菠菜SoNAC转录因子家族的全基因组鉴定与分析

采用生物信息学分析方法,从菠菜基因组中筛选鉴定了57个菠菜NAC转录因子,并对其基因结构、编码蛋白和系统进化进行了分析;通过荧光定量聚合酶链式反应qRT-PCR分析,研究了高温和盐处理后菠菜叶片中NAC基因的表达模式.研究结果显示:菠菜NAC转录因子可以被归入2组17个亚组,GroupⅠ包含10个亚组,GroupⅡ包含...     

热诱导表达的水稻OsBBX30基因克隆和表达分析

生物信息学分析表明:OsBBX30基因启动子含有与逆境相关的作用元件HSE.为进一步了解OsBBX30基因在生物体内受热诱导,通过OsBBX30基因克隆构建原核表达和实时定量PCR分析,证实OsBBX30基因表达受热胁迫诱导,能增强大肠杆菌的耐热能力,为深入了解该家族基因和挖掘水稻耐热基因奠定基础.     

转录因子OsNAC2对逆境下水稻产量性状的影响

以水稻OsNAC2过表达、RNAi转基因株系和野生型（日本晴）为材料,分别在苗期和生殖期进行干旱和盐胁迫处理,探索逆境条件下OsNAC2对水稻产量性状的影响。结果表明,不论是在苗期还是生殖期,OsNAC2-RNAi株系的叶相对含水量均比野生型更高,对干旱胁迫具有更强的适应能力;而OsNAC2过表达株系则相反。虽然苗期和生殖期遭遇盐胁迫的OsNAC2-RNAi株系相比野生型具有更高的叶相对含水量,但是OsNAC2的过量表达与RNAi株系的产量性状跟野生型相比并没有明显不同。生殖期干旱和盐胁迫下转基因株系的产量性状分析显示：干旱胁迫下,OsNAC2-RNAi株系的结实率与野生型相比显著提高了20.8%~29.2%,千粒重则无明显差异;而OsNAC2过表达株系每株粒数和千粒重相比野生型株系均显著降低。虽然盐胁迫下OsNAC2-RNAi株系的分蘖数和有效穗数明显比野生型高,但单株粒数和千粒重则无明显差异。上述结果表明,OsNAC2-RNAi株系具有更强的耐旱性,对于干旱胁迫下水稻的产量有显著的提高作用。     

上海市11种常规粳稻抗稻瘟病基因分子标记检测

利用分子标记检测技术,对9种参加2020年上海市水稻区域试验的品种和2种本课题组新培育的新品系的共10个抗稻瘟病基因位点进行了检测.结果显示,Pi37,Pi41,Pi-d23个基因在11种水稻中出现的频率达100％,Pi2,Pi5,Pi9,Pi36,Pikm和Pib抗性基因在11种水稻中出现的频率分别为18.18％,9...     

Cloning and expression profiles of 15 genes encoding WRKY transcription factor in wheat （Triticum aestivem L.）

WRKY proteins are involved in various physiological processes, including biotic and abiotic stress responses, hormone responses and development. However, no systematic identification, expression and function analysis of WRKY genes in wheat were reported. In this study, we isolated 15 wheat cDNAs with complete open reading frame （ORF） encoding putative WRKY proteins using in silico cloning. Phylogenetic analysis indicated that the 15 wheat WRKY genes belonged to three major WRKY groups. Expression analysis revealed that most genes expressed drastically in leaf, except Ta WRKYIO which expressed in crown intensively. Four genes were strongly up-regulated with the senescence of leaves. Eight genes were responsive to low temperature, high temperature, NaCl or PEG treatment. Moreover, differential expression patterns were also observed between wheat hybrid and its parents, and some genes were more responsive to PEG treatment in the hybrid. These results demonstrated that wheat WRKY genes are involved in leaf senescing and abiotic stresses. And the changed expression of these WRKY genes in hybrid might contribute to the heterosis by improving the stress tolerance in hybrids.     

水稻低温胁迫响应 ｍｉｃｒｏＲＮＡ 鉴定

水稻是世界三大粮食作物之一,然而低温胁迫会严重抑制水稻的生长发育。为了探究micoRNA在水稻低温胁迫中的作用,采用低温处理前,5℃低温处理24h和5℃低温处理48h的2~3叶期水稻整株,构建9个小RNA文库。通过高通量测序后,对9个小RNA文库的microRNA进行差异表达分析,一共筛选出21个与冷胁迫相关的microRNA,其中16个在冷胁迫下上调,5个在冷胁迫下下调。通过对这21个microRNA靶基因的CO富集结果表明,其靶基因广泛富集在包括信号转导,免疫系统和物质合成等细胞内过程中。这表明水稻可能通过多种micoRNA 介导,从各个方面来协同抵御低温胁迫。本研究为进一步阐明microRNA响应低温胁迫的分子机制提供了基础,且本研究所鉴定的microRNA为增强水稻对低温耐受性遗传改良提供了优异的miRNA资源。     

Transformation of japonica rice with<Emphasis Type="Italic">RHL</Emphasis> gene and salt tolerance of the transgenic rice plant

Overexpression of the yeastHAL2 gene increases salt tolerance of yeast and plant. RiceHAL2-like (RHL) gene was introduced into ajaponica rice cultivar HJ19 withAgrobacterium tumefaciens-mediated transformation. Transgenic plants in R₀ generation were selected on the principle of GUS-positive,RHL gene PCR-positive and normal growth. Hygromycin-resistant plants of some transgenic lines in R₁ generation increased salt tolerance during the seedling and booting stage, being less damaged in the cytomembrane and stronger in leaf tissue viability under salt stress during booting period. Southern analysis of transgenic lines tolerant to salt in R₁ generation showed that theRHL gene expression cassette had been successfully integrated into rice genome. Moreover, gene engineering breeding methodology and really salt-tolerant rice cultivar were discussed.     

菠菜抗坏血酸过氧化物酶基因家族的鉴定及表达分析

利用生物信息学分析方法,在菠菜全基因组中鉴定出了菠菜(Spinacia oleracea)抗坏血酸过氧化物酶(APX)家族成员,并对其理化性质、亚细胞定位、基因结构、保守基序、同源关系及基因表达进行了分析,发现菠菜中存在7个SoAPXs(SoAPX1～7)基因,并通过进化树分析将菠菜APX家族分为4类.基因结构分析发现该家族基因由5～9个外显子构成.亚细胞定位预测表明大部分菠菜APX蛋白定位在细胞质.实时荧光定量聚合酶链式反应(qRT-PCR)结果表明:SoAPXs在各个组织器官中呈组成型表达,其中SoAPX1和SoAPX3的组织表达模式相似,SoAPX4和SoAPX5相似,SoAPX2在新叶中表达最高,SoAPX7在雄花中表达最高.对经胁迫处理后的样品进行表达分析发现,低温胁迫与氧化胁迫对SoAPXs的表达均有诱导作用,盐胁迫与干旱胁迫也刺激了大部分SoAPXs的表达.这些结果表明:SoAPXs可能在菠菜的抗盐、耐寒、抗旱以及抗氧化过程中起作用,为后续深入鉴定APX家族成员的功能提供参考.     

High-quality reference genes for quantifying the transcriptional responses of Oryza sativa L. (ssp. indica and japonica) to abiotic stress conditions

Rice (Oryza sativa L.) is important to food security and is also an excellent model plant for numerous cereal crops. A functional genomics study in rice includes characterization of the expression dynamics of genes by quantitative real-time PCR (qPCR) analysis; this is a significant key for developing rice varieties that perform well in the face of adverse climate change. The qPCR analysis requires the use of appropriate reference genes in order to make any quantitative interpretations meaningful. Here, the new potential reference genes were selected from a huge public database of rice microarray experiments. The expression stability of 14 candidates and 4 conventional reference genes was validated by geNorm PLUS and NormFinder software. Seven candidates are superior to the conventionally used reference genes in qPCR and three genes can be used reliably for quantitating the expression of genes involved in abiotic stress responses. These high-quality references EP (LOC_Os05g08980), HNR (LOC_Os01g71770), and TBC (LOC_Os09g34040) worked very well in three indica genotypes and one japonica genotype. One of indica genotypes including the Jasmine rice, KDML105 developed in Thailand for which no reference genes have been reported until now.     

Structural and expressional analysis of a cDNA that expresses predominantly in rice stamens

The development of stamen is an important stage during the sexual reproduction in higher plants. Studying the genes that predominantly or even specifically expressed in stamens will aid in undestanding the molecular mechanism of stamen development. The structure of a rice CHS-like gene (D5) which predominantly expressed in anthers has been analysised, showing high structural similarity with two other anther-specific genes (A1 and BA42) fromBrassica napus. Northern analysis also reveals that the three genes share a similar expression pattern and that the expression of D5 may relate to the early development of pollens. It is concluded that D5, A1 and BA42 represent a group of diverged CHS-like genes which are related to stamen development.     

分子标记辅助选育含有抗稻瘟病基因和软米基因两系不育系水稻新品系

以"浦软粳S"为转育亲本,利用分子标记辅助常规育种技术成功培育出含有Pi9,Pita,Pib和Pigm稻瘟病抗性基因及软米基因(Wx~(mq)),同时表现柱头外露率高的两系不育系水稻新品系"2179S"."2179S"不育系茎秆粗壮,矮杆大穗,株高为63.8 cm,柱头外露率平均为60%.研究结果为今后培育具有稻瘟病抗性的优质两系杂交水稻新组合提供不育系亲本.     

A vacuole localized β-glucosidase contributes to drought tolerance in Arabidopsis

The phytohormone abscisic acid (ABA) plays a critical role in plant growth, development, and adaptation to various stress conditions. The cellular ABA level is constantly adjusted to respond to changing physiological and environmental conditions. To date, the mechanisms for fine-tuning ABA levels remain elusive. Here, we report that BGLU10, a member of a multigene family of β-glucosidases, contributes to drought tolerance in Arabidopsis. The T-DNA insertion mutant bglu10 exhibited a droughtsensitive phenoty...     

出芽酵母蛋白激酶TOS3的过量表达及其在热胁迫应答中的作用

出芽酵母SNF1蛋白激酶参与葡萄糖阻遏和细胞胁迫应答.TOS3可通过磷酸化激活SNF1,参与SNF1调控的信号途径.本研究利用PCR方法扩增tos3基因的蛋白质编码序列,克隆到多拷贝表达载体pYES2/NTA上构建真核表达载体,转化酵母细胞并诱导TOS3过量表达.带HIS6标签的重组TOS3蛋白通过免疫印迹得以鉴定.进一步研究了过量表达TOS3对细胞热胁迫耐受性的影响,发现在热胁迫处理条件下,TOS3过量表达可恢复△snf1突变体细胞的生长缺陷,表明TOS3可能通过不依赖SNF1的其他信号途径参与细胞对热胁迫应答的调控.     

An on-line multi-parameter analyzing optical biosensor for real-time and non-invasive monitoring of plant stress responses in vivo

Photosynthetic dysfunction and reactive oxygen species （ROS） production are the common features of plant stress responses. Based on quantitative measurement of ROS production and delayed fluorescence （DF） emission, which is an excellent marker for evaluating photosynthesis, an on-line multiparameter analyzing optical biosensor for detecting plant stress responses was developed. Performances of the proposed biosensor were tested in the wild type （WT） Arabidopsis and heat shock protein （Hsp） 101 T-DNA knockout mutant （hsp101） plants with different thermotolerance. Results demonstrated that DF intensity correlates with net photosynthesis rate （Pn） in response to elevated temperature in both the WT Arabidopsis and hsp101 mutant plants. The light response characteristics and the recovery dynamics of the DF intensity were also in line with those of Pn in both the WT Arabidopsis and hsp101 mutant plants after heat stress （HS, 40℃ for 30 min）, respectively. In all experiments discussed above, the hsp101 plant showed the worse photosynthetic performance than the WT plant. Moreover, after HS, more ROS production in the hsp101 mutant than in WT Arabidopsis, which was found to be mainly localized at chloroplasts, could be directly detected by using the proposed biosensor. In addition, the hsp101 mutant showed severer chloroplasts alterations than the WT plant within the first 1 h of recovery following HS. Nevertheless, pre-infiltration with catalase （CAT） reduced ROS production and prevented the declines of the DF intensity. Therefore, HS-caused declines of photosynthetic performance might be due to oxidative damage to photosynthetic organelle. To sum up, we conclude that Hsp101 plays an important role in preventing oxidative stress, and the proposed optical biosensor might be a powerful tool to determine plant stress responses and identify plant resistant difference.     

菠菜HD-ZIP III基因家族鉴定与表达分析

植物叶片在生长发育过程中,近-远轴极性的建立与维持是叶片正常行使光合效应、呼吸作用、蒸腾作用的结构基础. HD-ZIP III是重要的叶片近轴极性基因,本研究在菠菜（Spinacia oleracea）基因组中鉴定到3个HD-ZIP III基因家族成员,依次命名为SoHB1,SoHB2和SoHB3,并对其蛋白理化性质、保守结构域和进化关系进行了分析. 研究结果表明：3个HD-ZIP III蛋白都具有4个保守结构域. 共线性分析表明：菠菜SoHB1,SoHB2和SoHB3分别与拟南芥ATHB8,ATREV和ATHB15存在共线性关系. 启动子顺式作用元件预测结果表明：菠菜HD-ZIP III家族基因启动子上游包含光响应、逆境胁迫反应、激素响应等顺式作用元件. 在不同叶型菠菜种质中的表达模式分析结果表明：单戟型叶片的SoHBs表达量最高;此外,经干旱胁迫处理后,SoHB1,SoHB2和SoHB3的基因表达量普遍下降;对菠菜包括根、茎、薹叶、功能叶、薹叶柄、功能叶柄等部位进行表达分析,发现SoHB1在根部表达最为丰富,SoHB2和SoHB3在薹叶处表达最为丰富.     

菠菜AMT基因家族鉴定与表达分析

铵态氮转运蛋白（AMT）负责铵态氮的吸收与转运,对植物的生长和发育起重要的调节作用.对菠菜SoAMT基因家族进行了基因组鉴定、生物信息学分析、组织表达谱及氮素响应表达谱分析,结果表明：菠菜基因组中共存在6个AMT的基因,包括5个AMT1成员和1个AMT2成员,主要分布在1,4,5,6条染色体上,编码区长度为1 443~15 06 bp;6个SoAMT蛋白均包含AMT基因家族特有的保守结构域及保守基序,含有9~11个跨膜结构域;SoAMT启动子含有较多茉莉酸、厌氧胁迫响应元件.SoAMT基因在根、叶、柄中均有表达,大部分SoAMT1亚家族成员受缺氮、硝态氮或铵态氮诱导表达,而SoAMT2基因表达量受铵态氮抑制.两类亚家族成员不同的氮响应模式,可能与其在氮素响应中的不同作用有关.