The Bioinformatics Analysis Of HXK Gene Family In Jilin Ginseng(Panax Ginseng C.A.Mayer) And Genetic Transformation Into Tomato

Hexokinase gene family is one of the important families involved in plant basal metabolism. As one of the key enzymes in cellular respiration process, it supplies the basic energy for plant growth and development, and participates in the plant photosynthesis, anti-aging and anti-pathogen, as well as in the regulation of plant color and other biological processes. In recent years, the multiple sugar sensing mechanism relying on hexokinase pathway has been the major concern. This study extracted 23 Jilin HXK Unigenes from 248,993 Unigenes databases and established Jilin ginseng HXK Unigenes database. The functions of Jilin HXK Unigenes were systematically studied from the aspects of conserved domains, origin and evolution, the synergistic interactions of relationship between family members and the expression levels in different tissues and organs. The results showed that Jilin ginseng HXK family contains five members including PgHXK1, PgHXK2, PgHXK3, PgGK and PgHXK5; the GO functional annotation in level-2 of 23 Unigenes were annotated to 11 functions including cellular process, metabolic process, biological regulation, and catalytic activity, etc.; three conserved regions of Jilin ginseng HXK were predicted, which were Hexokinase₂ super family（cl08402）,NBD_sugar- kinase_HSP70_actin（cd00012） and PLN02405（PLN02405）. The function and expressing activity for the family members of Jilin ginseng HXK have a certain degree of differentiation. KEGG metabolic pathway targeting displays that Jilin ginseng HXK family is involved in 9 metabolic pathways, including Streptomycin biosynthesis, Starch and sucrose metabolism and so on. The members of the family exist in the form of multi-cluster, and the expression activities are related to each other, although the members of Jilin ginseng HXK family express differences in different tissues and organs.This study selected the PgGK which was expressed in all 14 tissues and organs in Jilin ginseng as the target gene to do the cloning. The plant expression vector pBI121 was successfully constructed and transformed to exogenous species of tomato. Meanwhile this study selected ginseng large fragments DNA C11（50kb） containing both β-AS and PgGK and ginseng large fragments DNA C12（80kb） containing both P450 and PgGK were transformed into tomato. The genetic transformation conversion conditions of tomato were determined, including the preliminary training time, the co-culture time, the concentration of bacteria liquid, and the size of the filter press. This study explored the impact on tomato transformation efficiency of exogenous gene size. Single gene PgGKand large fragments DNA transformed tomato to verify gene function, which established the foundation for further study of the impact of plant metabolites on the basis of secondary metabolites of lycopene.The result showed: the resistance bud induction rate of single gene PgGK is 5.45%, but the resistance plants have not been obtained; the resistance bud induction rate of large fragment DNA C11 is 4.79% and percent conversion is 0.95%; the resistance bud induction rate of large fragment DNA C12 is 4.47% and percent conversion is 0.68%. This result indicated that the transformation efficiency may change with the size of the exogenous gene.