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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (02): 362-368.doi: 10.3724/SP.J.1006.2011.00362

• RESEARCH ACTIVITIES • Previous Articles     Next Articles

Isolation and Expression Analysis of Two GhBlind Homologs in Upland Cotton (Gossypium hirsutum L.)

XIONG Guan-Jun1,XU Qin1,HUA Jin-Ping1,2,*   

  1. 1 College of Agronomy & Biotechnology, China Agricultural University, Beijing 100193, China；2 Key Laboratory of Crop Genetic Improvement and Genome of Ministry of Agriculture / Key Laboratory of Crop Heterosis and Utilization of Ministry of Education / Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing 100193, China
  • Received:2010-06-23 Revised:2010-09-27 Online:2011-02-12 Published:2010-11-16
  • Contact: HUS Jin-Ping, E-mail: jinping_hua@cau.edu.cn

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Abstract: In higher plants, Blind homologs play an important role in plant architectures. Two Blind orthologs, GhBlind1 and GhBlind2, were isolated from cDNA of the shoot apex of upland cotton (Gossypium hirsutum L.) by homologous cloning strategy. GenBank accession number is HQ115643 for GhBlind1, and HQ115644 for GhBlind2. Similar to other Blind homologs, GhBlind1 and GhBlind2 were consisted of three exons and two introns, and encoded 359 and 262 amino acids, respectively. Based on the RT-PCR analysis, it was found that GhBlind1 expressed in root, leaf, and tender fiber, and predominantly in shoot apical meristem (SAM) and shoot apex, but not in stem; while GhBlind2 expressed in root, stem, leaf, SAM and shoot apex, but not in tender fiber. Using recombinant PCR technology, expression vectors Psuper-gus-b1 and Psuper-gus-b2 were constructed and used for Agrobacterium-mediated transient expression by transformation of embryogenic callus of upland cotton cv. Coker 201. Histochemical assays of GUS activity showed that after co-cultivation of embryogenic callus for four days, positive events were more obvious in Psuper-gus-b2 than in Psuper-gus-b1. The alignments between GhBlind1, GhBlind2 and the domain of Blind homologs showed the identities of 94% and 91%, respectively. The results of phylogenetic tree showed that GhBlind1 and GhBlind2 were genetically divergent from most of the other R2R3-MYB members in Gossypium hirsutum L. We speculated that GhBlind1 and GhBlind2 are probably related with the development of lateral branch in upland cotton.

Key words: Upland cotton, Blind, Homologous isolation, Expression profile

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