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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (1): 85-91.doi: 10.3724/SP.J.1006.2010.00085

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

Molecular Cloning and Characterization of Two Fiber Elongation Genes Using a Cotton Fiber Developmental Mutant (Gossypium hirsutum L.)

WANG Lei,ZHU Yi-Chao**,CAI Cai-Ping,ZHANG Tian-Zhen,GUO Wang-Zhen*   

  1. National Key Laboratory of Crop Genetics and Germplasm enhancement,Cotton Research Institute,Nanjing Agricultural University,Nanjing 210095,China
  • Received:2009-04-15 Revised:2009-06-25 Online:2010-01-12 Published:2009-11-17
  • Contact: GUO Wang-Zhen, E-mail: moelab@njau.edu.cn; Tel: 025-84395311

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Abstract:

Cotton fibers are single-celled seed trichomes of major economic importance. Many important genes are expressed during cotton fiber development and fiber developmental mutants can be used to preferentially detect the genes controlling fiber development. The Ligon lintless mutant (Li1li1) is a fiber elongation developmental mutant with a dominant monogenetic mutation characterized by short fibers and distorted leaf, stem and flower growth, and the recessive pure line (li1li1) exhibits normal fiber developmental characteristics. The objectives of this study were to isolate genes preferentially or specifically expressed in fiber elongation stage by comparing gene expression differences between Li1li1 and li1li1. RNA isolated from 10 days post anthesis (DPA) fibers and ovules mixture in Li1li1 and li1li1 were used to screen differential gene expression in fiber development using differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Two differential expression cDNA segments were isolated, the corresponding full-length cDNAs were cloned and their primary function was analyzed. The two genes encoded 542 and 667 amino acid residues and functioned as glutamate decarboxylase (GhGAD) and vacuole-pyrophosphatase (GhVP1), respectively. Transcriptional level assays showed the two genes were constitutively expressed in tested tissues with higher expression levels during the fiber elongation stage. Further, a BC1 mapping population derived from hybridization between G. hirsutum acc. TM-1 and G. barbadense cv. Hai 7124, and TM-1 as the recurrent parent, was used for the location of GhGAD and GhVP1 on chromosomes 12 and 8, respectively, using cleaved amplified polymorphic sequences (CAPs).

Key words: Cotton, Fiber developmental mutant, DDRT-PCR, Glutamate decarboxylase(GhGAD), Vacuole-pyrophosphatase(GhVP1)

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