| Modern rose (Rosa hybrida L.) is one kind of the most important ornamental plants, which is popular because of the colorful , fragrant, concurrent flowers with a wide range of application. They are irreplaceable materials for landscaping in modern cities. Due to climate reasons in the north of China, there are few cultivars of roses can live through the winter.Modern roses have a narrow genetic base, and traditional breeding methods need to take a long term, which delays the development of rose breeding at present. Bioengineering offers a new approach for breeding work. Callus and somatic cell are good receptor materials for clone variation and genetic transformation, through the callus tissue regeneration and somatic training can obtain variable plants, and somatic fusion provides a potential new ways to solve distant hybridization infertility. But the establishment of modern rose callus regenerative and somatic cells culture still exists great difficulty, the relevant reports are still few. Therefore, this study has vital significance to promote the cold-tolerance and other traits breeding.This study discussed the factors that affected callus inducement and regeneration of modern rose, and acquired embryogenic callus, and established cell suspension based on the acquired callus, the fators that affected somatic cells culture also were discussed. The study is aimed to acquire regenerated plants from callus and somatic cells, and provide base for genetic manipulation such as cold-resistant species improvement, somatic cell clones variable selection and protoplast cultivation and fusion. The main results of this study are as follows:1. Establishment of micropropagation of five modern roses: The best medium for the germination of cultivar 1 axillary bud and shoot multiplication was: MS + 6-BA2.5mg·L-1 + NAA 0.05mg·L-1, the rate of proliferation can reach 4.2; The best medium for the germination of cultivars 2, 6 and 8 axillary bud and shoot multiplication was: MS + 6-BA 2.0mg·L-1 + NAA 0.05mg·L-1, the rate of proliferation can respectively reach 3.68, 4.07, 4.2; The best medium for the germination of cultivar 9 axillary bud and shoot multiplication was: MS+6-BA2.0mg·L-1 +NAA0.1mg·L-1, the rate of proliferation can reach 3.99; The best rooting mediums for five cultivars were 1/2MS+ NAA0.05 mg·L-1 and 1/2MS+ NAA0.1 mg·L-1, rooting rate can be up to 80%; The survival rate of sterile seedlings cultivated in the open can be up to 88% by domestication and transplanting.2. The leaves and young stems of five cultivars'sterile seedlings have a good callus induction ability. The best medium for leaves callus induction of cultivars 1 and 8 is: MS+2,4-D3.0 mg·L-1, the rates of inducation both reach 100.00%; The best medium for leaves callus induction of cultivars 2, 6 and 8 is: MS+2,4-D2.0 mg·L-1 + 6-BA0.5 mg·L-1, the rate of inducation can respectively reach 97.67%, 98.67%, 100.00%. The best medium for young stems callus induction of cultivars 1 and 6 is: MS+2,4-D3.0 mg·L-1, the rate of inducation can respectively reach 78.67%, 84.33%; The best medium for young stems callus induction of cultivar 2 is: MS+2,4-D2.0 mg·L-1, the rate of inducation can reach 73.67%; The best medium for young stems callus induction of cultivars 8 and 9 is: MS+2,4-D2.0mg·L-1+6-BA0.5mg·L-1, the rate of inducation can respectively reach 82.33%, 80.33%.3. The callus of five cultivars all can regenerate, except 2 cultivar without obtaining the regeneration seedlings, the other four cultivars can obtaine regeneration seedlings. The best medium for leaves callus differentiation of cultivar 1 is: MS+2,4-D0.2 mg·L-1 +TDZ9.0 mg·L-1 +GA30.1 mg·L-1, the way of differentiation is adventitious buds, the rate can reach 18.00%; The best medium for leaves callus differentiation of cultivar 6 is: MS+6-BA0.1 mg·L-1 +2,4-D1.0 mg·L-1 +TDZ8.0 mg·L-1 +GA30.1 mg·L-1, , the rate can reach 22.00%; The suitable mediums for leaves callus differentiation of cultivar 8 are: 1/2MS+6-BA0.5 mg·L-1+TDZ10.0 mg·L-1+GA30.1 mg·L-1 and MS+2,4-D0.2 mg·L-1 +TDZ9.0 mg·L-1 +GA30.1 mg·L-1, the way of differentiation on the first medium is somatic embryogensis which can form regeneration seedlings, the second is adventitious buds, the rates can respectively reach 18.67% and 21.33%, the best medium for young stems callus differentiation is MS+TDZ8.0mg·L-1, the way of differentiation is adventitious buds, the rate can reach 46.67%; The best medium for leaves callus differentiation of cultivar 9 is: MS+6-BA0.1 mg·L-1 +TDZ9.0 mg·L-1 +GA30.5 mg·L-1, the way of differentiation is adventitious buds, the best medium for young stems callus differentiation is MS+2,4-D0.2 mg·L-1 +TDZ9.0 mg·L-1 +GA30.1 mg·L-1, the way of differentiation is somatic embryogensis, the rate can reach 35.33%.4. The stable cell suspension of five cultivars can be founded on the liquid mediums which have the same composition of callus induction, the best time interval for subculture is 8-10d, cells has vigorous growth and strong activity after 4-8d culture, which are suitable for cell regeneration culture; There are no obvious differences in the growth characteristics between different rose genotypes.5. The suspended cells of cultivars 1, 8 and 9 can induce the callus, the ability of callus induction is low. The callus induction rate of cultivars 1 is 22.00% on the medium MS+6-BA0.1 mg·L-1 +2,4-D1.0 mg·L-1 +KT0.1 mg·L-1, the rate is 5.33% on the liquid medium MS+6-BA0.5 mg·L-1 +2,4-D0.2 mg·L-1 +KT0.5 mg·L-1; The callus induction rate of cultivars 8 is 27.33% on the medium MS+2,4-D1.0 mg·L-1 +NAA1.0 mg·L-1 +KT0.5 mg·L-1; The callus induction rate of cultivars 9 is 26.67% on the medium MS+2,4-D1.0 mg·L-1 +NAA1.0 mg·L-1 +KT0.5 mg·L-1, the rate is 2.67% on the liquid medium MS+6-BA0.5 mg·L-1 +2,4-D1.0 mg·L-1 +NAA0.2 mg·L-1.6. The callus induced by suspended cells of cultivars 1 can differentiate adventitious buds on the medium MS+2,4-D0.2 mg·L-1 +TDZ9.0 mg·L-1 +GA30.1 mg·L-1, the rate can reach 32.33%; The callus induced by suspended cells of cultivars 8 can differentiate somatic embryogensis on the medium MS+2,4-D0.2 mg·L-1 +TDZ9.0 mg·L-1 +GA30.1 mg·L-1, the rate can reach 26.67%; The callus induced by suspended cells of cultivars 9 can differentiate somatic embryogensis on the medium MS + 6-BA0.1mg·L-1 + TDZ 9.0mg·L-1 + GA3 0.5mg·L-1, the rate can reach 38.67%. |
PDF Full Text Request