草甘膦铵盐诱导谷子雄性不育的转录组分析

doi:10.13304/j.nykjdb.2022.0198

摘要/Abstract

摘要：

为促进谷子杂种优势的利用，提高杂交育种效率，采用 Illumina HiSeq 高通量测序技术对草甘膦铵盐处理后的谷穗进行转录组测序，分析筛选响应草甘膦铵盐的差异表达基因，同时测定谷穗抗性淀粉、可溶性糖等生理指标。结果表明，草甘膦铵盐处理后产生的雄性不育谷穗与未处理谷穗相比，共检测到797个差异表达基因，其中645个差异表达基因获得GO功能分类，主要集中在糖代谢与生物合成、激素代谢和生物合成以及细胞壁等方面，反映了谷穗对草甘膦铵盐处理响应基因的生物学功能。通过KEGG富集分析，共建立了138条通路，发现差异表达基因中涉及植物激素信号转导途径的最多，为19个；其次是淀粉和蔗糖代谢途径，为16个。经诱导产生雄性不育的谷穗样品中抗性淀粉、可溶性糖和赤霉素含量降低，生长素含量增加，与GO和KEGG富集分析得到的差异表达基因的表达模式相吻合。以上结果为筛选适宜的化学杀雄试剂提供了理论依据。

关键词: 谷子, 草甘膦铵盐, 诱导雄性不育, 差异表达基因, 生理指标

Abstract:

To promote the heterosis utilization of foxtail millet and improve the efficiency of cross breeding， transcriptome sequencing of the grain spike treated by ammonium glyphosate was performed using the Illumina HiSeq high-throughput sequencing technology， the differentially expressed genes were analytically screened among different treatments. The physiological indexes such as resistant starch and soluble sugar were determined. The results showed that the grain ears treated with glyphosate had 797 differentially expressed genes compared with untreated ears， among which 645 differentially expressed genes obtained GO functional classification， and they mainly focused on glucose metabolism and biosynthesis， hormone metabolism and biosynthesis and cell wall， reflecting the biological function of response genes to glyphosate treatment. A total of 138 pathways were established by KEGG enrichment analysis， including 19 differentially expressed genes involved in the plant hormone signaling pathway and 16 differentially expressed genes involved in the starch and sucrose metabolism pathways. In the grain spike treated by ammonium glyphosate， the contents of resistant starch， soluble sugar and gibberellin decreased， while the IAA content increased， which were consistent with the expression pattern of differeatly expressed genes by GO and KEGG enrichment analysis. These above results provided a theoretical basis for screening suitable chemical androcide reagents.

Key words: foxtail millet, glyphosate ammonium salt, induced male sterility, differentially expressed genes, physiological indicators

中图分类号:

S515

郭瑞锋, 任月梅, 杨忠, 刘贵山, 任广兵, 张绶, 朱文娟. 草甘膦铵盐诱导谷子雄性不育的转录组分析[J]. 中国农业科技导报, 2022, 24(10): 35-43.

Ruifeng GUO, Yuemei REN, Zhong YANG, Guishan LIU, Guangbing REN, Shou ZHANG, Wenjuan ZHU. Transcriptomic Analysis of Mechanism of Foxtail Millet Male Infertility Induced by Glyphosate Ammonium Salt[J]. Journal of Agricultural Science and Technology, 2022, 24(10): 35-43.

图/表 7

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样品编号 Sample code	高质量序列 Clean read		高质量序列碱基数 Base number of clean read		Q20/%	Q30/%	比对序列 Total mapped read
	数量 Number	占比 Percentage/%	数量 Number	占比 Percentage/%			比对序列 Total mapped read
	数量 Number	占比 Percentage/%	数量 Number	占比 Percentage/%			数量 Number	占比 Percentage/%
S1	42 672 248	98.99	6 424 467 638	98.69	96.12	90.79	36 768 433	86.16
S2	41 691 824	98.98	6 276 055 582	98.67	96.07	90.71	34 558 553	82.89
P1	44 638 924	99.18	6 719 144 472	98.87	96.36	91.24	35 018 847	78.45
P2	44 681 598	98.87	6 729 889 484	98.62	95.96	90.52	37 171 009	83.19
CK1	45 815 562	98.94	6 896 999 720	98.63	95.96	90.48	33 550 009	73.23
CK2	45 088 252	99.07	6 786 443 052	98.75	96.02	90.66	35 095 278	77.84

样品编号 Sample code	高质量序列 Clean read		高质量序列碱基数 Base number of clean read		Q20/%	Q30/%	比对序列 Total mapped read
	数量 Number	占比 Percentage/%	数量 Number	占比 Percentage/%			比对序列 Total mapped read
	数量 Number	占比 Percentage/%	数量 Number	占比 Percentage/%			数量 Number	占比 Percentage/%
S1	42 672 248	98.99	6 424 467 638	98.69	96.12	90.79	36 768 433	86.16
S2	41 691 824	98.98	6 276 055 582	98.67	96.07	90.71	34 558 553	82.89
P1	44 638 924	99.18	6 719 144 472	98.87	96.36	91.24	35 018 847	78.45
P2	44 681 598	98.87	6 729 889 484	98.62	95.96	90.52	37 171 009	83.19
CK1	45 815 562	98.94	6 896 999 720	98.63	95.96	90.48	33 550 009	73.23
CK2	45 088 252	99.07	6 786 443 052	98.75	96.02	90.66	35 095 278	77.84

样本对 Paired samples	差异表达基因数量 Number of differentially expressed genes
样本对 Paired samples	上调Up	下调Down	总计Total
S1/CK1	296	501	797
S1/P1	56	113	169
P1/CK1	1 513	1 559	3 072

样本对 Paired samples	差异表达基因数量 Number of differentially expressed genes
样本对 Paired samples	上调Up	下调Down	总计Total
S1/CK1	296	501	797
S1/P1	56	113	169
P1/CK1	1 513	1 559	3 072

类别 Catergory	GO条目 GO term	代谢节点编号 Metabolic node number	差异表达基因数量 Number of DEGs			P值（校正后） P-value （corrected）
类别 Catergory	GO条目 GO term	代谢节点编号 Metabolic node number	上调 Up-regulated	下调 Down-regulated	总计 Total	P值（校正后） P-value （corrected）
分子功能 Molecular function	水解酶活性，作用于糖基键 Hydrolase activity， acting on glycosyl bonds	GO∶0016798	6	31	37	0.000 25
	水解酶活性，水解O糖基化合物 Hydrolase activity， hydrolyzing O-glycosyl compounds	GO∶0004553	5	29	34	0.000 37
	氧化还原酶活性 Oxidoreductase activity	GO∶0016705	11	18	29	0.025 41
	铁离子结合 Iron ion binding	GO∶0005506	11	19	30	0.033 03
	糖跨膜转运蛋白活性 Sugar transmembrane transporter activity	GO∶0051119	6	4	10	0.036 38
	双加氧酶活性 Dioxygenase activity	GO∶0051213	6	9	15	0.044 21
	半乳糖苷酶活性Galactosidase activity	GO∶0015925	1	5	6	0.044 90
生物过程 Biological process	二糖生物合成过程 Disaccharide biosynthetic process	GO∶0046351	0	8	8	0.000 99
	海藻糖生物合成过程 Trehalose biosynthetic process	GO∶0005992	0	7	7	0.002 67
	海藻糖代谢Trehalose metabolic process	GO∶0005991	0	7	7	0.003 73
	低聚糖生物合成过程 Oligosaccharide biosynthetic process	GO∶0009312	0	8	8	0.006 66
	激素代谢过程 Hormone metabolic process	GO∶0042445	2	11	13	0.011 38
	激素生物合成过程 Hormone biosynthetic process	GO∶0042446	2	2	4	0.032 92
细胞组分 Cellular component	细胞壁Cell wall	GO∶0005618	6	20	26	0.008 61
	外部封装结构 External encapsulating structure	GO∶0030312	6	20	26	0.011 89
	植物型细胞壁Plant-type cell wall	GO∶0009505	2	9	11	0.358 56