水稻早熟多子房突变体加以5的遗传分析和基因定位

在水稻中花11的后代中筛选到1例花器官数目突变体,突变体主要表现为多雄蕊、多子房和早开花。遗传分析表明,该突变表型受1对隐性核基因控制。因为对花器官数目突变体曾有报道,如fon1、fon2、fon3和fon4,所以该突变体暂定名为fort5。利用fort5与籼稻品种华粳籼74构建的F2群体对fort5进行基因定位,发现其与第6染色体上的标记RM400和RM412连锁,遗传距离分别为10．5cM和1．6cM。通过在两标记间发展6个新的Indel标记,将该基因定位于116kb区间。     

水稻无内稃突变体的遗传分析和基因定位

花器官发育异常的突变体是研究植物花发育分子遗传机制的良好实验材料,以水稻无内稃突变体为父本,生47、N625和CDR22为母本配制杂交组合进行性状遗传分析,根据F2代表型及X^2测验结果表明,突变性状是由单隐性基因控制的,选用突变体为父本,生47为母本杂交的F2群体作定位群体,利用SSLP标记的和RFLP标记将与突变性状相关的基因定位在第6染色体短臂上RFLP标记C498和RZ450之间,暂定名为npa-1。为进一步的基因克隆及功能研究奠定了基础。     

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。     

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.     

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。     

水稻叶状颖壳突变体Oslh的遗传分析和OsLH基因的定位

通过γ射线诱变,从粳稻品种9522的M2代中筛选出一株具有叶状颖壳的突变体,定名Oslh(1h=leafy hull).Oslh突变体的开花时间要比野生型晚15 d左右,内外稃和浆片发育成了叶片状器官.Oslh突变体与粳稻品种9522回交结果表明Oslh突变性状可能由单核基因隐性突变造成.以Oslh突变体与籼稻品种广陆矮4号杂交的F2代群体为基因定位群体,利用SSR和InDel分子标记将Oslh突变位点定位在3号染色体上的SSR标记RM5475和InDel标记GY305之间,遗传距离分别为2.5 cM和1.9 cM.这些结果为克隆OsLH基因和研究花器官发育的调控机理奠定了基础.     

水稻多分蘖矮秆突变体htd1-2的遗传分析和基因定位

文章所采用的多分蘖矮秆突变体为htd1-2(high-tillering dwarf 1-2), 是野生型籼稻品种9311经350Gy的60Co- g射线辐射处理后产生的后代中选育出来的稳定多分蘖矮秆突变体。遗传分析表明, 突变体htd1-2多分蘖矮秆性状是由一对隐性核基因的突变造成的。文章利用简单重复序列(Simple sequence repeat, SSR)、酶切扩增多态性序列(Cleaved amplified polymorphic sequence, CAPS)和衍生型CAPS(derived CAPS, dCAPS)等分子标记的方法, 最终将多分蘖矮秆基因HIGH-TILLERING DWARF1-2(HTD1-2)定位在水稻第4号染色体116 kb的物理区间内。在该物理区间内有一个已经克隆的控制水稻分蘖的基因HIGH-TILLERING DWARF1(HTD1), 经过测序比对和dCAPS特异性分析, 认为HTD1就是HTD1-2基因。尽管突变体htd1与突变体htd1-2是等位基因的不同位点发生突变, 但是由于遗传背景的不同, 两者表型并不完全相同。此外, 通过去除分蘖芽的实验证明了突变体htd1-2的矮化部分是由于分蘖过多造成的。     

一个新的水稻花器官突变体的鉴定和基因定位

在水稻（Oryza sativa）品种台中65的组培后代中发现一个花器官发育异常突变体flower organ number6（fon6）,其主要表型为：双子房,多柱头,7-8枚雄蕊。遗传分析表明,该突变表型由一对隐性基因控制。以该突变体与籼稻3037杂交的F2代分离群体作为定位群体,利用STS标记将与突变性状相关的基因定位于第6染色体短臂上STS标记PL4和PL5之间约480kb的范围内。该研究结果为进一步的基因克隆及功能研究奠定了基础。     

水稻短根突变体Osksr6的遗传分析和基因定位

该研究对从甲基磺酸乙酯(EMS)诱变的水稻突变体库中筛选到的一个短根突变体Osksr6(Oryza sativa kasalath short root 6)进行了表型鉴定、遗传分析与基因定位。结果表明:(1)生长7d的突变体Osksr6与野生型相比,株高与不定根数量差异不明显,但主根变短61.98%、不定根变短46.42%,侧根的发生与根毛的伸长也受不同程度抑制;成熟期的Osksr6分蘖数明显减少,总穗长与结实率均较野生型差。(2)遗传分析结果显示,突变体Osksr6的短根性状受1对隐性基因控制。(3)利用图位克隆技术,将突变基因OsKSR6定位于3号染色体InDel标记28420k和28880k之间,物理距离约460kb,该区间没有已报道的与根系发育相关的基因。该研究为进一步研究水稻根系生长的分子机理奠定了基础。     

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     

Genetic analysis and gene mapping of a new rolled-leaf mutant in rice (Oryza sativa L.)

To understand the development of rice leaf blades,we identified a new rolled-leaf mutant,w32,from indica cultivar IR64 through EMS mutagenesis. The mutant showed a stable rolled-leaf phenotype throughout the life cycle. Two F2 populations were developed by crossing w32 to cultivar IR24 and PA64. Genetic analysis showed that the rolled-leaf phenotype was controlled by a single recessive gene. To determine the location of the gene,bulked segregant analysis was carried out using mutant and wild-type DNA pools ...     

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。     

Genetic mapping of hypervariable minisatellite sequences in rice (Oryza sativa L.)

Minisatellites, or DNA fingerprinting sequences, have been utilized in animal linkage studies for several years but have not been used as markers for plant genome mapping. In animal genome mapping they have resulted in limited success because they are evenly dispersed in some species but are often clustered near telomeric regions, as observed on human chromosomes. The purpose of the present study was to generate DNA fingerprints utilizing several rice-derived minisatellites containing different core sequences and numbers of repeat units, followed by assessing their potential for use as genetic markers when mapped to a rice recombinant inbred line (RIL) population. Sites of segregating minisatellite loci were mapped onto 11 of the 12 rice RIL linkage maps. The implications for the use of rice minisatellite core sequences as genetic markers on linkage maps in rice are discussed. Received: 1 March 1999 / Accepted: 22 June 1999     

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.     

Genetic analysis of plant regeneration in rice (Oryza sativa L.)

Summary A diallel study involving reciprocal crosses of four genotypes (IR8, 36, 54, and 64) was carried out to understand the genetic mechanism of plant regeneration from immature embryo-calli in rice. Regeneration frequency (% of calli that produced plants) varied from a high of 86% for IR54 to a low of 0% for IR36, while regenerated plants per embryo numbered from 0 to 7 when these same IR lines and the F₁ hybrids were compared. Combining ability analysis revealed that both nuclear (with both additive and dominant effects) and cytoplasmic genes are important in controlling regeneration in rice. Parental lines and F1 hybrids with high ability to regenerate were identified.     

Genetic analysis of waxy locus in rice (Oryza sativa L.)

Summary Inheritance of waxy locus was studied in crosses of a waxy variety with four non-waxy parents having high-, intermediate-, low- or very low-amylose content. The analysis for amylose content was done on a single grain basis in parents, F₁, F₂, B₁F₁, and B₂F₁ seeds. The waxy parent lacking synthesis of amylose content was found to differ from the ones having high-, intermediate-, low- or very low-amylose content by one gene with major effect. Dosage effects for amylose content were observed to have great influence on segregation pattern and efficiency of selection. Selection efficiency for amylose content can be enhanced by selecting for endosperm appearance in early segregating generations.     

Genetic analysis of salinity tolerance in rice (Oryza sativa L.)

Summary The genetics of salinity tolerance in rice was investigated by a nine-parent complete diallel including reciprocals. Test materials involved susceptible (IR28, IR29, and MI-48), moderately tolerant (IR4595-4-1-13, IR9884-54-3-1E-P1, and IR10206-29-2-1), and tolerant (Nona Bokra, Pokkali, and SR26B) parents. Twoweek-old seedlings were grown in a salinized (EC = 12 dS/m) culture solution for 19 days under controlled conditions in the IRRI phytotron. Typical characteristics of salinity tolerance in rice were found to be Na⁺ exclusion and an increased absorption of K⁺ to maintain a good Na-K balance in the shoot. Genetic component analysis (GCA) revealed that a low Na-K ratio is governed by both additive and dominance gene effects. The trait exhibited overdominance, and two groups of genes were detected. Environmental effects were large, and the heritability of the trait was low. Our findings suggest that when breeding for salt tolerance, selection must be done in a later generation and under controlled conditions in order to minimize environmental effects. Modified bulk and single-seed descent would be the suitable breeding methods. Combining ability analysis revealed that both GCA and specific combining ability (SCA) effects were important in the genetics of salt tolerance. Moderately tolerant parents — e.g., IR4595-4-1-13 and IR9884-54-3-1E-P1 — were the best general combiners. Most of the best combinations had susceptible parents crossed either to moderate or tolerant parents. The presence of reciprocal effects among crosses necessitates the use of susceptible parents as males in hybridization programs. Large heterotic effects suggest the potential of hybrid rice for salt-affected lands.     

Genetic analysis and molecular mapping of low amylose gene du12(t) in rice (Oryza sativa L.)

Key message

We obtained interesting results for genetic analysis and molecular mapping of the du12(t) gene.

Abstract

Control of the amylose content in rice is the major strategy for breeding rice with improved quality. In this study, we conducted genetic analysis and molecular mapping to identify the dull gene in the dull rice, Milyang262. A single recessive gene, tentatively designated as du12(t), was identified as the dull gene that leads to the low amylose character of Milyang262. To investigate the inheritance of du12(t), genetic analysis on an F₂ population derived from a cross between the gene carrier, Milyang262, and a moderate amylose content variety, Junam, was conducted. A segregation ratio of 3:1 (χ ² = 1.71, p = 0.19) was observed, suggesting that du12(t) is a single recessive factor that controls the dull character in Milyang262. Allelism tests confirmed that du12(t) is not allelic to other low amylose controlling genes, wx or du1. Recessive class analysis was performed to localize the du12(t) locus. Mapping of du12(t) was conducted on F₂ and F₃ populations of Baegokchal/Milyang262 cross. Linkage analysis of 120 F₂ plants revealed that RM6926 and RM3509 flank du12(t) at a 2.38-Mb region. To refine the du12(t) locus position, 986 F₂ and 289 F₃ additional normal plants were screened by the flanking markers. Twenty-six recombinant plants were identified and later genotyped with four additional adjacent markers located between RM6926 and RM3509. Finally, du12(t) was mapped to an 840-kb region on the distal region of the long arm of chromosome 6, delimited by SSR markers RM20662 and RM412, and co-segregated by RM3765 and RM176.