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作物学报 ›› 2021, Vol. 47 ›› Issue (9): 1779-1790.doi: 10.3724/SP.J.1006.2021.04151

• 研究论文 • 上一篇    下一篇

外源褪黑素对干旱胁迫下绥农26大豆鼓粒期叶片碳氮代谢调控的途径分析

曹亮(), 杜昕, 于高波, 金喜军, 张明聪, 任春元, 王孟雪*(), 张玉先*()   

  1. 黑龙江八一农垦大学农学院, 黑龙江大庆 163319
  • 收稿日期:2020-07-10 接受日期:2021-01-21 出版日期:2021-09-12 网络出版日期:2021-03-01
  • 通讯作者: 王孟雪,张玉先
  • 作者简介:E-mail: miss9877@126.com
  • 基金资助:
    国家重点研发计划项目“大田经济作物优质丰产的生理基础与调控”(2018YFD1000905);黑龙江省应用技术研究与开发计划项目(GA19B101-02);黑龙江省农垦总局重点科研计划项目(HKKY190206-01)

Regulation of carbon and nitrogen metabolism in leaf of soybean cultivar Suinong 26 at seed-filling stage under drought stress by exogenous melatonin

CAO Liang(), DU Xin, YU Gao-Bo, JIN Xi-Jun, ZHANG Ming-Cong, REN Chun-Yuan, WANG Meng-Xue*(), ZHANG Yu-Xian*()   

  1. College of Agronomy, Heilongjiang Bayi Agricultural University, Daqing 163319, Heilongjiang, China
  • Received:2020-07-10 Accepted:2021-01-21 Published:2021-09-12 Published online:2021-03-01
  • Contact: WANG Meng-Xue,ZHANG Yu-Xian
  • Supported by:
    National Key Research and Development Program of China “Physiological Basis and Agronomic Management for High-quality and High-yield of Field Cash Crops”(2018YFD1000905);Heilongjiang Application Technology Research and Development Projects(GA19B101-02);Heilongjiang Provincial Land Reclamation Bureau Key Research Project(HKKY190206-01)

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摘要：

鼓粒期是大豆碳氮代谢最复杂的阶段, 干旱胁迫必然限制鼓粒期大豆碳氮同化、分配和转移, 影响大豆产量的形成。在我们前期的研究中, 明确了外源褪黑素对干旱胁迫下鼓粒期大豆抗旱和碳氮代谢的生理调控效应。本研究通过转录组和代谢组分析来确定褪黑素对大豆干旱条件反应的一些重要的碳氮代谢基因和途径。转录组分析表明, 与干旱胁迫处理相比, 正常供水和干旱胁迫下喷施外源褪黑素处理的大豆叶片共同上调和下调的基因分别有37个和493个。上调的基因中存在着直接和间接参与碳氮代谢的功能基因, 包括正向调控的参与半胱氨酸合成、光合作用、碳水化合物代谢和葡萄糖代谢等途径关键基因。代谢组分析发现, 与干旱胁迫处理相比, 正常供水和干旱胁迫下喷施外源褪黑素处理的大豆叶片共同上调和下调的代谢物分别有17个和43个, 上调的代谢物中绝大部分(14/17)属于氨基酸、脂质、有机酸和碳水化合物, 进一步揭示了外源褪黑素能够提高大豆碳氮代谢与抗旱的能力。结合转录组和代谢组分析发现, 褪黑素通过调节氨基酸代谢和淀粉蔗糖代谢途径, 促进干旱胁迫下β-葡萄糖苷酶基因表达, 提高了L-天冬酰胺和6-磷酸葡萄糖代谢物的含量, 最终提高了大豆的抗旱性。

关键词: 褪黑素, 大豆, 鼓粒期干旱, 碳氮代谢, 代谢组和转录组

Abstract:

The grain-filling stage is the most complex stage of carbon and nitrogen metabolism. Drought stress inevitably inhibits the assimilation, distribution, and transition of carbon and nitrogen at grain-filling stage in soybean, resulting in less soybean yield. The objective of this study was to investigate the effects of exogenous melatonin on the carbon and nitrogen metabolism genes and pathways under drought stress in soybean. Transcriptome analysis showed that, compared with drought stress treatment, 37 and 493 genes were jointly up-regulated and down-regulated in soybean leaves treated with normal water supply and treated with exogenous melatonin under drought stress, respectively. The up-regulated genes included functional genes directly and indirectly involved in carbon and nitrogen metabolism, such as the key genes involved in the cysteine synthesis pathway, photosynthesis, carbohydrate metabolism, and glucose metabolism. Metabolomic analysis revealed that, compared with drought stress treatment, 17 and 43 metabolites were jointly up-regulated and down-regulated in soybean leaves treated with normal water supply and treated with exogenous melatonin under drought stress, respectively. Most (14/17) of up-regulated metabolites were amino acids, lipids, organic acids, and carbohydrates, which further indicated that exogenous melatonin could improve soybean carbon and nitrogen metabolism and drought resistance in soybean. Combined with transcriptome and metabolomic profile, melatonin promoted the relative expression level of β-D-Glucosidase gene due to regulate the pathway of amino acid metabolism and starch and sucrose metabolism, improved the contents of L-Asparagine and D-glucose-6P metabolites, and ultimately improves the ability of drought resistance in soybean.

Key words: melatonin, soybean, drought at grain-filling stage, carbon and nitrogen metabolism, metabolome and transcriptome

图1

WW/D和DM/D比较中差异表达基因图 灰点代表无明显差异的表达基因, 红色和蓝色点分别表示在WW/D和DM/D比较中显著上调和显著下调的基因。WW为正常供水处理, D为干旱处理, DM为干旱条件下喷施褪黑素处理。"

图2

外源褪黑素对干旱胁迫下大豆叶片转录组的影响 A: 韦恩图显示了WW/D和DM/D比较之间的共同差异表达基因。B: WW/D和DM/D共同上调和下调表达的差异基因的热图。处理同图1。 "

图3

外源褪黑素上调干旱胁迫下大豆叶片基因的生物学过程富集分析 A: 热图显示了WW/D和DM/D比较之间的共同表达上调的基因。B: WW/D和DM/D比较之间的共同表达上调基因GO生物学过程富集分析。处理同图1。 "

图4

WW/D和DM/D共同上调与下调基因的KEGG富集图 A: 上调。B: 下调。富集因子的计算方法是: 差异表达基因数除以任何给定途径的基因数, 点的大小表示基因数, 点的颜色表示调整后的P值范围。处理同图1。 "

图5

差异表达基因的 RT-qPCR 验证 白色的表示差异表达基因的转录组数据, 黑色的表示差异表达基因的qRT-PCR结果。处理同图1。 "

图6

代谢组主成分分析 处理同图1。 "

图7

干旱胁迫和外源褪黑素处理对代谢组的影响 A: 韦恩图显示了WW/D和DM/D比较中共同的差异累积代谢物。B: WW/D和DM/D共同上调和下调基因的KEGG富集图。横坐标是差异表达基因数。处理同图1。 "

图8

淀粉蔗糖代谢和L-天冬酰胺代谢的KEGG途径 A: 淀粉和蔗糖代谢。B: L-天冬酰胺代谢。红色和粉色分别表示与干旱胁迫处理相比, 正常供水处理和干旱胁迫下喷施外源褪黑素大豆叶片中共同上调的代谢产物和基因。"

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