Development and characterization of interspecific hybrids between Oryza sativa and O. latifolia by in situ hybridization

Oryza sativa and O. latifolia belong to the AA and CCDD genomes of Oryza, respectively. In this study, interspecific hybrids of these species were obtained using the embryo rescue technique. Hybrid panicle traits, such as long awns, small grain, exoteric large purple stigma, grain shattering and dispersed panicles, resemble that of the paternal parent, O. latifolia, whereas there is obvious heterosis in such respects as plant height, tillering ability and vegetative vigor. Chromosome pairing and the genomic components of the hybrid were subsequently investigated using genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH) analysis. Based on the mitotic metaphase chromosome numbers of the root tips investigated, the hybrid is a triploid with 36 chromosomes. The genomic constitution of the hybrid is ACD. In the meiotic metaphase I of the hybrid pollen mother cell, poor chro- mosome pairing was identified and most of the chromosomes were univalent, which resulted in com- plete male sterility in the hybrid.     

普通栽培稻与非洲栽培稻杂交的可交配性研究

以二倍体普通栽培稻(Oryza sativa，2n=2x=24)为母本，以非洲栽培稻(Oryza globerrima，2n=2x=24)为父本进行远缘杂交配组。研究结果表明，二倍体普通栽培稻与非洲栽培稻杂交表现出一定的可交配性，但结实率很低(0．32％～1．97％)。对其杂交结实的胚胎学观察发现，非洲栽培稻的花粉能在二倍体普通栽培稻的柱头上萌发，萌发率仅为0．18％～0．56％，花粉管在花柱内的伸长速度比较慢，发生的受精频率比较低。以同源四倍体水稻(Oryza sativa，2n=4x=48)为母本，以非洲栽培稻为父本杂交配组。研究结果表明，两间比较容易杂交，其结实率达到5．70％～8．86％。观察其胚胎学表现，非洲栽培稻的花粉在同源四倍体水稻的柱头上萌发频率高达12．81％～14．56％，花粉管在柱头内伸长速度较快，发生的受精频率比较高。利用同源四倍体水稻为桥梁，更容易将非洲栽培稻的遗传物质引入普通栽培稻。     

Species-specific aminoacylation of Oryza sativa mitochondrial tRNA^Trp

Accuracy in protein synthesis is essential to the survival of all organisms. Aminoacyl-tRNA synthetases catalyzed aminoacylation is the basis of the fidelity of protein synthesis[1]. In recent years, the identification of nucleotide determinants, specific…     

Fine mapping of locus S-b for F1 pollen sterility in rice （Oryza sativa L.）

Strong heterosis existed in the hybrid of the subspe-cies in rice[1,2]. However, the partial sterility of the hy-brid hinders the utilization of the heterosis[3,4]. Ikehashi et al.[5,6] considered the female gamete as the main ste-rility form and proposed…     

拟南芥多效性基因CPR5转化水稻中花11的研究

 利用根癌农杆菌介导法将拟南芥多效性基因CPR5转化水稻中花11,同时优化了其再生体系和遗传转化体系。结果表明：相比27 ℃、暗培养,32 ℃、持续光照培养可使诱导率提高到100 %,且缩短了诱导周期;〖JP2〗预分化培养后,在附加5 mg/L 6 BA和1 mg/L NAA的分化培养基上分化率和再生率可分别达到100 %和90.50 %。同时探索了较高转化频率的条件为 A600≈ 0.05~0.1的农杆菌菌液浸染5 min,共培养时间为3 d,同时添加1张用AAI培养液浸湿的无菌滤纸。对部分转化植株进行PCR、Southern blot和半定量RT PCR检测,结果表明AtCPR5基因已经整合到水稻基因组中,在所试验的转化植株中均为单拷贝,但表达程度存在差异。     

Identification of salt-tolerance QTL in rice (Oryza sativa L.)

Quantitative trait loci (QTLs) controlling salt-tolerance at the seedling stage in rice (Oryza sativa L.) were identified by interval mapping (SIM) and composite interval mapping (CIM) using a doubled haploid population ZJDH and its high resolution genetic linkage map. The population was derived from an inter-subspecific cross between an indica variety Zhaiyeqing8 (ZYQ8) and a japonica variety Jingxi17 (JX17). Analysis of survival days of seedlings treated with 0.7% NaCI revealed that a major salt-tolerance quantitative trait locus (QTL), Std, was present between markers RG612 and C131 on chromosome 1 when using both MAPMAKER/QTL 1.1 and PLABQTL 1.0 (SIM). Its allele which contributes to salt-tolerance was from ZYQ8. In addition, seven more QTLs which give additive effect on salt-tolerance are identified when using PLABQTL (CIM), and most of them were from JX17.     

Study on heterosis of intersubspecies between indica and japonica rice （Oryza sativa L.） using chromosome segment substitution lines

Great heterosis exists in the inter-subspecific crossesbetween indica and japonica rice cultivars[1,2] and the ex-ploitation of this heterosis has long been considered as apromising method to further increase rice yield potential[3].Previous studies indicated that there were large differencesbetween the two subspecies with respect to morphology,isozyme, polymorphism of molecular markers and DNAstructure due to long time genetic differentiation[4,5]. Thosedifferences were identified to be clos…     

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.     

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。     

水稻基因整合平台系统供体质粒pURKMT的构建

提高水稻产量,改良稻米品质是育种学家广泛研究的课题．随着现代生物技术的发展,水稻已成为植物基因工程的重要研究对象．许多实验室已成功地建立了一系列供外源基因转化水稻的系统．但是这些转化系统主要应用Ｔｉ质粒衍生的载体,通过Ｔ－ＤＮＡ左右两端的序列将目的基...     

Characterization and fine mapping of an early senescence mutant （es-t） in Oryza sativa L.

An es-t (early senescence-temporary) mutant, produced by ethylene methylsulfonate treatment of strain Nipponbare, was identified in rice. The leaves of es-t appeared yellow at the seedling stage, and had decreased chlorophyll content. Rust spots were found during growth in es-t, especially at the leaf margin and tip. The plants showed a typical early-senescence phenotype at the milky stage. The leaf surface of es-t appeared smoother than wild-type leaves under a scanning electron microscope, because the lea...     

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.     

栽宽杂种水稻不同发育时期幼穗离体培养的研究

随栽宽杂种幼穗发育过程的进程,其愈伤组织诱导率呈现由低→高→低的变化．通过愈伤组织阶段和幼穗直接分化培养均在第２期（第１次枝梗原基分化期）获得了最高的绿苗分化率．当第６期（花粉母细胞减数分裂期）幼穗诱导的愈伤组织完全丧失分化绿苗能力时,而幼穗直接分化仍能再生绿苗,并且此法具有操作简单,获得绿苗快的特点．     

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.     

高效的苜蓿组织培养再生体系的建立

以保定苜蓿为实验材料,从外植体的基因型、培养基的选择,激素(KT)、水解酪蛋白的浓度等方面探讨了影响保定苜蓿组织培养的因素.结果表明:下胚轴比子叶作为外植体获得的效果更好;改良的SH培养基相对于UM培养基对愈伤组织的诱导更有效;在含有0.2mg/L KT和2 g/L水解酪蛋白的UM培养基上,得到了最大的胚状体诱导率;水解酪蛋白的加入提高了胚状体的质量和成熟率.     

Genetic and physical finemapping of the Sc locus conferring indica-japonica hybrid sterility in rice （Oryza sativa L.）

Hybrid sterility is a major hindrance to utilizing the heterosis in indica-japonica hybrids. To isolate a gene Sc conferring the hybrid sterility, the locus was mapped using molecular markers and an F2 population derived from a cross between near isogenic lines. A primary linkage analysis showed that Sc was linked closely with 4 markers on chromosome 3, on which the genetic distance between a marker RG227 and Sc was 0.07 cM. Chromosome walking with a rice TAC genomic library was carried out using RG227 as a starting probe, and a contig of ca. 320 kb covering the Sc locus was constructed. Two TAC clones, M45EI4 and M90J01 that might cover the Sc locus, were partially sequenced. By searching the rice sequence databases with sequences of the TACs and RG227 a japonica rice BAC sequence, OSJNBb0078P24 was identified. By comparing the TAC and BAC sequences, six new PCR-based markers were developed. With these markers the Sc locus was further mapped to a region of 46 kb. The results suggest that the BAC OSJNBb0078P24 and TAC M45EI4 contain the Sc gene. Six ORFs were predicted in the focused 46-kb region.     

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…     

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.