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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (10): 2503-2514.doi: 10.3724/SP.J.1006.2024.31075

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

Salt tolerance analysis of HvMBF1c in barley

ZHAN Xiao-Xiao1,2(), FENG Ju-Ling1,2, ZHANG Zhen-Huan1,3, ZHANG Hong1,2, WANG Jun-Cheng1,2, LI Bao-Chun1,3, YANG Ke1,2, SI Er-Jing1,2, MENG Ya-Xiong1,2, MA Xiao-Le1,2, WANG Hua-Jun1,2, YAO Li-Rong1,2,*()   

  1. 1State Key Laboratory of Crop Science in Arid Habitat Jointly Established by the Ministry of Province and Ministry of Finance / Key Laboratory of Crop Genetic Improvement and Germplasm Innovation in Gansu Province, Lanzhou 730070, Gansu, China
    2Agronomy College, Gansu Agricultural University, Lanzhou 730070, Gansu, China
    3College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2023-11-30 Accepted:2024-06-20 Online:2024-10-12 Published:2024-07-11
  • Contact: *E-mail: ylr0384@163.com
  • Supported by:
    China Agriculture Research System of MOF and MARA(Barley, CARS-05-03B-03);China Agriculture Research System of MOF and MARA(CARS-05-03B-03);National Natural Science Foundation of China(30771331);National Natural Science Foundation of China(32160496);Gansu Agricultural University Student Research Training Program(202301047);Gansu Agricultural University Student Research Training Program(202210733008);Gansu Agricultural University Student Research Training Program(S202210733003);Gansu Youth Science and Technology Fund Program(20JR5RA010);Innovation Fund of Education Department of Gansu Province(2021A-055);Industry Support Plan of Education Department of Gansu Province(2021CYZC-12)

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Abstract:

The HvMBF1c gene in barley is implicated in the plant’s response to salt stress. To investigate the HvMBF1c response mechanism to salt stress, we conducted bioinformatics analysis of the HvMBF1 gene, qRT-PCR analysis, and physiological index determination in Nakagawa barley (salt-tolerant type), GN18 (salt-sensitive type), wild-type (WT) and transgenic HvMBF1c Arabidopsis thaliana treated with 200 mmol L-1 NaCl for 0, 6, 12, 24, 48, and 72 hours. The results showed that the barley HvMBF1c gene is closely related to the wheat TaMBF1c gene, and is located on chromosome 7H. The exon of HvMBF1c is larger than those of HvMBF1a and HvMBF1b, with HvMBF1a and HvMBF1b containing a typical motif 4 domain, while the HvMBF1c gene has a unique motif 5 domain. The promoter sequence of the HvMBF1c gene includes photoreactive, root-specific regulatory elements, and hypoxia-specific inducible enhancing elements, homologous to wheat chromosomes 2, 3, and 7. qRT-PCR analysis revealed significant upregulation of HvMBF1c expression in two barley genotypes at the seedling, jointing, heading, and filling stages, with higher expression in Nakagawa barley compared to GN18 as salt treatment duration increased. Similarly, the expression of HvMBF1c in Arabidopsis thaliana also increased with prolonged salt treatment time. Physiological indices of Nakagawa barley at each growth stage were lower than those in GN18, and transgenic Arabidopsis exhibited lower indices compared to WT under different salt stress conditions. These findings provide a foundation for further exploration of the HvMBF1c gene function.

Key words: barley, HvMBF1c, physiological indicators, collinearity analysis, the extension

Table 1

Sequence and purpose of primers"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 用途
Purpose
HvMBF1c-qF
HvMBF1c-qR
HvActin-F
HvActin-R		 CATCCAGAAGGCTCGCGT
CTCTCGTACTCCTGCACCAC
AAGCAGCCAGAATGTACAGCGAGAAC
GGTACAGACCAGCAAAGCCAGAAATG		 荧光定量PCR
Fluorescence quantitative PCR
内参基因引物
Internal reference gene primer

Fig. 1

Phylogenetic evolutionary tree of MBF1 gene families in barley, wheat, rice, maize, and Arabidopsis Hv: barley; Ta: wheat; Os: rice; Zm: maize; At: Arabidopsis."

Fig. 2

Distribution of barley HvMBF1 gene family members on barley chromosomes"

Fig. 3

Gene structure of barley HvMBF1 gene family members UTR: untranslated region; CDS: coding region sequence."

Fig. 4

Family members of barley HvMBF1 gene conserve motif Motif: conserved base sequence."

Fig. 5

Logo of conserved motif sequence of barley HvMBF1 gene"

Fig. 6

Cis-acting element in the promoter of barley HvMBF1 gene"

Fig. 7

Collinear analysis of HvMBF1 gene in barley and wheat"

Fig. 8

HvMBF1c expression of barley at various periods and Arabidopsis thaliana under salt stress Different letters indicate significant differences at P < 0.05."

Fig. 9

Changes of SOD, POD, MDA, CAT, and soluble protein activities in barley under salt stress Different letters indicate significant differences at P < 0.05."

Fig. 10

Changes in SOD, POD, MDA, CAT, and soluble protein activities in Arabidopsis thaliana under salt stress Different letters indicate significant differences at P < 0.05."

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