The Cloning And Expression Analysis Of GhSERKs Gene From Cotton (Gossypium Hirsutum L.)

Somatic embryogenesis conditions induced by the External conditions,the somatic cells of morphological and biochemical changes in the process of the formation of somatic embryos.Somatic embryogenesis is based on the pluripotency of the cells of higher plants.unlike zygotic embryos,somatic embryogenesis is very easily manipulated and controlled culture conditions.Some plants through tissue culture to form a regenerated plants in a short time,such as rice, tobacco,etc;the formation of some plants by tissue culture regenerated plants is more difficult and a long time.such as cotton.Through the use of somatic embryogenesis-related genes to improve plant regeneration rate and the rate of genetic transformation,shorten the time of tissue culture,it would be rich have access to transgenic plants.So the cloning GhSERK family genes and its expression pattern is very necessary too.The major findings of this study are as follows:1. By in silico cloning and RACE cloning of four SERK gene,GhSERK3ORF of1863bp encoding620amino acid residues,molecular weight of68574.3Da;GhSERK4ORFof1836bp encoding611amino acid residues,molecular weight67721.5Da;GhSERK5ORF of1833bp encoding610amino acid residues,molecular weight67608.2Da;GhSERKL1ORF of1791bp encoding596amino acid residues molecular weight66033.8Da,the existence of alternative splicing and the gene.2. Bioinformatics analysis shows that they are with a negative charge.water solubility.half-life>10the stability of the protein,a signal peptide. membrane protein.The GhSERK3,GhSERK4and GhSERK5a single transmembrane domain,and has the typical structure of plant receptor-like protein kinase,extracellular domain,a single transmembrane domain and intracellular kinase domain,while GhSERKL1transmembrane twice;These genes generally contain leucine-rich repeat sequences,protein kinase ATP binding site,the serine/threonine protein kinase active site,the proline-rich region,the protein kinase domain,is a plant Somatic Embryogenesis Receptor-like Kinase.The cluster analysis show that GhSERK1belong to the Arabidopsis SER1/2subfamily,GhSERK3,GhSERK4,GhSERK5,GhSERKL1A and GhSERKL1B belonging to the Arabidopsis SERK3/4/5subfamily.3. Expression analysis in different periods of ovules and callus can show that GhSERK1,GhSERK3GhSERK4and GhSERK5gene expression,in the third and sixth day of the ovule expression is almost unchanged;in the ninth day ovule GhSERK1,GhSERK3,GhSERK4and GhSERK5expression level is much higher than the third day of the expression level,while GhSERKL1the twelfth day of the ovule the highest expression level,GhSERK1expression in the embryonic calli much higher than the non-embryogenic callus-The GhSERK3,GhSERK5and GhSERKL1expression of non-embryogenic callus and embryogenic callus almost no change,GhSERK4in non-embryogenic callus the expression levels higher than the embryogenic callus.According to the above results suggest that GhSERK1,GhSERK3,GhSERK4and GhSERK5in the globular embryo was highly expressed,while GhSERKL1was highly expressed in immature embryo into the heart-shaped period;GhSERK1may be involved in the process of somatic embryos,GhSERK3,GhSERK5and GhSERKL1may not be involved in the body cell embryos process;while GhSERK4the may play an important role in the non-embryogenic.In summary.this study was cloned from cotton the GhSERK family genes,the expression of these genes in different ovule and callus.Foundation,but also lay the foundation for further study GhSERKs cotton somatic embryogenesis and zygotic embryo development in the function for application of GhSERKs somatic embryogenesis in cotton.