棉 花 学 报   Cotton Science    2010，22（5）：403-408

Cloning and Quantitative Analysis by Real-Time RT-PCR of GhF3´5´H in the Natural Green Cotton
TIAN Xin-hui¹, LI Yan-jun¹, ZHANG Xin-yu¹, WANG Hong-juan², SUN Jie^1*
（1.College of Agriculture / The key Oasis Eco-agriculture Laboratory of Xinjiang Production and Construction Group, Shihezi University, Shihezi, Xinjiang 832003, China; 2.College of Life Science, Shihezi University, Shihezi, Xinjiang 832003, China）

Abstract: cDNA-amplified fragment length polymorphism （cDNA-AFLP） was employed to analyze genes that are specifically or preferentially expressed in color cotton fiber cells. Using rapid amplification of cDNA ends(RACE), the complete cDNA sequence of GhF3´5´H gene was isolated from green cotton fiber at the 18 day post anthesis (DPA). The full-length of this cDNA clone was 1873 bp, and its largest open reading frame encoded 509 amino acids. The gene was designated as GhF3´5´H (GenBank accession: GU062184). Fluorescence real time quantitative PCR method were used to study the expression of GhF3´5´H gene in different development stages. Real-time PCR analysis showed that GhF3´5´H specially expressed in naturally green cotton fibers prior to white cotton fiber, its expression level was much higher in green cotton than that in white cotton, and it shrinked in proportion to fiber development. It is suggested that GhF3´5´H gene may involve in the formation of its precursor and play an important role in green pigment development in natural green cotton fibers.
Key words: cotton; natural green fiber; F3´5´H; real-time quantitative PCR   [Full Textk, 506KB]