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利用花粉管通道法将BcWRKY2抗旱基因导入玉米的研究 被引量:6
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作者 曹士亮 王成波 +5 位作者 史桂荣 祁永红 王巍 贺强 王明泉 林忠平 《作物杂志》 CAS CSCD 北大核心 2013年第1期32-36,共5页
利用花粉管通道法将含有PPT抗性筛选标记的BcWRKY2转录因子基因导入优良玉米自交系。采用正交设计对玉米品种、导入时间及DNA浓度等条件进行了优化。对T0代种子进行了除草剂抗性筛选,结果表明玉米品种龙抗11授粉后16h进行导入及DNA导入... 利用花粉管通道法将含有PPT抗性筛选标记的BcWRKY2转录因子基因导入优良玉米自交系。采用正交设计对玉米品种、导入时间及DNA浓度等条件进行了优化。对T0代种子进行了除草剂抗性筛选,结果表明玉米品种龙抗11授粉后16h进行导入及DNA导入浓度为200ng/μL时效果最佳。对获得的除草剂抗性植株进行PCR及PCR-Southern检测,得到PCR阳性植株71株,初步证明了外源目的基因已整合到玉米基因组中。 展开更多
关键词 玉米 花粉管通道法 Bcwrky2抗旱基因
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农杆菌介导的BcWRKY2基因原位转化玉米茎尖的初步研究 被引量:5
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作者 曹士亮 李文滨 +2 位作者 王石 林忠平 卢翠华 《作物杂志》 CAS CSCD 北大核心 2013年第4期51-56,I0002,共7页
利用农杆菌介导的原位转化方法将植物抗旱相关转录因子BcWRKY2基因导入优良玉米自交系郑58、K10、黄早四和Mo17。采用正交设计对玉米品种、菌液浓度、乙酰丁香酮浓度(AS)、真空处理时间和表面活性剂Silwet-77浓度进行了优化。研究结果表... 利用农杆菌介导的原位转化方法将植物抗旱相关转录因子BcWRKY2基因导入优良玉米自交系郑58、K10、黄早四和Mo17。采用正交设计对玉米品种、菌液浓度、乙酰丁香酮浓度(AS)、真空处理时间和表面活性剂Silwet-77浓度进行了优化。研究结果表明,各因素对转化率影响顺序依次是菌液浓度>品种>AS浓度>真空处理时间>Silwet-77浓度。确定了最优转化条件为品种郑58、真空处理时间20min、菌液浓度为OD600=0.9、AS浓度100μmol/L、Silwet-77浓度0.05%。PCR及RT-PCR分子检测试验说明,BcWRKY2基因已经整合到玉米基因组中,并且得到表达。 展开更多
关键词 玉米 农杆菌介导 原位转化 Bcwrky2基因
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小麦TaWRKY2基因组分析及在进化和育种材料中的检测 被引量:5
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作者 董聪 孟肖 +4 位作者 张正斌 徐萍 卫云宗 张文胜 刘新月 《麦类作物学报》 CAS CSCD 北大核心 2015年第5期585-590,共6页
小麦WRKY转录因子在抗旱抗逆等方面有重要作用。为小麦抗旱抗逆分子模块组装育种提供参考,本研究在已有小麦TaWRKY2基因cDNA序列(GenBank登录号:EU665425)的基础上,从小麦品种晋麦79号中克隆获得TaWRKY2基因全长序列,并对TaWRKY2的... 小麦WRKY转录因子在抗旱抗逆等方面有重要作用。为小麦抗旱抗逆分子模块组装育种提供参考,本研究在已有小麦TaWRKY2基因cDNA序列(GenBank登录号:EU665425)的基础上,从小麦品种晋麦79号中克隆获得TaWRKY2基因全长序列,并对TaWRKY2的基因结构、基因组信息进行研究,同时检测其在部分小麦亲缘种、黄淮麦区水旱地代表品种、本课题组选育的高代品系及部分水旱地杂交组合F1代中的分布。结果表明,克隆所得到的TaWRKY2基因全长DNA序列包含4个外显子和3个内含子,其中,3个内含子长度变化范围为111~172bp;外显子和内含子的交接处序列符合GT-AG原则;与乌拉尔图小麦(Triticum urartu,AA)和粗山羊草(Aegilops tauschii,DD)中的片段序列同源性分别为93%和99%,与中国春小麦(AABBDD)1AS、1BS和1DS染色体的片段序列同源性分别为93%、88%和99%。除野生一粒小麦外,在小麦进化祖先种、黄淮海水旱地推广的代表品种、课题组选育的高代品系及F1代共78份材料中,都能够检测出该基因的普遍存在,说明该基因可应用于小麦抗旱抗逆分子设计育种中。 展开更多
关键词 小麦 Tawrky2基因 基因结构 基因组比较 基因检测
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Cloning of WRKY2 Gene in Tomato 被引量:1
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作者 李娜 于涌鲲 +4 位作者 赵福宽 汤世坤 孙清鹏 于同泉 路苹 《Agricultural Science & Technology》 CAS 2011年第8期1125-1128,共4页
[Objective]The aim was to explore the function of WRKY transcription factor in tomato.[Method]The primers were designed in this study according to the obtained WRKY fragments,and the total RNA from tomato treated with... [Objective]The aim was to explore the function of WRKY transcription factor in tomato.[Method]The primers were designed in this study according to the obtained WRKY fragments,and the total RNA from tomato treated with 100 μmol/L of JA for 6 h was used as the template for RT-PCR.[Result]The 608 bp fragment was obtained from tomato with RT-PCR method.Sequence analysis indicated that this sequence contained WRKYGQK conservative domain and the similarity with Capsicum annuum WRKY-c and Nicotiana tabacum NtWRKY-7 were 79% and 74%,respectively.[Conclusion]WRKY gene sequence in tomato was cloned successfully. 展开更多
关键词 TOMATO WRKY transcription factor wrky2 gene CLONING
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番茄中WRKY2基因的克隆
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作者 李娜 于涌鲲 +4 位作者 赵福宽 汤世坤 孙清鹏 于同泉 路苹 《中国农学通报》 CSCD 北大核心 2011年第2期95-98,共4页
WRKY转录因子在植物应对生物或非生物胁迫的反应及生长和发育中起重要作用,为研究WRKY转录因子在番茄中的功能,本研究根据课题组已经克隆的番茄WRKY转录因子片段设计引物,采用RT-PCR方法,从JA诱导的番茄幼苗中,获得了608bp的cDNA片段。... WRKY转录因子在植物应对生物或非生物胁迫的反应及生长和发育中起重要作用,为研究WRKY转录因子在番茄中的功能,本研究根据课题组已经克隆的番茄WRKY转录因子片段设计引物,采用RT-PCR方法,从JA诱导的番茄幼苗中,获得了608bp的cDNA片段。序列分析表明,该序列含有WRKYGQK保守结构域,与Capsicum annuum WRKY-c序列相似性是79%,与Nicotiana tabacum NtWRKY-7序列相似性是74%,表明已经成功克隆了番茄WRKY2基因片段。 展开更多
关键词 番茄 WRKY转录因子 wrky2基因 克隆
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WRKY2 and WRKY10 regulate the circadian expression of PIF4 during the day through interactions with CCA1/LHY and phyB 被引量:3
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作者 Shulei Wang Qingbin Sun +2 位作者 Min Zhang Chengzhu Yin Min Ni 《Plant Communications》 SCIE 2022年第2期121-137,共17页
WRKY transcription factors are known mostly for their function in plant defense,abiotic stress responses,senescence,seed germination,and development of the pollen,embryo,and seed.Here,we report the regulatory function... WRKY transcription factors are known mostly for their function in plant defense,abiotic stress responses,senescence,seed germination,and development of the pollen,embryo,and seed.Here,we report the regulatory functions of two WRKY proteins in photomorphogenesis and PIF4 expression.PIF4 is a critical signaling hub in light,temperature,and hormonal signaling pathways.Either its expression or its accumulation peaks in the morning and afternoon.WRKY2 and WRKY10 form heterodimers and recognize their target site in the PIF4 promoter near the MYB element that is bound by CCA1 and LHY under red and blue light.WRKY2 and WRKY10 interact directly with CCA1/LHY to enhance their targeting but interact indirectly with SHB1.The two WRKY proteins also interact with phyB,and their interaction enhances the targeting of CCA1 and LHY to the PIF4 promoter.SHB1 associates with theWRKY2 andWRKY10 loci and enhances their expression in parallel with the PIF4 expression peaks.This forward regulatory loop further sustains the accumulation of the two WRKY proteins and the targeting of CCA1/LHY to the PIF4 locus.In summary,interactions of two WRKY proteins with CCA1/LHY and phyB maintain an optimal expression level of PIF4 toward noon and afternoon,which is essential to sketch the circadian pattern of PIF4 expression. 展开更多
关键词 CCA1 LHY light signaling PIF4 expression wrky2 WRKY10
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The WRKY46–MYC2 module plays a critical role in E-2-hexenal-induced anti-herbivore responses by promoting flavonoid accumulation 被引量:2
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作者 Xin Hao Shuyao Wang +5 位作者 Yu Fu Yahui Liu Hongyu Shen Libo Jiang Eric SMcLamore Yingbai Shen 《Plant Communications》 SCIE CSCD 2024年第2期228-242,共15页
Volatile organic compounds(VOCs)play key roles in plant–plant communication,especially in response to pest attack.E-2-hexenal is an important component of VOCs,but it is unclear whether it can induce endog-enous plan... Volatile organic compounds(VOCs)play key roles in plant–plant communication,especially in response to pest attack.E-2-hexenal is an important component of VOCs,but it is unclear whether it can induce endog-enous plant resistance to insects.Here,we show that E-2-hexenal activates early signaling events in Ara-bidopsis(Arabidopsis thaliana)mesophyll cells,including an H2O2 burst at the plasma membrane,the directedflow of calcium ions,and an increase in cytosolic calcium concentration.Treatment of wild-type Arabidopsis plants with E-2-hexenal increases their resistance when challenged with the diamond-back moth Plutella xylostella L.,and this phenomenon is largely lost in the wrky46 mutant.Mechanistically,E-2-hexenal induces the expression of WRKY46 and MYC2,and the physical interaction of their encoded proteins was verified by yeast two-hybrid,firefly luciferase complementation imaging,and in vitro pull-down assays.The WRKY46–MYC2 complex directly binds to the promoter of RBOHD to promote its expres-sion,as demonstrated by luciferase reporter,yeast one-hybrid,chromatin immunoprecipitation,and electrophoretic mobility shift assays.This module also positively regulates the expression of E-2-hexenal-induced naringenin biosynthesis genes(TT4 and CHIL)and the accumulation of totalflavonoids,thereby modulating plant tolerance to insects.Together,our results highlight an important role for the WRKY46–MYC2 module in the E-2-hexenal-induced defense response of Arabidopsis,providing new in-sights into the mechanisms by which VOCs trigger plant defense responses. 展开更多
关键词 E-2-hexenal early signaling events WRKY46–MYC2 RBOHD NARINGENIN
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