苹果TIFY家族基因鉴定及其在虫害胁迫下的表达分析

doi:10.16420/j.issn.0513-353x.2020-0350

摘要/Abstract

摘要：

利用生物信息学方法在苹果全基因组中鉴定了TIFY转录因子家族基因,对基因结构、蛋白保守基序和进化关系进行了分析,同时基于新疆野苹果（Malus sieversii）转录组数据分析,明确了苹果TIFY家族基因在虫害胁迫条件下的差异表达情况。共鉴定到了16个苹果TIFY基因,其蛋白质大小介于209 ~ 449 aa之间。16个TIFY基因分布于苹果8条染色体中,所有的MdTIFY蛋白都具有保守的TIFY结构域。其中8个苹果MdTIFY与新疆野苹果转录组测序拼接转录本对应,这些基因均含有保守的TIFY功能域,多物种进化分析表明该类蛋白可聚类成7个组。7个TIFY基因的表达量在不同处理间表现出显著差异,虫害侵染后,抗虫株系中MsTIFY10B-a表达量升高34倍,MsTIFY9-c上升5.2倍。同时抗虫株系在自然条件下茉莉酸—异亮氨酸含量显著高于敏感株系。新疆野苹果中重要抗虫响应基因MsTIFY10B-a和MsTIFY9-c位于预测关联网络节点中心位置,使它们在调节植物对生物胁迫的响应中发挥重要作用。

关键词: 新疆野苹果, 基因家族, TIFY, 表达分析, 茉莉素

Abstract:

The TIFY transcription factors in the apple were identified using bioinformatics methods. The gene structure,conserved motifs and evolutionary relationship were analyzed. Meanwhile,the differential expression of apple TIFY family member genes under insect Agrilus mari stress conditions was determined based on Malus sieversii transcriptome data analysis. A total of 16 TIFY transcription factors were identified through analysis,which encodes proteins between 209-449 aa. The 16 MdTIFY genes are located on the 8 chromosomes of apples,and all MdTIFY proteins have conserved TIFY domains. Among them,eight MdTIFY correspond to the spliced transcripts of M. sieversii transcriptome sequencing. These genes all contain conserved TIFY functional domains. Multi-species phylogeny analyses showed that these proteins can be clustered into 7 groups. The expression levels of the seven TIFY genes showed significant differences among different treatments. After pest infestation,the expression of MsTIFY10B-a increased by 34-fold in insect-resistant plants,and the expression of MsTIFY9-c increased by 5.2-fold. Meanwhile the contents of jasmonic acid-isoleucine（JA-Ile）in insect-resistant plants were significantly higher than that of sensitive plants. The important insect-resistance response genes,MsTIFY10B-a and MsTIFY9-c,are located in the key nodes of association network in M. sieversii,being as important regulators during the plant responses tobiotic stresses.

Key words: Malus sieversii, gene family, TIFY, expression analysis, jasmonate

中图分类号:

S661.1

梅闯, 张小燕, 闫鹏, 艾沙江·买买提, 冯贝贝, 马凯, 韩立群, 董连新, 王继勋. 苹果TIFY家族基因鉴定及其在虫害胁迫下的表达分析[J]. 园艺学报, 2021, 48(2): 233-242.

MEI Chuang, ZHANG Xiaoyan, YAN Peng, Aisajan Mamat, FENG Beibei, MA Kai, HAN Liqun, DONG Lianxin, WANG Jixun. Identification of TIFY Family in Apple and Their Expression Analysis Under Insect Stress[J]. Acta Horticulturae Sinica, 2021, 48(2): 233-242.

图/表 7

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物种 Species	基因名 Gene name	基因号 Gene ID	物种 Species	基因名 Gene name	基因号 Gene ID
拟南芥 Arabidopsis thaliana	AthTIFY11A	AT1G17380.1	番茄 Solanum lycoperiscon	SLTIFY5A	Solyc01g097060.2
	AthTIFY10A	AT1G19180.1		SLTIFY10A	Solyc01g103600.2
	AthTIFY11B	AT1G72450.1		SLTIFY10b	Solyc03g122190.2
	AthTIFY10B	AT1G74950.1		SLTIFY8	Solyc06g065650.2
	AthTIFY3A	AT3G43440.1		SLTIFY10b-A	Solyc07g042170.2
	AthTIFY8	AT4G32570.1		SLTIFY5A-a	Solyc08g036620.2
	AthTIFY3B	AT5G20900.1		SLTIFY5A-b	Solyc08g036640.2
水稻 Oryza sativa	OsTIFY11a	Os03t0180800-01		SLTIFY5A-c	Solyc08g036660.2
	OsTIFY11c	Os03t0180900-01		SLTIFY10b-B	Solyc12g009220.1
	OsTIFY11b	Os03t0181100-01		SLTIFY10b-C	Solyc12g049400.1
	OsTIFY11g	Os03t0396500-00	葡萄 Vitis vinifera	VvTIFY5A	VIT_04s0008g00110.t01
	OsTIFY10a	Os03t0402800-01		VvTIFY8	VIT_04s0008g04950.t01
	OsTIFY3	Os04t0653000-01		VvTIFY10A	VIT_09s0002g00890.t01
	OsTIFY5	Os07t0153000-00		VvTIFY5A-a	VIT_10s0003g03790.t01
	OsTIFY10b	Os07t0615200-01		VvTIFY5A-b	VIT_10s0003g03800.t01
	OsTIFY10c	Os09t0439200-01		VvTIFY5A-c	VIT_10s0003g03810.t01
	OsTIFY11e	Os10t0391400-01		VvTIFY5A-d	VIT_01s0011g05560.t01
	OsTIFY11d	Os10t0392400-01		VvTIFY10A-a	VIT_11s0016g00710.t01
				VvTIFY10A-b	VIT_12s0035g00900.t01

物种 Species	基因名 Gene name	基因号 Gene ID	物种 Species	基因名 Gene name	基因号 Gene ID
拟南芥 Arabidopsis thaliana	AthTIFY11A	AT1G17380.1	番茄 Solanum lycoperiscon	SLTIFY5A	Solyc01g097060.2
	AthTIFY10A	AT1G19180.1		SLTIFY10A	Solyc01g103600.2
	AthTIFY11B	AT1G72450.1		SLTIFY10b	Solyc03g122190.2
	AthTIFY10B	AT1G74950.1		SLTIFY8	Solyc06g065650.2
	AthTIFY3A	AT3G43440.1		SLTIFY10b-A	Solyc07g042170.2
	AthTIFY8	AT4G32570.1		SLTIFY5A-a	Solyc08g036620.2
	AthTIFY3B	AT5G20900.1		SLTIFY5A-b	Solyc08g036640.2
水稻 Oryza sativa	OsTIFY11a	Os03t0180800-01		SLTIFY5A-c	Solyc08g036660.2
	OsTIFY11c	Os03t0180900-01		SLTIFY10b-B	Solyc12g009220.1
	OsTIFY11b	Os03t0181100-01		SLTIFY10b-C	Solyc12g049400.1
	OsTIFY11g	Os03t0396500-00	葡萄 Vitis vinifera	VvTIFY5A	VIT_04s0008g00110.t01
	OsTIFY10a	Os03t0402800-01		VvTIFY8	VIT_04s0008g04950.t01
	OsTIFY3	Os04t0653000-01		VvTIFY10A	VIT_09s0002g00890.t01
	OsTIFY5	Os07t0153000-00		VvTIFY5A-a	VIT_10s0003g03790.t01
	OsTIFY10b	Os07t0615200-01		VvTIFY5A-b	VIT_10s0003g03800.t01
	OsTIFY10c	Os09t0439200-01		VvTIFY5A-c	VIT_10s0003g03810.t01
	OsTIFY11e	Os10t0391400-01		VvTIFY5A-d	VIT_01s0011g05560.t01
	OsTIFY11d	Os10t0392400-01		VvTIFY10A-a	VIT_11s0016g00710.t01
				VvTIFY10A-b	VIT_12s0035g00900.t01

基因 Gene	基因号 Gene ID	基因组位置 Genomic position	开放阅读框/bp ORF	蛋白大小/aa Protein size	外显子 Number of exons
MdTIFY1	MD02G1096100	Chr02:7600659..7602728+	843	280	5
MdTIFY2	MD15G1220400	Chr15:17843310..17845336+	837	278	5
MdTIFY3	MD14G1238100	Chr14:31732049..31736399-	1 134	377	7
MdTIFY4	MD16G1020800	Chr16:1492513..1504489+	1 128	375	7
MdTIFY5	MD06G1228900	Chr06:35860218..35864674-	1 188	395	7
MdTIFY6	MD15G1225800	Chr15:18345401..18348150+	621	206	5
MdTIFY7	MD02G1105900	Chr02:8591750 8594168	633	210	5
MdTIFY8	MD13G1030000	Chr13:2164003..2166858+	1 149	382	7
MdTIFY9	MD09G1178600	Chr09:15274688..15275998-	615	204	2
MdTIFY10	MD15G1434400	Chr15:53470255..53474071+	810	269	5
MdTIFY11	MD16G1199200	Chr16:18010085..18014820+	1 026	341	9
MdTIFY12	MD13G1199500	Chr13:17721679..17726448+	1 050	349	9
MdTIFY13	MD08G1137500	Chr08:13179781..13184749+	1 338	445	8
MdTIFY14	MD15G1116100	Chr15:8188285..8192990+	1 350	449	7
MdTIFY15	MD16G1127400	Chr16:9274777..9277069+	636	211	4
MdTIFY16	MD13G1127100	Chr15:9515820..9518542+	630	209	4

基因 Gene	基因号 Gene ID	基因组位置 Genomic position	开放阅读框/bp ORF	蛋白大小/aa Protein size	外显子 Number of exons
MdTIFY1	MD02G1096100	Chr02:7600659..7602728+	843	280	5
MdTIFY2	MD15G1220400	Chr15:17843310..17845336+	837	278	5
MdTIFY3	MD14G1238100	Chr14:31732049..31736399-	1 134	377	7
MdTIFY4	MD16G1020800	Chr16:1492513..1504489+	1 128	375	7
MdTIFY5	MD06G1228900	Chr06:35860218..35864674-	1 188	395	7
MdTIFY6	MD15G1225800	Chr15:18345401..18348150+	621	206	5
MdTIFY7	MD02G1105900	Chr02:8591750 8594168	633	210	5
MdTIFY8	MD13G1030000	Chr13:2164003..2166858+	1 149	382	7
MdTIFY9	MD09G1178600	Chr09:15274688..15275998-	615	204	2
MdTIFY10	MD15G1434400	Chr15:53470255..53474071+	810	269	5
MdTIFY11	MD16G1199200	Chr16:18010085..18014820+	1 026	341	9
MdTIFY12	MD13G1199500	Chr13:17721679..17726448+	1 050	349	9
MdTIFY13	MD08G1137500	Chr08:13179781..13184749+	1 338	445	8
MdTIFY14	MD15G1116100	Chr15:8188285..8192990+	1 350	449	7
MdTIFY15	MD16G1127400	Chr16:9274777..9277069+	636	211	4
MdTIFY16	MD13G1127100	Chr15:9515820..9518542+	630	209	4