桃叶片低温响应转录组分析

doi:10.7668/hbnxb.20194251

摘要/Abstract

摘要：

抗寒是桃育种工作重要的目标性状,受多个基因的调控,转录组测序是对特定生理条件下的转录本进行测序分析,用以深入探究桃叶片低温响应的分子机制。以熊岳巨桃的叶片为试验材料对4 ℃低温(LT)处理下和正常叶片(RT)进行转录组测序分析。利用DESeq2进行差异表达分析,获得72个上调差异表达基因和3个下调表达基因。KEGG代谢通路分析差异表达基因的代谢通路,并进行蛋白的互作预测。结果显示,低温处理的叶片呈现卷曲、变黄和失水的生理状态,差异代谢基因主要富集在植物病原体互作、MAPK信号通路和次生物质代谢积累等途径。MYC2、SPCH 和Pti4~6转录因子可能在响应冷胁迫中调节低温带来的伤害中起到重要作用,MYC2和CBF蛋白预测相互作用,且分别与多个蛋白互作。本研究表明,有多个代谢途径共同响应桃叶片的低温胁迫,还预测到了可能在冷反应调节途径中起到关键作用的相关转录因子,为探究桃响应冷胁迫的分子机制打下基础。

关键词: 桃, 叶片, 低温, 转录组分析

Abstract:

Cold resistance is an important target trait for peach breeding and is regulated by multiple genes.Transcriptome sequencing is used to sequence transcripts under specific physiological conditions to explore the molecular mechanism of low temperature response of peach leaves.Leaves of Xiongyuejutao under 4 ℃ low temperature(LT)treatment and normal leaves(RT)were analyzed by transcriptome sequencing to explore the molecular regulation mechanism of cold resistance.A total of 72 up-regulated differentially expressed genes and 3 down-regulated genes were obtained by DESeq2.KEGG analysis was used to analyze the functions of differentially expressed genes in MAPK signaling pathway and plant-pathogen interaction pathway,and protein interaction was predicted.The results showed that leaves under low temperature treatment showed physiological states of curled,yellow,and water loss.The differentially expressed genes were mainly enriched in plant-pathogen interaction,MAPK signaling pathway,and secondary biomass accumulation.MYC2,SPCH and Pti4—6 transcription factors may play an important role in the regulation of cold-induced injury in response to cold stress.MYC2 and CBF proteins were predicted to interact with each other and interacted with multiple proteins.The results showed that multiple metabolic pathways responded to low temperature stress in peach leaves,and related transcription factors that may play a key role in the regulation of cold response pathways were predicted,which provided research direction and basis for the exploration of molecular mechanism of cold response regulation in peach.

Key words: Peach, Leaf, Low temperature, Transcriptome analysis

时梦, 冀晓昊, 杨兴旺, 王莹莹, 王海波, 王孝娣. 桃叶片低温响应转录组分析[J]. 华北农学报, 2023, 38(S1): 49-57. doi: 10.7668/hbnxb.20194251.

SHI Meng, JI Xiaohao, YANG Xingwang, WANG Yingying, WANG Haibo, WANG Xiaodi. Transcriptome Analysis of Low Temperature Response in Peach Leaves[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(S1): 49-57. doi: 10.7668/hbnxb.20194251.

http://www.hbnxb.net/CN/Y2023/V38/IS1/49

图/表 9

图1 4 ℃处理和正常状态下的叶片对比图

Fig.1 Comparison of leaves under 4 ℃ treatment and normal condition

图2 样品聚类分析(A)和主成分分析(B)的叶片对比图 A:LT-1、 LT-2、 LT-3.处理组,RT-1、 RT-2、 RT-3.对照组;B:t.试验组,ut.对照组。

Fig.2 Leaf comparison of sample cluster analysis (A) and principal component analysis (B) A: LT-1, LT-2, and LT-3.The treatment group, RT-1, RT-2, and RT-3.The control group;B: t.The experimental group,ut.The control group.

图3 差异基因的表达热图 RT.对照组叶片;LT.低温胁迫下的处理组叶片。

Fig.3 Expression heatmap of differential genes RT.Leaves of control group; LT.Leaves of treatment groups under low temperature stress.

图4 差异基因的KEGG功能富集

Fig.4 KEGG function enrichment of DEGs

图5 差异基因的KEGG代谢通路富集

Fig.5 KEGG metabolic pathway enrichment of DEGs

表1 差异表达基因的数目及注释到的代谢通路

Tab.1 Number of differentially expressed genes and annotated metabolic pathways

序号 No.	通路 Pathway	ko_ID	差异基因数 DEGs	P-value
1	植物病原体互作	ko04626	12	2.20E-03
2	脂肪酸伸长	ko00062	1	2.08E-02
3	植物激素信号转导	ko04075	5	2.12E-02
4	吞噬体	ko04145	2	4.15E-02
5	不饱和脂肪酸的生物合成	ko01040	1	6.62E-02
6	MAPK信号通路-植物	ko04016	5	1.08E-01
7	昼夜节律-植物	ko04712	2	1.25E-01
8	各种植物次生代谢物的生物合成	ko00999	1	1.60E-01
9	异黄酮的生物合成	ko00943	1	1.64E-01
10	光合作用	ko00195	1	1.69E-01
11	丙氨酸、天冬氨酸和谷氨酸的代谢	ko00250	1	1.73E-01
12	SNARE在水泡运输中的相互作用	ko04130	1	2.03E-01
13	脂肪酸代谢	ko01212	1	2.28E-01
14	色氨酸代谢	ko00380	1	2.40E-01
15	抗坏血酸和藻酸盐代谢	ko00053	1	2.72E-01
16	谷胱甘肽代谢	ko00480	1	2.76E-01
17	RNA降解	ko03018	1	3.73E-01
18	氨基酸的生物合成	ko01230	1	5.05E-01
19	内吞作用	ko04144	1	6.56E-01
20	次生代谢物的生物合成	ko01110	4	7.88E-01
21	代谢途径	ko01100	5	9.98E-01

图6 MAPK信号通路

Fig.6 MAPK signaling pathway

图7 植物病原体互作代谢通路

Fig.7 Plant-pathogen interaction pathway

图8 蛋白互作预测圆球.预测蛋白;线条.互作关系;不同颜色的线条.预测互作的证据来源。

Fig.8 Protein interaction prediction The spheres.Prediction proteins; The lines.Interaction relationships;The different colored lines.Sources of evidence for prediction interactions.

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