孙宇 刘志鑫 叶子 罗睿雄 刘晓妹 蒲金基 张贺

摘要： 為了揭示RAV家族基因在杧果中的序列特征及其表达特性，采用生物信息学方法对杧果RAV家族基因进行序列分析，并通过qRT-PCR技术研究杧果胶孢炭疽菌和细菌性黑斑病菌侵染过程中该家族基因的相对表达量。结果表明，从杧果基因组中鉴定出6个RAV基因家族成员，其编码的蛋白质均具有AP2和B3超家族保守域结构，命名为MiRAV1～MiRAV6。MiRAV1和MiRAV5为不稳定亲水碱性蛋白质，MiRAV2和MiRAV3为不稳定亲水酸性蛋白质，MiRAV4为稳定亲水酸性蛋白质，MiRAV6为稳定亲水碱性蛋白质。杧果RAV基因编码的蛋白质主要定位于细胞核中，无规则卷曲和α-螺旋是6个MiRAVs蛋白二级结构的主要元件。系统进化树结合基序分析结果表明，在杧果与拟南芥、烟草、苹果、菠萝和毛果杨中，RAV家族基因具有多样性，在进化上结构具有保守性。qRT-PCR结果显示，胶孢炭疽菌侵染过程中，杧果RAV基因的相对表达量均显著下调;细菌性黑斑病菌侵染过程中，MiRAV1～ MiRAV6的相对表达量在3 h时均显著下调，MiRAV1、MiRAV2、MiRAV3和MiRAV6的相对表达量在6 h时均显著上调，MiRAV1、MiRAV5和MiRAV6的相对表达量分别在12 h、24 h、48 h、72 h时均显著上调。以上发现将为研究杧果RAV基因家族成员的功能和作用机制奠定基础。

关键词： 杧果;RAV基因家族;全基因组鉴定;表达分析

中图分类号： S667.7   文献标识码： A   文章编号： 1000-4440（2021）04-0957-11

Genome-wide analysis of the RAV gene family in mango

SUN Yu1，2， LIU Zhi-xin2， YE Zi1，2， LUO Rui-xiong3， LIU Xiao-mei1，2， PU Jin-ji1，2， ZHANG He1，2

（1.College of Plant Protection， Hainan University， Haikou 570228， China;2.Environment and Plant Protection Institute， Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Integrated Pest Management on Tropical Grops， Ministry of Agriculture and Rural Affairs， Haikou 571101， China;3.Tropical Crops Genetic Resources Institute， Chinese Academy of Tropical Agricultural Sciences， Haikou 571101， China）

Abstract： In order to reveal the sequence and expression characteristics of RAV gene in Mangifera indica， the sequence of the RAV gene family was analyzed using bioinformatics methods， and the relative expression level of RAV gene family was studied by qRT-PCR during the infection of Colletotrichum gloeosporioides and Xanthomonas campestris pv. mangiferaeindicae. The results showed that six RAV gene family members were identified from the mango genome， and the encoded proteins had the conserved domains of the AP2 and B3 superfamily， named MiRAV1-MiRAV6. MiRAV1 and MiRAV5 were unstable hydrophilic basic proteins， MiRAV2 and MiRAV3 were unstable hydrophilic acidic proteins， MiRAV4 was a stable hydrophilic acidic protein， and MiRAV6 was a stable hydrophilic basic protein. The proteins encoded by RAV gene of mango were mainly located in the nucleus， and random coil and α-helix were the main elements of secondary structure of six MiRAVs proteins. Phylogenetic tree construction combined with motif analysis showed that， among mango and Arabidopsis thaliana， tobacco， apple， pineapple and Populus tomentosa， the RAV family genes were diverse and their structure was conservative in evolution. The results of qRT-PCR indicated that the relative expression of RAV gene in mango was significantly down regulated during the infection of Colletotrichum gloeosporioides. During the infection of Xanthomonas campestris pv. mangiferaeindicae， the relative expression levels of MiRAV1-MiRAV6 were significantly down regulated at 3 h， and the relative expression levels of MiRAV1， MiRAV2， MiRAV3 and MiRAV6 were significantly up-regulated at 6 h， and the relative expression levels of MiRAV1， MiRAV5 and MiRAV6 were significantly up-regulated at 12 h， 24 h， 48 h and 72 h， respectively. These results will lay a foundation for further study on the function and mechanism of RAV gene family members in mango.

Key words： mango;RAV gene family;genomic identification;expression analysis

转录因子，又称反式作用因子，其具有特殊的结构可调控基因表达，是植物中重要的调控因子。植物转录因子在胁迫反应中，对调控基因表达发挥重要作用[1-3]，当植物受到逆境胁迫时，转录因子与相应的顺式作用元件结合，可以调控下游相关基因的表达 [4]。……