一个新的水稻叶绿素缺失黄叶突变体的特征及基因分子定位

从粳稻"嘉花1号"60Coγ射线辐照的后代中筛选到一个叶绿素缺失黄叶突变体(yl11),与野生型"嘉花1号"相比该突变体表现为全生育期植株叶片呈黄色,叶绿素含量以及净光合速率明显下降,叶绿体发育不完善,并且伴随着株高等主要农艺性状的变化。遗传分析表明,该突变性状受一对隐性核基因(yl11)控制。该突变体与籼稻"培矮64S"杂交生产的F2、F3群体中的分离出突变体型920个单株作为定位群体,利用SSR和InDel分子标记将yl11基因定位在水稻第11染色体长臂上的MM2199和ID21039分子标记之间,其物理距离约为110kb,目前该区域内没有发现与水稻叶绿素合成/叶绿体发育相关已知功能基因。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。     

一个新的水稻矮秆突变体的遗传分析和基因定位

新的水稻矮秆基因的发掘,对深入研究植物株高的调控途径及株型育种有非常重要的作用。我们报道了从日本特早熟粳稻品种Kitaake的组织培养后代获得的一个矮秆突变体dm,该突变体植株细小,紧凑,机械强度降低,结实率下降,籽粒变窄,千粒重降低等。利用分离群体中的矮秆株,最终将目标基因定位在第4染色体长臂末端InDel标记EL-72和L-1之间,物理距离为168 Kb的区间内,该区间内无已报道的水稻矮秆基因,该基因可能是一个尚未被克隆的新的株高决定基因。     

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。     

一个新的水稻小穗梗弯曲突变体的形态特征及基因定位

稻穗小穗梗的发育与产量有着密切关系。文章利用⁶⁰Co g 射线辐照籼稻品种“浙农7号”, 获得一个性状能稳定遗传的小穗梗弯曲突变体bpb1 (bent pedicel branch 1), 表现为小穗梗弯曲, 并伴有小穗梗长度增长、穗长缩短和植株矮化等特点。扫描电镜观察显示, bpb1突变体小穗梗的表皮毛及气孔变小, 外表皮细胞和厚壁细胞排列不规则, 接近弯曲部位的细胞变小、排列更为紧密。bpb1突变体小穗梗横切面观察表明, 小维管束排列结构发生明显变化。遗传分析表明该突变表型受隐性单基因控制。利用bpb1突变体与粳稻品种“浙农大104”杂交构建的F₂群体进行基因定位, 将bpb1基因定位于水稻第7号染色体长臂SSR标记RM21537和RM21552之间, 该区间的物理距离为343 kb, 该区域内未发现与水稻小穗梗发育相关的已知基因。文章为bpb1基因的克隆和功能研究奠定了重要基础。     

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)。     

水稻温敏叶绿素突变体叶片超微结构的研究

对温敏转绿型叶绿素突变体1103S和武金4B“斑马叶”性状表达过程中叶绿素含量、叶绿体超微结构的变化进行了比较研究。结果表明,在一定条件下,叶片的失绿、复绿与叶绿素含量的下降、上升变化趋势一致;叶绿体结构在失绿区表现为严重退化,基粒和基粒片层减少,淀粉粒和嗜锇粒增多;复绿后,其叶绿体结构重建和恢复     

水稻高温敏感侧根缺失突变体k209的鉴定和基因定位

该研究以甲基磺酸乙酯（EMS）诱变得到的1个水稻高温敏感侧根缺失突变体k209及其野生型Kasalath为材料,在常温（白天32 ℃/夜晚22 ℃）和高温（34 ℃恒温）培养条件下,对其7 d龄幼苗进行表型比较鉴定,并采用亚甲基蓝染色观察侧根原基形成;以突变体k209为母本,分别与野生型Kasalath和粳稻品种日本晴杂交构建2个F₂群体进行遗传分析和基因定位,确定基因所属染色体以及在该染色体上的位置。结果表明：（1）在正常温度培养下,突变体k209的7 d龄幼苗株高、主根长和不定根长均与野生型Kasalath相似,但侧根长度变短,数量也减少;在高温条件下,k209幼苗株高变矮,主根和不定根的长度变短,表现出无侧根表型。（2）亚甲基蓝染色发现,野生型和k209幼苗主根在正常温度和高温条件下均可以观察到侧根原基,但在高温下k209的侧根原基数目明显少于Kasalath,约为Kasalath的58.03%,且不能突破表皮长出侧根。（3）遗传分析表明,k209的突变表型受隐性单基因控制,利用图位克隆技术将K209基因定位于4号染色体的InDel标记7522K和11524K之间,物理距离约4 002 kb。该研究结果为K209基因的克隆和解析水稻侧根的发生机制奠定了基础。     

植物叶绿素缺失突变体分子机制研究进展

植物叶绿素缺失突变体在自然界中广泛存在,是研究叶绿素形成和叶绿体发育等代谢途径的良好材料.该文主要从分子层面上阐述了叶绿素缺失突变体产生的原因,如叶绿素合成受阻、叶绿体光合蛋白合成或输入受阻、叶绿体RNA转录物未被编辑、过量光损伤和卟啉循环各物质之间的相互抑制,并归纳了近年来鉴定出来的一些叶绿素缺失突变基因,简要介绍了叶绿素和叶绿体之间的关系以及叶绿素缺失突变体的应用.     

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 theF ₁ hybrid between anindica-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 aBC ₂F₂ population. Genetic analysis in theBC ₂F₂ 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 inBC ₂F₂, was constructed to screen polymorphism with simple sequence repeat (SSR) markers in comparison with the02428 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 C424 and S13984 with a genetic distance of 2.4 cM and 0.6 cM, respectively. The gene is designatedft1.     

Genetic analysis and molecular mapping of a novel gene for zebra mutation in rice （Oryza sativa L.）

A novel zebra mutant, zebra-15, derived from the restorer line JinhuilO （Oryza sativa L. ssp. indica） treated by EMS, displayed a distinctive zebra leaf from seedling stage to jointing stage. Its chlorophyll content decreased （55.4%） and the ratio of Chla/Chlb increased （90.2%） significantly in the yellow part of the zebra-15, compared with the wild type. Net photosynthetic rate and fluorescence kinetic parameters showed that the decrease of chlorophyll content significantly influenced the photosynthetic efficiency of the mutant. Genetic analysis of F2 segregation populations derived from the cross of XinonglA and zebra-15 indicated that the zebra leaf trait is controlled by a single recessive nuclear gene. Ninety-eight out of four hundred and eighty pairs of SSR markers showed the diversity between the XinonglA and the zebra-15, their F2 population was then used for gene mapping. Zebra-15 （Z-15） gene was primarily restricted on the short arm of chromosome 5 by 150 F2 recessive individuals, 19.6 cM from marker RM3322 and 6.0 cM from marker RM6082. Thirty-six SSR markers were newly designed in the restricted location, and the Z-15 was finally located between markers nSSR516 and nSSR502 with the physical region 258 kb by using 1,054 F2 recessive individuals.     

水稻雄性不育突变体D63的育性基因遗传分析与精细定位

利用化学诱变剂甲基磺酸乙酯(EMS)处理籼稻品种冈46B获得雄性不育突变体D63,并对该突变体进行表型鉴定、遗传分析和基因定位。结果显示D63突变体花药瘦小呈乳白色,花药内完全无花粉粒,属于无花粉型雄性不育。与野生型亲本冈46B相比,D63突变体成熟期株高降低了13.7%,穗伸出度减少了266.7%,自交结实率为0,其他农艺性状无显著差异。遗传分析表明该不育性状受1对隐性核基因控制,该突变基因定位于第2号染色体长臂靠近着丝粒区域In Del标记J2和J4之间,与J2和J4的遗传距离分别为0.2 c M和0.1 c M,该定位区间的物理距离为105.8 kb。候选基因分析结果表明,D63突变体在编码分泌性成束糖蛋白基因LOC_Os02g28970编码区第1580位碱基A突变为C,使编码蛋白的氨基酸序列第527位组氨酸(His)突变为脯氨酸(Pro)。D63突变体与已报道的mtr1突变体表型上不同之处主要是后者花药含有败育花粉粒,二者表型上的差异可能是由于LOC_Os02g28970基因序列突变位点不同,以及它们分别属于籼、粳亚种2个不同遗传背景所致。     

水稻黄叶少分蘖突变体yllt1的鉴定及基因定位

采用⁶⁰Co-γ射线诱变籼稻（Oryza sativa subsp.indica）保持系‘T98B’获得一份兼具黄叶和少分蘖表型的突变体yllt1（yellow leaf and less tillering 1）,利用色素含量测定、构建显隐性混池和基因表达量测定等方法从表型和遗传层面对其遗传特征进行分析。结果显示：yllt1苗期叶绿素a和叶绿素b含量为野生型水稻品种‘T98B’的77.78%和60.00%,叶绿体发育异常,缺乏功能性叶绿体类囊体片层;其分蘖盛期的单株分蘖数为野生型的21.43%。遗传分析发现,在突变体yllt1与‘T98B’的杂交F₂群体中,黄叶与少分蘖性状的重组率为0.00%,表明yllt1同时控制叶色与分蘖表型;yllt1呈隐性遗传,受一个细胞核基因独立控制。该研究进一步采用连锁分析法将yllt1精细定位到第11染色体上,经测序分析推断发生了突变的登录号为LOC_Os11g05552的基因是yllt1的目的基因;该基因编码叶绿体前体信号识别颗粒54 kD（cpSRP54）蛋白,其第1外显子的第29位碱基C发生了缺失,将造成其蛋白产物从N-端至C-端氨基酸组成的严重破坏。RT-qPCR分析结果显示,yllt1叶中叶绿素合成基因OsCAO1、OsCAO2与OsNOL等的表达量明显下调;茎中分蘖正向调控基因OsTAC1受到显著抑制,而负调控基因OsTB1与OsDLT的表达量明显增强。研究结果表明cpSRP54同时参与了水稻叶色和分蘖的调控。     

Molecular mapping of split rice spikelet mutant srs-1 and analysis of its homeotic function in rice

srs-1, a new floral organ identity gene in rice, was mapped using RAPD and RFLP markers. Firstly, the cross was made between "ZhaiYeQing 8" (ZYQ8, indica) and split rice spikelet (SRS, japonica) mutant. The ratio of wild-type individuals and mutant plants in F2 population is 3:1, which indicates that the mutant characteristics are controlled by single recessive gene, srs-1. Consequently, BSA method was adopted and 520 random 10-mer primers were used to screen polymorphic bands between two bulks. Six primers could amplify polymorphic bands, of which S465 generates the most stable RAPD patterns. Then, S465 that cosegregates in F2 population has been converted into an RFLP marker successfully. Furthermore, srs-1 gene was mapped on chromosome 3 using DH mapping population. The effect of srs-1 gene results in the mutant of split rice spikelet. The mutant has longer and softer palea/lemma than those of wild-type, and two small palea/lemma-like organs between palea and lemma. In addition, there is a flower with three stamens and carpel in the axil of lemma. Thus, there are nine stamens and two carpels in the spikelet of mutant. srs-1 gene may belong to homeotic gene of class A according to the mutant characteristics and ABC model.     

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     

Isolation,characterization, and mapping of the stay green mutant in rice

Leaf color turns yellow during senescence due to the degradation of chlorophylls and photosynthetic proteins. A stay green mutant was isolated from the glutinous japonica rice Hwacheong-wx through N-methyl-N-nitrosourea mutagenesis. Leaves of the mutant remained green, while turning yellow in those of the wild-type rice during senescence. The stay green phenotype was controlled by a single recessive nuclear gene, tentatively symbolized as sgr(t). All the phenotypic characteristics of the mutant were the same as those of the wild-type lines except for the stay green trait. The leaf chlorophyll concentration of the mutant was similar to that of the wild-type before heading, but decreased steeply in the wild-type during grain filling, while very slowly in the mutant. However, no difference in photosynthetic activity was observed between the stay green mutant and the yellowing wild-type leaves, indicating that senescence is proceeding normally in the mutant leaves and that the mutation affects the rate of chlorophyll degradation during the leaf senescence. Using phenotypic and molecular markers, we mapped the sgr(t) locus to the long arm of chromosome 9 between RFLP markers RG662 and C985 at 1.8- and 2.1-cM intervals, respectively. Received: 29 April 2001 / Accepted: 17 July 2001     

水稻Ds插入淡绿叶突变体的鉴定和遗传分析

Ac/Ds插入突变是水稻基因功能鉴定的有力工具之一。文章从水稻中花11 Ds-T-DNA转化纯合体与Ac-T-DNA 转化纯合体的杂交群体中筛选到一个淡绿叶突变体。该突变体在三叶期由绿苗转为淡绿叶苗, 自然光照下突变体迅速焦枯, 但是在弱光照条件下, 突变体能缓慢生长至开花结实; 突变体光合作用特性研究表明该突变是典型的光抑制突变体。遗传分析表明该突变为Ds插入导致的隐性突变。     

水稻矮秆突变体dtl1的分离鉴定及其突变基因的精细定位

文章通过对所构建的水稻突变体库进行大规模筛选,获得一个稳定遗传的矮秆突变体,与野生型日本晴相比,该突变体表现为植株矮化、叶片卷曲、分蘖减少和不育等性状,命名为dtl1(dwarf and twist leaf 1)。dtl1属于nl型矮秆,激素检测表明,矮秆性状与赤霉素和油菜素内酯无关。遗传分析显示,突变性状受单一隐性核基因控制。利用dtl1与籼稻品种Taichung Native 1杂交构建F2群体,将该突变基因DTL1定位于水稻第10染色体长臂2个SSR标记RM25923和RM6673之间约70.4 kb区域内,并与InDel标记Z10-29共分离,在该区域预测有13个候选基因,但未见调控水稻株高相关基因的报道,因此,认为DTL1基因是一个新的控制水稻株高的基因。