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杧果CPP转录因子家族基因的鉴定及表达分析
杨楠1, 孙瑞青1, 孙宇1, 夏煜琪1, 刘志鑫2, 高庆远1, 蒲金基1, 党志国3, 张贺1
0
(1.海南大学热带作物学院;2.中国热带农业科学院环境与植物保护研究所/农业农村部热带作物有害生物综合治理重点实验室;3.中国热带农业科学院热带作物品种资源研究所)
摘要:
为了探索杧果(Mangifera indica)CPP(cysteine-rich polycomb-like protein)基因家族的序列特征与表达特征,通过生物信息学的方法对杧果CPP基因家族进行序列分析,并通过qRT-PCR技术研究杧果细菌性黑斑病菌(Xanthomonas campestris pv. mangiferaeindicae,Xcm)和胶孢炭疽菌(Colletotrichum gloeosporioides,Cg)侵染过程中杧果CPP家族基因的转录水平和组织特异性表达水平。结果表明,从杧果基因组中鉴定了10个CPP基因家族成员,均含有保守的CRC结构域,均为不稳定疏水的酸性蛋白,与梨、长春花、胡杨、水稻、葡萄和拟南芥等6个物种69个CPP转录因子成员构建系统进化树,分为7个进化分支,杧果的10个CPPs基因分布在5个进化分支中,多和梨的CPP家族成员聚类在一起。qRT-PCR分析表明,在杧果细菌性黑斑病菌(Xcm)侵染过程中,MiCPP1和MiCPP2在侵染3 h、6 h、12 h后上调表达;杧果胶孢炭疽菌(Cg)侵染的0-72 h时,MiCPP8呈上调表达,而MiCPP9则为下调表达;为后续开展杧果CPP基因家族成员响应病原菌侵染的机制研究奠定基础。qRT-PCR分析表明,MiCPP8在幼叶、花、芽的相对表达量均很高而MiCPP7在幼叶、花、芽中的相对表达很低,MiCPP4与MiCPP9在幼叶、花中的相对表达量高而在芽中的表达量却很低,MiCPP6在花中的相对表达量很低,说明杧果MiCPPs基因家族成员在不同组织中具有表达的特异性。
关键词:  杧果  CPP转录因子  杧果细菌性黑斑病菌  杧果胶孢炭疽菌  qRT-PCR
DOI:10.13430/j.cnki.jpgr. 20210319001
投稿时间:2021-03-19修订日期:2021-05-08
基金项目:国家重点研发专项(2019YFD1000504,2019YFD1001103);中国热带农业科学院基本科研业务费专项资金(1630042017019);农业农村部项目“滇桂黔石漠化地区特色作物产业发展关键技术集成示范”“杧果病虫害监测与防治项目”
Identification and Expression Analysis of CPP Transcription Factor Family Genes in Mango
YANG Nan1, SUN Rui-qing1, SUN Yu1, XIA Yu-qi1, LIU Zhi-xin2, GAO Qing-yuan1, PU Jin-ji1, DANG Zhi-guo3, ZHANG He1
(1.College of Tropical Crops, Hainan University;2.Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture and Rural Affairs;3.Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences)
Abstract:
The sequence analysis of CPP (cysteine-rich polycomb-like protein) gene family in mango (Mangifera indica) was carried out by bioinformatics methods, and the transcription level and tissue-specific expression of the CPP family genes during the infection of Xanthomonas campestris pv. mangiferaeindicae (Xcm) and Colletotrichum gloeosporioides (Cg) were studied using qRT-PCR. Ten members of the CPP gene family were identified from the mango genome, all of which contained conserved CRC domains, and they were all unstable hydrophobic proteins. A phylogenetic tree was built with 69 members of the CPP gene family from mango, Pyrus bretschneideri, periwinkle, Populus euphratica, rice, grape, and Arabidopsis thaliana, which were divided into 7 evolutionary branches. The 10 MiCPPs genes of mango were distributed in 5 of the evolutionary branches, most of which were clustered with members of the CPP gene family of Pyrus bretschneideri. qRT-PCR analysis showed that the relative expression levels of MiPP1 and MiCPP2 were up-regulated at 3 h, 6 h and 12 h after the Xcm infection, while the relative expression level of MiCPP8 was up-regulated and that of MiCPP9 was down-regulated at 0-72 h after the Cg infection. qRT-PCR analysis also showed that the relative expression levels of MiCPP8 were very high in young leaves, flowers and buds infected by Colletotrichum gloeosporioides, while the relative expression levels of MiCPP7 were very low in those parts, the relative expression levels of MiCPP4 and MiCPP9 were high in young leaves and flowers but very low in buds, and the relative expression of MiCPP6 was very low in flowers. These results indicated the expression specificity of the mango MiCPPs gene family members in different tissues. This research would provide a basis for further studies on the response mechanism of mango CPP gene family members to pathogen infection and help breed for disease resistance in mango.
Key words:  Mangifera indica  CPP transcription factor  Xanthomonas campestris pv. mangiferaeindicae  Colletotrichum gloeosporioides  qRT-PCR

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