小豆VaDREB1A基因克隆及其应答锈菌侵染的表达分析
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黑龙江八一农垦大学/国家杂粮工程技术研究中心/黑龙江省作物-有害生物互作生物学及生态防控重点实验室

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黑龙江自然科学基金(YQ2020C034);黑龙江省应用技术研究与开发计划(GA19B104);黑龙江省普通高等学校青年创新人才培养计划(UNPYSCT-2016201、UNPYSCT-2017113);黑龙江省农垦总局重点科研计划(HKKY190602)


Cloning of VaDREB1A Gene from Adzuki Bean and Expression Analysis in Response to Rust Infection
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Heilongjiang Bayi Agricultural University/National Coarse Cereals Engineering Research Center/Heilongjiang Provincial Key Laboratory of Crop-Pest Interaction Biology and Ecological Control

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Natural Science Foundation of Heilongjiang Province (YQ2020C034); Research and Development Plan of Applied Technology in Heilongjiang Province (GA19B104); University Nursing Program for Young Scholars with Creative Talents in Heilongjiang Province (UNPYSCT-2016201, UNPYSCT-2017113); Research Project of Heilongjiang Provincial Land Reclamation Bureau (HKKY190602)

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    摘要:

    为明确小豆VaDREB1A在抗锈菌侵染过程中的作用,本研究分别从小豆基因组DNA及cDNA中克隆了该基因,测序结果表明VaDREB1A无内含子,全长660 bp编码219个氨基酸,氨基酸序列包含1个AP2结构域,序列特征及系统发育分析表明VaDREB1A属于DREB/CBF亚家族A1亚类。启动子顺式元件分析发现,VaDREB1A启动子包含乙烯、水杨酸及生物和非生物胁迫等响应元件。对该基因响应豇豆单胞锈菌(Uromyces vignae)侵染的表达分析发现,与不接种对照相比,VaDREB1A在抗病品种中于接种后24 h和120 h显著下调,而在感病品种中于接种后120 h和192 h显著上调。在1-氨基环丙烷-1-羧酸(ACC)诱导小豆抗锈病过程中,VaDREB1A的表达与其在抗病品种中的表达相似,分别于ACC处理并接种后24 h和192 h显著下调。上述结果说明VaDREB1A可能参与小豆对锈病的抗性。

    Abstract:

    To clarify the role of VaDREB1A that interplays with rust fungus infection, the gene VaDREB1A from genomic DNA (gDNA) and cDNA were isolated from adzuki bean. VaDREB1A was found without intron and contained a 660-bp full-length coding sequence that encodes 219 amino acids. VaDREB1A was annotated with one AP2 conserved domain, and this belonged to the A1 subclass of DREB/CBF subfamily using amino acid sequence and phylogenic analysis. The promoter of VaDREB1A harbored several cis-elements that are involved in responsive to ethylene, salicylic acid, and biotic and abiotic stresses. RT-qPCR showed that in resistant cultivar the relative expression of VaDREB1A was significantly reduced at 24 h and 120 h post inoculation (hpi) with the rust fungus Uromyces vignae, whereas in the susceptible cultivar the gene was significantly up regulated at 120 and 192 hpi following infection. Moreover, expression of VaDREB1A was significantly inhibited at 24 and 192 hpi in susceptible cultivar if pretreated with 1-aminocyclopropane-1-carboxylic acid (ACC) pretreatment, as observed from the resistant cultivar. Collectively, these results provided a preliminary evidence that supports an involvement of VaDREB1A underlying adzuki bean rust resistance.

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柯希望,徐菁,张金鹏,等.小豆VaDREB1A基因克隆及其应答锈菌侵染的表达分析[J].植物遗传资源学报,2021,22(2):493-501.

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  • 收稿日期:2020-08-17
  • 最后修改日期:2020-10-07
  • 录用日期:2020-10-19
  • 在线发布日期: 2021-03-09
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