Developing Functional Markers Of PI4K And TPS Genes Related Cotton Fiber Development

Cotton as the most important economic crops in the world, its product fiber is an important textilematerial. As people’s growing needs of material and cultural, traditional cotton breeding is graduallyreplaced by molecular marker assisted breeding. In this study, functional markers is developed ofPI4K and TPS genes related cotton fiber quality,based on the former research of transcriptional QTL ofthe quality of cotton fiber and cloning genes related cotton fiber development. The main results are asfollows:1. Based on the fiber development gene, Phosphatidylinositol-4-kinase gene (PI4K), develop of itscorresponding functional marker successfully.（1）The PI4K gene is located at Ca5and Cd13chromosomes. with the full length sequence ofPI4K cDNA based on the database of diploid ancestors’genome of G. arboreum and G.raimondii.（2）Design three nested primers K1, K2, K3, based on the PI4K gene sequence of G. Raimondii.Four allele of PI4K gene, gGbPI4K1, gGbPI4K2, gGhPI4K1and gGhPI4K2,sequenced in thegenome of CCRI8(G. hirsutum) and Pima90-53(G. barbadense)..（3）Develop of specific primers for four allele of PI4K gene, including two INDEL markers(K1-ID-1/K1-ID-12) and two SNP markers (K2-5-1/K2-11-1). K1-ID-1and K1-ID-12amplified of PI4K1specifically;K2-5-1and K2-11-1amplified of PI4K2specifically.K1-ID-1have a amplified fragment of324bp in Pima90-53’s genome and a amplifiedfragment of350bp in CCRI8’s genome;K1-ID-12have a amplified fragment of259bp inPima90-53’s genome and a amplified fragment of293bp in CCRI8’s genome;K2-5-1have noamplified fragment in CCRI8’s genome and a amplified fragment of280bp in Pima90-53’sgenome; K2-11-1have no amplified fragment in CCRI8’s genome and a amplified fragmentof280bp in Pima90-53’s genome.（4）Analysis the genetic linkage of gene specifical markers in the BC1population of (Pima90-53×CCRI8)×CCRI8contained95individuals and the gene PI4K1and PI4K2is located atlinkage group D13(c18) and A13(c13).2. Trehalose phosphate synthase (Trehalose-6-phosphate Synthase, TPS), the fiber developmentrelated gene, was developed functional markers successfully.（1）The TPS gene is located at Ca4and Cd2chromosomes. with the full length sequence of TPScDNA based on the database of diploid ancestors’genome of G. arboreum and G. raimondii. （2）Design fourteen nested primers T1to T14, based on the TPS gene sequence of G. Raimondii.Four allele of TPS gene, gGbPI4K1, gGbPI4K2, gGhPI4K1and gGhPI4K2, sequenced in thegenome of CCRI8(G. hirsutum) and Pima90-53(G. barbadense).（3）Develop of specific primers for four allele of TPS gene, including four INDEL markers(T2-ID-1/T1-ID-2/T2-ID-3/T2-ID-4). T1-ID-2amplified of PI4K1specifically andT2-ID-1/T2-ID-3and T2-ID-4amplified of PI4K2specifically. T1-ID-2have two amplifiedfragment of259bp and207bp in Pima90-53’s genome and CCRI8’s genome respectively;T2-ID-1have two amplified fragment of351bp and341bp in Pima90-53’s genome andCCRI8’s genome respectively; T2-ID-3have two amplified fragment of284bp and269bp inPima90-53’s genome and CCRI8’s genome respectively; T2-ID-4have two amplifiedfragment of576bp and551bp in Pima90-53’s genome and CCRI8’s genome respectively.（4）Analysis the genetic linkage of gene specifical markers in the BC1population of (CCRI8×Pima90-53)×CCRI8contained95individuals and the gene TPS1and TPS2is located atlinkage group D1(c15) and A1(c1).3. Use the sequence amplified by SSR markers from tetraploid genetic linkage in tetraploid cottongenome and the sequence of PI4K and TPS gene BLAST in the diploid genome database to makethe tetraploid cotton’s genetic linkage group mapped onto the diploid chromosome.The linkagegroup of D13(c18), A13(c13), D1(c15) and A1(c1), mapped to the diploid’s chromosome Cd13,Ca5, Cd2, and Ca4respectively.