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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (8): 2001-2013.doi: 10.3724/SP.J.1006.2024.33059

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

Functional analysis of maize N-acetyltransferase ZmNAT1 gene in response to abiotic stress

GUO Si-Yu1,2(), ZHAO Ke-Yong2(), DAI Zheng-Gang1,2, ZOU Hua-Wen1,*(), WU Zhong-Yi2,*(), ZHANG Chun2,*()   

  1. 1College of Agriculture, Yangtze University, Jingzhou 434025, Hubei, China
    2Institute of Biotechnology, Beijing Academy of Agriculture and Forestry Sciences / Beijing Key Laboratory of Agricultural Gene Resources and Biotechnology, Beijing 100097, China
  • Received:2023-10-17 Accepted:2024-01-31 Online:2024-08-12 Published:2024-02-28
  • Contact: * E-mail: spring2007318@163.com;E-mail: zouhuawen@yangtzeu.edu.cn;E-mail: zwu22@126.com
  • About author:** Contributed equally to this work
  • Supported by:
    National Natural Science Foundation of China(32001430);National Natural Science Foundation of China(32171952);National Natural Science Foundation of China(31971839)

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Abstract:

The GNAT (Gcn5-related N-acetyltransferase) family proteins play a crucial role in regulating plant growth and development, and responding to stress. At present, the biological functions of GNAT family genes have been reported in many species, but there are few studies on its function validation in maize (Zea mays L.). Exploring the functions of maize GNAT family genes not only enriches the genetic resources for maize breeding in China, but also provides an important basis for the creation of new germplasm resources of maize. In this study, the ZmNAT1 gene (Gene ID: 541936, GRMZM2G123159) was cloned. Bioinformatics analysis showed that the CDS of this gene was 519 bp, encoding 172 amino acids. ZmNAT1 contained a conserved domain unique to GNAT family. The relative expression level of ZmNAT1 gene in different tissues of maize at different stages under different stress conditions showed that the expression level of ZmNAT1 was the highest in mature roots, and the relative expression of ZmNAT1 gene could be induced in different degrees under different abiotic adversity stress conditions. Three independent transgenic Arabidopsis (Arabidopsis thaliana L.) pure lines with higher expression were obtained by heterologous expression, and phenotypic characterisation experiments under different adversity stress treatments were carried out on them, the results showed that transgenic Arabidopsis had a better phenotype relative to wild-type Arabidopsis, and the roots of transgenic plants under salt stress, osmotic stress, and drought were significantly longer than those of the wild-type,and the plants showed higher green leaf rate and chlorophyll content, and lower malondialdehyde content than the wild-type plants, which were significant difference. We speculated that ZmNAT1 gene may be involved in response to abiotic stress such as drought and salt in maize. This study provides an important reference for further analysis of the biological functions of ZmNAT1 in maize.

Key words: maize (Zea mays L.), ZmNAT1, root system, growth and development, adversity stress

Table 1

Information of primers"

引物名称Primer name 引物序列Primer sequence (5'-3')
pZmNAT1-F CCCGGGCTGCAGAATTCATGACGACAATCCGTCGGTTCTG
pZmNAT1-R CCATGGTACCCTCGAGAAGCTTGTTCAAAAATAAACTAGTGG
pZmNAT1RT-F AAGGGTGCTCCGGTACTACT
pZmNAT1RT-R AGTGGACGACAAATATTCGAGA
pGAPDHRT-F CCCTTCATCACCACGGACTAC
pGAPDHRT-R AACCTTCTTGGCACCACCCT
pActinRT-F GGTAACATTGTGCTCAGTGGTGG
pActinRT-R AACGACCTTAATCTTCATGCTGC

Fig. 1

Bioinformatics and evolutionary analysis of ZmNAT1 A: the conserved domain analysis; B: transmembrane structure prediction; C: promoter analysis of ZmNAT1; D: phylogenetic tree of HAT family proteins in Arabidopsis, rice, and maize."

Fig. 2

Analysis of the relative expression of ZmNAT1 in different tissues at different developmental periods in maize Different lowercase letters are significantly different at the 0.05 probability level."

Fig. 3

Relative expression of ZmNAT1 in maize above-ground and roots under different abiotic stresses A: dehydration treatment; B: high salt treatment; C: osmotic treatment; D: low temperature treatment. *: P < 0.05; **: P < 0.01."

Fig. 4

Positive identification and relative expression analysis of ZmNAT1 transgenic Arabidopsis thaliana in T3 generation A: PCR identification of the T3 generation transgenic Arabidopsis thaliana ZmNAT1 gene; B: RT-qPCR identification of the T3 generation transgenic Arabidopsis thaliana ZmNAT1 gene. M: DL2000 marker; P: positive control; N: negative control; W: water control; WT: wild-type Arabidopsis thaliana; L-1-L-6: T3 generation ZmNAT1 transgenic Arabidopsis thaliana; **: P < 0.01."

Fig. 5

Growth development and root length statistics of ZmNAT1 transgenic Arabidopsis thaliana under different salt concentration treatments A-E: growth and development of ZmNAT1 transgenic Arabidopsis thaliana and wild-type Arabidopsis thaliana under 0, 0.10, 0.12, 0.15, and 0.18 mol L-1 salt treatments; F: statistics and comparison of mean primary root lengths of transgenic and wild-type Arabidopsis thaliana under different concentrations of salt treatments; WT: wild type Arabidopsis thaliana; L-2, L-5, and L-6: Arabidopsis thaliana transgenic for ZmNAT1; Bar: 1.5 cm. *: P < 0.05; **: P < 0.01."

Fig. 6

Growth development and root length statistics of ZmNAT1 transgenic Arabidopsis thaliana treated with different mannitol concentrations Abbreviations are the same as those given in Fig. 5. A-D: growth and development of ZmNAT1 transgenic Arabidopsis thaliana and wild-type Arabidopsis thaliana under 0, 0.15, 0.20, and 0.30 mol L-1 mannitol treatments; E: statistics and comparison of mean primary root lengths of transgenic and wild-type Arabidopsis thaliana treated with different concentrations of mannitol. Bar: 1.5 cm. *: P < 0.05; **: P < 0.01."

Fig. 7

Growth and development of ZmNAT1 transgenic Arabidopsis thaliana in drought treatment and analysis of physiological indices A: growth of wild-type Arabidopsis thaliana and transgenic Arabidopsis thaliana in soil under normal conditions, drought treatment, and after rehydration, respectively; B: MDA content of WT and transgenic Arabidopsis thaliana in control and experimental groups; C: total chlorophyll contents of WT and transgenic Arabidopsis thaliana in control and experimental groups; D: statistics on green leaf rate of WT and transgenic Arabidopsis thaliana after drought treatment; E: statistics on green leaf rate of WT and transgenic Arabidopsis thaliana after 2 d of rehydration treatment. Abbreviations are the same as those given in Fig. 5. *: P < 0.05; **: P < 0.01."

Fig. 8

Growth and development of ZmNAT1 transgenic Arabidopsis thaliana in salt treatment and analysis of physiological indices A: growth of wild-type Arabidopsis thaliana and transgenic Arabidopsis thaliana in soil under normal conditions, salt treatment, respectively; B: MDA content of WT and transgenic Arabidopsis thaliana in control and experimental groups; C: total chlorophyll contents of WT and transgenic Arabidopsis thaliana in control and experimental groups; D: statistics on green leaf rate of WT and transgenic Arabidopsis thaliana after salt treatment. Abbreviations are the same as those given in Fig. 5. *: P < 0.05; **: P < 0.01."

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