Study On Methods For Chromosome Doubling And Ploidy Identification In Brassica Napus L.

In this study with 2 homozygous rapeseed (B. napus L.) varieties, (Zhongshuangll and F009),2 F1 hybrids (3828 x 0009,0009 x 3828), and 2 F1 hybrids between B.napus rape and B. campestris rape (0470 x 3383,0009 x 3383), the induction of chromosome doubling by colchicine and some other main chemical reagent was studied. Microscopical observation through traditional root tip cells, morphological observation and flow cytometry detection method, the double handle materials are compared, and ploidy identification and rape chromosome ploidy of flow cytometry detection method for further research. The main results are as follows:1. Five levels of gradient colchicine concentrations,0,20,40,60,80 mg/L were set, in the MS culture medium to double the receptor material seeds, record and calculate the seed germination rate, seedling survival rate, success rate, double experiment found that the experimental materials with the increasing of concentration of colchicine, seed germination rate and seedling survival rate gradually decreased, and double success rate rise gradually;To determine the concentration of 60 mg/L for rape seed the optimal concentration in culture medium on the double.2. This experiment adopts the homozygous B.napus rape, hybrid B.napus rape, B.napus and Brassica campestris hybrid rape,three kinds of material, through the experiment found that the B.napus and Brassica campestris hybrid rape strongest tolerance ability of colchicine, have the highest germination rate, survival rate and success rate, is the most suitable for double material.3. Article by traditional root tablet counting chromosome number method and flow cytometry assay to detect chromosome ploidy double handling material, counting tablet chromosome method for reference to the apex and determine the rapeseed its reliability of flow cytometry detection, and detect the triploid GanBai F1 generation of hybrid rape double success of hexaploid canola and tetraploid hybrid F1 generation of brassica napus 1. were used successfully to get double eight times of rape..By studying the flow cytometry detection rapeseed chromosomal method, explore a set of suitable for rape ploidy detection technology system:MOPS 20 mmol/L, sodium citrate 30 mmol/L, MgC12 45 mmol/L, TritonX-1000.2%(V/V), β-mercaptoethanol 20 ul/ml;0.1 g test sample the best dosage of young leaf;Specific DNA concentration of dyeing liquid volume is best for the 400 ul, best to 100 ug/ml, dyeing time best for 15 min.4. Based on the double offspring plant morphological observation found that successful plant performance for double stem stout, plant dwarf, leaves shrivel, bud, the petals, stigma and anther are bigger, double success rape flowering delay;Can’t normal strong triploid B.napus and Brassica campestris hybrid rape by double for six times can strong body, tetraploid after double to eight times the body the seed setting rate is reduced, but seed kernels increased by about 70%.