panicle (fzp) was discovered in the progeny of a cross between Oryza sativa ssp. indica cv. V20B and cv. Hua1B. The mutant exhibits normal plant morphology but has apparently fewer tillers. The most striking change in fzp is that its spikelet differentiation is completely blocked, with unlimited subsequent rachis branches generated from the positions where spikelets normally develop in wild-type plants. Genetic analysis suggests that fzp is controlled by a single recessive gene, which is temporarily named fzp (t). Based on its mutant phenotype, fzp (t) represents a key gene controlling spikelet differentiation. Some F2 mutant plants derived from various genetic background appeared as the "middle type", suggesting that the action of fzp (t) is influenced by the presence of redundant, modifier or interactive genes. By using simple sequence repeat (SSR) markers and bulked segregant analysis (BSA) method, fzp (t) gene was mapped in the terminal region of the long arm of chromosome 7, with RM172 and RM248 on one side, 3.2 cM and 6.4 cM from fzp (t), and RM18 and RM234 on the other side, 23.1 cM and 26.3 cM from fzp(t), respectively. These results will facilitate the positional cloning and function studies of the gene.     

一份水稻矮杆小粒突变体的形态特征和基因定位

从水稻T-DNA插入突变体库中鉴定出一个矮杆小粒突变体t129,该突变体与野生型植株相比,植株明显矮化,籽粒粒长明显缩短,千粒重下降。遗传分析表明,t129的突变性状由一对隐性核基因控制,该基因(T129)经图位克隆定位于水稻第5染色体长臂上,引物InDel43和InDel57之间,物理距离为430 kb,并与标记InDel51共分离。本研究明确了该矮杆小粒突变体的表型特征及遗传规律,为进一步研究调控水稻株高和粒型基因奠定基础。     

水稻msp1-4突变体的鉴定及其UDT1和GAMYB基因的表达分析

通过对粳稻‘9522’辐射诱变,得到一隐性雄性不育突变体msp1-4（MULTIPLE SPOROCYTE）,用遗传定位方法将该基因座位定位在分子标记WY-4和WY-8之间,相距0．8cM,物理距离247kb。测序分析证明这247kb区间中的MSP1基因的编码区在第758bp到767bp之间发生了10个碱基的缺失。形态学观察结果表明该突变体和已经报告过的msp1突变体的表型基本一致。为分析水稻其它与花药发育相关的基因在msp1-4中的表达变化,用半定量RT-PCR技术检测到影响绒毡层和花粉发育的重要基因UDT1和GAMYB的表达在突变体中比在野生型水稻中低,说明这2个基因可能位于MSP1基因的下游。