Functional Analysis of ZmNF-YB13 Responding to Drought and Salt Stress

doi:10.13560/j.cnki.biotech.bull.1985.2022-0066

Abstract

Abstract:

Maize NF-Y family is a class of important transcription factors，they play an important role in regulating plant development and stress response. Exploring the gene function of this family will provide important gene resources for maize stress-resistant breeding. In this study，ZmNF-YB13 gene was cloned，and its basic characteristics，the relative tissue-specific expressions and expression patterns under different stresses were analyzed by bioinformatics and real-time quantitative PCR. The gene was 537 bp in length and encoded 178 amino acids. The molecular weight of ZmNF-YB13 protein was 18.9 kD，and the theoretical isoelectric point was 6.83. ZmNF-YB13 protein had a unique conserved domain of NF-Y family. qPCR analysis showed that ZmNF-YB13 gene expression was the highest in maize silk；meanwhile ZmNF-YB13 gene expression was up-regulated under different abiotic stress and hormone stress. On 1/2 MS medium containing mannitol，NaCl，ABA and JA，the root length of ZmNF-YB13 transgenic Arabidopsis thaliana was significantly longer than that of the wild type. Under drought and high salt treatments，transgenic A. thaliana grown in soil had more green leaves and lower MDA content than wild-type. Concurrently，the POD activity of transgenic plants was significantly higher than that of wild type under drought treatment. It is speculated that ZmNF-YB13 gene may be involved in response to drought and salt stress in maize.

Key words: maize, transcription factor, heterologous expression, drought stress, salt stress

ZHANG Tong-tong, ZHENG Deng-yu, WU Zhong-yi, ZHANG Zhong-bao, YU Rong. Functional Analysis of ZmNF-YB13 Responding to Drought and Salt Stress[J]. Biotechnology Bulletin, 2022, 38(10): 115-123.

Figures/Tables 7

Table 1 Primers used in this study

引物名称 Primer name	引物序列 Primer sequence（5'-3'）
pZmNF-YB13-FP	ATGGCGGAAGCTCCGGCGAGCC
pZmNF-YB13-RP	TTAGTTTGAGATATCCCCGTTA
pZmNF-YB13RT-FP	AATGGCGACGATCTGCTGTGG
pZmNF-YB13RT-RP	GCTATCACCCTGCACCTCTCTG
pGAPDH-FP	CCCTTCATCACCACGGACTAC
pGAPDH-RP	AACCTTCTTGGCACCACCCT
pZmNF-YB13T-FP	GAGAAGCGGAAGACCATCA
pZmNF-YB13T-RP	GTATAATTGCGGGACTCTAATC

Fig.1 Phylogenetic analysis of gene ZmNF-YB13

Fig.2 Tissue-specific expression pattern of gene ZmNF-YB13 in maize The error line in the figure refers to the standard deviation. Different letters indicate significant differences at the 0.05 level. The same below

Fig.3 Expressions of ZmNF-YB13 in response to abiotic and hormone stress in maize A：Dehydration treatment；B：cold（4℃）treatment；C：high salt（200 mmol/L NaCl）treatment；D：PEG treatment. The error bar represents± SD of triplicate experiments

Fig.4 Identification of T3 transgenic A. thaliana A：Identification of T3 generation A. thaliana by PCR. B：Identification of T3 generation A. thaliana by qPCR

Fig.5 Phenotypic analysis of ZmNF-YB13 transgenicA. thaliana under different stress treatments A：Phenotypic observation of A. thaliana under stress. B：Root length statistics of A. thaliana under stress treatment. *indicates significant difference at the 0.05 level. ** indicates very significant difference at the 0.01 level, the same below

Fig.6 Phenotypic analysis of A. thaliana under high salt and drought treatment in soil A：Analysis of phenotype of ZmNF-YB13 transgenic A. thaliana after high salt，drought and rewatering. B and C：Statistical analysis of green leaf number of transgenic and wild-type A. thaliana under high salt and drought treatment. D：Measurement of MDA content. E：Measurement of POD activity

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