In Silico Cloning Of TRIPTYCHON In Cotton (gossypium Hirsutum L.) And Preliminary Confirmation Of TRIPTYCHON Gene Function

The length of cotton fiber is one important standard to evaluate the quality of cotton fibre. And the quantity of seed cotton fiber is direct factor to affect ginned cotton harvest.The study reveals that the growth of plant epidermal cell be regulated by many genes and the development of plant epidermal cell has similar molecule regulating pattern. The study reveals that cotton fiber is epidermal hair, epidermal cell of ovule outer integument from cotton fiber. And cotton fiber cell possesses the similar molecule regulating mode as the growth of epidermal hair cell in Arabidopsis. GLABROUS1 (GL1) / GLABRA2 (GL2) / GLABRA3 (GL3) / TRANSPARENT TESTA GLABRA1 (TTG1) /TRANSPARENT TESTA GLABRA2 (TTG2) to promot development of epidermal hair and TRIPTYCHON(TRY)/CAPRICE(CPC) inhibit development of epidermal hair in Arabidopsis. The study reveals homologous genes of TTG1/GL1/GL2 to promot development of cotton fiber in cotton, but no report about homologous genes of TRY/CPC in cotton.In silico cloning and RT-PCR technique was performed to isolate a TRIPTYCHON(TRY) gene in Xuzhou 142 no fiber mutants. After RT-PCR cloning, the DNA fragment was sequenced, and an ORF (Open read frame) of TRY which was 270bp in length, encoding 89 amino acids was found. The Protein is composed of 18 different amino acids. There are only one locus difference between the actual DNA sequence of ORF and the DNA sequence of ORF by in silico cloning. Both of their protein had only one animol acid difference, which indicated that the gene got from in silico cloning is more authentic.Through bioinformatics analysis, the protein molecular weight is 10638 Da, and theoretical pI:8.66, atomic composition: C461H734N140O136S7, extinction coefficients measured at 280 nm: 19605, the instability index: 75.26, aliphatic index: 61.35, grand average of hydropathicity: -0.899. This protein is same as Arabidopsis TRY protein and located in nucleolus. It belongs to instability, hydrophobic proteins and transcription factors of MYB family, and without signal peptide, glycosylation sites, trans-membrane structure, and coiled coil structure. Function prediction demonstrated that the protein coded by cotton TRY gene plays the same function as the protein does, coded by Arabidopsis TRY gene.Cotton TRY gene was recombined to pBI121 vector.. And integrate the cotton TRY gene to tobacco genome by the ways of Agrobacterium tumefaciens mediating. Masculine transgene plant could be got by real time fluorescence PCR detection. It is initially indicated that cotton TRY gene possess the function to restrain the growth of epidermal hair cell by observing and analyzing the development of Masculine transgene tobacco leaf hair and roots hair.