Analysis Of Bolting Inheritance Character And Related Gene Molecular Markers In Mustard (Brasscia Juncea Coss.)

Mustard (Brasscia juncea Coss.) is an important vegetable crop with a long cultivation history in China. Mustard intoleranced heat and cold, appropriate cultivation with cool humid. However the development of mustard is not strict with the low temperature and long day, vernalization and long day will complement each other, as long as completed the vegetative growth, it will bolting and flowering. In the breeding of the mustard, we need the premature bolting to add generation, propagation, shorten breeding period, and accelerated breeding process. In the seeding production, regulate the parents to flowering simultaneously. Therefore, it is necessary to study the bolting and flowering of mustard, this research studied the mechanism from the physiology and the molecular two different aspects which the leaf mustard reproduction growth transforms.This experiment used the mustard for the material, with the method of bulked sergeant analysis, divided bolting characters of F2 groups from one coup parents into two sections and established DNA pool. In order to find the gene linked together, further appraise the characters of bolting with 76 materials. The results:1. Mustard bolting inheritanceParent material by early bolting No.3 after vernalization was early 30days than parent material by late bolting No.41. As a reference, division vernalization time of F2 plants, with thex2 test showed, the mustard bolting accord 1:2:1, Reasoned the bolting gene may be have one major gene.2. ISSR Maker of mustard bolting geneAn ISSR analysis system is established with mustard by optimizing some factors of ISSR technological system. The results showed that the dNTPs should be 0.8 mmol/L, TaqDNA polymerase should be 1U, primer should be 1.0 pmol/mL. Find the ISSR maker-UBC826 which showed a band near 1.9kbp was identified to be linked with the bolting gene. The genetic distance was 4.0cM. 3. Comparison of two electrophoresis methodsAn PAGE analysis system is established with mustard, the results showed that the 30% Acr/Bis should be 6.20 mL,5xTBE should be 7.50 mL,10%Aps should be 270μL, TEMED should be 50μL.The reaction results for ISSR-PCR by mustard electrophoresis for 1% agarose gel and 5% polyacrylamide, the result showed that 5% polyacrylamide gel electrophoresis is more effected for the PCR products；especially the bands between 1kbp-2kbp can significantly improve resolution and reduce the analysis difficult.4. Analysis of bolting related proteinUsing 5% stacking gel and 10% separating gel on the soluble protein for F2 group; finally find a band near 35kDa should be the soluble protein band with the mustard bolting.