| Isolated microspore culture technique has been widely used in practice.With its obvious advantages, it will have a wide prospect. Athough there were some reports on microspores culture of cabbage,but there were lower rate of embryoid induction,narrow application range of genotypes and lower seedling rate. Therefore, this experiment takes 13 superior cabbage varieties as test materials for microspore culture.Study on the effects of different genotypes, bud plant growth state,the environmental maxium temperature, adult plants and half adult plants, adding colchicine in NLN-17 culture medium on embryoid induction, and the effects of induced buds, rooting medium supplemented with hormones and paclobutrazol on embryoids seedling induction. The main results are as follows:1. Cabbage plant genotype plays a major role in the microspore culture for the high rate of embryo induction. four materials were not induced embryoid and nine materials were induced embryoids of all the 13 cabbage materials. The number of embryoids induction for Q4 was the largest of all the nine material, and the highest induction rate was 30.25 embryos in one bud. induced the number of embryoids induction for HP72 was least, and the lowest induction rate was 2.00 embryos in one bud.2. Cabbage plant growth temperature is too low or too high will result in differences of microspore embryo induction. In Q4 plant growth environment, when the day's highest temperature was 28℃, for the next day, the consistency of bud development was high, and the number of embryoid induction for microspore was most, the rate of embryo induction reach 30.3 embryos in one bud.3. The embryo induction in half of cabbage adult flower buds is easier than that of cabbage adult. The rate of embryo induction of the half adult was higher significantly than that of the adult for Q2, Q4 and Q5, and up to 9.6 embryos ~ 18.7 embryos in one bud.4. Adding colchicine can promote swelling and equal division of cabbage microspore in the heat shock culture, and is helpful for the improvement of embryo induction. The rate of embryo induction which adding 1,10,50 mg / L colchicine added 0.59 to 22.2 embryo in one bud than that of not adding colchicine. In which the generation of hybrid added 1 mg / L or 10 mg / L colchicine, inbred added 10 mg / L or 50 mg / L colchicine can better induce embryo. The rate of embryo induction for Q4 and Q5 were more than 15.30 embryos in one bud and 29.5 embryos in one bud, for HP63 and HP72 were more than 7.96 embryos in one bud and 1.98 embryo in one bud.5. The best medium and hormone combination for adventitious buds of cabbage microspore embryos is B5 +6- BA 0.1 mg / L + NAA0.01 mg / L, in such differentiation medium embryoids callus and browning occurred less , the rate of adventitious buds and differentiation were higher, can reach 86.7% and 330.8%.6. Add 0.2mg / L paclobutrazol MS +0.01 mg / LNAA regenerated plantlets rooting medium up to 96.7% survival rate of regenerated shoots thick stems and roots. |
