Construction of a Full-Length cDNA Library of Gossypium hirsutum L. and Identification of Two MADS-Box Genes

• Published in: Agricultural sciences in China Vol. 10; no. 1; pp. 28 - 40
• Main Authors: WANG, Li-na, WU, Dong, YU, Shu-xun, FAN, Shu-li, SONG, Mei-zhen, PANG, Chao-you, LIU, Jun-jie
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01-01-2011
• Subjects: cDNA libraries; clones; complementary DNA; corolla; cotton; developmental stages; d基因; EST; ESTs; expressed sequence tags; flowering; Gossypium hirsutum; MADS; MADS-box gene; normalized cDNA library; ovules; phylogeny; screening; stamens; unigenes; 全长cDNA文库; 分子调控机制; 基因鉴定; 表达序列标签; 陆地棉; normalized cDNA library; cotton; MADS-box gene; EST
• ISSN: 1671-2927
• DOI: 10.1016/S1671-2927(11)60304-0

A full-length normalized cDNA library for the flower development stages of short-season cotton （Gossypium hirsutum L.） （CCRI36） was constructed. A total of 3 421 clones were randomly selected for sequencing, with a total of 3 175 effective sequences obtained after removal of empty-carriers and low-quality sequences. Clustering the 3 175 high-quality expressed sequence tags （ESTs） resulted in a set of 2 906 non-redundant sequences comprised of 233 contigs and 2 673 singletons. Comparative analyses indicated that 913 （43.6%） of the unigenes had homologues with function-known genes or functionassumed genes in the National Center for Biotechnology Information. In addition, 763 （36.4%） of the unigenes were functionally classified using Gene Ontology hierarchy. Through EST alignment and the screening method, the full-length cDNA of two MADS-box genes viz., GhMADSll and GhMADS12 were acquired. These genes may play a role in flower development. Phylogenetie analysis indicated that GhMADS11 and GhMADS12 had high homology and close evolutionary relationship with AGL2/SEP-type and PI-type genes, respectively. The expression of both GhMADSll and GhMADS12, genes was high in reproductive organs. In floral organs, GhMADSll expression was high in petals （whor12） and ovules, while GhMADS12 expression was high in petals （whor12） and stamens （whor13）. Results show that the EST strategy based on a normalized cDNA library is an effective method for gene identification. The study provides more insights for future molecular research on the regulation mechanism of cotton flower development.