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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (4): 1080-1090.doi: 10.3724/SP.J.1006.2024.31053

• RESEARCH NOTES • Previous Articles    

Alleviative effect of salicylic acid on wheat seedlings with stripe rust based on transcriptome and differentially expressed genes

QI Xue-Li1(), LI Ying2(), LI Chun-Ying3, HAN Liu-Peng1, ZHAO Ming-Zhong1, ZHANG Jian-Zhou3,*()   

  1. 1Crops Molecular Breeding Academy of Henan, Zhengzhou 450002, Henan, China
    2Editorial Department of Journal of Henan Agricultural University, Zhengzhou 450002, Henan, China
    3Wheat Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
  • Received:2023-09-17 Accepted:2024-01-12 Online:2024-04-12 Published:2024-02-23
  • Contact: * E-mail: zhangjianzhou00@163.com
  • About author:**Contributed equally to this work
  • Supported by:
    China Agriculture Research System of MOF and MARA (Wheat, CARS-3-7);Independent Innovation Project of HAAS(2022ZC03)

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Abstract:

In order to explore the mechanism of exogenous salicylic acid in improving the resistance of wheat stripe rust, using, the untreated wheat seedlings (control), stripe rust wheat seedlings (SR), and salicylic acid treated stripe rust wheat seedlings (SA-SR) as the experimental materials were investigated using Zhoumai 18. After 15 days of treatment, the wheat seedlings were identified for disease resistance, amino acid detection, transcriptome sequencing, and fluorescence quantification. The results indicated that the exogenous salicylic acid could significantly reduce the pathogenicity of stripe rust in wheat seedlings. The content of amino acids in leaves in the SR group was generally reduced, while there was no significant difference between the SA-SR group and the control group pretreated with salicylic acid, indicating that the content of amino acids in leaves was significantly reduced due to wheat stripe rust. GO enrichment indicated that the differentially expressed genes of SR and SA-SR groups were enriched in biological processes related to photosynthesis involving cell components such as chloroplasts and thylakoids. KEGG pathway enrichment showed that both SR and SA-SR subgroups were significantly enriched in the MAPK signaling pathway and the biosynthesis of secondary metabolites played an important role in plant disease resistance or stress resistance.

Key words: wheat stripe rust, salicylic acid, amino acid, transcriptom

Fig. 1

Grading standard of wheat stripe rust at seedling stage (grades 0-9)"

Table 1

Primers for real-time quantitative PCR"

基因名称
Gene ID		 正向引物
Forward sequence (5'-3')		 反向引物
Reverse sequence (5'-3')
TraesCS2D02G433300 CGCCGTGCGTGAGTAATTC CGTCGGGTTCGACACTCTTA
TraesCS2D02G014800 CGACCTCAACTCCAACTCCATA ACGGATTCCCCTCTCCGAA
TraesCS6B02G078400 TGGGAGCAATACTTCGCCTG ATAGCCCATCACCCCCGTAT
TraesCS1B02G038100 AGTTAGACCAAATGGCCCCTC CCCCCTGAGGTATAGACCGA
TraesCS5D02G099000 AAGCCTTTGCTGTCCTACGG AATGGGCCTGTAATCGGTGG
TraesCS2A02G378900 CCAGTCGGAGCACGTTATCA GATGGTGCCGTAGATCCGTT
TraesCS2D02G375200 TCCATTGTAGGCTTGCTGGA CCGGGTGATAACCCACAAGTA
TraesCS5D02G531600 CAGCCCTTTATAGGCCCACG TAGGCCGAAAGCTAACACGG
TraesCS1D02G290100 CGACGCCTATAGCAAACGGA GCCTGAAGCCAACGAAACTC
TraesCS1B02G011100 CGTATGCGCTATTCGTTGGC GGCGAGGATTCAGGAACTGT

Fig. 2

Response types of SR and SA-SR to CYR32 at seedling stage"

Table 2

Hydrolyzed amino acid content of wheat leaves in three groups"

氨基酸
Amino acid		 对照Control SR SA-SR
C_1 C_2 C_3 SR_1 SR_2 SR_3 SA-SR_1 SA-SR_2 SA-SR_3
天冬氨酸Asp 3.302 3.421 3.411 2.659 2.723 2.502 3.761 3.613 3.559
谷氨酸Glu 6.076 5.078 5.295 5.492 5.333 5.880 5.878 5.295 6.492
丝氨酸Ser 1.213 1.473 1.354 1.022 0.988 0.813 1.073 1.054 1.122
组氨酸His* 0.574 0.525 0.578 0.571 0.501 0.402 0.525 0.578 0.671
甘氨酸Gly 1.500 1.621 1.548 1.447 1.413 1.500 1.521 1.548 1.747
苏氨酸Thr* 0.664 0.617 0.649 0.512 0.551 0.643 0.617 0.649 0.670
精氨酸Arg 3.233 3.701 3.623 2.612 2.813 2.233 2.791 2.820 3.609
丙氨酸Ala 1.063 1.111 1.262 1.200 0.961 1.063 1.011 0.962 1.212
酪氨酸Tyr 0.771 0.752 0.729 0.831 0.689 0.771 0.652 0.679 0.831
胱氨酸Cys-s 0.115 0.112 0.099 0.029 0.016 0.109 0.120 0.199 0.129
缬氨酸Val* 1.153 1.011 1.121 1.119 1.115 1.153 1.011 1.121 1.319
甲硫氨酸Met* 0.169 0.155 0.177 0.128 0.134 0.129 0.155 0.157 0.188
苯丙氨酸Phe* 1.378 1.216 1.313 1.209 1.008 1.018 1.216 1.113 1.209
异亮氨酸Ile* 0.984 0.851 0.894 0.727 0.696 0.683 0.851 0.894 0.827
亮氨酸Leu* 1.645 1.555 1.599 1.360 1.402 1.345 1.519 1.581 1.580
赖氨酸Lys* 0.803 0.792 0.832 0.860 0.803 0.703 0.850 0.842 1.060
脯氨酸Pro 0.912 0.902 0.927 0.878 0.820 0.790 0.960 0.938 1.268
总量 Total 24.118 24.626 24.113 21.772 19.713 20.346 24.236 22.765 26.349

Fig. 3

Comparison of 17 hydrolyzed amino acids in wheat leaves in the three groups"

Table 3

Statistics of all the samples of sequencing data"

样品
SA sample		 原始序列数
Raw reads		 过滤后序列数
Clean reads		 碱基错误率
Error rate (%)		 GC含量
GC content (%)		 比对率
Total mapped
C_1 96129064 95045666 0.02 55.57 92039992 (96.84%)
C_2 89832494 88505872 0.02 55.44 85551890 (96.66%)
C_3 79838786 78959024 0.02 55.61 76422986 (96.79%)
SR_1 97621350 96607070 0.02 55.67 93194776 (96.47%)
SR_2 96303100 95196330 0.02 55.72 91799894 (96.43%)
SR_3 95914068 94943432 0.02 55.72 91563514 (96.44%)
SA-SR_1 101359676 100284658 0.02 54.67 96353104 (96.08%)
SA-SR_2 109261866 108220880 0.02 54.75 104057256 (96.15%)
SA-SR_3 93098314 92227658 0.02 54.71 88651196 (96.12%)

Fig. 4

Correlation analysis of transcriptome sequencing data between different samples a: Pearson correlation analysis within and between groups; b: PCA analysis of gene expression levels between groups."

Fig. 5

Volcano plot of differentially expressed genes"

Fig. 6

Analysis of differentially expressed genes in different combinations a: clustering of genes that share differences. b: Venn diagram showed the number of DEGs between SA-SR vs SR and SR vs Control."

Table 4

Top 10 significantly enriched GO term in SA-SR vs SR"

GO条目
GO_accession		 注释描述
Description		 P值
P-value		 上调
Up		 下调
Down		 类别
Term_type
GO:0009536 质体Plastid 2.28E-95 44 681 细胞组分Cell component
GO:0009507 叶绿体Chloroplast 2.03E-91 40 663 细胞组分Cell component
GO:0009579 类囊体Thylakoid 3.11E-61 34 362 细胞组分Cell component
GO:0042651 类囊体膜Thylakoid membrane 5.71E-56 14 222 细胞组分Cell component
GO:0015979 光合作用Photosynthesis 3.09E-51 34 263 生物过程Cell component
GO:0034357 光合作用膜Photosynthetic membrane 2.99E-49 31 265 细胞组分Cell component
GO:0009534 叶绿体类囊体Chloroplast thylakoid 3.60E-47 8 200 细胞组分Cell component
GO:0031976 质体类囊体Plastid thylakoid 3.60E-47 8 200 细胞组分Cell component
GO:0009532 质体基质Plastid stroma 1.56E-42 6 215 细胞组分Cell component
GO:0009535 叶绿体类囊体膜Chloroplast thylakoid membrane 2.67E-42 8 172 细胞组分Cell component

Table 5

Top 10 significantly enriched GO term in SA-SR vs Control"

GO条目
GO_accession		 注释描述
Description		 P值
P-value		 上调
Up		 下调
Down		 类别
Type
GO:0009800 肉桂酸生物合成过程
Cinnamic acid biosynthetic process		 1.19E-70 35 0 生物过程
Cell bioprocess
GO:0009803 肉桂酸代谢过程
Cinnamic acid metabolic process		 1.19E-70 35 0 生物过程
Cell bioprocess
GO:0045548 苯丙氨酸解氨酶活性
Phenylalanine ammonia-lyase activity		 1.19E-70 35 0 分子功能
Molecular function
GO:0016301 激酶活性
Kinase activity		 1.19E-66 1260 263 分子功能
Molecular function
GO:0006468 蛋白质磷酸化
Protein phosphorylation		 9.25E-66 1163 226 生物过程
Molecular function
GO:0004672 蛋白激酶活性
Protein kinase activity		 1.48E-65 1163 230 分子功能
Molecular function
GO:0016773 磷酸转移酶活性, 醇基作为受体 4.14E-65 1230 247 分子功能
Molecular function
GO:0016310 磷酸化
Phosphotransferase activity, alcohol group as acceptor		 1.76E-58 1208 249 生物过程
Cell bioprocess
GO:0006796 含磷酸盐化合物代谢过程
Phosphate-containing compound metabolic process		 9.77E-56 1399 361 生物过程
Cell bioprocess
GO:0006793 磷代谢过程
Phosphorus metabolic process		 2.37E-55 1401 361 生物过程
Cell bioprocess

Fig. 7

GO enrichment analysis of differentially expressed genes in different combinations a: SA-SR vs SR; b: SR vs Control."

Fig. 8

Two KEGG pathways comparing combining differential genes a: KEGG pathway enrichment analysis of differentially expressed genes in SA-SR vs SR. b: KEGG pathway enrichment analysis of differentially expressed genes in SR vs Control."

Fig. 9

Spearman correlation analysis of RNA seq and qRT- PCR"

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