Cloning and characterization of the Gossypium hirsutum major latex protein gene and functional analysis in Arabidopsis thaliana
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The major latex protein (MLP) gene in Gossypium hirsutum was cloned and designated Gh-MLP. Expression in cotton root was induced by salt stress and Verticillium dahliae toxin, and bioinformatic analysis showed that Gh-MLP encodes a 157-amino acid protein that is similar to members of the MLP subfamily in the Bet v 1 family. Although the structure of MLP is similar to Bet v 1 family proteins, the sequence identity to other subfamilies of Bet v 1 proteins is less than 20%. The Gh-MLP promoter contains potential cis-acting elements for response to salt stress and fungal elicitor. RT-PCR analysis showed that Gh-MLP expression was rapidly induced by NaCl and V. dahliae toxin, and induction was maintained over 72 h. However, Gh-MLP transgenic Arabidopsis thaliana did not show resistance to V. dahiae, salt tolerance was significantly enhanced. In contrast to the wild type, the Gh-MLP transgene allowed plants to germinate normally after treatment with 75 mM NaCl. Total flavonoid was twofold higher in transgenic Arabidopsis than in the control, suggesting that Gh-MLP might be involved in altering flavonoid content. We hypothesize Gh-MLP, like other Bet v 1 family proteins, participates in the binding or transport of ligands through its specific three-dimensional structure, and takes part in defensive responses to biotic and abiotic stresses.
KeywordsFlavonoid Induce expression Major latex proteins Salt stress Verticillium wilt resistance
Cytokinin-specific binding proteins
Major latex protein
Pathogenesis-related proteins family 10
Rapid amplification of cDNA ends
This work was funded by the National Natural Science Foundation of China (No. 30571210). We thank Rong-Qi Xu, Ting-Hui Zhou, Jia-Ni Wang, Feng-Xuan Zhao and Yu Weng for their technical assistance.
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