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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (7): 1882-1894.doi: 10.3724/SP.J.1006.2023.24172

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Evaluation of salt tolerance in Cyperus esculentus and transcriptomic analysis of seedling roots under salt stress

WANG Hui-Wei(), ZHANG Xiang-Ge, LI Chun-Xin, XU Xin-Ran, HU Hai-Yan, ZHU Ya-Jing, WANG Yan, ZHANG Xin-You*()   

  1. Industrial Crop Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
  • Received:2022-08-01 Accepted:2022-10-10 Online:2023-07-12 Published:2022-10-24
  • Contact: *E-mail: haasz@126.com E-mail:whuiweiw@163.com;haasz@126.com
  • Supported by:
    The National Key Research and Development Project(2019YFD1002600);The Henan Major Science and Technology Project(211100110100);The Excellent Youth Science and Technology Fund of Henan Academy of Agricultural Sciences(2022YQ19)

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Abstract:

Cyperus esculentus is a new industrial crop with the high comprehensive utilization value, which is a strongly resistant to stresses and has a great potential to grow in saline soil. In order to clarify its salt tolerance, five NaCl concentrations (0, 0.3%, 0.6%, 0.9%, and 1.2%) were set in this study to analyze the effects of NaCl stress on morphological and physiological indexes during germination and seedling growth. The results showed that the germination percentage, root length, and seedling height were less affected under 0.3% and 0.6% NaCl stresses. Meanwhile, the contents of indicator of membrane damage degree, malondialdehyde (MDA), and oxidative stress substance, hydrogen peroxide (H2O2), did not increase significantly. However, the contents of osmoregulation substances [glycine betaine (GB), and proline (Pro)] and the activities of antioxidant enzymes [superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)] were significantly increased, and the damage of salt stress was less on the whole. However, under 0.9% and 1.2% NaCl stresses, the salt stress injury was very obvious and the germination and seedling growth of Cyperus esculentus were severely inhibited. In order to further identify the genes related to salt tolerance, RNA-seq technology was used to detect the gene expression in roots under 0, 0.3%, and 0.6% NaCl stresses. 24 GO terms mainly related to oxidoreduction, transmembrane transport, chitin hydrolysis was significantly enriched, among which 15 significantly up-regulated genes were involved, through gene differential expression analysis, weighted gene co-expression network analysis (WGCNA), and GO enrichment analysis. Among them, DN23985_c0_g1, DN2960_c0_g1, and DN8384_c1_g1 encoded zeaxanthin epoxidase, L-ascorbate peroxidase, and glutathione S-transferase, respectively, which had antioxidant effects and participated in antioxidant regulation. Both DN21785_c1_g1 and DN6596_c0_g1 encoded amino acid transporters, which may enhance osmoregulation by accumulating small amino acid molecules such as Pro. DN14393_c0_g1 encoded chitinase, which hydrolyzed chitin and promoted plant response to stress. In this study, it was confirmed that Cyperus esculentus had a good salt tolerance under 0.6% or less NaCl stress, and the salt-tolerance related genes were further screened out, which provided an important reference for the cultivation in saline soil and the breeding of salt-tolerant varieties.

Key words: Cyperus esculentus, salt tolerance, NaCl stress, transcriptomics, salt-tolerance related gene

Fig. 1

Effects of different NaCl concentrations on germination of Cyperus esculentus tubers A: germination morphology of Cyperus esculentus tubers under NaCl stress with different concentrations. B: the germination rate and salt tolerance index of Cyperus esculentus tubers under NaCl stress with different concentrations. Bar: 2 cm. YYS2: Yuyousha 2; YYS3: Yuyousha 3."

Fig. 2

Effects of different NaCl concentrations on seedling growth of Cyperus esculentus A: the characteristics of seedling growth of YYS2 under NaCl stress with different concentrations. B: the characteristics of seedling growth of YYS3 under NaCl stress with different concentrations. Bar: 5 cm. YYS2: Yuyousha 2; YYS3: Yuyousha 3."

Table 1

Effects on seedling height and root length of two Cyperus esculentus varieties under NaCl stress"

NaCl胁迫处理
NaCl stress treatment		 苗高Seedling height (cm) 根长Root length (cm)
豫油莎2号
Yuyousha 2		 豫油莎3号
Yuyousha 3		 豫油莎2号
Yuyousha 2		 豫油莎3号
Yuyousha 3
0 (CK) 18.00 ± 1.16 a 15.75 ± 1.25 a 16.56 ± 1.37 a 19.28 ± 1.68 a
0.3% 15.26 ± 1.08 b 13.53 ± 1.17 b 14.16 ± 1.12 b 17.54 ± 1.34 b
0.6% 14.65 ± 0.96 b 13.56 ± 0.85 b 13.53 ± 1.03 c 14.56 ± 1.23 c
0.9% 12.92 ± 1.12 c 11.50 ± 1.23 c 12.85 ± 0.93 d 13.26 ± 1.08 c
1.2% 10.93 ± 0.84 d 9.75 ± 0.99 d 8.92 ± 0.78 e 9.58 ± 0.93 d

Fig. 3

Variation of physiological indexes of Cyperus esculentus roots under different NaCl concentrations * represents significant difference at P < 0.05. YYS2: Yuyousha 2; YYS3: Yuyousha 3."

Table 3

Statistic analysis of transcriptome sequencing data"

处理
Treatment		 测序总读数
Total reads number		 质控后读数
Clean reads number		 比对率
Mapped ratio (%)		 Q20碱基百分比
Q20
(%)		 Q30碱基百分比
Q30
(%)		 非冗余基因数
Unigene
number
CK-1 46,067,738 42,556,954 92.37 97.55 93.77 30,191
CK-2 42,308,034 39,254,802 92.78 97.71 94.06
CK-3 50,830,434 47,129,824 92.71 97.59 93.79
0.3%-1 57,271,072 53,011,182 92.56 97.70 93.93
0.3%-2 43,325,228 40,182,946 92.74 97.76 94.00
0.3%-3 52,217,758 48,377,604 92.64 97.48 93.43
0.6%-1 43,209,368 40,018,352 92.61 97.59 93.73
0.6%-2 46,496,392 43,155,836 92.81 97.52 93.58
0.6%-3 41,707,820 38,775,376 92.96 97.39 93.37

Fig. 4

Correlation analysis of gene relative expression levels among duplicate samples Red box represents three replicates under the same treatment."

Fig. 5

Differential relative expression level of genes under different concentrations of NaCl stress A: the number of DEGs in different treatment groups (0.3% vs CK and 0.6% vs CK); B: Venn analysis of DEGs between different treatment groups."

Fig. 6

Screening of salt tolerance-related genes A: the WGCNA based on superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glycine betaine (GB), and proline (Pro). Each row represents a module and each column represents a trait; the correlation coefficient and the corresponding P-value are shown in each rectangular box. B: Venn analysis between the overlapped DEGs and the salt tolerance-related yellow module genes."

Fig. 7

GO enrichment analysis of salt tolerance-related genes A: the bubble diagram of GO enrichment based on MF (molecular function). B: the relative expression level and functional annotation of all genes in significantly enriched GO terms."

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