水稻小穗特征基因FZP的图位克隆

FZP是水稻中控制小穗分化的一个关键基因,先前已将它定位在第7染色体上。通过进一步对该基因进行精细定位和图位克隆,找到2个SSR标记NRM6和NRM8,将该基因锁定在一个遗传距离为1．2cM的范围内(两标记与目标基因的遗传距离分别为0．2cM和1．0cM),相应的物理距离为144kb。发现在预期的目标基因位置,存在一个具有类似AP2结构域的基因。已知AP2是一个控制植物花发育的重要基因。因此,这个基因应是FZP的一个候选基因。PCR扩增结果显示,突变体中该基因有一个大约4kb的插人片段,与向共分离。由此可以初步认为,该基因就是FZP。     

水稻矮化相关基因的研究进展

矮秆水稻的培育成功是20世纪农业最主要的成就之一.本文综述了水稻矮化相关基因的遗传学、激素对矮化基因突变体的调控以及矮化相关基因的克隆等方面的研究进展,并分析了利用基因工程手段控制农作物生长的巨大潜能.     

水稻体细胞无性系变异

水稻体细胞无性系变异研究取得了很大进展 ,获得了大量抗病、抗逆、优质、矮杆等突变体。对这些突变体遗传分析表明 ,大多数突变性状由 1对或 2对基因控制。水稻体细胞无性系变异的发生与基因型、性状、继代时间、培养方式等有关 ,并具有内在的机制 ,点突变和反转录转座子插入可能是引起水稻无性系变异的两个重要原因。     

Coleoptile Senescence in Rice (Oryza sativa L.)

We investigated the cellular events associated with cell deathin the coleoptile of rice plants (Oryza sativa L.). Seeds germinatedunder submergence produced coleoptiles that were more elongatedthan those grown under aerobic conditions. Transfer of seedlingsto aerobic conditions was associated with coleoptile opening(i.e. splitting) due to death of specific cells in the sideof the organ. Another type of cell death occurred in the formationof lysigenous aerenchyma. Senescence of the coleoptile was alsonoted, during which discolouration of the chlorophyll and tissuebrowning were apparent. DNA fragmentation was observed by deoxynucleotidyltransferase-mediateddUTP nick end labelling (TUNEL) assay, and further confirmedby the appearance of oligonucleosomal DNA ladders in senescentcoleoptile cells. Two nucleases (Nuc-a and Nuc-b) were detectedby in-gel-assay from proteins isolated from coleoptiles. Nuc-a,commonly observed in three cell death phases required eitherCa²⁺or Mg²⁺, whereas Nuc-b which appeared during senescencerequired both Ca²⁺and Mg²⁺. Both nucleases were strongly inhibitedby Zn²⁺. Copyright 2000 Annals of Botany Company Aerenchyma, rice, cell death, coleoptile, fragmentation, nuclease, Oryza sativa, senescence, split, submergence, TUNEL     

Genome-Wide Survey and Expression Analysis of Amino Acid Transporter Gene Family in Rice (Oryza sativa L.)

Background

Amino acid transporters (AATs) that transport amino acids across cellular membranes are essential for plant growth and development. To date, a genome-wide overview of the AAT gene family in rice is not yet available.

Methodology/Principal Findings

In this study, a total of 85 AAT genes were identified in rice genome and were classified into eleven distinct subfamilies based upon their sequence composition and phylogenetic relationship. A large number of OsAAT genes were expanded via gene duplication, 23 and 24 OsAAT genes were tandemly and segmentally duplicated, respectively. Comprehensive analyses were performed to investigate the expression profiles of OsAAT genes in various stages of vegetative and reproductive development by using data from EST, Microarrays, MPSS and Real-time PCR. Many OsAAT genes exhibited abundant and tissue-specific expression patterns. Moreover, 21 OsAAT genes were found to be differentially expressed under the treatments of abiotic stresses. Comparative analysis indicates that 26 AAT genes with close evolutionary relationships between rice and Arabidopsis exhibited similar expression patterns.

Conclusions/Significance

This study will facilitate further studies on OsAAT family and provide useful clues for functional validation of OsAATs.     

Environmental Parameters Affecting Dark Response of Rice Seedlings (Oryza sativa L.) to Triacontanol

Triacontanol applied to IR-8 rice (Oryza sativa L.) seedlings in nutrient solution caused an increase in dry weight during a 6-hour dark period. This increase was altered by atmospheric CO₂ and O₂ concentrations. The largest growth response occurred from 200 to 350 μliters/liter CO₂ with 5% O₂. The treated seedlings did not fix atmospheric CO₂ in the dark, and the immediate products of photosynthesis were not involved in the dry weight increase. The growth response was characterized by an increase in soluble and insoluble Kjeldahl-N, and soluble carbohydrates. The response curve for dry weight increase was a linear function of log presentation time of triacontanol. The response exhibited an apparent K_dose of 25 minutes in 10 μg/liter triacontanol in the dark and 18 minutes in the light. Concentrations of 50 μg/liter and higher inhibited growth.     

水稻质体葡萄糖-6-磷酸脱氢酶基因的克隆与表达研究

戊糖磷酸途径是高等植物中重要的代谢途径,主要生理功能是产生NADPH以及供核酸代谢的磷酸戊糖。葡萄糖-6-磷酸脱氢酶（G6PDH）是戊糖磷酸途径的关键酶,广泛存在于高等植物细胞的细胞质和质体中。木研究首次从水稻（Oryza sativa L.）幼苗中分离了核编码的质体G6PDH基因OsG6PDH2,序列分析表明OsG6PDH2编码一个具有588个氨基酸残基的多肽,等电点为8．5,分子量66kDa。OsG6PDH2的N端有1个70个氨基酸的信号肽,推测的裂解位点为Gly55和Val56,表明OsG6PDH2编码产物可能定位于质体。多序列比较的结果表明OsG6PDH2与拟南芥、烟草、马铃薯质体G6PDH的一致性分别达81％、87％、83％。进化关系说明水稻OsG6PDH2与拟南芥（AtG6PDH3）、马铃薯（StG6PDH1）处于高等植物P2型质体G6PDH分支上,暗示了OsG6PDH2可能是一个P2型的质体蛋白。Matinspector程序分析表明,OsG6PDH2在起始密码子上游含有一个bZIP转录因子识别位点、一个ABA应答元件、一个CRT／DRE元件和1个W-box元件。半定量RT-PCR分析表明,OsG6PDH2在水稻根、茎、叶和幼穗组织中都呈低丰度组成型表达,在根部表达较高,在水稻幼苗中的表达显著受暗处理的诱导。将OsG6PDH2的完整开放阅读框构建到大肠杆菌表达载体pET30a（＋）中,pET30a（＋）-OsG6PDH2在大肠杆菌中得到了有效表达。酶活性测定证明,OsG6PDH2的编码产物具有葡萄糖-6-磷酸脱氢酶的功能。     

水稻叶色突变体叶绿体发育规律研究

从温敏核不育系水稻'810S'中筛选出一个生长发育正常的淡黄绿叶色自然突变株'标810S',其叶绿素含量约为'810S'的50%,光合速率比野生型高.以'810S'为对照,对'标810S'进行叶片形态、叶肉细胞和叶绿体超微结构以及叶绿体蛋白研究.结果显示,'标810S'的叶长、宽和面积与'810S'相似;叶肉细胞和叶绿体发育稍迟缓,片层结构减少;叶绿体蛋白约为对照的55%,并初步鉴定出与光合作用相关的差异蛋白点13个,其中4个缺失蛋白,包括1个RuBP大亚基缺失.推测该水稻突变体叶色变浅与叶绿体基粒片层减少有关.     

水稻低温发芽力QTL定位和遗传分析

以Kinmaze(粳稻)／DV85(籼稻)的重组自交系F10世代群体检测了影响水稻低温发芽力性状的数量性状基因座(QTL)。通过测定不同时期的低温发芽率,确定了15℃低温、第10d为检测低温发芽率的最适处理温度和时间,该条件下能够充分检测到品种的差异和分离群体的变异。通过设置对照,证明所检测的低温发芽率不受休眠及二次休眠的影响。15℃低温、第10d时,Kinmaze的发芽率达35％,DV85的发芽率只有7％,两亲本之间存在明显差异,该群体81个家系的低温发芽率变幅在0％～99％之间。QTL分析结果检测到5个与低温发芽力相关的基因座,分别位于第2、6、7、11和12染色体上。位于第2、6和11染色体上的qLTG-2、qLTG-6和qLTG-11贡献率分别为27．1％、17．1％和15．0％,对低温发芽力性状的增效基因来自DV85;位于第7、12染色体上qLTG-7和qLTG-12的贡献率分别为22．9％和8．8％,增效基因来自Kinmaze。其中,qLTG-6和qLTG-11在染色体上的位置与已报道的有关低温发芽力QTL位置相似,而qLTG-2、qLTG-7和qLTG-12为新检测的低温发芽力基因座。上位性分析结果显示,第3与第5染色体上存在影响低温发芽力的互作位点,其互作可以提高低温发芽力,而第7染色体上的两位点之间的互作降低了低温发芽力。     

水稻三系不育系育性与温度关系研究

通过自然条件下分期播种实验,对生产上应用的12个三系不育系花粉育性与温度的关系进行比较分析。结果表明:自然高温条件下不育系金23A、T98A、丰源A及冈46A花粉不育性稳定;隆398A、隆香634A、4302A、优IA及D62A出现少量可染花粉;高温诱导天丰A、五丰A和粤泰A花粉可育作用明显。此外,对可染花粉率与水稻不同发育时期的日均温相关性的分析表明,五丰A、天丰A和粤泰A可染花粉率均与抽穗前13~18 d的日均温显著相关(P0.05),高温诱导五丰A、天丰A和粤泰A育性转换敏感期为雌雄蕊原基形成末期至减数分裂期,育性转换临界日均温度分别为27.11℃、26.48℃和27.30℃。     

Release of Nitrogen From Chloroplasts During Leaf Senescence in Rice (Oryza sativa L.)

In order to ascertain the possibility that nitrogen associatedwith chloroplasts serves as a major source of nitrogen redistributedfrom senescent leaves, chloroplasts were isolated from riceleaves and changes with leaf age in total leaf nitrogen andchloroplast nitrogen were examined. Results presented here showthat decrease in total leaf nitrogen during leaf senescencewas closely correlated with decrease of chloroplast nitrogenand roughly 85–95 per cent of leaf nitrogen released fromsenescent leaves during the experimental period could be accountedfor by a loss of chloroplast nitrogen. By dividing chloroplastnitrogen into two fractions, i.e. lamellar and stroma fractions,the question of which fraction was more deeply concerned withthe loss of leaf nitrogen was clarified. Results suggested thatin the vegetative stage of plant growth the stroma was mainlyresponsible for the loss of leaf nitrogen. On the other hand,nitrogen was released from lamellar and stromal fractions atalmost the same rate during the reproductive stage. Oryza sativa L., rice, chloroplasts, nitrogen, leaf senescence     

Why Nitrogen Use Efficiency Decreases Under High Nitrogen Supply in Rice (Oryza sativa L.) Seedlings

Hydroponic experiments were conducted in a greenhouse to examine the effects of different nitrogen (N) supply (low, 20 mg L⁻¹; intermediate, 40 mg L⁻¹; and high, 100 mg L⁻¹) on the growth, nitrogen use efficiency, and photosynthetic characteristics of rice seedlings (Oryza sativa L., cv. “Shanyou 63” hybrid indica. China). Leaf gas exchange was conducted to identify the photosynthetic-limiting factors in plants with high N supply. The results showed that (1) shoot biomass, leaf area, and tiller numbers per plant under low N were lower than under intermediate and high N supplies. No significant differences were observed between plants supplied with intermediate and high N. (2) About a 35% increase in leaf N content in plants fed by high N resulted in about a 15% increase in carboxylation efficiency (CE) and photosynthetic rate. (3) The noncorresponding increases in photosynthetic rate in rice seedlings fed by high N relative to low N resulted from Rubisco activity and/or CE. (4) The decreased Rubisco activity was induced by a relatively insufficient CO₂ supply under high N supply. These results indicated that insufficient CO₂ supply under high N supply accounted for the decreased Rubisco activity and the noncorresponding increases in photosynthetic rate to leaf N content, and as a result, decreased (photosynthetic) nitrogen use efficiency.     

Genome-Wide Identification, Classification, and Expression Analysis of Autophagy-Associated Gene Homologues in Rice (Oryza sativa L.)

Autophagy is an intracellular degradation process for recycling macromolecules and organelles. It plays important roles in plant development and in response to nutritional demand, stress, and senescence. Organisms from yeast to plants contain many autophagy-associated genes (ATG). In this study, we found that a total of 33 ATG homologues exist in the rice [Oryza sativa L. (Os)] genome, which were classified into 13 ATG subfamilies. Six of them are alternatively spliced genes. Evolutional analysis showed that expansion of 10 OsATG homologues occurred via segmental duplication events and that the occurrence of these OsATG homologues within each subfamily was asynchronous. The Ka/Ks ratios suggested purifying selection for four duplicated OsATG homologues and positive selection for two. Calculating the dates of the duplication events indicated that all duplication events might have occurred after the origin of the grasses, from 21.43 to 66.77 million years ago. Semi-quantitative RT–PCR analysis and mining the digital expression database of rice showed that all 33 OsATG homologues could be detected in at least one cell type of the various tissues under normal or stress growth conditions, but their expression was tightly regulated. The 10 duplicated genes showed expression divergence. The expression of most OsATG homologues was regulated by at least one treatment, including hormones, abiotic and biotic stresses, and nutrient limitation. The identification of OsATG homologues showing constitutive expression or responses to environmental stimuli provides new insights for in-depth characterization of selected genes of importance in rice.     

水稻(Oryza sativa L.)苹果酸酶(OsNADP-ME3)基因在逆境下的表达特性研究

水稻苹果酸酶（NADP-ME）是多基因家族,由3个胞质型NADP-ME和1个质体型NADP-ME构成。本研究针对水稻胞质型成员（命名为NADP-ME3）（NM₀01061367）进行初步的功能解析。克隆获得的NADP-ME3基因的cDNA序列全长为2240bp,其中5’非翻译区为151bp,3’非翻译区为376bp,开放读码框（ORF）长1713bp,编码570个氨基酸。为研究NADP-ME3在逆境胁迫下的表达量变化,Northern blot检测结果显示,在NaCl、NaHCO3和PEG胁迫条件下,NADP-ME3随胁迫处理时间的不同表达量呈现不同程度的变化,推断NADP-ME3可能与非生物胁迫有应答关系,将NADP-ME3转入拟南芥中并通过观察转基因拟南芥在非生物胁迫下表型变化,发现NADP-ME3能够在一定程度上提高植物对非生物胁迫的耐受性。     

脆茎基因在水稻亚种间杂种后代中的遗传

脆茎基因在水稻粳型品种间杂种后代中按孟德尔遗传比例传递,而在亚种间杂种后代中呈脆茎(bcl bcl)频率显著增加和显著降低两种偏离孟德尔比例的异常分离型。Kamairazu／／Ketan Nangka／Kamairazu F1呈脆茎频率增加型,F2群体则呈正常分离,但F3和F4仍出现少数增加型和降低型的株系。IR36／Kamairazu的F2呈降低型分离,F3和F4株系中有正常型和降低型两种,F4还出现个别增加型株系。Ketan Nangka／IR36／／Kamairazu的F2中除了有增加型和降低型外,出现部分正常型。水稻bcl基因的异常分离与花粉育性无关,而是由雄配子选择性授精引起,与位于第3染色体的配子体基因ga2、ga3和ga14有关。     

Response of Tonoplast and Plasma Membrane ATPases in Chilling-Sensitive and -Insensitive Rice (Oryza sativa L.) Culture Cells to Low Temperature

Tonoplast and plasma membrane vesicles were prepared from chilling-sensitive(CS) and chilling-insensitive (CI) cultured cells of rice (Oryzasativa L.) to examine how they would respond to low temperature.With CS cells, the specific activity of ATPase in tonoplastvesicles was relatively higher than that of plasma membraneATPase. Tonoplast ATPase activity was decreased by low temperaturetreatment, and a slight decrease in plasma membrane ATPase activitywas also observed. The decrease in the specific activity ofthe tonoplast ATPase by low temperature may reflect a decreasein V_max. However, no change was noted in K_m. The break pointof the Arrhenius plots of the tonoplast ATPase was ca. 32?C,this value being ca. 9?C higher than that of the plasma membraneATPase. With CI cells, the specific activity of tonoplast ATPasewas somewhat less than that of the plasma membrane ATPase. TonoplastATPase activity was decreased by low temperature at 5?C, whereasan increase in plasma membrane ATPase activity was observed.The break point of the tonoplast ATPase activity was ca. 22?C,which was 3?C higher than that of the plasma membrane ATPase.Using ATPase solubilized from the plasma membrane or tonoplast,the Arrhenius plots of log ATPase activity against the reciprocalof absolute temperature gave a straight line fit from 5?C to45?C with no obvious break point. The break point appeared onadding a phospholipid mixture (asolectin) to a reaction mixturecontaining solubilized enzyme. The slope of the curve of theArrhenius plot was very different between the CS and CI cells.The plasma membrane and tonoplast ATPases from the CS cellshad a higher E_a above 20?C, whereas that from the CI cells hada lower one. These findings indicate that the tonoplast ATPase in a riceplant is more sensitive to low temperature than the plasma membraneATPase, with this response possibly being due to interactionsbetween the proteins and phospholipids. (Received January 6, 1988; Accepted July 5, 1988)     

水稻苗期生长对缺磷响应的QTL定位(英文)

缺磷是抑制全球水稻产量主要因素之一。本研究利用Asomonori(粳型)/IR24(籼型)杂交重组自交株系,对5个水稻苗期性状(相对苗高、相对根长、相对根重、相对苗重以及相对总重)在缺磷条件下的响应QTL进行定位。共检测到20个水稻苗期生长对缺磷响应的QTL位点,分别位于第1(4QTLs)、第4(4QTLs)、第5(2QTLs)、第7、第8(4QTLs)、第9(2QTLs)、第11(2QTLs)和第12号染色体上,其中13个QTLs位于与C3029C、XNpb302、C621B、C621C、R2976和C1263分子标记紧密连锁的6个基因组区域上。另外,每个性状均能从双亲中检测到正负效应QTL位点,这些能解释重组自交系群体中出现超亲和连续分布的现象。本文主要报道了水稻第5、第7和第11染色体上存在水稻苗期生长对缺磷响应的QTL位点。研究表明,该结果及其中检测到的QTLs两侧的连锁分子标记可用于水稻苗期耐低磷性分子育种。     

水稻rab5B基因在大肠杆菌中的表达、纯化和GTP结合分析

Rab5B类蛋白因为其编码产物的N端具有特殊结构而被认为是一类特殊的蛋白质.水稻rab5B基因Osrab5B是这类蛋白质基因在单子叶植物中的首例发现.将Osrab5B基因的编码序列按正确读码框重组到具有谷胱甘肽硫转移酶(glutathione S-transferase, GST)融合标签的pGEX-4T1表达载体中,转化大肠杆菌,获得了稳定表达目标融合蛋白的菌株,经GSTrapTM柱纯化,获得了纯化的目标融合蛋白.GTP结合试验表明,在原核细胞中表达出的GST-OsRab5B融合蛋白具有体外结合GTP的能力.