The Separation And Expression Analysis Of Poplar Crown Type Regulation Factor

Populus is currently more extensive cultivation of one of the important species. The Popular leuopyramidalis is by ring Ye Yang and the hybrid hybrid new breed, with fast-growing, root deep, narrow crown, no flying catkins, good material, flank light excellent characteristics, is focused on the promotion of the national excellent poplar varieties. Has been widely used in Shandong, Hebei and other places.This experiment to Popular leuopyramidalis single bud stem segments as experimental materials, of Popular leuopyramidalis callus induction, bud differentiation induction, rooting induction of study; Popular leuopyramidalis and Populus tomentosa as experimental material of poplar crown type gene cloning, separation and expression analysis; To investigate the relationship between the polar auxin transport and narrow crown type. The results of this study are as follows:?1? The culture meadium for tissue culture adaptation of Popular leuopyramidalis was determined.Callus induction: MS+6-BA1.0mg/L+2,4-D4.0 mg/L; Bud differentiation inducing medium: MS + 6-BA0.3mg/L +ZT0.25mg/L+IAA0.25mg/L; Rooting induction medium: MS + 6-BA0.01 mg/L+ IBA0.5 mg/L +NAA0.1 mg/L.?2? Cloning the related genes by PCR technique successfully, Named as the gene PtTAC and PlTAC. PtTAC in Populus tomentosa Carr of the full-length cDNA sequence of 1028 bp,with a 813 bp consisting of a complete reading frame, 3'-UTR fragment length was 180 bp,5'-UTR fragment length 35bp; Pl TAC in Popular leuopyramidalis of the full-length cDNA sequence of 976 bp has a are 723 bp composition complete reading code, 3'-UTR fragment long 218 bp and 5'-UTR fragment length 35 bp. Through the comparative analysis found that the nucleotide sequence deletion fragment is the reason for the narrow crown.?3? The protein expression of PtTAC is C₁₃₄₉H₂₁₆₃N₃₈₃O₄₂₉S₇, molecular weight is30835.7 Da,Instability index is 41.04, PI value is 5.39, hydrophilic, hydrophobic proteins have no obvious relationship, no signal peptide and transmembrane domain, which belongs to the non secretory protein, a total of 6 Ser phosphate site, 3 Thr phosphorylation sites, 4 Tyr phosphorylation sites, Alpha helix is 40.37%, Beta turn is 8.15%, Extended strand is15.56%,Random coil is 35.93%; The protein expression of PlTAC is C1193H1887N325O385S5, molecularweight is 27103.3Da and PI is 4.92, hydrophilic instability index is 38.14, no signal peptide and transmembrane domain, which belongs to the non secretory protein, a total of 6 Ser phosphorylation sites, 3Thr phosphorylation sites and 4 Tyr phosphorylation sites, Alpha helix is 44.58%, Beta turn is 8.33%, Extended strand is16.25%, Random coil is 30.83%.?4? In order to test the expression of TAC in different organs, the expression of in different organs such as stem and was analyzed with the test materials of Popular leuopyramidalis and Populus tomentaso. The results of the test showed that expression of TAC in Populus tomentosa in stem, leaf and bud organ respectively is: the expression of leaf,stem and bud expression quantity is small; in the Popular leuopyramidalis in the expression of stem and leaf respectively is: the expression of stem, bud expression of minimum leaf expression in the middle. The overall expression of TAC was higher than that of the Popular leuopyramidalis.?5? The stems and buds for the analysis of gene expression of auxin transport test materials. PopPIN and PoptrPIN-formed6 expression in tems was lower than the bud, in the Popular leuopyramidalis bud is higher than in the Populus tomentosa bud; The expression of PopAUX2 in bud is higher than in the stem, the expression in Populus tomentosa bud is higher than in Popular leuopyramidalis bud.?6? The difference with Popular leuopyramidalis and Populus tomentosa branching and leaf buds as the experimental material, through the observation of measuring the leaf size,toward the buds, lateral branches and branchlets form to compare Popular leuopyramidalis and Populus tomentosa. The test results show that Popular leuopyramidalis branches erect,thick and easy to form competitive branch, branch node swelling and cracking. Branching angle between 45-90 deg, collateral branch 90 deg bending upward growth almost and branch in parallel, the leaf buds to grow, volume is small. Branchlets not smooth. Populus tomentosa branch branch angle greater than 90 degrees, leaf level or down growth, large volume.Branchlets smooth.