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作物学报 ›› 2023, Vol. 49 ›› Issue (1): 225-238.doi: 10.3724/SP.J.1006.2023.24020

• 耕作栽培·生理生化 • 上一篇    下一篇

氮素缓解花生干旱胁迫的生理和转录调控机制

丁红(), 张智猛, 徐扬, 张冠初, 郭庆, 秦斐斐, 戴良香()   

  1. 山东省花生研究所, 山东青岛 266100
  • 收稿日期:2022-01-15 接受日期:2022-05-05 出版日期:2023-01-12 网络出版日期:2022-05-13
  • 通讯作者: 戴良香
  • 作者简介:E-mail: dingpeanut@163.com
  • 基金资助:
    国家自然科学基金项目(31971854);国家自然科学基金项目(31971856);山东省花生产业技术体系(SDAIT-04-06);山东省重大科技创新工程项目(2019JZZY010702)

Physiological and transcriptional regulation mechanisms of nitrogen alleviating drought stress in peanut

DING Hong(), ZHANG Zhi-Meng, XU Yang, ZHANG Guan-Chu, GUO Qing, QIN Fei-Fei, DAI Liang-Xiang()   

  1. Shandong Peanut Research Institute, Qingdao 266100, Shandong, China
  • Received:2022-01-15 Accepted:2022-05-05 Published:2023-01-12 Published online:2022-05-13
  • Contact: DAI Liang-Xiang
  • Supported by:
    National Natural Science Foundation of China(31971854);National Natural Science Foundation of China(31971856);Shandong Peanut Industry Technology System(SDAIT-04-06);Major Scientific and Technological Innovation Projects in Shandong Province(2019JZZY010702)

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摘要：

干旱胁迫下氮素施用对植物生长发育具有重要的影响。为明确氮素提高花生抗旱性的生理和转录调控机制, 本研究对施氮、干旱及旱氮同存处理下的花生生理指标和根系转录组进行了测定。结果表明, 旱氮同存处理提高了干旱胁迫下花生生物量和叶片相对含水量。施用氮肥增加了干旱胁迫下花生根系的总酚和类黄酮含量, 提高其过氧化物酶(POD)和过氧化氢酶(CAT)活性, 降低其丙二醛(MDA)含量, 提高花生抗旱性。转录组分析表明, 施用氮肥产生5396个差异表达基因, 这些基因主要参与谷胱甘肽代谢、氮代谢和碳代谢相关过程及应激和防御反应。干旱处理和旱氮同存处理下, 次生代谢物生物合成、运输和分解代谢及碳水化合物运输和代谢这两类功能差异表达基因富集。旱氮同存处理下酚类代谢物质相关的3种途径中有51个差异基因上调表达, 207个基因下调表达。由此表明, 施用氮肥通过调控花生次生产物代谢、碳水化合物代谢等途径提高干旱胁迫下花生植株的抗氧化能力, 从而提高花生的抗旱性。

关键词: 花生, 干旱, 氮素, 抗氧化系统, 转录组

Abstract:

Nitrogen application has an important effect on plant growth and development under drought stress. The aim of this study is to clarify the physiological and transcriptional regulation mechanism of nitrogen for improving drought resistance in peanut. The physiological indexes and root transcriptome of peanut under nitrogen application, drought stress and drought and nitrogen application simultaneously exist treatments were determined. The results showed that the drought and nitrogen application simultaneously exist treatment increased peanut biomass and the relative water content of leaves under drought stress. The content of total phenols and flavonoids in peanut roots were increased by nitrogen application under drought stress. Meanwhile, the activities of POD and CAT were increased, the content of MDA was decreased, which improved the drought resistance of peanut. Transcriptome analysis showed that the relative expression of 5396 genes changed due to nitrogen application. These genes were mainly involved in glutathione metabolism, nitrogen metabolism, and carbon metabolism, as well as stress and defense responses. Under drought stress, the drought and nitrogen application simultaneously exist treatments, the differentially expressed genes of secondary metabolite biosynthesis, transportation and catabolism and carbohydrate transport and metabolism were enriched. Among the three pathways related to phenolic metabolites, 51 differential genes were up-regulated and 207 genes were down-regulated under the drought and nitrogen application simultaneously exist treatment. In conclusion, the application of nitrogen fertilizer could enhance the antioxidant capacity of peanut plants under drought stress through regulating secondary metabolites and carbohydrate metabolism, and thus improve the drought resistance of peanut.

Key words: peanut, drought stress, nitrogen, antioxidant system, transcriptome analysis

表1

qRT-PCR引物信息"

基因名称
Gene ID		 GO注释
GO annotation		 引物序列
Primer sequence (5°-3°)
Actin F: TTGGAATGGGTCAGAAGGATGC
R: GCTTCTTACTGAGGCACCACT
Arahy.HGTY5Y UDP葡萄糖6-脱氢酶活性, NAD结合
UDP-glucose 6-dehydrogenase activity (GO:0003979), NAD binding (GO:0051287)		 F: TATCACCCAACCGTGTCAGC
R: CATCCCTACAGCAACCGGAG
Arahy.ZL9QUM O-甲基转移酶活性, 甲基化
O-methyltransferase activity (GO:0008171), Methylation (GO:0032259)		 F: TTGATCTTCCCCAGGTTGTGG
R: GTGATGTCATGCTTGTCCTGC
Arahy.5I1IFA 催化活性, 氧化应激反应的调节
Catalytic activity (GO:0003824), regulation of response to oxidative stress (GO:1902882)		 F: GTGGACTTGGTGGTGCTCAA
R: TGTTTTCGGCTTCACTCGGA
Arahy.JA1EPG 水解酶活性, 水解邻糖基化合物, 碳水化合物代谢
Hydrolase activity, hydrolyzing O-glycosyl compounds (GO:0004553), carbohydrate metabolic process (GO:0005975)		 F: ACGCGCAGTCGTATAACAGT
R: TGGAAGAGTCCCCAGTTCCT
Arahy.WS4P7I 谷氨酸5-激酶活性, 谷氨酸-5-半醛脱氢酶活性
Glutamate 5-kinase activity (GO:0004349), Glutamate-5-semialdehyde dehydrogenase activity (GO:0004350)		 F: AATGACAGTTTGGCGGGTCT
R: CCCCTGCCCAATCTTGACTT
Arahy.QWX0SS 蛋白激酶活性, 膜的整体组成
Protein kinase activity (GO:0004672), integral component of membrane (GO:0016021)		 F: GTTAAGGGCGTCGCGAATG
R: TTCTGGAGCAGCATAGCCAA

表2

不同处理对花生农艺性状的影响"

处理
Treatment		 生物量 Biomass (g plant-1) 根冠比Root/shoot ratio 叶片相对含水量Leaf relative water content (%)
2020 2021 2020 2021 2020 2021
WWNN 5.86 ab 5.07 b 0.14 a 0.12 a 92.54 a 96.71 a
WWNA 6.78 a 5.43 a 0.12 a 0.11 a 93.24 a 96.00 a
DSNN 5.04 b 3.71 d 0.15 a 0.12 a 72.93 c 64.71 b
DSNA 6.33 ab 4.64 c 0.13 a 0.10 a 84.39 b 84.93 a
W 0.120 0 ** 0.397 0.536 0** 0.001**
N 0.016* 0.001** 0.028* 0.046* 0.002** 0.037*
W×N 0.630 0.052 0.667 0.826 0.005** 0.028*

表3

不同处理对花生酚类物质含量的影响"

处理
Treatment		 总酚含量Total phenol content (mg g-1) 类黄酮含量Flavonoid content (mg g-1)
地上部
Shoot		 根系
Root		 整株
Whole plant		 地上部
Shoot		 根系
Root		 整株
Whole plant
2020 2021 2020 2021 2020 2021 2020 2021 2020 2021 2020 2021
WWNN 7.49 ab 7.41 a 5.02 a 4.62 a 7.22 ab 7.10 a 8.79 a 8.93 a 3.96 a 4.56 a 8.26 a 8.48 a
WWNA 6.97 c 6.73 b 4.96 a 4.19 b 6.72 c 6.49 b 7.19 b 7.67 b 4.09 a 4.94 a 6.80 b 7.67 b
DSNN 7.27 bc 6.81 b 4.46 a 3.41 c 6.99 bc 6.49 b 6.90 b 7.52 b 3.58 a 4.34 a 5.95 b 7.22 b
DSNA 7.83 a 7.25 a 4.87 a 4.14 b 7.40 a 6.92 a 8.87 a 7.99 ab 3.86 a 5.80 a 8.22 a 7.76 ab
W 0.212 0.668 0.183 0** 0.338 0.325 0.863 0.106 0.380 0.529 0.235 0.142
N 0.930 0.001** 0.319 0** 0.837 0** 0.764 0.020* 0.548 0.097 0.274 0.015*
W×N 0.052 0.301 0.464 0.133 0.076 0.334 0.016* 0.224 0.829 0.306 0.001** 0.315

图1

不同处理对花生根系抗氧化酶活性及丙二醛含量的影响 标以不同小写字母的柱值表示处理间在0.05水平上差异显著。处理同表2。"

图2

不同处理间样本相关性分析及花生根系差异表达基因维恩图 处理同表2。"

图3

施氮处理下差异表达基因的功能注释分析"

图4

干旱处理下差异表达基因的功能注释分析 AP2/ERF: AP2/ERF转录因子; GRAS: GRAS转录因子; WRKY: WRKY转录因子; G2-like: G2-like转录因子; LOB: 侧生器官边界结构域转录因子; bHLH: 碱性螺旋-环-螺旋转录因子; C2H2: 锌指结构转录因子; NAC: NAC转录因子; B3-ARF: B3-ARF转录因子; MYB: MYB类转录因子; MYB-related: MYB-related转录因子; B3: B3转录因子; C2C2-CO-like: C2C2-CO-like转录因子; bZIP: 碱性亮氨酸拉链类转录因子; SRS: SRS转录因子。"

图5

旱氮同存处理差异表达基因的KEGG的富集分析"

图6

不同处理下酚类物质相关途径DEGs表达分析 处理同表2。"

图7

转录组测序结果的qRT-PCR 验证 A~F为6个基因的表达趋势图; G为相关性分析图。处理同表2。**, P < 0.01。"

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