《植物生理学报》 2021, 57(4): 862-878
杧果RAV基因家族全基因组鉴定及表达分析
通信作者：马武强；E-mail: wuqiangma@hainanu.edu.cn
摘 要：
RAV蛋白是一类植物特有的转录因子, 在植物生长发育和抗逆过程中发挥重要作用, 目前在杧果(Mangifera indica)中尚未报道。本研究对杧果MiRAV进行了全基因组鉴定, 并利用已公布的转录组数据和实时荧光定量PCR (qPCR)实验分析了它们在不同组织(包括成熟叶片、成熟树皮、种子、根、花、果肉和果皮)、不同发育阶段果肉和果皮中, 以及采后低温和热水处理下的表达情况。结果表明, 杧果中共有6个RAV基因家族成员。基因组定位发现MiRAV分布于5条不同的染色体。联合拟南芥(Arabidopsis thaliana)、番茄(Solanum lycopersicum)、辣椒(Capsicum annuum)、酸橙(Citrus × aurantium)的RAV蛋白进行系统发育分析, 发现共可分为5个亚组。 MiRAV蛋白分布在第II和IV亚组, 同一亚组间有相似的保守基序和保守结构域分布。共线性分析发现, MiRAV存在6对片段重复基因, 推测MiRAV家族的扩张可能与片段复制事件有关。选择压分析发现, 共线性基因对非同义替换/同义替换(Ka/Ks)值远小于1, 说明MiRAV在进化过程中可能主要受到纯化选择作用。顺式作用元件分析显示, MiRAV含有光响应元件数量最多,其次是激素响应元件, 以及部分生物与非生物胁迫和生长发育相关元件。表达分析发现, MiRAV在‘Alphonso’杧果不同组织中的表达具有偏好性, MiRAV2和MiRAV3主要在果皮中发挥作用, MiRAV4主要果肉中发挥作用。 MiRAV2、 MiRAV3和MiRAV4与杧果果实发育和成熟以及应答逆境胁迫有关, MiRAV3还与‘肯特杧’应对低温造成的损伤应激和热水处理‘Ataulfo’杧果果实软化有关。本研究较为全面地对MiRAV家族进行了鉴定和分析, 为进一步探究RAV基因在杧果果实发育过程中的调控机制提供参考。
关键词：杧果; RAV基因家族; 果实发育; 低温和热水胁迫; 表达分析
收稿：2020-12-23 修定：2021-03-27
资助：海南省基础与应用基础研究计划(自然科学领域)高层次人才项目(2019RC082)和青年项目(320QN191)、海南大学引进高层次人才 启动基金(KYQD(2R)1970)和海南省研究生创新课题(Hys2019-90)。
Corresponding author: MA Wuqiang; E-mail: wuqiangma@hainanu.edu.cn
Abstract:
RAV protein is a type of plant-specifc transcription factor, which plays an important role in plant
growth, development and stress resistance. It has not been reported in mango (Mangifera indica) currently. Here, RAV gene in mango (MiRAV) was identifed at the whole genome level, and their expression in different tissues (including mature leaf, mature bark, seed, flower, flesh and peel), fleshes and peels of different developmental stages, and peels treated by hot water and low temperature after harvest was analyzed by published transcriptome data and real-time quantitative PCR (qPCR) experiments. The result show that six members of MiRAV gene family were obtained. Genome localization analysis showed that MiRAV is distributed on fve chromosomes. Phylogenetic analysis combined with RAV proteins of Arabidopsis thaliana, Solanum lycopersicum, Capsicum annuum and Citrus × aurantium showed that all of these proteins can be divided into fve classes. MiRAV proteins belong to II and IV subgroups, and similar conserved motifs and domains are displayed in the same subgroup. Collinearity analysis revealed that there are six pairs of fragment duplication genes in MiRAV, indicating that the expansion of gene family may be involved in fragment replication events. Selection pressure analysis found that the non-synonymous substitution/synonymous substitution (Ka/Ks) value of collinearity gene is much less than one, suggesting that MiRAV may be mainly affected by purifcation selection during the evolution process. Cis-regulatory elements analysis showed that MiRAV genes contain the largest number of light-responsive elements, followed by hormone-response elements, biological and abiotic stress elements, and growth and development related elements. Expression analysis showed that MiRAV has a preferential expression in different tissues of ‘Alphonso’ mango. MiRAV2 and MiRAV3 mainly play a role in the peel, while MiRAV4 mainly function in the flesh. MiRAV2, MiRAV3 and MiRAV4 are related to fruit development, maturation and response to adversity stress. MiRAV3 is also related to the damage stress of ‘Kent’ mango caused by low temperature and softening of ‘Ataulfo’ mango after hot water treatment. In this study, the MiRAV family was identifed and analyzed comprehensively. This study may provide a reference for further exploring the regulatory mechanism of RAV gene in mango fruit development.
Key words: mango (Mangifera indica); RAV gene family; fruit development; low temperature and hot water stress; expression analysis
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