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.     

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 TaWRKY10 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.     

Cloning and expression pro?les 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 identi?cation, 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 TaWRKY10 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. 2007 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.     

Cloning and expression pro?les 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 identi?cation, 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 TaWRKY10 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. 2007 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.     

水稻转录因子WRKY39的克隆表达及DNA结合活性分析

根据水稻转录因子WRKY39的cDNA基因序列设计引物,克隆水稻日本晴(Oryza sativa L.)WRKY类转录因子的cDNA基因并进行原核表达,通过凝胶阻滞实验分析融合蛋白与DNA的结合活性。结果克隆了1个新的WRKY基因OsWRKY39,其编码的蛋白OsWRKY39能够与顺式元件W盒特异结合,Os-WRKY39具有转录因子的最基本特征。     

AhBG1转基因拟南芥的ABA敏感性和抗旱性研究

为探究花生基因AhBG1对拟南芥ABA敏感性和抗旱性的影响,以过表达AhBG1拟南芥为材料,检测其ABA敏感性及脱水处理下ABA质量分数、叶片失水率、干旱存活率及ABA稳态相关基因表达变化.结果表明:AhBG1是编码花生β-葡萄糖苷酶的家族成员,定位于细胞质;与野生型相比,AhBG1过表达拟南芥植株在干旱条件下体内AB...     

Molecular characterization of GmNHX2, a Na＋/H＋ antiporter gene homolog from soybean,and its heterolosous expression to improve salt tolerance in Arabidopsis

Na＋/H＋ antiporters have been well documented to enhance plant salt tolerance by regulating cellular ion homeostasis. Here, a putative Na＋/H＋ antiporter gene homolog GmNHX2 from soybean was cloned and predicted to encode a protein of 534 amino acids with 10 putative transmembrane domains. GmNHX2 was expressed in all soybean plant tissues but enriched in roots and its expression was induced by NaCI and polyethylene glycol （PEG） treatments. GmNHX2 exhibits greater sequence similarity with LeNHX2 and AtNHX6 than that of AtNHX1 and AtSOS1. Although phylogenetic analysis clustered GmNHX2 with organellar （tonoplast and vesicles） antiporters, the GmNHX2-EGFP （enhanced green fluorescent protein） fusion protein was possibly localized in the plasma membrane or organelle membrane of transgenic plant cells, Furthermore, transgenic Arabidopsis plants expressing GmNHX2 were more tolerant to high NaCl concentrations during germination and seedling stages when compared with wild-type plants. These results suggest that GmNHX2 is a membrane Na＋/H＋ antiporter and may function to regulate ion homeostasis under salt stress.     

Overexpression of OsRAA1 promotes flowering and hypocotyls elongation in Arabidopsis

Previously, OsRAA1, an AtFPF1 homologue gene, was found to play an important role in modulating rice root development. In the current study, OsRAA1 was overexpressed in Arabidopsis, and the transgenic plants showed early flowering and elongated hypocotyl phenotypes as compared with the wild-type under white-light conditions. The hypocotyls of transgenic lines were twice as long as those of wild-type plants under red-light conditions but were indistinguishable from those of the wild-type under blue and far-red light and darkness. In addition, the phenotypes of AtFPF1 transgenic lines were similar to those of OsRAA1 transgenic lines. These results suggested that OsRAAI/AtFPF1 protein is involved in regulating flowering time and plays an important role in the inhibition of hypocotyl elongation under continuous red light. The functions were preserved during the evolution.     

Overexpression of rice OsLOL2 gene confers disease resistance in tobacco to Pseudomonas syringae pv. Tabaci

LSD1-related proteins of Arabidopsis with LSDl-like zinc finger domains regulate disease resistance and programmed cell death （PCD）. We cloned a rice OsLOL2 gene, orthologous to LSD1 of Arabidopsis and expressed it in a tobacco plant. Transgenic tobacco lines displayed enhanced disease resistance to a virulent bacterium Pseudomonas syringae pv. tabaci （Pst）. RT-PCR analysis showed that overexpression of OsLOL2 in transgenic tobacco lines resulted in upregulation of two pathogenesis-related （PR） protein genes, PR2 and PR5. Our results suggest that overexpression of OsLOL2 in transgenic tobacco enhances the resistance through the induction of PR proteins and hypersensitive response-like reaction.     

The foxtail millet Si69 gene is a Wali7 (wheat aluminum-induced protein 7) homologue and may function in aluminum tolerance

We isolated a clone, named Si69, from a foxtail millet immature seed cDNA library. The protein encoded by Si69 contains a conserved Wali7 (wheat aluminum induced protein 7) domain and shares high-level homology with aluminum-induced proteins from other species including rice and Arabidopsis. The Si69 gene presents as a single locus in foxtail millet genome and is globally expressed in all tissues examined. Its expression is up-regulated by aluminum. The sequence feature and expression pattern suggest that the Si69 gene is involved in aluminum tolerance or detoxification. To confirm its biological functions, Si69 controlled by the CaMV35S promoter was introduced into Arabidopsis. Transgenic plants did not show any visible morphological changes compared to wild-type plants under normal growth conditions. However, when treated with 20 or 50 μmol/L Aluminum (Al), the root apices of wild-type plants were heavily stained by hematoxylin, whereas those of Si69 transgenic plants were not stained when treated with 20 μmol/L Al and slightly stained when treated with 50 μmol/L Al. Scanning electron microscopy (SEM) results further demonstrated that the damage of the root apices was severer in wild-type plants than in transgenic plants. Inhibition of root growth and accumulation of malondialdehyde (MDA), an indicator of lipid peroxidation, were lower in transgenic plants than in wild-type plants. The results show that overexpression of Si69 may increase Al tolerance in transgenic plants, indicating that a series of Wali7-containing genes may play similar roles in Al tolerance/detoxification.     

棉花DELLA蛋白GhGAl4a基因在拟南芥中功能初步分析

为了探究棉花DELLA蛋白基因GhGAI4a在拟南芥中的功能,构建植物表达载体pBP35S：GhGAI4a和pBP35S：Ghgai4a,利用农杆菌介导的花滴法将其转入Col野生型拟南芥。结果显示,2个载体各自获得6个独立的转基因拟南芥纯合株系。分别统计48—96h种子萌发率,测量生长7d后幼苗的主根长度,与非转基因植株相比,过量表达GhGAI4a、Ghgai4a对拟南芥种子萌发及主根生长具有明显的抑制作用。1μmol／LGA处理转基因植株,GhGAI4a转基因植株主根长和萌发率均增大,而Ghgai4a转基因植株主根长度和萌发率均无显著变化。     

Complexes of MADS-box proteins are sufficient to convert leaves into floral organs

Genetic studies, using floral homeotic mutants, have led to the ABC model of flower development. This model proposes that the combinatorial action of three sets of genes, the A, B and C function genes, specify the four floral organs (sepals, petals, stamens and carpels) in the concentric floral whorls. However, attempts to convert vegetative organs into floral organs by altering the expression of ABC genes have been unsuccessful. Here we show that the class B proteins of Arabidopsis, PISTILLATA (PI) and APETALA3 (AP3), interact with APETALA1 (AP1, a class A protein) and SEPALLATA3 (SEP3, previously AGL9), and with AGAMOUS (AG, a class C protein) through SEP3. We also show that vegetative leaves of triply transgenic plants, 35S::PI;35S::AP3;35S::AP1 or 35S::PI;35S::AP3;35S::SEP3, are transformed into petaloid organs and that those of 35S::PI;35S::AP3;35S::SEP3;35S::AG are transformed into staminoid organs. Our findings indicate that the formation of ternary and quaternary complexes of ABC proteins may be the molecular basis of the ABC model, and that the flower-specific expression of SEP3 restricts the action of the ABC genes to the flower.     

Comparative mapping of QTLs for Al tolerance in rice and identification of positional Al-induced genes

Aluminum (A1) toxicity is the major factor limiting crop productivity in acid soils. In this study, a recombinant inbreed line (RIL) population derived from a cross between an A1 sensitive lowland indica rice variety IR1552 and an A1 tolerant upland japonica rice variety Azucena, was used for mapping quantitative trait loci (QTLs) for A1 tolerance. Three QTLs for relative root length (RRL) were detected on chromosome 1,9, 12, respectively, and 1 QTL for root length under A1 stress is identical on chromosome 1 after one week and two weeks stress. Comparison of QTLs on chromosome 1 from different studies indicated an identical interval between C86 and RZ801 with gene(s) for A1 tolerance. This interval provides an important start point for isolating genes responsible for A1 tolerance and understanding the genetic nature of Al tolerance in rice. Four A1 induced ESTs located in this interval were screened by reverse Northern analysis and confirmed by Northern analysis. They would be candidate genes for the QTL.     

Over-expression of an Arabi-dopsisd δ-OAT gene enhances salt and drought tolerance in transgenic rice

δ-OAT, ornithine-δ-aminotransferase, is the key enzyme involved in proline biosynthesis. In this study the Arabidopsis δ-OAT gene was transferred into rice (Oryza sativa L. ssp japonica cv. Zhongzuo 321), whose successful integration was demonstrated by PCR and Southern blot analysis. The over-expression of the gene in transgenic rice was also confirmed. Biochemical analysis showed that, under salt or drought stress conditions, proline contents in the leaves and roots in transgenic rice plants were 5- to 15-fold of those in non-transgenic controls. Under stress conditions, germinating rate of transgenic lines is higher than that of controls. Although the growth of rice plants tested were more and more retarded with the increasing of NaCI concentration, the transgenic plants grow faster compared to the controls under the same stress condition. Meanwhile, the resistance to KCl and MgSO4 stresses was also found enhanced in transgenie rice. Furthermore, the over-expression of δ-OAT also improved the yield of transgenic plants under stress conditions. The average yield per plant of transgenic lines increases about 12%--41% more than that of control line sunder 0.1 mol/L NaCI stress. These data indicated that the over-expression of δ-OAT, with the accumulation of proline, resulted in the enhancement of salt and drought tolerance and an increase of rice yield, which is of significance in agriculture.     

拟南芥SnRK2.2/2.3激酶参与调节镉胁迫响应的机制探究

为探究拟南芥SnRK2.2和SnRK2.3基因对Cd胁迫响应的分子机制. 以野生型（WT）、双突变体SnRK2.2/2.3、过表达SnRK2.2和过表达SnRK2.3的转基因植物为材料,研究SnRK2.2和SnRK2.3基因与Cd胁迫响应的关系.发现过表达两个基因可以提高拟南芥对Cd的耐受性,表现为可以减少Cd、丙二醛（MDA）及活性氧（ROS）的累积量,增加抗氧化酶CAT、POD和SOD的活性. qRT PCR结果显示在Cd胁迫下,两种过表达植株中铁转运蛋白IRT1和转录因子FIT、bHLH038和bHLH039表达水平受到明显抑制,ABA合成相关基因AAO3和NCED3的表达量显著上调.在Cd胁迫下,两种过表达植株中ABA含量显著高于WT和双突变体. 以上结果表明：拟南芥遭受Cd胁迫时,SnRK2.2和SnRK2.3基因通过下调IRT1基因表达从而减少植物对Cd的吸收,同时通过增加内源ABA含量来缓解Cd对植物的毒害.     

脱落酸对水稻根系生长素合成与运输的调控

利用一系列不同浓度的脱落酸(ABA)处理水稻幼苗根系,观察水稻初生根根长、冠根数目、侧根数目及长度、DR5::GUS转基因材料GUS活性、生长素的合成及运输相关基因表达水平等变化.结果表明,ABA可以抑制水稻初生根的伸长、冠根和侧根的发生和伸长,并且抑制效应呈现剂量效应.DR5::GUS活性在整个根系中随ABA处理浓度升高而减弱,但根尖部位的DR5::GUS活性经过ABA处理后表现增强;生长素合成相关基因YUCCA2、YUCCA4、YUCCA7,生长素运输载体基因AUX3、AUX5、PiN1b、PiN2、PiN5a、PiN9、PiN10a均显著下降表达.