Cloning and characterization of the Gossypium hirsutum major latex protein gene and functional analysis in Arabidopsis thaliana
- 775 Downloads
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.
- Marković-Housley Z, Degano M, Lamba D, von Roepenack-Lahaye E, Clemens S, Susani M, Ferreira F, Scheiner O, Breiteneder H (2003) Crystal structure of a hypoallergenic isoform of the major birch pollen allergen Bet v 1 and its likely biological function as a plant steroid carrier. J Mol Biol 325:123–133CrossRefPubMedGoogle Scholar
- Park SY, Fung P, Nishimura N, Jensen DR, Fujii H, Zhao Y, Lumba S, Santiago J, Rodrigues A, Chow TF, Alfred SE, Bonetta D, Finkelstein R, Provart NJ, Desveaux D, Rodriguez PL, McCourt P, Zhu JK, Schroeder JI, Volkman BF, Cutler SR (2009) Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL family of START proteins. Science 324:1068–1071PubMedGoogle Scholar
- Pozueta-Romero J, Klein M, Houlné G, Schantz ML, Meyer B, Schantz R (1995) Characterization of a family of genes encoding a fruit-specific wound-stimulated protein of bell pepper (Capsicum annuum): identification of a new family of transposable elements. Plant Mol Biol 28:1011–1025CrossRefPubMedGoogle Scholar
- Suyama T, Yamada K, Mori H, Takeno K, Yamaki S (1999) Cloning cDNAs for genes preferentially expressed during fruit growth in cucumber. J Am Soc Hort Sci 124:136–139Google Scholar
- Wang LH, Dai XF, Fang XJ, Rong JK, Paterson AH (2004) Cloning and mapping of cDNA from cotton induced by toxin of V. dahliae. Scientia Agricultura Sinica 37:1474–1480Google Scholar