| The study used young leaves sheath of three sugarcane varieties G35, G21and ROC22as the experimental materials. The technique system for induction, multiplication, differentiation and plantlets regeneration of sugarcane callus and embryonic cells cluster was set up. We have studied that mutagenic effects of multiplication, differentiation and plantlets regeneration after EMS treatment to the embryonic cells cluster. We also have studied that the effect of transplanting survival rate after EMS soak sugarcane seedling roots, and physiological identification for drought resistance of sugarcane mutagenic strain. The researches showed that:1、Test results of sugarcane callus induction, multiplication, differentiation and embryonic cells cluster multiplication, differentiation and bud proliferation and rooting:(1)Sugarcane young leaves sheath cultured in MS medium were suitable to callus induction among MS, B5, N6, Miller medium. The induction rate of G35reached the highest when sampled in spring, it was68.75%.(2) Compared with other seasons, the callus induction rate of three sugarcane varieties reached the highest when sampled in summer(in August). The induction rate of G35、G21、 ROC22were82.5%,79.17%and75.42%respectively.(3) The callus induction rate of three varieties reached the highest when the basic Medium and hormone combination was MS+2,4-D2.5mg/L+NAA0.2mg/L+KT1.0mg/L. The induction rate of G35、G21、ROC22were93.33%,91.67%and87.08%respectively.(4) When callus subcultured was the5th generation, the callus multiplication multiple of three varieties reached the highest. The callus multiplication multiple of G35、G21、ROC22were4.08multiple,3.97multiple and3.85multiple respectively; The incidence rate of ROC22embryonic cells cluster was the highest in the3th generation and the rate was46.25%, the incidence rate of G35embryonic cells cluster was the highest in the4th generation and the rate was45.00%, the incidence rate of G21embryonic cells cluster was the highest in the5th generation and the incidence rate was40.00%.(5) When the callus and embryonic cell cluster subculture was the4th generation, the budding differentiation of three sugarcane varieties were the best. The bud differentiation rate of callus were G35(83.00%)>G21(79.58%)>ROC22(75.00%) and the bud differentiation rate of embryonic cell cluster were G35(92.08%)>ROC22(90.00%)>G21(89.17%)(6) Bud growth potential was the best when Medium and hormone combination were MS+6-BA3.0mg/L+NAA0.2mg/L with TDZ0.05mg/L or PP3330.5mg/L. The effect of bud multiplication multiple was the best when additional TDZ0.05mg/L:G35(5.00)>ROC22(4.88)>G21(4.50)(7) The rooting effect was the best when medium and hormone combination were l/2MS+NAA0.2mg/L+IBA1.0mg/L+PP3330.6mg/L. Rooting rate of three sugarcane varieties reached100%, the average root number was G35(13.95)>ROC22(14.30)>G21(11.00) and the average root length was G35(2.87cm)>G21(2.53cm)>ROC22(2.17cm).2^The test results of EMS treatment on embryonic cells cluster mutagenesis:(1) As the extension of EMS soak time and the increasing of EMS soak concentration, death rates in brown of embryonic cells cluster were increased, while the multiplication multiple and bud differentiation rate of embryonic cells cluster were reduced.(2) Some mutation strains that were excellent in bud multiplication were got after EMS treatment, they are G21E1(0.2%EMS treatment3h),G35B2(0.05%EMS treatment5h) and ROC22C2(0.10%EMS treatment5h). When the medium was MS+6-BA3.0mg/L+NAA0.2mg/L+TDZ0.05mg/L, the bud growth potential of all strains showed the best and the multiplication multiple reached the highest. The bud multiplication multiple was G35B2(4.86)>G21E1(4.66)>ROC22C2(3.90)(3) The rooting rate was close to100%when selecting the optimum rooting medium1/2MS+NAA2.0mg/L+IB A1.0mg/L+PP3330.6mg/L to induce the mutant rooting. Integrated average root length, root number, seedling growth potential,G21E1(0.2%EMS treatment3h), G35E2(0.2%EMS treatment5h), ROC22C2(0.1%EMS treatment5h) and ROC22D2(0.15%EMS treatment5h) take root quality were better. 3、The physiological analysis results of EMS treatment on sugarcane mutant strain for drought resistance:By measuring mda content, proline content, CAT content,soluble protein content, soluble sugar content, chlorophyll content of three sugarcane mutant strains, the membership method according to function in fuzzy mathematics comprehensive evaluation was carried on. Subordinate function value is the weighted value. The greater the weighted value was, the better the drought toleranc was. Aseptic seedling rounding out the top five of seven indexes of subordinate function weighted value size order were ROC22Y2>ROC22X2>G35Y2>G21>G21X1>G21Z1>G35W2.Transplanting seedling rounding out the top six of seven indexes of subordinate function weighted value size order were G35Y2>ROC22Y2>G21X2>ROC22X2>ROC22Z1>G35W2>G35Z1.4、The test results of EMS second treatment on transplanting survival rate of sugarcane mutant strains:As the extension of EMS soak root seedling time and the increasing of EMS soak concentration, transplanting survival rates of three sugarcane varieties mutant strains were reduced. |
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