2种黑籽南瓜响应枯萎病菌侵染的转录组学研究

doi:10.11924/j.issn.1000-6850.casb2023-0088

摘要/Abstract

摘要：

为了筛选白皮黑籽南瓜和绿皮花纹黑籽南瓜抗枯萎病的差异表达基因，挖掘参与抗性相关的代谢通路。以尖孢镰刀菌侵染2 d后和未侵染的2种幼苗叶片作为材料，利用Illumina Hiseq 2500测序技术进行转录组测序分析。以未侵染的幼苗作为对照，白皮和绿皮花纹黑籽南瓜差异表达基因分别有2082和1116个。其中141个基因在2种黑籽南瓜中均呈现出差异表达，62个基因具有相同表达趋势。对差异表达基因进行GO功能注释，结果显示2种黑籽南瓜差异表达基因主要富集在生物学过程中。KEGG代谢通路分析发现白皮有22个差异表达基因显著富集在植物激素信号通路，绿皮花纹有30个差异表达基因富集在苯丙烷生物合成通路。本研究挖掘出黑籽南瓜抗枯萎病差异表达基因，预测了抗性通路，为瓜类抗病机制的深入研究奠定了理论基础。

关键词: 黑籽南瓜, 尖孢镰刀菌, 抗病机制, 转录组测序, 差异表达基因

Abstract:

To screen the differentially expressed genes for resistance to Fusarium wilt in two cultivated varieties of black-seeded pumpkin (Cucurbita ficifolia) named ‘White Peel’ and ‘Green Peel with White’, and explore metabolic pathway involved in resistance, two varieties seedling leaves infected by Fusarium oxysporum for 2 days and uninfected by Fusarium oxysporum were used as materials, and transcriptome sequencing was performed by using Illumina Hiseq 2500 sequencing technology. Compared to the uninfected seedlings, there were 2082 and 1116 differentially expressed genes in ‘White Peel’ and ‘Green Peel with White’, respectively. Among them, 141 genes showed differential expression, and 62 genes had the same expression trend. Go functional annotation to differentially expressed genes showed that they were mainly enriched in biological processes. KEGG metabolic pathway analysis revealed that 22 differentially expressed genes were significantly enriched in the plant hormone signaling pathway in the ‘White Peel’, while 30 differentially expressed genes were enriched in the phenylpropane biosynthesis pathway in ‘Green Peel with White’. In this study, differentially expressed genes for resistance to Fusarium wilt in Cucurbita ficifolia were identified and the resistance pathway was predicted, which laid a theoretical foundation for further research on the resistance mechanism of melons.

Key words: Cucurbita ficifolia, Fusarium oxysporum, disease resistance mechanism, transcriptome sequencing, differentially expressed genes

张浩, 李海玉, 施松梅, 张忠润, 唐榕梓, 弓月芳, 邓文鑫, 杨正安. 2种黑籽南瓜响应枯萎病菌侵染的转录组学研究[J]. 中国农学通报, 2023, 39(18): 107-116.

ZHANG Hao, LI Haiyu, SHI Songmei, ZHANG Zhongrun, TANG Rongzi, GONG Yuefang, DENG Wenxin, YANG Zhengan. Two Varieties of Cucurbita ficifolia: Transcriptomic Studies in Response to Fusarium oxysporum Infection[J]. Chinese Agricultural Science Bulletin, 2023, 39(18): 107-116.

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参考文献 28

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基因名称	引物序列
AOC3	F：CCACCATGATTGTCCGTCCA，R：TGGAATTGCGCTCTCAGTGT
TAT	F：TGCAGAAACGGCAGAAAAGC，R：AGTCACGAACCACCCAAGTC
gpmI	F：TCTGGTGGAGGCCGTATGTA，R：ACTTGTATGGTGCTTCGCCA
GLDC	F：CCGTTTCCGTTGAGGCTTTG，R：TCAACAAGCGAATCGAGGCT
GCSH	F：GCGGTAGAAAGTGTCAAGGC，R：TGGGCTTGAGTTCACCAGAC
Actin	F：AGCCATCTCTCATCGGTAT，R：CATGGTTGAACCACCACTG

基因名称	引物序列
AOC3	F：CCACCATGATTGTCCGTCCA，R：TGGAATTGCGCTCTCAGTGT
TAT	F：TGCAGAAACGGCAGAAAAGC，R：AGTCACGAACCACCCAAGTC
gpmI	F：TCTGGTGGAGGCCGTATGTA，R：ACTTGTATGGTGCTTCGCCA
GLDC	F：CCGTTTCCGTTGAGGCTTTG，R：TCAACAAGCGAATCGAGGCT
GCSH	F：GCGGTAGAAAGTGTCAAGGC，R：TGGGCTTGAGTTCACCAGAC
Actin	F：AGCCATCTCTCATCGGTAT，R：CATGGTTGAACCACCACTG

基因名	注释	基因id	差异表达倍数（上调/下调）		P值
基因名	注释	基因id	b2d1-vs-b2ck	h2d1-vs-h2ck	P值
AOC3	伯胺氧化酶	csv:101222468 cmo:103491401 cmo:103502378 csv:101222468	0.10157(Down) 0.06889(Down) --- 0.30612(Down) ---	--- --- 0.32215(Down) --- 4.63593(Down) 0.45864(Up)	8.34E-06 1.11E-05 0.00152 0.00501 8.92E-05 0.00283
TAT	酪氨酸转氨酸	csv:101212407 cmo:103490501	0.15352(Down) 0.20640(Down) ---	--- --- 0.38787(Down)	9.49E-06 0.00020 0.00077

基因名	注释	基因id	差异表达倍数（上调/下调）		P值
基因名	注释	基因id	b2d1-vs-b2ck	h2d1-vs-h2ck	P值
AOC3	伯胺氧化酶	csv:101222468 cmo:103491401 cmo:103502378 csv:101222468	0.10157(Down) 0.06889(Down) --- 0.30612(Down) ---	--- --- 0.32215(Down) --- 4.63593(Down) 0.45864(Up)	8.34E-06 1.11E-05 0.00152 0.00501 8.92E-05 0.00283
TAT	酪氨酸转氨酸	csv:101212407 cmo:103490501	0.15352(Down) 0.20640(Down) ---	--- --- 0.38787(Down)	9.49E-06 0.00020 0.00077

基因名	注释	基因id	差异表达倍数（上调/下调）		P值
基因名	注释	基因id	b2d1-vs-b2ck	h2d1-vs-h2ck	P值
gpmI	2,3-二磷酸甘油酸非磷酸甘油酸突变酶	csv:101217925	0.27390(Down) ---	--- 3.10421(Up)	0.00030 0.00099
SerA	D-3-磷酸甘油酸脱氢酶/2-羟戊二酸还原酶	csv:101219910	---	2.75970(Up)	0.00023
AGXT	丙氨酸-乙醛酸转氨酶	csv:101207975	---	0.41561(Down)	0.03110
GLDC	甘氨酸脱氢酶	cmo:103487785	0.42547(Down) 0.42796(Down) 0.28923(Down)	--- --- --- 2.86282(Up)	0.01453 0.04926 0.00052 0.00255
GCSH	甘氨酸裂解系统蛋白	csv:101222346	0.10886(Down) ---	--- 0.43892(Down)	0.04916 0.00900
AGXT2	丙氨酸乙醛酸转氨酶/(R)3氨基2甲基丙酸丙酮酸转氨酶	cmo:103494397	2.30218(Up)	---	0.03533
gcvT	氨基甲基转移酶	csv:101209820 cmo:103485188	0.452126(Down) 0.155741(Down)	--- ---	0.00945 0.002682