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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (1): 225-238.doi: 10.3724/SP.J.1006.2023.24020

• TILLAGE & CULTIVATION · PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

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 Online:2023-01-12 Published:2022-05-13
  • Contact: DAI Liang-Xiang E-mail:dingpeanut@163.com;liangxiangd@163.com
  • 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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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

Table 1

Information of qRT-PCR primers"

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

Table 2

Effects of different treatments on agronomic characters of peanut"

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*

Table 3

Effects of different treatments on the content of secondary metabolites in peanut"

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

Fig. 1

Effects of different treatments on antioxidant enzyme activities and MDA contents of peanut roots Bars superscripted by different letters are significantly different at the 0.05 probability level. Treatments are the same as those given in Table 2."

Fig. 2

Correlation analysis of samples between different treatments and Venn diagram of differential gene expression of peanut roots Treatments are the same as those given in Table 2."

Fig. 3

Function annotations of differentially expressed genes in nitrogen application treatment"

Fig. 4

Function annotations of differentially expressed genes in drought treatment AP2/ERF: apetala 2/ethylene-responsive element binding factor; GRAS: GRAS transcription factor; WRKY: WRKY transcription factor; G2-like: G2-like transcription factor; LOB: lateral organ boundaries domain transcription factor; bHLH: basic Helix-Loop-Helix transcription facto; C2H2: zinc finger domain transcription factor; NAC: NAC transcription factor; B3-ARF: B3-ARF transcription factor; MYB: MYB (v-myb avian myeloblastosis viral oncogene homolog) transcription factor; MYB-related: MYB (v-myb avian myeloblastosis viral oncogene homolog) related transcription factor; B3: B3 domain transcription factor; C2C2-CO-like: C2C2-CO-like transcription factor; bZIP: basic region/leucine zipper motif ; SRS: SRS transcription factor."

Fig. 5

KEGG analysis of differentially expressed genes in drought and nitrogen application simultaneously exist treatment"

Fig. 6

Relative expression profiles of DEGs phenolic related pathways under different treatments Treatments are the same as those given in Table 2."

Fig. 7

qRT-PCR validation of transcriptome sequencing A-F are the relative expression profiles of six validated genes, respectively; G is the correlation diagram of validated gene. Treatments are the same as those given in Table 2. **, P < 0.01."

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