摘要
【目的】探究干旱胁迫下葡萄AQP的转录调控网络,为后续研究葡萄抗旱基因功能和转录调控机制提供理论依据。【方法】通过生物信息学方法分析葡萄AQP基因家族成员的理化性质、系统进化和染色体定位,进行共线性分析和转录因子调控预测,通过转录组数据分析该基因家族在干旱胁迫和脱落酸(abscisic acid,ABA)处理下的表达情况。【结果】37个VvAQP基因主要分为PIP、TIP、NIP、SIP四类,分别分布在17条染色体上,复制分析结果表明,共有5对节段复制和2对串联复制。蛋白序列分析表明,每个VvAQP蛋白序列中均有1个植物AQP主要内在蛋白(major intrinsic proteins,MIP)的保守结构域特征。通过转录调控网络预测发现,靶向VvAQP基因的转录调控因子主要分为16类。干旱胁迫下,根和叶中大部分VvAQP基因表达持续下调,VvPIP2-3、VvPIP2-6在根中持续上调,7个VvAQP基因受ABA显著诱导。干旱胁迫下15个差异表达的转录因子可能与13个VvAQP基因存在调控关系。【结论】鉴定并提供了葡萄AQP基因家族成员的基本信息,通过转录组数据发现可能参与干旱胁迫的VvAQP基因,并预测了这些基因的转录调控网络。
【Objective】Grape is one of the oldest fruit crops in the world. Most of the grapes in the world are cultivated in arid and semi-arid areas. Facing with the increasingly severe environment on the earth, the grape industry will face a serious impact in the future, drought will become one of the key factors restricting grape production. The aim of this study was to identify members of the AQPs gene family in the whole grape genome. In addition, the transcriptional regulatory network of VvAQPs under drought stress was explored to provide theoretical basis for the subsequent research on the function and the transcriptional regulation mechanism of drought resistance genes in grape.【Methods】Through BLAST Arabidopsis thaliana AQP protein sequences and the HMM file(PF00230), 37 VvAQPs were identified based on the grape genome database. The phylogenetic analysis, gene structure and protein sequence analysis, chromosome location and series replication analysis of grape AQP genes family were performed by using bioinformatics method, interspecific collinear analysis, targeted VvAQPs transcription factor regulatory network analysis. Through drought stress and ABA treatment to further identify the VvAQPs involved in drought, and explore possible transcriptional regulatory relationships.【Results】Thirty-seven VvAQPs members were identified from the grape genome. The sequence lengths of VvAQPs varied from 65 to 354, the molecular weights of VvAQPs varied from 6.79 to 37.33 kDa, and the isoelectric points of VvAQPs varied from 4.33 to 9.79. Most of the VvAQPs protein had 6 transmembrane domains. Phylogenetic analysis showed that the family was divided into PIPs, TIPs, NIPs and SIPs. These VvAQPs were distributed among 17 chromosomes, one VvAQP was distributed on 4,11, 12, 18random chromosome, and two on 2, 5, 9, 10, 16, 7random, Un chromosome. There were 3 VvAQPs on 3, 6, 13, 14, and 15, 4 VvAQPs on the chr8 chromosomes. VvAQPs were accompanied with5 pairs of segmental copies and 2 pairs of tandem copies. There were 27 AQP homologous genes between grape and Arabidopsis thaliana, 31 AQP homologous genes in tomato, 36 AQP homologous genes in peach, 55 AQP homologous genes in poplar and 44 AQP homologous genes in kiwifruit, respectively. In addition, the number of homologous genes on the 6, 8 and 13 chromosomes of grape were large, 5 VvAQPs were direct homologous genes with other 6 species, and 7 VvAQPs were homologous with other 5 dicotyledons. Gene structure and protein sequence analysis showed that there was a conserved domain characteristic(MIP) of plant aquaporin in each grape AQP protein sequence and genes from the same subgroup had similar structures. The prediction of transcriptional regulatory network revealed that the transcriptional regulatory factors targeting VvAQPs were mainly divided into 16 types(HD-ZIP, MYB, ERF, bZIP etc.). Combined with transcriptome data, most VvAQPs were significantly down-regulated in roots and leaves under drought stress, and VvAQPs in most roots or leaves were not differentially expressed during the T1 period of drought stress. In roots, VvPIP3-1, VvNIP3-1, VvTIP1-2 and VvTIP2-1 were significantly down-regulated, while VvPIP2-3 showed the opposite trend and was significantly up-regulated. VvNIP2-3, VvPIP1-3, VvNIP1-1 and VvTIP4-1 were significantly down-regulated in the later stage of drought stress, and VvTIP2-2 was only significantly down-regulated in the M4 drought-tolerant genotype. In leaves, VvTIP1-4, VvTIP2-1 and VvPIP3-2 were significantly down-regulated, VvTIP1-2 and VvNIP3-1 were up-regulated first and then down-regulated, and most genes of grape PIP subfamily members were significantly down-regulated in later stages, such as VvPIP1-3,VvPIP1-5, VvPIP2-1, VvPIP2-2, VvPIP2-4 and VvPIP2-5. VvAQPs were induced by ABA in different tissues. In roots, 4 VvAQPs were significantly up-regulated after being induced by ABA, such as VvPIP3-1, VvPIP1-6, VvPIP2-2 and VvTIP3-1, and VvNIP4-2 was down-regulated after ABA treatment.In leaves, VvNIP1-1, VvPIP3-1 and VvTIP1-3 were induced to be up-regulated, while VvPIP2-2 was induced to be down-regulated. Combined transcriptome expression data with transcriptional regulatory network prediction results, 15 differentially expressed transcription factors may regulate 13 VvAQPs under drought stress. Under drought stress, HD-ZIP(VIT16 s0098 g01170, VIT04 s0023 g01330 and VIT15 s0048 g02870) showed an up-regulation trend, and their target genes VvPIP2-1, VvPIP2-4 and VvTIP1-1 basically showed a trend of up-regulation first and then down-regulation. bZIP(VIT01 s0010 g00930, VIT18 s0001 g10450 and VIT15 s0046 g01440) was all up-regulated. Their target genes VvTIP1-4, VvNIP1-1, VvPIP1-5, VvPIP1-4, VvPIP2-5, VvTIP1-3 and VvTIP1-1 basically showed a downward-regulation trend in roots and leaves under drought stress. VvTIP1-2 was first up-regulated and then down-regulated in the leaves, while it was continuously down-regulated in roots. NAC(VIT01 s0026 g02710 and VIT19 s0014 g03290) all showed an up-regulation trend, and they jointly targeted the VvTIP2-1, which was continuously down-regulated in roots and leaves under drought stress. In addition, VIT11 s0016 g02410(MYBrelated) and target gene VvNIP3-1 continued to be down-regulated in roots under drought stress, VIT16 s0013 g01000(ERF) and target gene VvPIP1-7 were continuously down-regulated in roots under drought stress, and VIT07 s0197 g00060(G2-like) and target gene VvTIP2-1 showed opposite trends in roots and leaves under drought stress.【Conclusion】The basic information about grape AQPs family members was identified and provided, and the VvAQPs involved in drought stress and the predicted transcriptional regulatory networks were further identified, which provided a theoretical basis for further study on grape AQPs involved in drought stress and transcriptional regulation mechanism.
作者
杨盛迪
郭大龙
裴茂松
刘海楠
韦同路
余义和
YANG Shengdi;GUO Dalong;PEI Maosong;LIU Hainan;WEI Tonglu;YU Yihe(College of Horticulture and Plant Protection,Henan University of Science and Technology,Luoyang 471023,Henan,China;Henan Engineering Technology Research Center of Quality Regulation and Controlling of Horticultural Plants,Luoyang 471023,Henan,China)
出处
《果树学报》
CAS
CSCD
北大核心
2021年第10期1638-1652,共15页
Journal of Fruit Science
基金
国家自然科学基金(31701893)
河南省高校科技创新人才计划(21HASTIT035)
河南省高校科技创新团队支持计划(21IRTSTHN021)
河南省高等学校青年骨干教师培养计划(No.81)
洛阳市科技发展计划项目(2101102A)。
关键词
葡萄
AQP
干旱胁迫
转录因子
调控
Grapevine
AQP
Drought stress
Transcription factor
Regulation
作者简介
杨盛迪,男,在读硕士研究生,研究方向为园艺植物生长发育和逆境胁迫。Tel:18438615773,E-mail:yangshengdi2050@163.com;通信作者:余义和.E-mail:yuyihe@haust.edu.cn。
