玉米自交系籽粒响应高温胁迫的转录组分析

doi:10.7668/hbnxb.20193984

摘要/Abstract

摘要：

玉米籽粒发育时期对高温胁迫十分敏感,严重影响玉米的产量和品质。为研究不同耐高温玉米自交系籽粒响应高温胁迫的基因表达差异,在基因水平解析不同玉米种质响应高温胁迫的分子机制,以前期筛选的耐高温自交系XN202和敏感自交系CT110为材料,利用RNA-Seq技术挖掘正常和高温胁迫下授粉后15 d玉米籽粒的差异表达基因(DEG)。与对照相比,XN202和CT110分别检测到1 517,1 012个DEGs,共同的DEGs有142个,包含7个转录因子。不同耐高温玉米自交系的籽粒对高温胁迫的响应存在明显差异,通过基因本体(GO)功能注释和全基因组及代谢途径数据库(KEGG)信号通路富集分析筛选出DNA复制、核小体、微小染色体维持复合物、DNA引物酶-多聚酶α复合体、营养库活性、丙氨酸、天冬氨酸和谷氨酸代谢等共同参与响应高温胁迫的应答过程。通过生物信息学分析,共有374个DEGs位于已报道的玉米耐高温相关性状QTL区间,其中42个DEGs为已报道的耐高温相关基因。综上所述,高温胁迫下玉米籽粒会形成复杂的细胞保护和防御系统,AP2/ERF、MYB、bHLH、NAC、HSF、HSP等耐高温相关的DEGs可能在分子调控网络中发挥重要作用。

关键词: 玉米, 耐热性, 转录组测序, 差异表达基因, 数量性状位点(QTL)

Abstract:

Maize grain development stage is very sensitive to high temperature stress,which seriously affects the yield and quality of maize.To study the difference of gene expression in grain and analyze the gene molecular mechanism of different high-temperature tolerant maize inbred lines in response to high-temperature stress,RNA-Seq technology was used to analysis the differentially expressed genes(DEGs)of XN202(heat tolerant)and CT110(heat susceptible)in maize grains 15 days after pollination under normal and high temperature stress.Compared with the control,XN202 and CT110 detected 1 517 and 1 012 DEGs,respectively,with 142 common DEGs,including 7 transcription factors.There were significant differences in grain response to high temperature stress among different heat tolerant inbred lines.DNA replication,nucleosome,minichromosome maintenance complex,alpha DNA polymerase:primase complex,nutrient reservoir activity,alanine,aspartate and glutamate metabolism were involved in response to high temperature stress through gene ontology(GO)function annotation and kyoto encyclopedia of genes and genomes(KEGG)signal pathway enrichment analysis.According to bioinformatics analysis,a total of 374 DEGs were located in the reported QTL interval of heat tolerance related traits in maize,and 42 DEGs were putative heat tolerance candidate genes.In summary,maize grains could form complex cellular protection and defense system under high temperature stress,and DEGs related to high temperature tolerance,such as AP2/ERF,MYB,bHLH,NAC,HSF and HSP,might play an important role in the molecular regulatory network.

Key words: Zea mays L., Heat stress, Transcriptome sequencing, Differentially expression genes, Quantitative trait loci(QTL)

汤彬, 耿存娟, 曾强, 郭欢乐, 李涵, 曹钟洋, 邓力超, 彭明, 周虹, 陈志辉. 玉米自交系籽粒响应高温胁迫的转录组分析[J]. 华北农学报, 2023, 38(4): 11-19. doi: 10.7668/hbnxb.20193984.

TANG Bin, GENG Cunjuan, ZENG Qiang, GUO Huanle, LI Han, CAO Zhongyang, DENG Lichao, PENG Ming, ZHOU Hong, CHEN Zhihui. Kernel Transcriptome Analysis of Maize Inbred Lines in Response to High Temperature Stress[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(4): 11-19. doi: 10.7668/hbnxb.20193984.

http://www.hbnxb.net/CN/Y2023/V38/I4/11

图/表 7

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样品编号 Sample ID	过滤碱基数 Clean bases	Q30/%	GC/%	过滤序列 Clean reads	唯一配对序列(百分比/%) Unique mapped reads (Percentage)
SCK1	7 594 613 558	92.75	52.69	50 921 642	39 665 200 (77.89)
SCK2	5 917 021 526	93.52	54.13	39 571 676	32 000 572 (80.87)
SCK3	6 338 318 024	92.95	54.29	42 378 368	33 835 907 (79.84)
SHT1	5 947 773 382	93.38	52.58	39 770 066	32 253 255 (81.10)
SHT2	7 044 679 918	95.05	53.42	47 102 524	38 184 631 (81.07)
SHT3	6 332 099 890	93.34	52.80	42 462 144	34 644 680 (81.59)
TCK1	7 201 535 242	93.39	53.14	48 260 672	38 285 653 (79.33)
TCK2	7 629 111 632	92.59	52.87	51 143 088	40 059 957 (78.33)
TCK3	6 474 674 720	93.80	53.66	43 320 570	34 364 902 (79.33)
THT1	7 368 185 900	93.72	51.20	49 405 022	36 757 593 (74.40)
THT2	8 119 313 960	93.80	52.30	54 559 458	42 932 212 (78.69)
THT3	7 094 672 054	93.86	52.32	47 526 744	36 606 182 (77.02)

样品编号 Sample ID	过滤碱基数 Clean bases	Q30/%	GC/%	过滤序列 Clean reads	唯一配对序列(百分比/%) Unique mapped reads (Percentage)
SCK1	7 594 613 558	92.75	52.69	50 921 642	39 665 200 (77.89)
SCK2	5 917 021 526	93.52	54.13	39 571 676	32 000 572 (80.87)
SCK3	6 338 318 024	92.95	54.29	42 378 368	33 835 907 (79.84)
SHT1	5 947 773 382	93.38	52.58	39 770 066	32 253 255 (81.10)
SHT2	7 044 679 918	95.05	53.42	47 102 524	38 184 631 (81.07)
SHT3	6 332 099 890	93.34	52.80	42 462 144	34 644 680 (81.59)
TCK1	7 201 535 242	93.39	53.14	48 260 672	38 285 653 (79.33)
TCK2	7 629 111 632	92.59	52.87	51 143 088	40 059 957 (78.33)
TCK3	6 474 674 720	93.80	53.66	43 320 570	34 364 902 (79.33)
THT1	7 368 185 900	93.72	51.20	49 405 022	36 757 593 (74.40)
THT2	8 119 313 960	93.80	52.30	54 559 458	42 932 212 (78.69)
THT3	7 094 672 054	93.86	52.32	47 526 744	36 606 182 (77.02)

样本分组 Sample groups	差异表达基因的个数 DEG number	上调的差异表达基因个数 Up-regulated DEG number	下调的差异表达基因个数 Down-regulated DEG number
SCK-1-2-3 vs SHT-1-2-3	1 012	386	626
TCK-1-2-3 vs THT-1-2-3	1 517	801	716
共同响应基因	142	74	44
Common response genes

样本分组 Sample groups	差异表达基因的个数 DEG number	上调的差异表达基因个数 Up-regulated DEG number	下调的差异表达基因个数 Down-regulated DEG number
SCK-1-2-3 vs SHT-1-2-3	1 012	386	626
TCK-1-2-3 vs THT-1-2-3	1 517	801	716
共同响应基因	142	74	44
Common response genes

编号 Identity	描述 Description	CT110		XN202
编号 Identity	描述 Description	校正后P值 Corrected P value	基因个数 Number of genes	校正后P值 Corrected P value	基因个数 Number of genes
ko00230	嘌呤代谢	4.22E-02	14
ko00480	谷胱甘肽代谢	4.70E-02	12
ko03430	错配修复	3.88E-02	11
ko03440	同源重组	9.25E-03	16
ko04141	内质网蛋白质加工	2.59E-15	52
ko00250	丙氨酸、天冬氨酸和谷氨酸代谢	4.22E-02	8	1.20E-04	16
ko03030	DNA复制	8.11E-11	25	4.37E-05	24
ko00220	精氨酸生物合成			1.14E-02	10
ko00630	乙醛酸和二羧酸代谢			1.64E-02	17
ko03008	真核生物核糖体的生物合成			2.36E-02	20
ko03010	核糖体			5.90E-20	89

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