桃TGA家族鉴定及BABA诱导的抗病表达分析

doi:10.16420/j.issn.0513-353x.2020-1000

摘要/Abstract

摘要：

以高度保守的BRLZ（PF07716）和DOG1（PF14144）结构域为种子序列,通过生物信息分析共鉴定得到15个桃TGA（TGACG motif-binding factor）家族基因,这些基因分布于桃的4条染色体上,其编码蛋白大小介于333 ~ 546 aa,分子量介于37.07 ~ 61.47 kD,等电点介于6.01 ~ 8.59,均定位于细胞核上。根据系统进化关系,拟南芥、大豆、番茄、水稻和桃的 TGA 家族分为5 个亚族,其中桃 TGA 家族成员主要分布于第Ⅰ、Ⅱ和Ⅳ亚族。对桃TGA家族成员启动子区域的顺式调控元件进行预测分析,其启动子区含有至少1个激素或逆境胁迫响应元件。经RNA-seq 数据分析可知,β-氨基丁酸（β- aminobutyric acid,BABA）处理和匍枝根霉（Rhizopus stolonifer）侵染能诱导桃TGA成员表达,其中PpTGA1-1在处理后12 h内上调最为显著;显著表达的PpTGA1-1通过与PpNPR1蛋白相互作用赋予PpNPR1蛋白DNA结合功能,启动一系列病程相关（pathogenesis-related,PR）基因的表达,从而诱导果实敏化（priming）抗性。桃 TGA 家族成员（尤其PpTGA1-1）可直接响应激发子诱导和病原菌侵染,并通过修饰PpNPR1蛋白从而在防卫反应中发挥重要调控作用。

关键词: 桃, TGA, 转录因子, β-氨基丁酸, 诱导抗病, 敏化, 软腐病

Abstract:

The highly conserved amino acid sequences of BRLZ（PF07716）and DOG1（PF14144）domains were used to identify the TGA family members,and the results showed that 15 members of the TGA family from the peach genome could be identified through bioinformatics analysis. The TGAs were unevenly distributed on the 1st,2rd,6th and 7th Prunus persica chromosomes. The protein parameters of the determined TGA TFs listed as follows：the sizes ranging from 333 to 546 aa,molecular weights ranging from 37.07 to 61.47 kD,the isoelectric points ranging from 6.01 to 8.59,respectively. And all members of the TGA family were located in the nucleus. According to the phylogenetic relationship, the TGA family of Arabidopsis thaliana,Glycine max,Solanum lycopersicum,Oryza sativa and Prunus persica was divided into five subfamilies,among which the peach TGA family members were mainly distributed inⅠ,Ⅱ and Ⅳ clades. Moreover,the promoter regions of the TGA members contained at least one plant hormone or stress response-related elements. In addition,the RNA-sequencing data exhibited that the expressions of the 15 TGA family members were potentiated by BABA treatment and/or Rhizopus stolonifer infection,and PpTGA1-1 expression was rapidly up-regulated within 12 h after stimulation. Further investigation indicated that the physical interaction between PpTGA1-1 and PpNPR1 provided the DNA binding capacity required by PpNPR1 for its activation of pathogenesis-related genes（PR）and consequently induced the priming resistance in harvested peaches. Collectively,a distinct member of TGA gene family in Prunus persica,particularly in PpTGA1-1,directly modified the PpNPR1 and thus exerted a crucial regulatory function for defensive response.

Key words: Prunus persica, TGA, transcription factor, β-aminobutyric acid, induced resistance, priming, Rhizopus rot

中图分类号:

S662.1

黎春红, 汪开拓, 雷长毅, 许凤, 季娜娜, 蒋永波. 桃TGA家族鉴定及BABA诱导的抗病表达分析[J]. 园艺学报, 2022, 49(2): 265-280.

LI Chunhong, WANG Kaituo, LEI Changyi, XU Feng, JI Nana, JIANG Yongbo. Identification of TGA Gene Family in Peach and Analysis of Expression Mode Involved in a BABA-Induced Disease Resistance[J]. Acta Horticulturae Sinica, 2022, 49(2): 265-280.

图/表 13

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所用载体 Vector	引物名称 Primer name	寡核苷酸引物对 Oligonucleotide primers
pGADT7	PpNPR1-SmaI-F PpNPR1-BamHI-R	GGCCAGTGAATTCCACCCGGGCCCGGGATGGAATTCAAAGCCGGAGTC CAGCTCGAGCTCGATGGATCCGGATCCTTGATTGAGGGTGAGCATTCCA
pGBKT7	PpTGA1-1-SmaI-F PpTGA1-1-BamHI-R	CATGGAGGCCGAATTCCCGGGATGAATTCTCCATCCACCCAGTT ATGCGGCCGCTGCAGGTCGACGGCAGGCTCACGAGGACG

所用载体 Vector	引物名称 Primer name	寡核苷酸引物对 Oligonucleotide primers
pGADT7	PpNPR1-SmaI-F PpNPR1-BamHI-R	GGCCAGTGAATTCCACCCGGGCCCGGGATGGAATTCAAAGCCGGAGTC CAGCTCGAGCTCGATGGATCCGGATCCTTGATTGAGGGTGAGCATTCCA
pGBKT7	PpTGA1-1-SmaI-F PpTGA1-1-BamHI-R	CATGGAGGCCGAATTCCCGGGATGAATTCTCCATCCACCCAGTT ATGCGGCCGCTGCAGGTCGACGGCAGGCTCACGAGGACG

基因名称 Gene name	蛋白ID（NCBI） Protein ID	编码序列长度/bp CDS length	蛋白特性 Protein property
基因名称 Gene name	蛋白ID（NCBI） Protein ID	编码序列长度/bp CDS length	氨基酸数 Number of amino acids	分子量/kD MW	等电点 pI	不稳定系数 Instability index	平均亲水性 GRAVY
PpTGA1-1 PpTGA1-2 PpTGA2.2 PpTGA7-1 PpTGA7-2 PpTGA7-3 PpTGA9-1 PpTGA9-2 PpTGA9-3 PpTGA10-1 PpTGA10-2 PpTGA10-3 PpHBP-1b-1 PpHBP-1b-2 PpHBP-1b-3	XP_007205406.1 XP_020421616.1 XP_007218367.2 XP_020411363.1 XP_020411364.1 XP_007222089.2 XP_007205044.2 XP_020422804.1 XP_020422805.1 XP_007204568.1 XP_020423301.1 XP_020423302.1 XP_020423805.1 XP_007226716.1 XP_020413849.1	1 092 1 089 1 002 1 131 1 116 1 104 1 596 1 515 1 467 1 641 1 638 1 641 1 359 1 395 1 221	363 362 333 376 371 367 531 504 488 546 545 546 452 464 406	40.99 40.86 37.07 42.78 42.12 41.67 58.59 55.55 53.87 61.47 61.39 61.47 49.95 51.67 45.14	6.41 6.41 8.59 6.51 6.51 6.37 6.45 6.58 6.32 6.64 6.64 6.64 7.84 6.01 6.38	49.13 48.70 58.49 52.03 51.20 50.29 56.81 56.70 56.54 67.80 67.75 67.80 62.22 40.31 41.33	-0.468 -0.459 -0.570 -0.594 -0.587 -0.584 -0.538 -0.555 -0.520 -0.757 -0.757 -0.757 -0.616 -0.539 -0.509

基因名称 Gene name	蛋白ID（NCBI） Protein ID	编码序列长度/bp CDS length	蛋白特性 Protein property
基因名称 Gene name	蛋白ID（NCBI） Protein ID	编码序列长度/bp CDS length	氨基酸数 Number of amino acids	分子量/kD MW	等电点 pI	不稳定系数 Instability index	平均亲水性 GRAVY
PpTGA1-1 PpTGA1-2 PpTGA2.2 PpTGA7-1 PpTGA7-2 PpTGA7-3 PpTGA9-1 PpTGA9-2 PpTGA9-3 PpTGA10-1 PpTGA10-2 PpTGA10-3 PpHBP-1b-1 PpHBP-1b-2 PpHBP-1b-3	XP_007205406.1 XP_020421616.1 XP_007218367.2 XP_020411363.1 XP_020411364.1 XP_007222089.2 XP_007205044.2 XP_020422804.1 XP_020422805.1 XP_007204568.1 XP_020423301.1 XP_020423302.1 XP_020423805.1 XP_007226716.1 XP_020413849.1	1 092 1 089 1 002 1 131 1 116 1 104 1 596 1 515 1 467 1 641 1 638 1 641 1 359 1 395 1 221	363 362 333 376 371 367 531 504 488 546 545 546 452 464 406	40.99 40.86 37.07 42.78 42.12 41.67 58.59 55.55 53.87 61.47 61.39 61.47 49.95 51.67 45.14	6.41 6.41 8.59 6.51 6.51 6.37 6.45 6.58 6.32 6.64 6.64 6.64 7.84 6.01 6.38	49.13 48.70 58.49 52.03 51.20 50.29 56.81 56.70 56.54 67.80 67.75 67.80 62.22 40.31 41.33	-0.468 -0.459 -0.570 -0.594 -0.587 -0.584 -0.538 -0.555 -0.520 -0.757 -0.757 -0.757 -0.616 -0.539 -0.509