玉米盐胁迫和MeJA处理下的转录组联合分析

doi:10.19586/j.2095-2341.2024.0183

生物技术进展 ›› 2025, Vol. 15 ›› Issue (2): 263-275.DOI: 10.19586/j.2095-2341.2024.0183

• 研究论文 • 上一篇

玉米盐胁迫和MeJA处理下的转录组联合分析

刘卓颖¹(), 周晓今²(), 黄燕丽³, 逄森¹()

^1.中国农业大学理学院，北京 100193
^2.中国农业科学院生物技术研究所，北京 100081
^3.菏泽学院创新创业学院，山东菏泽 274000

收稿日期:2024-11-24 接受日期:2025-01-21 出版日期:2025-03-25 发布日期:2025-04-29
通讯作者: 周晓今,逄森
作者简介:刘卓颖E-mail：zz9629@126.com
基金资助:
国家自然科学基金面上项目(32472217)

Joint Transcriptome Analysis of Maize Under Salt Stress and MeJA Treatment

Zhuoying LIU¹(), Xiaojin ZHOU²(), Yanli HUANG³, Sen PANG¹()

^1.College of Science，China Agricultural University，Beijing 100193，China
^2.Biotechnology Research Institute，Chinese Academy of Agricultural Sciences，Beijing 100081，China
^3.School of Innovation and Entrepreneurship，Heze University，Shandong Heze 274000，China

Received:2024-11-24 Accepted:2025-01-21 Online:2025-03-25 Published:2025-04-29
Contact: Xiaojin ZHOU,Sen PANG

摘要/Abstract

摘要：

玉米是重要的粮饲兼用作物，盐胁迫严重影响其生长发育，导致产量和品质下降。茉莉酸及其衍生物（jasmonates, JAs）是与植物防御相关的天然植物激素，基于模式植物的研究表明，JAs在响应盐胁迫中发挥重要作用。为探究玉米中JAs介导的盐胁迫响应的具体机制，分别对100 μmol·L^-1 MeJA和200 mmol·L^-1 NaCl处理6 h玉米幼苗的地上和地下部组织进行转录组测序，交叉分析得到JA诱导且响应高盐胁迫的差异表达基因（differential expression genes, DEGs），并挑选8个DEGs通过RT-qPCR进行验证。结果发现地上和地下部组织中分别共有362和803个基因差异表达，GO和KEGG富集分析显示这些基因涉及糖的合成转运、防御性次生代谢物合成、抗氧化酶合成以及脱落酸和乙烯信号通路。研究结果提示JA信号通路诱导玉米高盐胁迫响应的潜在关键基因和代谢途径，为进一步挖掘JA信号通路调控抗盐性的具体分子机制提供了线索。

关键词: 盐胁迫, 茉莉酸, 转录组, 玉米, 基因表达分析

Abstract:

Maize， an important food and feed crop， faces severe growth inhibition， yield reduction， and quality deterioration under salt stress. Jasmonic acid and its derivatives （JAs）， crucial phytohormones involved in plant defense mechanisms， have been shown through studies in model plants to play essential roles in salt stress responses. To investigate JAs-mediated salt stress adaptation mechanisms in maize， we subjected seedlings to combined treatments of 200 mmol·L^-1 NaCl and 100 μmol·L^-1 methyl jasmonate （MeJA） for six hours. Transcriptomic analysis of shoots and roots identified differentially expressed genes （DEGs） associated with both JA signaling and salt stress response. Eight overlapping DEGs from shoot-root comparisons were subsequently validated using RT-qPCR. The study revealed 362 and 803 stress-responsive DEGs in overground and subterranean tissues， respectively. Functional enrichment analyses （GO and KEGG） demonstrated these genes participate in carbohydrate metabolism/transport， defensive secondary metabolite biosynthesis， antioxidant enzyme production， along with abscisic acid and ethylene signaling pathways. These findings indicated that JA signaling activates specific genetic and metabolic networks underlying salt stress adaptation of maize， providing critical insights for further elucidating the molecular mechanisms of JA-mediated salt tolerance regulation.

Key words: salt stress, jasmonic acid （JA）, transcriptome, maize, gene expression analysis

中图分类号:

Q943.2

刘卓颖, 周晓今, 黄燕丽, 逄森. 玉米盐胁迫和MeJA处理下的转录组联合分析[J]. 生物技术进展, 2025, 15(2): 263-275.

Zhuoying LIU, Xiaojin ZHOU, Yanli HUANG, Sen PANG. Joint Transcriptome Analysis of Maize Under Salt Stress and MeJA Treatment[J]. Current Biotechnology, 2025, 15(2): 263-275.

图/表 9

表1 qPCR所用引物序列

Table 1 Primer sequences used for qPCR analysis

基因名称（ID）	上游引物序列（5'→3'）	下游引物序列（5'→3'）
ZmActin (Zm00001d013410)	5'-ATGTTTCCTGGGATTGCCGAT-3'	5'-CCAGTTTCGTCATACTCTCCCTTG-3'
SWEET6a (Zm00001d044421)	5'-CGAGACGTGCTTTACCCACA-3'	5'-GAATGGCAAACACAGCCACA-3'
CKX4 (Zm00001d043293)	5'-CGGCCGACTCGTAACGTAAT-3'	5'-CACGCGATTAACAACCCCAC-3'
POD64 (Zm00001d040705)	5'-GTGCCCAACTCCTACTCTGG-3'	5'-TGACACACGAACTACGCACT-3'
EREB113 (Zm00001d010175)	5'-GCCAAGGCAGCAGCAGTCA-3'	5'-GGCACAGAAGCCACGGTAA-3'
WRKY114 (Zm00001d036726)	5'-CTAGCTCTAGCACGTACGCC-3'	5'-GAGCGGTACAAACTCGGTCA-3'
ZIM27 (Zm00001d027900)	5'-AGGAAGGTGTCGCTAAAGAG-3'	5'-CTGCCGTCGATGAGATTG-3'
TPS11 (Zm00001d018342)	5'-CGGCGTGGTGTAGGATTGAT-3'	5'-ACCATGCGGGCATATACAGG-3'
LOX10 (Zm00001d053675)	5'-TTCCAAACAGCATCTCCATT-3'	5'-GCCTTATTACAACAGTCCTCACG-3'

表2 测序数据质量分析和图谱统计结果

Table 2 The quality analysis and mapping statistics of the RNA sequencing data

样本	原始Reads	过滤后Reads	Q20	Q30	GC含量	Uniquely mapped
CK-S-1	43678100	43047426	98.30 %	95.06 %	55.69 %	95.74 %
CK-S-2	50852996	50083122	98.18 %	94.75 %	55.37 %	95.44 %
CK-S-3	43444624	42661464	98.27 %	94.99 %	55.15 %	95.64 %
CK-S-4	51020748	50012420	98.19 %	94.90 %	56.15 %	95.19 %
CK-R-1	47340142	46607418	98.23 %	94.92 %	55.15 %	93.16 %
CK-R-2	42318602	41636470	98.28 %	95.09 %	55.11 %	95.27 %
CK-R-3	42916752	42228274	98.32 %	95.14 %	54.54 %	93.34 %
CK-R-4	47196988	46286054	98.24 %	94.94 %	55.29 %	93.58 %
NaCl-S-1	44346810	43693600	98.41 %	95.37 %	55.97 %	95.90 %
NaCl-S-2	44482508	43736948	98.13 %	94.64 %	56.02 %	95.60 %
NaCl-S-3	45904696	45070904	98.23 %	94.89 %	55.94 %	96.16 %
NaCl-S-4	55913340	54947042	98.20 %	94.86 %	55.90 %	95.91 %
NaCl-R-1	47132296	46300150	98.25 %	95.00 %	55.83 %	93.94 %
NaCl-R-2	42765116	42001334	98.09 %	94.64 %	55.81 %	93.33 %
NaCl-R-3	44430198	43679556	98.17 %	94.79 %	56.33 %	93.88 %
NaCl-R-4	44579594	43831320	98.28 %	95.08 %	56.02 %	94.16 %
MeJA-S-1	53707412	52824432	98.23 %	94.92 %	54.87 %	94.11 %
MeJA-S-2	43478130	42516802	98.24 %	94.97 %	54.42 %	94.95 %
MeJA-S-3	40559722	39809098	97.95 %	94.15 %	54.50 %	95.22 %
MeJA-S-4	46413454	45685218	98.37 %	95.28 %	54.46 %	95.43 %
MeJA-R-1	42939854	41593670	98.30 %	95.02 %	54.49 %	88.28 %
MeJA-R-2	43131000	42474978	97.98 %	94.26 %	54.20 %	90.04 %
MeJA-R-3	40744494	40164434	98.36 %	95.20 %	54.64 %	90.73 %
MeJA-R-4	48265060	47530412	98.31 %	95.09 %	54.41 %	93.33 %

图1 样本组间的相关系数（R2）热图A：CK组地上部和地下部样品间相关系数热图；B：高盐胁迫组地上部和地下部样品间相关系数热图；C：MeJA处理组地上部和地下部样品间相关系数热图

Fig. 1 Heatmaps of the correlation coefficient between sample groups

图2 处理组与对照组间差异表达基因火山图A：高盐胁迫组地上部与对照组差异表达基因火山图；B：高盐胁迫组地下部与对照组差异表达基因火山图；C：MeJA处理组地上部与对照组差异表达基因火山图；D：MeJA处理组地下部与对照组差异表达基因火山图。横坐标表示基因在不同样本中表达倍数变化（log2 Fold Change）；纵坐标表示基因表达量变化的统计学显著性（-log10 Padj），红色点表示有显著性差异表达的上调基因，绿色点表示有显著性差异表达的下调基因，蓝色点表示表达变化不显著的基因。

Fig. 2 Volcano plots showing DEGs between the treatment and control groups

图3 高盐胁迫和MeJA激素处理组中差异表达基因韦恩图

Fig. 3 Venn diagrams showing DEGs in high salt stress and hormone treatment groups

图4 处理组与对照组间差异表达基因GO富集柱状图A：高盐胁迫组地上部差异表达基因的GO富集柱状图；B：高盐胁迫组地下部差异表达基因的GO富集柱状图；C：MeJA处理组地上部差异表达基因的GO富集柱状图；D：MeJA处理组地下部差异表达基因的GO富集柱状图

Fig. 4 GO enrichment of DEGs between the treatment group and the control group

图5 处理组与对照组间差异基因KEGG富集散点图A：高盐胁迫组地上部差异表达基因KEGG富集散点图；B：高盐胁迫组地下部差异表达基因KEGG富集散点图；C：MeJA处理组地上部差异表达基因KEGG富集散点图；D：MeJA处理组地下部差异表达基因KEGG富集散点图。纵坐标表示通路名称，横坐标表示富集因子（rich factor），点的大小表示此通路中DEGs个数，点的颜色对应于不同q值范围。

Fig. 5 Scatter plot of KEGG enrichment of DEGs between the treatment group and the control group

图6 候选基因的FPKM热图A：茉莉酸相关的基因；B：脱落酸相关的基因；C：乙烯相关的基因；D：氧化还原相关的基因；E：转录因子相关的基因；F：糖代谢相关的基因。右侧标尺表示不同颜色对应的FPKM值，红色下划线标记的为后续荧光实时定量PCR验证的候选基因。

Fig. 6 FPKM heatmap of candidate genes

图7 候选基因的荧光实时定量PCR检测注：按2-ΔΔCt计算相对表达量，ns表示数据间无统计学差异（P>0.05），*，**，***，****分别表示数据在P<0.05、P<0.01、P<0.001、P<0.000 1水平有统计学差异。

Fig. 7 RT-qPCR analysis result of candidate genes

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玉米盐胁迫和MeJA处理下的转录组联合分析

Joint Transcriptome Analysis of Maize Under Salt Stress and MeJA Treatment

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