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  • 高鹏华,张 颢,晏慧君,等.月月粉TGAs基因结构及RcTGA2抗灰葡萄孢菌功能分析[J].植物遗传资源学报,2021,22(4):1157-1166.    [点击复制]
  • GAO Peng-hua,ZHANG Hao,YAN Hui-jun,et al.Analysis of Gene Structure of TGAs and Function of RcTGA2 in Rosa chinensis ‘Old Blush’ Against Botrytis cinerea[J].植物遗传资源学报,2021,22(4):1157-1166.   [点击复制]
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月月粉TGAs基因结构及RcTGA2抗灰葡萄孢菌功能分析
高鹏华1, 张 颢2, 晏慧君2, 陈宇春3, 范元兰3, 鄢 波1, 邱显钦2
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(1.西南林业大学/国家林业局西南风景园林工程技术研究中心/云南省功能性花卉资源及产业化技术工程研究中心;2.云南省农业科学院花卉研究所;3.云南大学生命科学学院)
摘要:
TGA(TGACG motif-binding factor)转录因子是bZIP转录因子家族中重要的一组,对植物病原菌侵染具有广谱抗性。本研究鉴定了‘月月粉’月季(Rosa chinensis ‘Old Blush’)TGA家族成员,并对其理化性质、亚细胞定位、进化特征和表达模式进行分析。鉴定获得7个RcTGAs,均包含bZIP1(cl21462)和DOG1(pfam14144)的保守结构域,氨基酸大小为324~545aa,分子量在36.68~60.17KD之间,等电点在5.52~8.96之间,二级结构主要为α-螺旋,均为亲水性蛋白,主要位于细胞核内。进化分析将来自‘月月粉’月季、拟南芥(Arabidopsis thaliana)、野草莓(Fragaria vesca)、水蜜桃(Prunus persica)、苹果(Malus domestica)的TGAs分为6个亚组。qPCR实验表明,灰葡萄孢菌侵染月季花瓣RcTGA4表达量无显著变化,RcTGA6随侵染时间的延长表达量呈下降趋势。RcTGA1/2表达呈显著上升的趋势,RcTGA3/5/7呈现先上升后下降的趋势,表明RcTGA1/2/3/5/7可能在月季-灰霉菌互作过程中发挥了重要作用,这5个基因可作为进一步抗病研究和功能分析的候选基因。通过VIGS沉默RcTGA2基因,灰葡萄孢菌侵染48 h后,月季花瓣病斑直径显著增大,表明RcTGA2与月季对灰葡萄孢菌的抗性有关。
关键词:  TGAs  月季灰霉病  灰葡萄孢菌  生物信息学分析  实时荧光定量PCR  VIGS
DOI:10.13430/j.cnki.jpgr.20201212001
投稿时间:2020-12-12修订日期:2021-01-22
基金项目:国家重点研发计划(2018YFD1000400);云南省科技厅科技人才引进与培养专项“科技领军人才”(2016HA005);云南省“万人计划”青年拔尖人才和云岭产业技术领军人才培养项目;云南省高校科技创新团队支持计划
Analysis of Gene Structure of TGAs and Function of RcTGA2 in Rosa chinensis ‘Old Blush’ Against Botrytis cinerea
GAO Peng-hua1, ZHANG Hao2, YAN Hui-jun2, CHEN Yu-chun3, FAN Yuan-lan3, YAN Bo1, QIU Xian-qin2
(1.Southwest Forestry University/ State Forestry Administration Southwest Landscape Gardening Engineering Technology Research Center/ Functional Flower Resources and Industrialization Technology Engineering Research Center of Yunnan Province;2.Flower Research Institute, Yunnan Academy of Agricultural Sciences;3.School of Life Sciences, Yunnan University)
Abstract:
TGA (TGACG motif binding factor) transcription factors are an important group of the bZIP transcription factor family, which have broad-spectrum resistance to plant pathogens. Rosa chinensis ‘Old Blush’ TGA family members were identified in this study, with their physicochemical properties, subcellular localization, evolutionary characteristics and expression patterns analyzed. Seven RcTGAs were identified, all containing the conserved domains of bZIP1 (cl21462) and DOG1 (pfam14144), with the amino acid size of 324 to 545 aa, the molecular weight of 36.68 to 60.17 KD, the isoelectric point of 5.52 to 8.96, and the secondary structure of mainly α-helix, all being hydrophilic proteins mainly located in the nucleus. TGAs from Rosa chinensis ‘Old Blush’, Arabidopsis thaliana, Fragaria vesca, Prunus persica, and Malus domestica were divided into six subgroups through evolutionary analysis. The qPCR test showed that the expression of RcTGA4 did not significantly change in the petals after being infected by Botrytis cinerea, and the expression of RcTGA6 decreased after the infection, whereas the expression of RcTGA1/2 increased significantly, and the expression of RcTGA3/5/7 increased first and decreased later, indicating that RcTGA1/2/3/5/7 may play an important role in the resistance to B. cinerea, and could be candidate disease-resistance gene for further research and functional analysis. After silencing the expression of RcTGA2 by VIGS, the lesion diameter in rose petals increased significantly 48 h after infection by B. cinerea, indicating that RcTGA2 may be closely related to the resistance of rose to B. cinerea.
Key words:  TGAs  rose gray mold  Botrytis cinerea  bioinformatics analysis  real time fluorescent quantitative PCR  VIGS

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