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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (03): 390-396.doi: 10.3724/SP.J.1006.2014.00390

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

Cloning and Functional Analysis of GhVacInc2a Encoding Vacuolar Invertase in Cotton

XU Wen-Ting,WANG Cheng,XU Xiao-Yang,NIU Er-Li,CAI Cai-Ping,GUO Wang-Zhen*   

  1. Engineering Research Center, Ministry of Education, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2013-07-03 Revised:2013-10-29 Online:2014-03-12 Published:2014-01-16
  • Contact: 郭旺珍, E-mail: moelab@njau.edu.cn

Abstract:

Cotton fiber development depends on a large supply of sugar. Invertase plays a central role in sucrose metabolism. Structural and functional analysis of gene encoding invertase will conductively reveal the complex molecular mechanism of fiber development, and also provide elite gene resources for improvement of cotton fiber quality. In the study, a novel gene encoding vacuolar invertase was obtained by sillico cloning, based on a differentially expressed EST (GenBank accession number EY196825) between G. hirsutum acc. TM-1 and im mutant, combined with reconfirmation of ORF information in transcriptional and genomic level. BlastN showed that the gene shared 99% identity of GhVacInv2 reported previously (GenBank accession number FJ864677), and designated as GhVacInv2a (GenBank accession number KF305322). GhVacInv2a contained an open reading frame of 1857 bp that encoded a polypeptide of 618 amino acids, with seven exons and six introns at genome sequence. GhVacInv2a comprised one copy in diploid cotton species G. herbaceum and G. raimondii, and two copies in tetraploid cotton species G. hirsutum acc. TM-1 and G. barbadense cv. Hai 7124, with GhVacInv2a in A-subgenome and GhVacInv2 in D-subgenome, respectively. Q-PCR expression analysis showed that the set of homoelogous genes performed the highest expressive abundance in anther in comparison with other tissues. In developing fibers, it was predominantly expressed from fiber rapid elongation stage, and there existed significant difference between TM-1 and im mutant during 13 to 19 days post anthesis (DPA) in fiber tissues. Further, GhVacInv2a was located on chromosome 3 by developing subgenome-specific SNP marker, and the association analysis showed that there was a significant correction (P=0.0087) between GhVacInv2a and fiber strength, suggesting that GhVacInv2a probably plays a key role in fiber qualities formation.

Key words: Cotton, Fiber development, Vacuolar invertase 2, Cloning, Functional analysis

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