大理野茄M239响应黄萎病病菌侵染的差异表达蛋白分析

doi:10.7668/hbnxb.20194359

摘要/Abstract

摘要：

为了探索植物响应黄萎病病原菌侵染的作用机制,以黄萎病高感野茄材料大理野茄(M239)为研究对象,通过人工接种黄萎病病原菌大丽轮枝菌的方法,首先测定M239在接种大丽轮枝菌后0,6,12,24,48 h的根部4个关键生理指标,即过氧化物酶(POD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)的活性和可溶性蛋白(SP)含量的变化。结果确定接种大丽轮枝菌后12,24 h为蛋白组分析取样的关键时间点;在此基础上,利用iTRAQ技术分析M239在接种大丽轮枝菌后根部蛋白的变化。结果发现,与清水接种(CK)相比,M239接种大丽轮枝菌后12 h有463个差异表达蛋白(DEPs),其中305个下调,158个上调;接种后24 h,有456个DEPs被病原菌诱导表达,包括296个下调,160个上调。这些蛋白主要富集在碳代谢、乙醛酸和二羧酸代谢、次生代谢物的生物合成、丙酮酸代谢、新陈代谢等途径。通过与已报道的黄萎病抗性材料的研究结果进行对比分析发现,M239在响应大丽轮枝菌的侵染过程中,参与防御响应的蛋白数量和代谢通路较少,未能形成有效的防御网络,最终表现为感病。

关键词: 野茄, 黄萎病, 大丽轮枝菌, iTRAQ技术, 差异表达蛋白

Abstract:

In order to explore the mechanism of plant in response to Verticillium wilt pathogen infection,Dali wild eggplant (M239) was took as the material.By the artificial inoculation of Verticillium dahliae,four key physiological indicators of M239 root were determined at 0,6,12,24,and 48 h after inoculation with V.dahliae, which namely the peroxidase (POD) activity,catalase (CAT) activity,phenylalanine ammonilyase (PAL) activity and the soluble protein (SP) content.The result showed that 12 and 24 h after inoculation with V.dahliae were determined as the proper sampling time points for proteome analysis.On this basis,iTRAQ technique was used to analyze the protein changes of M239 root after inoculation with V.dahliae.Compared with CK,463 differentially expressed proteins (DEPs) were found in M239 root at 12 h after inoculation with V.dahliae,among which 305 DEPs were down-regulated and 158 were up-regulated.At 24 h after inoculation with V.dahliae in M239,456 DEPs were induced to express by pathogens,including 296 down-regulated and 160 up-regulated proteins.These proteins were mainly enriched in carbon metabolism,glyoxylate and dicarboxylic metabolism,biosynthesis of secondary metabolites,pyruvate metabolism,and metabolic pathways.Compared with the reported Verticillium wilt resistant materials,it was found that the number of proteins and metabolic pathways involved in the defense response of M239 in the process of responding to the infection of V.dahliae was less.Therefore,it failed to form an effective defense network,and finally manifested as susceptible to the disease.

Key words: Wild eggplant, Verticillium wilt, Verticillium dahliae, Isobaric tag for relative and absolute quantitation (iTRAQ), Differentially expressed proteins

蔚亚楠, 黎志彬, 龚亚菊, 鲍锐, 桂敏, 杜光辉, 刘家迅, 吴丽艳. 大理野茄M239响应黄萎病病菌侵染的差异表达蛋白分析[J]. 华北农学报, 2024, 39(1): 165-174. doi: 10.7668/hbnxb.20194359.

YU Yanan, LI Zhibin, GONG Yaju, BAO Rui, GUI Min, DU Guanghui, LIU Jiaxun, WU Liyan. Analysis of Differentially Expressed Protein of M239 in Response to Verticillium Wilt Pathogen Infection[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(1): 165-174. doi: 10.7668/hbnxb.20194359.

http://www.hbnxb.net/CN/Y2024/V39/I1/165

图/表 10

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编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	差异表达蛋白DEPs		P值 P value
编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	上调Up	下调Down	P值 P value
1	碳代谢(sly01200)	14	13	3.09×10^-7
2	乙醛酸盐和二羧酸代谢(sly00630)	8	4	5.16×10^-6
3	次生代谢物的生物合成(sly01110)	28	35	1.86×10^-6
4	丙酮酸代谢(sly00620)	6	6	7.71×10^-5
5	新陈代谢(sly01100)	41	47	2.41×10^-3
6	氨酰生物合成(sly00970)	4	5	2.42×10^-3
7	柠檬酸循环(sly00020)	4	3	2.78×10^-3
8	氨基酸的生物合成(sly01230)	5	11	5.56×10^-3
9	蛋白酶体(sly03050)	2	4	6.21×10^-3
10	糖解与糖代谢合成(sly00010)	4	6	1.18×10^-3
11	缬氨酸、亮氨酸和异亮氨酸的生物合成(sly00290)	0	4	1.19×10^-2
12	光合生物中的碳固定(sly00710)	2	5	1.64×10^-2
13	有机含硒化合物代谢(sly00450)	0	3	1.80×10^-2
14	α亚麻酸代谢(sly00592)	4	1	2.36×10^-2
15	脂肪酸代谢(sly01212)	5	2	2.80×10^-2
16	脂肪酸降解(sly00071)	5	0	2.96×10^-2
17	c5支链二元酸代谢(sly00660)	0	2	4.25×10^-2
18	单菌霉素生物合成(sly00261)	0	2	4.25×10^-2

编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	差异表达蛋白DEPs		P值 P value
编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	上调Up	下调Down	P值 P value
1	碳代谢(sly01200)	14	13	3.09×10^-7
2	乙醛酸盐和二羧酸代谢(sly00630)	8	4	5.16×10^-6
3	次生代谢物的生物合成(sly01110)	28	35	1.86×10^-6
4	丙酮酸代谢(sly00620)	6	6	7.71×10^-5
5	新陈代谢(sly01100)	41	47	2.41×10^-3
6	氨酰生物合成(sly00970)	4	5	2.42×10^-3
7	柠檬酸循环(sly00020)	4	3	2.78×10^-3
8	氨基酸的生物合成(sly01230)	5	11	5.56×10^-3
9	蛋白酶体(sly03050)	2	4	6.21×10^-3
10	糖解与糖代谢合成(sly00010)	4	6	1.18×10^-3
11	缬氨酸、亮氨酸和异亮氨酸的生物合成(sly00290)	0	4	1.19×10^-2
12	光合生物中的碳固定(sly00710)	2	5	1.64×10^-2
13	有机含硒化合物代谢(sly00450)	0	3	1.80×10^-2
14	α亚麻酸代谢(sly00592)	4	1	2.36×10^-2
15	脂肪酸代谢(sly01212)	5	2	2.80×10^-2
16	脂肪酸降解(sly00071)	5	0	2.96×10^-2
17	c5支链二元酸代谢(sly00660)	0	2	4.25×10^-2
18	单菌霉素生物合成(sly00261)	0	2	4.25×10^-2

编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	差异表达蛋白DEPs		P值 P value
编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	上调Up	下调Down	P值 P value
1	糖酵解/糖异生(sly00010)	8	7	8.65×10^-5
2	新陈代谢(sly01100)	34	68	1.49×10^-4
3	次生代谢物的生物合成(sly01110)	23	41	3.33×10^-4
4	氨基酸的生物合成(sly01230)	8	11	1.18×10^-3
5	碳代谢(sly01200)	7	13	2.87×10^-3
6	果糖和甘露糖代谢(sly00051)	2	6	4.66×10^-3
7	柠檬酸循环(sly00020)	2	5	4.97×10^-3
8	蛋白酶体(sly03050)	4	2	1.02×10^-2
9	苯丙类生物合成(sly00940)	4	10	1.69×10^-2
10	嘌呤代谢(sly00230)	6	5	2.41×10^-2
11	丙酮酸代谢(sly00620)	4	4	2.64×10^-2
12	氰胺酸代谢(sly00460)	2	3	2.75×10^-2
13	非同源末端连接(sly03450)	0	2	3.36×10^-2
14	丙氨酸、天冬氨酸和谷氨酸代谢(sly00250)	1	4	4.36×10^-2

编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	差异表达蛋白DEPs		P值 P value
编号 No.	KEGG富集通路(ID) Enriched KEGG pathways	上调Up	下调Down	P值 P value
1	糖酵解/糖异生(sly00010)	8	7	8.65×10^-5
2	新陈代谢(sly01100)	34	68	1.49×10^-4
3	次生代谢物的生物合成(sly01110)	23	41	3.33×10^-4
4	氨基酸的生物合成(sly01230)	8	11	1.18×10^-3
5	碳代谢(sly01200)	7	13	2.87×10^-3
6	果糖和甘露糖代谢(sly00051)	2	6	4.66×10^-3
7	柠檬酸循环(sly00020)	2	5	4.97×10^-3
8	蛋白酶体(sly03050)	4	2	1.02×10^-2
9	苯丙类生物合成(sly00940)	4	10	1.69×10^-2
10	嘌呤代谢(sly00230)	6	5	2.41×10^-2
11	丙酮酸代谢(sly00620)	4	4	2.64×10^-2
12	氰胺酸代谢(sly00460)	2	3	2.75×10^-2
13	非同源末端连接(sly03450)	0	2	3.36×10^-2
14	丙氨酸、天冬氨酸和谷氨酸代谢(sly00250)	1	4	4.36×10^-2

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