Study On Microspore Culture Technology Of Single Flower Bud In Cabbage Untwisted Ball DH Strain Axillary Bud Regeneration

To explore a new idea of high-efficiency embryogenesis by microspore culture of cabbage,and compare the culture conditions of culture dish,6-BA concentration in medium,centrifugation speed and time of free microspore,heat shock time of single flower bud,etc.The effect was to establish a microspore culture technology system for single-flower buds of cabbage.The previous year,the micro-spore regeneration DH strain could not be bred and flowering could not achieve reproductive growth and complete seed reproduction.In order to prevent the loss of excellent DH strain resources,the DH strain axillary bud regeneration plant method was carried out.Exploring,from the DH strain,the best disinfection method,medium and hormone concentration,axillary buds and other factors were studied;the main conclusions are as follows:1.Cabbage single flower bud microspore culture Adding 3 mL medium in a diameter Φ30 mm culture dish is most conducive to high embryo rate microspore culture;test cabbage hybrid generation F17-A and F17-B at 4.7-5.1 mm The flower bud length range can be cultured in single flower buds,and the average embryo yield is 22.1 embryos/buds and 23.0 embryos/buds,respectively.2.F17-B flower buds of hybrids of Brassica oleracea L.,single-flower bud microspores cultured in NLN-14+NAA 0.5 mg/L+6-BA 0.5 mg/L medium,the highest rate of emergence was 24.5 compared with other mediums.Embryo / bud.3.In the single flower bud microspore culture centrifugation and heat shock reaction,the centrifugal speed was 2000 rpm,the centrifugation time was 5min,the heat shock time was 48 h,and the average embryo yield was higher than 25.0 embryo/bud.4.Using the non-twisting DH strain axillary bud regenerated plants,compared with 75% alcohol 20 s disinfection axillary bud tissue culture,the lowest pollution rate was 6.6%,the plant regeneration rate was 86.7%;MS+6-BA 2.0mg/L was selected.+NAA 0.5mg/L solid medium for axillary bud regeneration with a regeneration rate of up to 90.0%.5.Compared with different axillary bud regeneration plants in different parts of DH strain,the browning rate of axillary buds was the lowest in the middle of the ball,the highest rate of regenerated plants was 83.3%;the lowest rate of regenerated plants of rosettes was 70.0%;the highest survival rate of transplanted plants was the external axillary buds of the leaves.Up to 87.0%,the lowest is the internal axillary bud of the ball,which is 72.7%.The results showed that the optimal parts of the axillary bud regeneration plants of DH strain were the outer and middle axillary buds of the leaf.6.Compared with the axillary bud regeneration plants in different compactness of DH strains,the greater the compactness,the least the number of regenerated plants;the smaller the compaction,the higher the regeneration rate,and the better the axillary bud growth.