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.     

Identification of a novel tillering dwarf mutant and fine mapping of the TDDL（T） gene in rice （Oryza sativa L.）

Rice plant architecture is an important agronomic trait that affects the grain yield. To understand the molecular mechanism that controls plant architecture, a tillering dwarf mutant with darker-green leaves derived from an indica cultivar IR64 treated with EMS is characterized. The mutant, designated as tddl（t）, is nonallelic to the known tiilering dwarf mutants. It is controlled by one recessive nuclear gene, TDDL（T）, and grouped into the dn-type dwarfism according to Takeda＇s definition. The dwarfism of the mutant is independent of gibberellic acid based on the analyses of two GA-mediated processes. The independence of brassinosteroid （BR） and naphthal-3-acetic acid （NAA） of the tddl（t） mutant, together with the decreased size of parenchyma cells in the vascular bundle, indicates that the TDDL（7） gene might participate in another hormone pathway. TDDL（T） is fine mapped within an 85.51 kb region on the long arm of rice chromosome 4, where 20 ORFs are predicted by RiceGAAS （http：//ricegaas.dna.affrc. go.jp/rgadb/）. Further cloning of TDDL（T） will benefit both marker assisted selection （MAS） of plant architecture and dissection of the molecular mechanism underlying tillering dwarf in rice.     

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.     

Genetic analysis and mapping of rice （Oryza sativa L.） male-sterile （OsMS-L） mutant

A rice male-sterile mutant OsMS-L of japonica cultivar 9522 background, was obtained in M4 population treated with ^60Co γ-Ray. Genetic analysis indicated that the male.sterile phenotype was controlled by a single recessive gene. Results of tissue section showed that at microspore stage, OsMS-L tapetum was retarded. Then tapetal calls expanded and microspores degenerated. No matured pollens were observed in OsMS-L anther locus. To map OsMS-L locus, an F2 population was constructed from the cross between the OsMS-L (japonica) and LongTeFu B(indica). Firstly, the OsMS-L locus was roughly mapped between two SSR markers, RM109 and RM7562 on chromosome 2. And then eleven polymorphic markers were developed for further fine fine-mapping. At last the OsMS-L locus was mapped between the two lnDel markers, Lhsl0 and Lhs6 with genetic distance of 0.4 cM, respectively. The region was delimited to 133 kb. All these results were useful for further cloning and functional analysis of OsMS-L.     

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 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…     

Morphological and physiological analysis of narrow and striped leaf 1 （nsl1） mutant of rice （Oryza sativa L.） and the gene mapping

The shape and color of rice leaves are impor- tant agronomic traits that directly influence the proportion of sunlight energy utilization and ultimately affect the yield and quality. A new mutant exhibiting stable inheritance was identified as derived from ethyl methane sulfonate （EMS）-treated restorer Jinhui 10, tentatively named as narrow and striped leaf 1 （nsll）. The nsll displayed pale white leaves at the seeding stage and then white striped leaves in parallel to the main vein at the jointing stage. Meanwhile, its leaf blades are significantly narrower than the control group of Jinhui 10. The chloroplast structures of cells in the white striped area of the nsll mutant break down, and the photosynthetic pigments are significantly lower than that of the wild type. Moreover, fluorescence parameters, such as Fo, Fv/Fm, ФpsⅡ, qP, and ETR, in the nsll mutant are significantly lower than those of the wild type, and the photosynthetic efficiency is also significantly decreased. These changes in leaf color and shape, together with physiological changes in the nsll, result in smaller plant height and a decrease in the most important agro- nomic traits, such as the number of grains per panicle, grain weight, etc. Genetic analysis shows that the narrow and striped traits of the nsll mutant are controlled by a single recessive nuclear gene, which is located between InDel 16 and InDel 12 in chromosome 3. The physical distance is 204 kb. So far, no similar genes of such leaf color and shape in this area have been reported, This study has laid asolid foundation for the gene cloning and function analysis of NSL 1.     

Fine mapping of an incomplete recessive gene for leaf rolling in rice （Oryza sativa L.）

Genetic analysis and fine mapping of genes controlling leaf rolling were conducted using two backcrossed generations （BC4F2, BC4F3） derived from a cross between QMX, a non-rolled leaf cultivar as a recurrent parent, and JZB, a rolled leaf NIL of ZB as a donor parent. Results indicated that leaf rolling was mainly controlled by an incompletely recessive major gene, namely rl（t）, and at the same time, affected by quantitative trait loci （QTLs） and/or the environment. A genetic linkage map was constructed using MAPMAKER/EXP3.0 with eight polymorphic markers on chromosome 2, which were screened by BAS method from 500 SSR markers and 15 newly developed insertion/deletion （InDel） markers. The position of rl（t） was estimated with composite interval mapping （CIM） method using WinQTLcart2.5. Gene rl（t） was mapped between markers InDel 112 and RM3763, and 1.0 cM away from InDel 112 using 241 plants in BC4F2 population. To fine map r（t）, one BC4F3 line with 855 plants was generated from one semi-rolled leaf plant in BC4F2. Four new polymorphic InDel markers were developed, including InDel 112.6 and InDel 113 located between markers InDe1112 and RM3763. Based on the information of recombination offered by 191 rolled leaf plants and 185 non-rolled leaf plants from the BC4F3 line ,we mapped r（t） to a 137-kb region between markers InDel 112.6 and InDel 113. Homologous gene analysis suggested that r（t）was probably related to the process of leaf development regulated by microRNA.     

Fine mapping of a semidwarf gene sd-g in indica rice（Oryza sativa L.）

The semidwarf gene sd-g which has been usedin indiea rice breeding in southern China is a new one, non-allelic to sd-1. To map sd-g, an F2 population derived fromthe cross between Xinguiaishuangai and 02428 was con-structed. The sd-g was roughly mapped between two mi-crosatellite markers RM440 and RM163, with genetic dis-tances of 0.5 and 2.5 cM, respectively. Then nine new poly-morphic microsatellite markers were developed in this region.The sd-g was further mapped between two microsatellitemarkers SSR5-1 and SSR5-51, with genetic distances of 0.1and 0.3 cM, respectively, while cosegregated with SSR418. ABAC contig was found to span the sd-g locus, the region be-ing delimited to 85 kb. This result was very useful for cloningof the sd-g 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.     

水稻花粉Profilin基因启动子的克隆及序列分析

作者使用两次染色体步移（Genome walking)法，从灿稻（Oryza sativa subsp.indica)IR36中克隆到长度为569bp的花粉proiflin基因的启动子片段，并进行了全序列测定和分析。结果表明：在该段序列中含有3个TATA box，3个CAAT box和2个G－box。通过EPD（The Eukaryotic Promoter Database)的比较发现，序列的＋2－-71区域与番茄（Lycopersicon esculentum)花粉特异基因LAT52，向日葵（Helianthus annuus)花药特异基因SF2启动子关键序列的同源性为50％左右。     

水稻OsNAP基因的功能分析及其在育种价值方面的研究

NAC转录因子是植物特异的转录因子.本研究通过从水稻日本晴(Oryza sativa L.)的基因组中克隆到一个与拟南芥衰老相关的基因AtNAP(Atlg69490)的同源基因OsNAP.OsNAP的ORF为1 179bp,推测其编码蛋白含有392个氨基酸,等电点为8.55,分子质量为42.195ku.为进一步研究OsNAP的功能,利用显性抑制元件SRDX构建其pCAMBIA1304-35S:OsNAP-SRDX表达载体,通过农杆菌浸染方法将其转化水稻中花11品种.T1代转基因植株通过潮霉素、RealtimePCR、和Western blot鉴定,筛选出4个OsNAP高表达的阳性植株.在种子成熟期,观察到T1代转基因4号植株较野生型相比有明显的延缓衰老的表型,其他农艺性状如单株分蘖数、单株总粒数、单穗饱和穗粒数、千粒重、结实率等都优于野生型.实验证明抑制OsNAP基因的表达能够延缓水稻叶片衰老并可以提高产量,具有潜在的育种价值.     

Morphology and mapping analysis of rice (Oryza sativa L.) clustered spikelets( Cl) mutant

The rice clustered spikelets (Cl) mutant exhibits a phenotype that most of branch apical have 2-3 spikelets clustered together,SEM (scanning electron microscope )observation suggested that the Cl gene controlled branch apical development,and influenced the terminal spikelets elongation,The spikelet number was reduced in mutant,indicating that Cl may also have an effect on spikelet number,To map Cl locus,two F2 mapping populations derived from the crosses between the Cl and ZhongHua11,and Cl and ZheFu802 were constructed ,respectively,The Cl locus was roughly mapped between two CAPS markers CK0214 and SS0324,A further fine mapping analysis showed that the Cl locus was mapped between makers R0674E and Cl12560,with genetic distances of 0.2 and 2.1 cM,respectively ,Then we found a PAC conting spanning Cl locus,the region was delimited to 196 kb.This results was useful for cloning of the Cl gene,Allelism test demonstrated that Cl was allelic to Cl2 another rice clustered spikelets mutant.     

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.     

Genetic structure and diversity of Oryza sativa L. in Guizhou, China

Preserving many kinds of rice resources and rich variations, Guizhou Province is one of the districts with the highest genetic diversity of cultivated rice （Oryza sativa L.） in China. In the current research, genetic diversity and structure of 537 accessions of cultivated rice from Guizhou were studied using 36 microsetellite markers and 39 phenotypic characters. The results showed that the model-based genetic structure was the same as genetic-distance-based one using SSRs but somewhat different from the documented classification （mainly based on phenotype） of two subspecies. The accessions being classified into indica by phenotype but japonica by genetic structure were much more than that being classified into japonica by phenotype but indica by genetic structure. Like Ding Ying＇s taxonomic system of cultivated rice, the subspecific differentiation was the most distinct differentiation within cultivated rice. But the differentiation within indica or japonica population was different： japonica presented clearer differentiation between soil-watery ecotypes than indica, and indica presented clearer differentiation between seasonal ecotypes than japonica. Cultivated rices in Guizhou revealed high genetic diversity at both DNA and phenotypic levels. Possessing the highest genetic diversity and all the necessary conditions as a center of genetic diversity, region Southwestern of Guizhou was suggested as the center of genetic diversity of O. sativa L. from Guizhou.