The Post-meiotic Deficicent Anther1 （PDA1） gene is required for post-meiotic anther development in rice

To understand the molecular mechanism of male reproductive development in the model crop rice,we isolated a complete male sterile mutant post-meiotic deficient anther1 (pda1) from a γ-ray-treated rice mutant library.Genetic analysis revealed that the pda1 mutant was controlled by a recessive nucleus gene.The pda1 mutant anther seemed smaller with white appearance.Histological analysis demonstrated that the pda1 mutant anther undergoes normal early tapetum development without obvious altered meiosis.However,the pda1 mutant displayed obvious defects in postmeiotic tapetal development,abnormal degeneration occurred in the tapetal cells at stage 9 of anther development.Also we observed abnormal lipidic Ubisch bodies from the tapetal layer of the pda1 mutant,causing no obvious pollen exine formation.RT-PCR analysis indicated that the expression of genes involved in anther development including GAMYB,OsC4 and Wax-deficient anther1 (WDA1) was greatly reduced in the pda1 mutant anther.Using map-based cloning approach,the PDA1 gene was finely mapped between two markers HLF610 and HLF627 on chromosome 6 using 3,883 individuals of F2 population.The physical distance between HLF610 and HLF627 was about 194 kb.This work suggests that PDA1 is required for post-meiotic tapetal development and pollen/microspore formation in rice.     

Activation of gibberellin 2-oxidase 6 decreases active gibberellin levels and creates a dominant semi-dwarf phenotype in rice （Oryza sativa L.）

Gibberellin (GA) 2-oxidase plays a key role in the GA catabolic pathway through 2β-hydroxylation.In the present study,we isolated a CaMV 35S-enhancer activation tagged mutant,H032.This mutant exhibited a dominant dwarf and GA-deficient phenotype,with a final stature that was less than half of its wild-type counterpart.The endogenous bioactive GAs are markedly decreased in the H032 mutant,and application of bioactive GAs (GA3 or GA4) can reverse the dwarf phenotype.The integrated T-DNA was detected 12.8 kb upstream of the OsGA2ox6 in the H032 genome by TAIL-PCR.An increased level of OsGA2ox6 mRNA was detected at a high level in the H032 mutant,which might be due to the enhancer role of the CaMV 35S promoter.RNAi and ectopic expression analysis of OsGA2ox6 indicated that the dwarf trait and the decreased levels of bioactive GAs in the H032 mutant were a result of the up-regulation of the OsGA2ox6 gene.BLASTP analysis revealed that OsGA2ox6 belongs to the class III of GA 2-oxidases,which is a novel type of GA2ox that uses C20-GAs (GA12 and/or GA53) as the substrates.Interestingly,we found that a GA biosynthesis inhibitor,paclobutrazol,positively regulated the OsGA2ox6 gene.Unlike the over-expression of OsGA2ox1,which led to a high rate of seed abortion,the H032 mutant retained normal flowering and seed production.These results indicate that OsGA2ox6 mainly affects plant stature,and the dominant dwarf trait of the H032 mutant can be used as an efficient dwarf resource in rice breeding.     

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.     

Mapping of qGL7-2,a grain length QTL on chromosome 7 of rice

A residual heterozygous line(RHL)carrying a heterozygous segment between two SSR loci RM11 and RM134 on the rice chromosome 7 was selected from a set of recombinant inbred lines from the cross D50(javanica)/HB277(indica).The former parent produces much longer grains than the latter.Selfed progenies of this selection were analyzed genotypically(SSRs)and phenotypically(grain length).Grain length was discontinuously variable in the mapping populations,allowing for the placement of this QTL qGL7-2 within a～4.8 cM interval defined by RM351 and RM234.A set of new markers within this region were developed,which narrowed the QTL to a 278 kb region defined by the markers Indel1 and RM21945.This region contains 49 predicted genes.The results also suggest that the novel allele for grain length will be used for the application of marker assisted selection for the improvement of grain length.     

日本在隔离苗圃开始转基因水稻栽插试验

日本农业食品产业技术综合研究机构（NARO）作物研究所（NICS）,于2009年8月3日,在含转基因高色氨酸饲料用水稻的隔离苗圃开始栽插试验。配合该试验同时召开了插秧观摩会,参加观摩会的一般见习人员有10多个,报道有关人员4人。     

生物技术将成为发展中国家粮食自给自足的重要手段

2009年11月,中国为转基因抗虫水稻和植酸酶玉米颁发了生物安全证书,对于推广转基因作物商业化来说,这是一项里程碑式的决策。水稻是全球最重要的粮食作物,养育全世界近半数人口,而玉米是最重要的饲料作物。对二者的生物安全认证对未来转基因作物在中国、亚洲乃至全世界的推广有巨大的影响。     

外源一氧化碳对干旱胁迫下水稻幼苗抗氧化系统的影响

以水稻(Oryza sativa L.) 品种‘D奇宝优1号'幼苗为材料,采用20%聚乙二醇(PEG-6000)模拟水分胁迫,研究外源一氧化碳(CO)对干旱胁迫下水稻幼苗抗氧化能力的影响,以探索CO提高水稻幼苗抗旱性的机制.结果显示,CO供体高铁血红素(Hematin,H)显著降低干旱胁迫下水稻幼苗叶片质膜相对透性和丙二醛(MDA)含量,提高脯氨酸和可溶性蛋白的含量,不同程度地促进叶片中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的活性,而CO清除剂血红蛋白(Hemoglobin,Hb)则逆转CO供体对干旱胁迫下水稻幼苗氧化损伤的缓解效应.由此表明,外源CO能通过调整保护酶活性和渗透调节物质含量来提高水稻幼苗的抗氧化能力,有效增强其抗旱性.     

葱胞质雄性不育花蕾生化物质含量和能量代谢酶活性的动态变化特征

以葱胞质雄性不育系CA及其同核异质保持系CB为试材,研究了花蕾发育过程中IAA、GA3、ZR、ABA含量以及细胞色素氧化酶(COD)和ATP酶(ATPase)活性、可溶性糖、游离氨基酸和可溶性蛋白质含量的动态变化规律.结果显示:(1)葱不育系花蕾的IAA和GA3含量在败育过程中显著低于保持系,而ZR和ABA含量则较保持系有不同程度的盈积.不育系花蕾COD和ATPase活性以及游离氨基酸含量、可溶性蛋白质含量在中蕾期后显著低于保持系,可溶性糖在不同发育时期均基本低于保持系.(2)不育系花蕾IAA和GA3含量与可溶性糖含量变化均呈显著正相关,不育系ZR和保持系ABA含量则分别与COD、ATPase活性以及可溶性糖、可溶性蛋白质含量存在负相关关系.研究表明,葱胞质雄性不育系花蕾中IAA和GA3亏缺,而ZR和ABA盈余,各种内源激素含量和营养物质含量与能量代谢有关酶活性相关性不尽相同.     

水稻OsNRT1-d启动子的分离和功能分析

采用PCR技术,从水稻基因组中分离到OsNRT1-d读码框上游2 019 bp序列.序列分析表明:在起始密码ATG上游-189 bp和-127 bp处分别存在CAAT-box和TATA-box,具有典型的启动子结构.推测的转录起始位点CAC位于起始密码ATG上游-93 bp处.将OsNRT1-d启动子5′-端系列缺失后,分别与GUS报告基因融合,获得的NRT2019∷GUS、NRT1196∷GUS 和NRT719∷GUS转基因载体.农杆菌介导转化水稻,获得的转基因水稻均能启动下游GUS报告基因在水稻的根、叶、花颖和种子中表达;将转基因水稻幼苗放在滤纸上紧急干旱处理和用15% PEG6000进行模拟干旱处理,GUS基因的表达量明显升高,且干旱应答元件在-719 bp～-1 bp的范围内;GUS活性分析表明:OsNRT1-d启动子对ABA、NaCl、(NH4)2SO4、KNO3和Gln等信号没有应答反应.     

麻花艽和管花秦艽(龙胆科)之间自然杂交类型的分子验证

野外考察发现麻花艽(Gentiana stramineaMaxim.)和管花秦艽(G.siphonantha Maxim.ex Kusn.)同域分布时存在大量形态位于二者之间的个体。经形态变异研究后发现它们可能是这两个物种之间的杂交后代。对两个亲本种以及假设杂交群体共55个个体的核糖体ITS序列和叶绿体trnS-G序列的分析结果表明:中间形态个体是麻花艽和管花秦艽的自然杂交后代。此外分析了两个亲本种以及杂交群内个体间trnS-G和ITS序列的变异状况以及分子标记结果与形态鉴定不一致的可能原因;指出可能是杂交诱导的叶绿体基因组重组以及早期物种分化中的谱系筛选不彻底等原因造成了亲本种群体内序列变异的多样化。