Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (17): 3523-3533.doi: 10.3864/j.issn.0578-1752.2013.17.001

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

Molecular Cloning and Expression Analysis of Three Chain Extension Genes Related to Fatty Acid Synthesis in Upland Cotton (Gossypium hirsutum L.)

 LIU  Li-12, ZHAO  Peng-1, WANG  Dan-1, LIU  Zheng-Jie-1, WANG  Yu-Mei-3, HUA  Jin-Ping-1   

  1. 1.College of Agronomy and Biotechnology, China Agricultural University/Key Laboratory of Crop Heterosis and Utilization of Ministry of Education/Beijing Key Laboratory of Crop Genetic Improvement, Beijing 100193
    2.Cotton Institute, Xinjiang   Academy Agricultural and Reclamation Science/Northwest Inland Region Key Laboratory of Cotton Biology and Genetic   Breeding (Xinjiang), Ministry of Agriculture, Shihezi 832000, Xinjiang 
    3.Institute of Cash Crops, Hubei Academy of  Agricultural Sciences, Wuhan 430064)
  • Received:2013-02-26 Online:2013-09-01 Published:2013-06-06

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Abstract: 【Objective】Anabolism of fatty acid controls the synthesis of fat and affects responses of plants to abiotic stress. The temporal-spatial expression of related genes directly decides the content of oil in seeds, and affects stress responses of plants associated with fatty acids. The objectives of this study are to clone the important genes related to fatty acid synthesis and to analyze the expression of genes in the process of fatty acid accumulation and the responses of gene expression to stress with the aim to provide candidate genes for high-oil cotton breeding and stress-resistance cotton breeding.【Method】Full length cDNA of GhKAR, GhHAD and GhENR were cloned by means of homologous cloning. Bioinformatics analyses were performed on the obtained genes to describe the characteristics of encoded proteins. The expression model of target genes in tissues, the levels of their temporal-spatial expression and the expression characteristics under abiotic stress were analyzed through RT-PCR.【Result】The cDNA of GhKAR, GhHAD and GhENR were 1 119 bp, 906 bp and 1 318 bp in length, respectively, and encoded 283, 221 and 394 amino acids. GhKAR and GhENR belonged to NADB Rosemann supergene family and GhHAD belonged to hotdog supergene family. The genes expressed variously in different tissues, such as root, stem, leaf and young ovule at different development stages. The expression level of young ovules in high-oil cotton material was slightly higher than that in low-oil cultivars. The oil accumulated majorly from 20DPA to 30DPA in both high-oil and low-oil cotton cultivars, and differed much after 30DPA to maturation. That is, all the 3 genes in high-oil cultivar increased steadily in expression after 20 DPA, and reached the maximum at 30DPA, and expressed at a low value in low-oil cultivars meanwhile. The expression level of GhHAD and GhENR in high-oil cultivars was twice more than that in low-oil ones. The expression level of GhKAR, GhHAD and GhENR induced differently by low temperature. And GhKAR and GhHAD expressed in up-regulation when induced by ABA, and could not be induced by MeJA. The expression level of GhENR was up-regulated at 2h after induction by MeJA, and went down-regulated gradually thereafter, but could not be induced by ABA.【Conclusion】Full length cDNA of GhKAR, GhHAD and GhENR were cloned. Three genes played an important role in oil accumulation in cotton seeds. The characteristics of gene expression inferred these 3 genes involved in responding to physiological stress.

Key words: upland cotton , fatty acid synthesis , gene clone , expression level , abiotic stress

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