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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (7): 1029-1037.doi: 10.3724/SP.J.1006.2019.83070

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

Cloning and functional analysis of ZmGRAS31 gene in maize

YIN Long-Fei1,2,WANG Zhao-Yang1,WU Zhong-Yi2,ZHANG Zhong-Bao2,*(),YU Rong1,*()   

  1. 1 College of Life Sciences, Capital Normal University, Beijing 100048
    2 Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences / Beijing Key Laboratory of Agricultural Gene Resources and Biotechnology, Beijing 100097, China
  • Received:2018-11-07 Accepted:2019-01-19 Online:2019-07-12 Published:2019-03-22
  • Contact: Zhong-Bao ZHANG,Rong YU E-mail:zhangzhongbao@baafs.net.cn;yurong@mail.cnu.edu.cn
  • Supported by:
    This study was supported by the Beijing Natural Science Foundation(6172007);the National Natural Science Foundation of China(31871351);the Beijing Academy of Agricultural and Forestry Sciences(KJCX20180404)

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Abstract:

The GRAS gene family is a kind of plant-specific transcription factor reported in many plants. To figure out the function of maize GRAS family under stress, we obtained ZmGRAS319 (AC: NC_024462) gene from maize (Zea mays) root. Sequence analysis showed that the coding sequence (CDS) of ZmGRAS31 was 1422 bp encoding a protein of 473 amino acids with molecular weight of 51,700.38 Da and an isoelectric point of 4.73. Bioinformatics analysis revealed that ZmGRAS31 had a conserved domain unique to the GRAS transcription factor family and poor hydrophilicity, but no transmembrane structure. Transient expression of maize protoplasts indicated that ZmGRAS31 was localized in the nucleus. Real-time quantitative PCR analysis showed that the expression of ZmGRAS31 was up-regulated in maize seedlings under low temperature, dehydration, high salinity and drought conditions. The root length of the ZmGRAS31 overexpression transgenic Arabidopsis plants was longer than that of wide type under different concentrations of NaCl treatments. Therefore, our data suggested that ZmGRAS31 might be involved in abiotic stress response.

Key words: maize, ZmGRAS31, transcription factor, protoplast, qPCR, abiotic stress

Table 1

Primers used in this study"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)
pZmGRAS31-F AAAGCCGATGGAGACCATTTCAT
pZmGRAS31-R CTTTCATACAGGAGTCCAAGCA
pZmGRAS31SL-F CATATCGAATTCATGGAGACCATTTCATATCC
pZmGRAS31SL-R CCTAGCGGTACCTACAGGAGTCCAAGCAGATAT
pZmGRAS31RT-F CTTCGACACCATTGCTATTGG
pZmGRAS31RT-R CTTGAATCTCCAGCCTCAAC
pGAPDHRT-F CCCTTCATCACCACGGACTAC
pGAPDHRT-R AACCTTCTTGGCACCACCCT
pZmGRAS31At-F CCTATCGGCGCCATGGAGACCATTTCATATCC
pZmGRAS31At-R CCTAGCACCGGTTCATACAGGAGTCCAAGCAG

Fig. 1

Conservative domain prediction of maize ZmGRAS31 gene"

Fig. 2

Subcellular localization of ZmGRAS31 in protoplasts GFP: subcellular localization of the GFP-ZmGRAS31 fusion protein in maize protoplast; DAPI: DAPI-labeled protoplast nucleus; Bright: protoplast under the same field; Merged: merge of GFP, DAPI, and bright; CK: protoplast with empty vector; ZmGRAS31: protoplast with destination vector. Bar=10 μm."

Fig. 3

Relative expression of ZmGRAS31 in different maize tissues"

Fig. 4

Real-time quantitative PCR analysis of expression level of ZmGRAS31 gene under different treatments in shoots and roots A: dehydration treatment; B: cold (4°C) treatment; C: high salt (200 mmol L-1 NaCl) treatment; D: drought treatment. The error bar represents ± SD of triplicate experiments 2.5 转ZmGRAS31基因拟南芥的耐盐性分析"

Fig. 5

Root length of transgenic Arabidopsis thaliana under gradient NaCl concentrations A-D was seedling growth under the NaCl concentrations of 0 mol L-1, 0.10 mol L-1, 0.15 mol L-1, and 0.20 mol L-1; E: the average main root length of Arabidopsis at different NaCl concentrations. WT: wild type; OE: overexpression Arabidopsis thaliana. Bar = 1.5 cm."

Fig. 6

Root length of transgenic Arabidopsis thaliana under gradient mannitol concentrations A-D was seedling growth under the mannitol concentrations of 0 mol L-1, 0.15 mol L-1, 0.30 mol L-1, and 0.40 mol L-1; E: the average main root length of Arabidopsis at different mannitol concentrations. WT: wild type; OE: overexpression Arabidopsis thaliana. Bar = 1.5 cm."

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