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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (12): 3144-3154.doi: 10.3724/SP.J.1006.2024.43017

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Salt tolerance evaluation and transcriptome analysis of maize mutant caspl2b2

ZHANG Jin-Hui1(), XIAO Zi-Yi1, LI Xu-Hua3, ZHANG Ming3, JIA Chun-Lan3, PAN Zhen-Yuan2,*(), QIU Fa-Zhan1,*()   

  1. 1National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University / Hubei Hongshan Laboratory, Wuhan 430070, Hubei, China
    2College of Agriculture, Shihezi University / Key Laboratory of Oasis Ecological Agriculture Corps, Shihezi 832003, Xinjiang, China
    3Shandong Denghai Seed Industry Co., Ltd, Laizhou 261448, Shandong, China
  • Received:2024-04-20 Accepted:2024-08-15 Online:2024-12-12 Published:2024-08-23
  • Contact: *E-mail: panzhenyuandawood@163.com; E-mail: qiufa
  • Supported by:
    National Natural Science Foundation of China(U2106230);Key Research and Development Program of Shandong Province(2022CXPT014)

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Abstract:

Salt stress-sensitive mutants are crucial genetic materials for studying the genetic basis and molecular mechanisms of salt tolerance in crops. In this study, the maize inbred line LY8405 and the mutant caspl2b2 were used to investigate phenotype, physiological, and biochemical indices at the seedling stage under normal growth and salt stress conditions. The results showed that, compared to LY8405, the survival rate of caspl2b2 significantly decreased under salt stress, and the growth of aboveground parts was notably inhibited. The content of Na+ ions and malondialdehyde (MDA) in aboveground parts significantly increased, while transpiration rate, stomatal conductance, and intercellular CO2 concentration also increased significantly. Conversely, the net photosynthetic rate significantly decreased. To uncover the molecular basis of the phenotypic differences under salt stress, transcriptome analysis was performed on leaf tissues of LY8405 and caspl2b2 under normal growth and salt stress conditions. The results revealed that differentially expressed genes (DEGs) were mainly concentrated in glutathione transferase activity, glutathione metabolism, REDOX enzyme activity, and cell homeostasis, with glutathione metabolism being the most significant. This study not only provides important germplasm resources for the genetic basis analysis of crop salt tolerance, but also lays a foundation for the identification of salt tolerance genes and the analysis of genetic regulatory networks.

Key words: Zea mays L., salt stress, mutant, transcriptome analysis, differentially expressed genes

Fig. 1

Salt concentration screening of caspl2b2 mutant A: the phenotype of WT and caspl2b2 mutant before salt treatment; B-D: the phenotype of WT and caspl2b2 mutant after 100 mmol L-1, 150 mmol L-1, and 200 mmol L-1 NaCl treated for 7 days; E: survival rate of caspl2b2 mutant treated with different salt concentrations."

Fig. 2

Phenotype of WT and caspl2b2 mutant after salt treatment A: the phenotype of WT and caspl2b2 mutant before salt treatment; B, D: the phenotype of WT and caspl2b2 mutant after 200 mmol L-1 NaCl treated for 3 days, 7 days, and 14 days; E: the survival rates of WT and caspl2b2 mutant after salt treatment for different days."

Fig. 3

Phenotypic traits of WT and caspl2b2 mutant after salt stress A-D: phenotypic traits of seedling height, primary root length, shoot fresh weight, and root fresh weight of WT and caspl2b2 mutant after 200 mmol L-1 NaCl treatment. E-G: the total phenotype, leaf phenotype, and root phenotype of WT and caspl2b2 mutant after 200 mmol L-1 NaCl treated 3 days. The above data are presented as mean ± SD. Student’s t-test was used for significance analysis. NS: no significant difference. *: P < 0.05; **: P < 0.01."

Fig. 4

Na+ and K+ content of WT and caspl2b2 mutant after salt stress A: Na+ content of WT and caspl2b2 in shoot; B: K+ content; C: Na+/K+; D: MDA content of WT and caspl2b2 mutant. The above data are presented as mean ± SD. Student’s t-test was used for significance analysis. NS: no significant difference. *: P < 0.05; **: P < 0.01."

Fig. 5

Photosynthetic indicators of WT and caspl2b2 mutant after salt stress A-D: photosynthetic indices of WT and caspl2b2 mutant treated with 200 mmol L-1 NaCl after 3 d; A: transpiration rate; B: stomatal conductivity; C: intercellular CO2 concentration; D: net photosynthetic rate. The above data are presented as mean ± SD. Student’s t-test was used for significance analysis. NS: no significant difference. *: P < 0.05; **: P < 0.01."

Table 1

Transcriptome sequencing data"

样本
Sample		 测序总读数
Total reads number		 质控后读数
Clean reads number		 错误率
Error rate
(%)		 总比对率
Mapped ratio (%)		 Q20 碱基百分比
Q20 base percentage (%)		 Q30 碱基百分比
Q30 base percentage (%)
WT-1 21,635,978 20,957,262 0.02 96.86 98.22 94.71
WT-2 24,644,756 23,996,460 0.02 97.37 98.21 94.81
WT-3 22,386,955 21,537,487 0.02 96.21 98.12 94.55
Caspl2b2-1 25,381,514 24,489,459 0.02 96.49 98.20 94.72
Caspl2b2-2 23,662,504 22,826,123 0.02 96.47 98.14 94.68
Caspl2b2-3 23,551,867 22,839,156 0.02 96.97 97.94 94.32

Fig. 6

Distribution and correlation analysis of gene expression in samples A: distribution of gene expression in each sample; B: correlation heat map between samples."

Fig. 7

Volcano plot of differentially expressed genes Up: up-regulated gene; Down: down-regulated gene; Not signifi-cant: not significant difference gene."

Fig. 8

Gene enrichment analysis with significant differences A: GO enrichment of differentially expressed genes; B: KEGG enrichment of genes with significant differences."

Table 2

Differentially expressed genes associated with ion transport"

基因编号
Gene ID		 基因名称
Gene name		 描述
Description		 log2
(Fold Change)		 P值
P-value
Zm00001d003861 ZmHAK1 Potassium high-affinity transporter -0.15 0.44
Zm00001d042244 ZmHAK5 Potasium ion uptake permease 1 -0.61 0.55
Zm00001d049987 ZmHAK4 Probable potassium transporter 4 -0.31 0.71
Zm00001d040627 ZmHKT1 Sodium transporter HKT1 0.92 0.01
Zm00001d030955 ZmSOS3 CBL11; calcineurin B-like11: cbl11 -0.07 0.77
Zm00001d031232 ZmSOS1 Sodium/hydrogen exchanger 0.14 0.49
Zm00001d002428 ZmNSA1 EF hand family protein 0.46 3.00E-03
Zm00001d024587 ZmNPF6.4 Protein NRT1/PTR FAMILY 6.3 0.38 0.03

Fig. 9

qRT-PCR was used to detect the expression of ion transport-related genes A: expression level of ZmHKT1 gene; B: expression level of ZmHKT2 gene; C: expression level of ZmHAK1 gene; D: expression level of ZmHAK5 gene."

Table 3

Differentially expressed genes associated with the formation of Casparian strip"

基因编号
Gene ID		 基因名称
Gene name		 描述
Description		 log2
(Fold Change)		 P值
P-value
Zm00001d038762 ZmRBOHF RBOH3; respiratory burst oxidase3 -0.32 0.11
Zm00001d027636 ZmESB1 Dirigent protein 16 0.42 0.87
Zm00001d040807 ZmCASPL1B2 CASPL4; CASP-like protein 4 0.10 0.60
Zm00001d005158 ZmCASPL1D1 CASPL3; CASP-like protein 3 -0.27 0.52
Zm00001d019457 ZmCASPL1D2 CASPL1D2; CASP-like protein 1D2 -0.44 0.05

Table 4

Glutathione-related differentially expressed genes"

基因编号
Gene ID		 基因名称
Gene name		 描述
Description		 log2
(Fold Change)		 P值
P-value
Zm00001d049657 ZmGSTU1 GST26; glutathione transferase 26 -0.31 0.34
Zm00001d038192 ZmGSTU2 GST41; glutathione transferase 41 1.08 0.37
Zm00001d016860 ZmGSTU4 GST17; glutathione transferase 17 0.31 0.07
Zm00001d036951 ZmGSTU5 GST19; glutathione transferase 19 1.28 3.28E-09
Zm00001d043787 ZmGSTU6 GSTU6; glutathione S-transferase 6 0.85 0.003
Zm00001d043795 ZmGSTU10 GSTU10; glutathione S-transferase 10 0.98 8.47E-05
Zm00001d043789 ZmGSTU11 GSTU11; glutathione S-transferase 11 2.40 0.01
Zm00001d029706 ZmGSTU12 GST39; glutathione transferase 39 1.10 3.50E-07
Zm00001d043344 ZmGSTF3 GST8; glutathione transferase 8 0.80 0.20
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