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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (9): 2433-2445.doi: 10.3724/SP.J.1006.2023.23071

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Response of maize transcriptional factor ZmEREB211 to abiotic stress

AI Rong1,2(), ZHANG Chun2(), YUE Man-Fang2, ZOU Hua-Wen1,*(), WU Zhong-Yi2,*()   

  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:2022-10-21 Accepted:2023-02-21 Online:2023-09-12 Published:2023-03-01
  • About author:First author contact:**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);Beijing Academy of Agricultural and Forestry Sciences(KJCX20220402)

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Abstract:

AP2/ERF (APETALA2/ethylene-responsive factor) transcription factor is one of the largest transcription factor families in plants, which plays an important role in regulating plant growth and development and responding to various stresses. Exploring the function of AP2/ERF family genes in maize (Zea mays L.) will provide the important genetic resources for the creation of new maize germplasm. In this study, ZmEREB211 (Gene ID: 103647485) gene was cloned and its basic characteristics, tissue expression characteristics, and the relative expression patterns in response to stress were analyzed by bioinformatics and qRT-PCR. The transgenic Arabidopsis lines were subjected to corresponding stress treatment and phenotypic identification. The results showed that the gene contained only one exon and the full-length cDNA was 792 bp which encoding 263 amino acids. The ZmEREB211 protein had a molecular weight of 27.9 kD and a theoretical isoelectric point of 6.01. It had a conserved domain unique to the AP2/ERF family. The relative expression level of ZmEREB211 gene was the highest in maize root system and the relative expression level in young roots was higher than mature roots. At the same time, the gene had different degrees of induced expression under dehydration, high salt, drought, and low temperature treatment conditions. On 1/2 MS medium containing different concentrations of NaCl, mannitol, and jasmonic acid (JA), root length of ZmEREB211 transgenic Arabidopsis lines was significantly longer than wild type. Under drought and high salt treatments, transgenic Arabidopsis lines had stronger tolerance than wild type, and the number of green leaves at seedling stage was significantly higher than wild type and the peroxidase (POD) activity and chlorophyll content were significantly higher than wild type. ZmEREB211 may be involved in the regulation of root growth and development in maize, and can play a positive regulatory role in high salt, drought, osmotic stress, and JA hormone treatments. This study provides an important reference for further analysis of the biological function of ZmEREB211 in maize.

Key words: maize, ZmEREB211, transcription factor, heterologous expression, root system, adversity stress

Table 1

Primers used in this study"

引物名称
Primer name		 引物序列
Primer sequence (5'-3')
pZmEREB211-F CTGAAATCACCAGTCGGTACCATGACCAAGAAGCTCATCT (Kpn I)
pZmEREB211-R GCCCTTGCTCACCATGGTACCTCAAAAACTAGACGCGGCG (Kpn I)
pZmEREB211RT-F AGGAGGACGCCATCATCGT
pZmEREB211RT-R GGTACATGTTCCTGTTCCTACG
Actin-F
Actin-R		 TACGAGATGCCTGATGGTCAGGTCA
TGGAGTTGTACGTGGCCTCATGGAC
pGAPDHRT-F
pGAPDHRT-R		 CCCTTCATCACCACGGACTAC
AACCTTCTTGGCACCACCCT

Fig. 1

Bioinformatics analysis of ZmEREB211 A: the conserved domain analysis; B: the transmembrane structure prediction; C: protein tertiary structure prediction; D: the analysis of cis-acting elements in promoter region."

Fig. 2

Relative expression level of ZmEREB211 genes in different maize tissues Different lowercase letters are significantly different at the 0.05 probability level."

Fig. 3

Relative expression level of ZmEREB211 genes in different abiotic stress A-D: the relative expression level of ZmEREB211 genes after dehydration, high salt, osmotic, and low temperature treatments."

Fig. 4

Relative expression level of ZmEREB211 genes after different phytohormone treatments *: P < 0.05; **: P < 0.01."

Fig. 5

PCR detection and relative expression level of T4 generation Arabidopsis A: PCR detection of T4 generation Arabidopsis; B: qPCR detection of T4 generation Arabidopsis; M: DL2000 marker; N: negative control; W: water control; WT: wild-type Arabidopsis; L-1-L-6: T4 transgenic Arabidopsis lines; *: P < 0.05; **: P < 0.01."

Fig. 6

Root length of transgenic Arabidopsis lines on gradient NaCl concentrations A-D indicate the seedling growth under the NaCl concentrations of 0, 0.1, 0.15, and 0.18 mol L-1, respectively. E: the average main root length of Arabidopsis under different salt concentrations; WT: wild type; L-3, L-4, and L-5: ZmEREB211 transgenic Arabidopsis lines; Bar: 1.5 cm; *: P < 0.05; **: P < 0.01."

Fig. 7

Root length of transgenic Arabidopsis lines on gradient mannitol concentrations A-D indicate the seedling growth under the mannitol concentrations of 0, 0.15, 0.2, and 0.3 mol L-1, respectively. E: the average main root length of Arabidopsis under different mannitol concentrations. Abbreviations are the same as those given in Fig. 6; Bar: 1.5 cm; *: P < 0.05; **: P < 0.01."

Fig. 8

Root length of transgenic Arabidopsis lines on gradient JA and ABA concentrations A: growth under 0, 50, 75, and 100 μmol L-1 JA treatment; B: the average main root length of Arabidopsis under different JA concentrations; C: growth under 0, 10, 25, and 50 μmol L-1 ABA treatment; D: the average main root length of Arabidopsis under different ABA concentrations. Abbreviations are the same as those given in Fig. 6; Bar: 1.5 cm; *: P < 0.05; **: P < 0.01."

Fig. 9

Phenotypic analysis of Arabidopsis lines under drought treatment in soil A: the phenotype of Arabidopsis; B: the rate of green leaves after drought treatment; C: the rate of green leaves after re-water treatment; D: the determination of POD activity; E: the determination of chlorophyll content. Abbreviations are the same as those given in Fig. 6; *: P < 0.05; **: P < 0.01."

Fig. 10

Phenotypic analysis of Arabidopsis lines under high salt treatment in soil A: the phenotype of Arabidopsis; B: the rate of green leaves after high salt treatment; C: the determination of POD activity; D: the determination of chlorophyll content. Abbreviations are the same as those given in Fig. 6; *: P < 0.05; **: P < 0.01."

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