杧果转录因子BES1s家族全基因组鉴定及生物信息学分析

doi:10.3969/j.issn.1004-1524.2022.05.13

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (5): 984-994.DOI: 10.3969/j.issn.1004-1524.2022.05.13

杧果转录因子BES1s家族全基因组鉴定及生物信息学分析

夏煜琪¹^,²(), 孙宇¹^,², 刘志鑫², 孙瑞青¹^,², 杨楠¹^,², 蒲金基²^,³, 张贺¹^,²^,³^,^*()

1.海南大学植物保护学院热带作物学院,海南海口 570228
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,海南海口 571101
3.中国热带农业科学院热带生物技术研究所,海南海口 571101

收稿日期:2021-03-15 出版日期:2022-05-25 发布日期:2022-06-06
作者简介:* 张贺, E-mail: atzzhef@163.com
夏煜琪(1997—),女,河北张家口人,硕士研究生,研究方向为植物保护。E-mail: xiayuqi16@163.com
通讯作者: 张贺
基金资助:
国家重点研发计划(2019YFD1000504);中国热带农业科学院基本科研业务费(1630032022007);杧果病虫害监测与防治项目

Genome-wide identification and bioinformatics analysis of BES1 transcription factor family in mango

XIA Yuqi¹^,²(), SUN Yu¹^,², LIU Zhixin², SUN Ruiqing¹^,², YANG Nan¹^,², PU Jinji²^,³, ZHANG He¹^,²^,³^,^*()

1. College of Plant Protection, College of Tropical Crops, 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. Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China

Received:2021-03-15 Online:2022-05-25 Published:2022-06-06
Contact: ZHANG He

摘要/Abstract

摘要：

BES1/BZR1是植物中特有的一类转录因子且是油菜素内酯(BR)信号转导途径的唯一转录因子,为探讨其在植物抗病抗逆反应中的作用,本研究通过运用生物信息学研究方法分析了杧果BES1s家族成员的理化性质、结构域、蛋白质结构和在不同胁迫条件下的基因表达。研究表明,杧果BES1s家族理化性质预测结果显示,成员外显子个数相差较大;结构域预测结果显示,杧果BES1s成员均具有BES1_N结构域;系统进化分析和二级结构表明,成员可分成3组,不同组之间成员存在较大差异、蛋白质三级结构预测结果显示,存在3种结构。在qRT-PCR法测定基因表达量中发现,在胶孢炭疽菌(Cg)侵染过程中MiBES1.1和MiBES1.5持续上调表达;MiBES1.12和MiBES1.13持续下调表达。在细菌性黑斑病菌(Xcm)侵染过程中,在12 h时除MiBES1.7和MiBES1.9,其他成员上调表达;在3 h时MiBES1.1~MiBES1.5和MiBES1.11下调表达。在茉莉酸甲酯(MeJA)处理过程中,MiBES1.7、MiBES1.9和MiBES1.2持续下调表达;在48 h时,除MiBES1.5和MiBES1.11~MiBES1.13其他成员下调表达。本研究结果可为杧果BES1s家族成员基因的功能研究提供依据。

关键词: 转录因子, 杧果, 生物信息学

Abstract:

BES1/BZR1 is a unique transcription factor in the plant and the only transcription factor in the brassinolide (BR) signal transduction pathway. To explore the role BES1 plays on the stress resistance and diseases resistance in mango, we used bioinformatics methods to analyze the physicochemical properties, domains, protein structures and gene expression of BES1s family in mango. The results showed that in mango that the number of exons varies greatly, and all BES1s family members have the BES1_N domain. Evolution analysis and protein secondary structures showed that three groups were divided in BES1s family of mango, and there were significant differences among different groups. The qRT-PCR assay’s results showed that during the infection process of Cg, MiBES1.1 and MiBES1.5 were continuously up-regulated expression, and MiBES1.12 and MiBES1.13 lasted down-regulated expression; during the infection process of Xcm, mango’s BES1s family up-regulated expression at 12 h, except MiBES1.7 and MiBES1.9, and MiBES1.1~MiBES1.5 and MiBES1.11 down-regulated expression at 3 h; during the MeJA treatment process, MiBES1.7、MiBES1.9 and MiBES1.2 lasted down-regulation at 48 h, mango’s BES1s gene family down-regulated expression, except MiBES1.5 and MiBES1.11~MiBES1.13. A scientific basis is provided for the further study of the BES1s family function in mango.

Key words: transcription factor, mango, bioinformatics

中图分类号:

S667.7

夏煜琪, 孙宇, 刘志鑫, 孙瑞青, 杨楠, 蒲金基, 张贺. 杧果转录因子BES1s家族全基因组鉴定及生物信息学分析[J]. 浙江农业学报, 2022, 34(5): 984-994.

XIA Yuqi, SUN Yu, LIU Zhixin, SUN Ruiqing, YANG Nan, PU Jinji, ZHANG He. Genome-wide identification and bioinformatics analysis of BES1 transcription factor family in mango[J]. Acta Agriculturae Zhejiangensis, 2022, 34(5): 984-994.

图/表 9

表1 杧果BES1s家族成员理化性质预测

Table 1 Identification of BES1s gene family in mango

基因 Gene	氨基酸数 Number of amino acids/aa	蛋白质分子量 MW/ku	蛋白等电点 pI	亲水性平均系数 GRAVY	脂溶指数 Aliphatic index	不稳定指数 Instability index	外显子 Exon
MiBES1.1	629	71.63	5.98	-0.400	72.23	38.79	2
MiBES1.2	722	81.87	6.12	-0.502	70.11	44.91	11
MiBES1.3	697	77.81	5.60	-0.411	74.40	46.53	11
MiBES1.4	697	77.94	5.66	-0.414	74.68	46.41	10
MiBES1.5	303	33.38	9.06	-0.624	62.18	65.57	10
MiBES1.6	303	33.42	9.20	-0.669	62.18	66.06	2
MiBES1.7	326	35.26	8.38	-0.588	57.24	62.79	2
MiBES1.8	1 278	142.67	7.57	-0.434	78.42	47.88	21
MiBES1.9	321	34.63	8.62	-0.551	62.02	67.39	2
MiBES1.10	441	47.89	8.70	-0.375	64.85	55.19	6
MiBES1.11	322	34.83	8.86	-0.570	63.70	66.21	2
MiBES1.12	345	36.84	7.98	-0.482	59.19	66.70	4
MiBES1.13	652	73.06	6.59	-0.337	76.41	41.30	9

图1 杧果与番茄、辣椒、甘蓝、拟南芥、黄瓜、水稻BES1s家族成员的系统进化分析 BES1,杧果;Solyc,番茄;CA,辣椒;XP,甘蓝;AT,拟南芥;Cucsa,黄瓜;LOC,水稻。

Fig.1 Phylogenetic analysis of BES1s family members among mango and tomato, pepper, cabbage, Arabidopsis, cucumber, and rice BES1, mango;Solyc,tomato;CA, chili; XP, cabbage; AT, Arabidopsis; Cucsa, cucumber; LOC, rice.

图2 杧果BES1s家族蛋白保守结构域分析

Fig.2 Conserved domain analysis of BES1s protein in mango

图3 杧果BES1s基因家族系统进化分析和基因结构分析 A,杧果BES1s家族成员系统发育树;B,保守基序预测;C,二级结构预测。

Fig.3 phylogenetic and gene structure analysis of BES1s gene family in mango A, A phylogenetic tree of BES1s family members; B, Conservative motif prediction; C, Secondary structure prediction.

图4 杧果BES1s家族成员motif 1多序列比对

Fig.4 motif 1 of BES1s family members in mango

图5 杧果BES1s家族成员蛋白质三级结构预测

Fig.5 Prediction of protein structure of BES1s family members in mango

表2 qRT-PCR引物序列

Table 2 qRT-PCR primer sequences

基因 Gene	正向引物 Forward primer(5'→3')	反向引物 Reverse primer (5' →3')
MiBES1.1	GATGACTTGTTTGTGGCGGG	ATGTCCCCGCAATCTAGCAG
MiBES1.2	AGGATGGGATGGAGGTATCC	CCCTCGTTCACAGAAGAAGC
MiBES1.3	GCTGAATGCTGCATGGGATG	CAACAACTGCTTCCCCATGC
MiBES1.4	TCCTGATGGAACCACCTTTC	GCTGAAGATTGGGACTGAGC
MiBES1.5	AGCACCTCAACCAGGATCAC	ATCCGGAGAGGAGGAAGAGA
MiBES1.6	TTTTCCACTCCTCCGAAATG	CAACTGACGATGCATTGGAC
MiBES1.7	AAGAGACGAGAGCGCAGAAG	TAGCTGATGCAGACCCTCCT
MiBES1.8	CTACACAACGATGGGCAATG	TAGCTGACTGAGGCCTGGTT
MiBES1.9	TCTTCATTTCCAAGCCCAAC	TTACCCAACGACCAGAGTCC
MiBES1.10	AGGAAGAAGTTATAGCTGGTGGTG	ACATTCTTCATGAATCCTCTCTCC
MiBES1.11	CCACCTCTATCATCTCCAACTTCT	ATCACATTCTGGTATTGTGACAGG
MiBES1.12	TTAGAGATGAGGTTCTGTCAGGTG	AACTGGTTCTAGAGTTTCCCAGTG
MiBES1.13	TGTAGACCTTTGGAAGAGAAGGAG	CAGGAGAGATTCCTCTTTGAGAAG
Miactin	GGCAAGTCTGGTGCCAG	ACGGTATCTATCTCTTCG

图6 产物扩增结果

Fig.6 Results of product amplification 1-13, MiBES1.1-MiBES13; 14, Miactin; M, DL2 000 marker。

图7 qRT-PCR中杧果BES1s基因家族成员相对表达分析 “*”表示显著上调,“#”表示显著下调。A,胶孢炭疽菌侵染下杧果BES1s基因家族成员相对表达分析; B,细菌性黑斑病菌侵染下杧果BES1s家族成员相对表达分析; C,施用茉莉酸甲酯条件下杧果BES1s家族成员相对表达分析。

Fig.7 Relative expression analysis of BES1s gene family members of mango with qRT-PCR ‘*’ the relative expression level of table was significant up-regulated,“#” the relative expression level of table was significant down-regulated. A, relative expression analysis of BES1 gene family members under the infection of Cg in mango, B, relative expression analysis of BES1s family members under the infection of Xcm in mango; C, relative expression analysis of BES1s gene family members under the infection of MeJA in mango.

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杧果转录因子BES1s家族全基因组鉴定及生物信息学分析

Genome-wide identification and bioinformatics analysis of BES1 transcription factor family in mango

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