| Cotton(Gossypium L.) is an important economic crops in the world and cotton fiber is the main raw material of natural textile industry. With the improvement of the development of textile technology and people’s living standards, the textile industry on cotton fiber quality rising demand, particularly the need for higher fiber strength of cotton fiber. Cotton fiber quality improvement has become a hot issue important cotton biology research.Cotton fiber development process can be divided into four stages: fiber cell initiation, elongation, secondary wall thickening and dewatering of maturity, in which the relation between strength of fiber and secondary wall thickening is close. The main structure of mature cotton fiber is cell wall, cytoskeleton in plants direct the synthesis of the cell wall. BTB protein family is one of Kuppel zinc finger protein families, able to guide the formation and development of the cytoskeleton and thus affect cell wall synthesis, thereby affect the entire cotton fiber quality development.This study is based on the identification and analysis of the system from upland cotton genome BTB protein family.In this study,the fracture of friber high strength(69307) and low strength(69362) were selected as materials,GhBTB1 was cloned, and the function of GhBTB1 was initial verification. Initially revealed the function of GhBTB1 in upland cotton fiber during development. The results of this study are as follows:1 Phylogenetic analysis of BTB gene family from upland cotton. This study identified 10 BTBs genes by using bioinformatics methods from tetraploid cotton G. hirsutum L.,then divided them into 3 subfamilies; and the genetic structure of family members, conserved protein domains, physical and chemical properties and secondary structure were systematic analysis, results showed that BTB protein family is highly conserved in evolution,this laid a foundation for the further study of protein function BTB.2 GhBTB1 was cloned. Using homologous cloning cloned GhBTB1 full-length cDNA, and the basic physical and chemical properties showed that theory of relative molecular weight was 49.488 kD,Theoretical isoelectric point was 5.73, belonged to hydrophobic protein,it may be stable.Secondary structure was mainly composed of alpha helix,extending chain and random coil.Sianal peptide prediction analysis found that the protein did not contain signal peptide,nor transmembrane,and speculated that the protein may be a scretory protein.The analysis of evolutionary tree found that the protein may be belong to a same branch with cocoa proteins.The analysis of multiple sequence alignment found that the protein compared with Arabidopsis and rice was similarity 77.42%, indicating protein structure of GhBTB1 was highly conserved.3 Expression pattern analysis of GhBTB1. Real-time quantitative RT-PCR analysis showed that,the peake of 69307 was 20 DPA, the peake of 69362 was 30 DPA.The results showed that GhBTB1 had an advantages of expression during secondary wall thickening in the fiber development.4 Functions of GhBTB1. The connection with the gene vector PBI121 placed downstream of a strong promoter 35 S eukaryotic expression vector had successfully transformed wild-type Arabidopsis thaliana. We got transgenic Arabidopsis by kanamycin resistance screening and PCR amplification.And we observed the phenotype difference of T2 generation transgenic Arabidopsis,and found that the transgenic Arabidopsis growed slowly compared with wide-type Arabidopsis, performance was late flowers,which laid foundation for subsequent research. |
PDF Full Text Request