Crops ›› 2016, Vol. 32 ›› Issue (4): 47-55.doi: 10.16035/j.issn.1001-7283.2016.04.008

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Cloning and Function Analysis of Resistance Gene ZmqLTG3-1 in Maize

Han Zanping1,2,Chen Yanhui2,Guo Shulei2,Zu Xiaofeng2,Wang Shunxi2,Zhao Xiyong2   

  1. 1 Agricultural College,Henan University of Science and Technology,Luoyang 471003,Henan,China
    2 Agronomy College of Henan Agricultural University,Zhengzhou 450002,Henan,China
  • Received:2016-04-02 Revised:2016-05-11 Online:2016-08-15 Published:2018-08-26

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Abstract:

By the method of orthologus gene cloning,the ZmqLTG3-1 gene in maize was successfully cloned, which was homologous to the qLTG3-1 gene in rice. The sequence analysis indicated that the full-length cDNA was composed of 606bp, including an open reading frame (ORF) of 441bp, which encoded 147 amino acids. The protein contained LTP conservative structure of the extra protein family AAI-LTSS, and protein molecular weight and isoelectric point were 14.16kDa and 8.3, respectively, with the similarity of amino acid sequence for ZmqLTG3-1 with sorghum and rice being 92% and 88%. The results of quantitative RT-PCR showed that the highest expression level was in the embryo, then in the stem tip, lower was in the leaf and root. The highest expression abundance at 24h and then regular down along with germination time. The result of subcell location revealed that the product of ZmqLTG3-1 gene encoding is positioned on the cell membrane and it was a trans-membrane protein. To carry the preliminary validation of its function,transgenic T3 generation, trans-empty vector and wild type Arabidopsis thaliana as materials were studied. Results showed that in different abiotic conditions, such as drought, high temperature and salt, compared with control group, the performance and the survival rate of transgenic plant are significantly higher. All these suggest that ZmqLTG3-1 gene plays an important role in resisting abiotic stress.

Key words: Maize (Zea mays L.), ZmqLTG3-1, Cloning, Function analysis

Fig.1

Detection of RNA M．DL2000,1-3.Sample"

Fig.2

PCR amplification of 18S M．DL2000,1-3.Sample"

Fig.3

PCR amplification of ZmqLTG3-1 cDNA"

Fig.4

Domain prediction of ZmqLTG3-1"

Fig.5

The similarity analysis of amino acid sequences"

Fig.6

Phylogenetic analysis of ZmqLTG3-1 and other selected plant species"

Fig.7

Relative abundance of the ZmqLTG3-1 transcripts in different maize tissues"

Fig.8

Relative abundance of the ZmqLTG3-1 transcripts in different embryo germination time"

Fig.9

Restriction enzyme digestion of pMD18-T-ZmqLTG3-1"

Fig.10

Restriction enzyme digestion of pCAMBIA1304-ZmqLTG3-1-GFP"

Fig.11

Restriction enzyme digestion of OE-ZmqLTG3-1"

Fig.12

PCR detection of OE-ZmqLTG3-1 plasmid M.DL2000,1-10.Monoclone,11.Recombination plasmid"

Fig.13

Performance of transgenic plants under different abiotic stress processing in Arabidopsis ** indicate the significant differences at 0.01 level"

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