| The tender leaf sheath of four sugarcane varieties G21, G28, ROC22 and RB86-7515 (B8) were cultured in vitro to study the effects of different hormone treatment on the induction, multiplication, differentiation and root regeneration of the and embryogenic cell cluster During the process of culture in vitro, the microscopoc results of sugarcane callus and embryogenic cell cluster were observerd and compared in the test. The main test reasults were as follow:1. The test results of sugarcane callus induction:(1) The best hormone combination of sugarcane callus induction was MS+2,4-D3.0mg/L+NAA0.2mg/L+KT1.0mg/L. The induction rate of sugarcane varieties G21, G28, ROC22 and B8 reached to 85.22%,79.21%,83.67% and 78.63% respectively.(2) MS medium was more suitable than White medium and Nf, medium on the callus induction of sugarcane explants.(3) Adding 5% coconut milk to the induction of sugarcane explants could improve the callus induction rate. The callus induction rate of G21, G28, ROC22 and B8 were 87.33%, 83.08%,84.13% and 82.14% respectively.(4) Adding Cys could improve its induction rate of sugarcane explants, the rate of G21, G28, ROC22 and B8 were 85.17%,82.14%,84.10% and 80.79% respectively.(5) It could improve the rate of callus induction when it used the dark culture firstly and the used the linght culture in the early days of the explant induction, the induction rate of G21, G28, ROC22 and B8 was 86.95%,81.11%,83.60% and 77.67% respectively.(6) The rate of callus induction of four sugarcane varieties reached the highest value when the cultural temperature was 30℃, the inductions rate of G21, G28, ROC22 and B8 was 88.33%,87.33%,86.27% and 81.24% respectively.(7) Compared with other seasons, the rate of callus induction of four sugarcane varieties reached the best when sampled in summer. The induction rate of G21, G28, ROC22 and B8 reached to 84.00%,76.19%,83.59% and 75.45% respectively.(8) The effect of callus induction could be reduce by the pretreatment with low temperature to the explants of sugarcane.(9) The order of early and late time of the callus induction between the varieties of sugarcane in different seaaons was G21<ROC22<G28<B8. The explant browning rate of the same variety showed difference when it was in the different seasons, the high and low order of the browning rate was witer> spring> autumn> summer.(10) Using MS+2,4-D 3.0mg/L + NAA 0.2 mg/L+KT 1.0 mg/L+0.03 AC+1.5 PVF hormone combination could effectively reduce the browing rate of sugarcane callus. The browing rate of G21, G28, ROC22 and B8 was 10.11%,17.74%,14.31% and 20.04% respectively.2. The test reasults of subculturing multiplication of sugarcane callus and embryogenic cell cluster.(1) The multiplication effect of four sugarcane varieties embryogenic cell cluster showed the best in the hormone combinations MS+2,4-D 3.0 mg/L+NAA 0.2 mg/L+KT 0.5 mg/L, the multiplication multiple of embryogenic cell cluster of G21, G28, ROC22 and B8 reached to 41.47 times,32.28 times,38.80 times,30.35 times respectively.(2) When the cuture time of embryogenic cell cluster was at the age of 15 days and subculturing time was the 4th generation, the multiplication multiple of embryogenic cell cluster of four sugarcane varieties reached the biggest value. The multiplication multiple of embryogenic cell cluster of G21, G28, ROC22 and B8 reached to 46.23 times,41.22 times, 44.80 times,37.33 times respectively.(3) Compared to other treatments of illumination cultivation, the treatment used the dark culture firstly and then used the light culture showed the hightest multiple of multiplication of the callus and the embryogenic cell cluster. The multiplication multiple of the callus of G21, G28, ROC22 and B8 was 4.45 times,3.33 times,4.13 times and 3.13 times respectively, but the multiplication multiple of the embryogenic cell cluster reached to 37.45 times,30.00 times, 35.05 times and 28.89 times.3.The test reasults of differentiation of sugarcane callus and embryogenic cell cluster(1) The differentiation effect of the medium and hormone combination MS+ +NAA 0.2 mg/L+6-BA 2.0 mg/L+TDZ 0.04 mg/L to the callus and enmbryogenic cell cluster of four sugarcane varieties was the best, the differentiation rate of G21, G28, ROC22 and B8 reached 86.83%,82.44%,88.89% and 80.00% respectively.(2) When the culture time of embryogenic cell cluster was 15 days and subculture time was the 4th generation, the differentiaion rate of the callus and embryogenic cell cluster of four sugarcane varieties reached the biggest value. The differentiation rate of callus was 88.11%,84.70%,90.97% and 80.15% respectively; but the differentiation rate of embryogenic cell cluster reached to 92.28%,89.00%,95.72% and 87.98% respectively.4. The test reasults of sugarcane buds multiplication:The medium and hormone combination MS+NAA 0.2mg/L+6-BA 2.5 mg/L+ PP3330.5 mg/L was benificial to the buds multiplication and growth. The buds multiplication of four sugarcane varieties G21, G28, ROC22 and B8 was 5.03 times,4.35 times,5.32 times and 3.78 times respectively.5. The test reasult of sugarcane plantlet rooting:Three factors hormone combination 1/2MS+NAA 0.2mg/L+6-BA 2.5 mg/L+ PP3330.5 mg/L was beneficial on rooting of sugarcane plantlets. The rooting rate of four sugarcane varieties all reached 100%.6.Analysis of microscopoc results of sugarcane callus and embryogenic cell cluster:Embryogenic cell cluster showed the following features:cell structure was compact, small cell, big nucleus, dyeing deeply; callus cell structure showed the following features:cell structure was loose, big cell, small nucleus,dyeing lightly. |
PDF Full Text Request