Studies On Plant Regeneration From Leaf And Polyploid Induction Of In Vitro Ginger (Zingiber Officinale Rosc.)

Ginger is difficult to use conventional means of breeding new varieties of germplasm and cultivation for which is the vegetative propagation of crops. Therefore, in this paper,we studied different explants of "Laiwu" ginger and "Shannong No.1" ginger plantlets as materials for callus induction and plant regeneration, discussed the effect of different concentrations of colchicine on polyploid induction, and the relationship between light quality and the regenerated plants growth as well as the rhizomes induced of ginger, in order to improve an efficiency ginger tissue culture and new germplasm, the main results are as follows:1. Young leaves/sheaths/adventitious roots were as explants for callus induction, and laves were cultured as the best explants. The effect on leaf's callus inducation of different basic media, hormone levels, time of dark culture and the relation between hormone combination and callus differentiation and plant regeneration were studied in the expeiment. The results showed that leaf-derived callus induction on N6 was better than MS, MN and 1/2 MS. Appropriate concentrations of 2,4-D were beneficial to caluss inducation. The effect of 2,4-D and 6-BA on callus induction was better than 2,4-D and KT, and the time of dark culture was an important factor. The best medium for "Laiwu" ginger leaf-derived callus induction was N6 supplemented with 1.0 mg/L 2,4-D+0.5 mg/L 6-BA+3% sucrose+0.6% agar, and the rate of callus formation was 77.78% when leaf explants were cultureed 30 days in the dark; the best medium for "Shannong No.1" ginger leaf-derived callus induction was N6 supplemented with 1.0 mg/L 2,4-D+1.0 mg/L 6-BA+3% sucrose+0.6% agar, and the rate of callus formation was 78.89% cultureed 30 days in the dark.2. We obtained suitable·culture conditions for callus proliferation and somatic embryogenesis by selecting factors of callus for proliferation. The results showed that calluses cultured on MS and N6 media under white light are experiencing three stages of slow growth, rapid growth and the growth of smooth, showing a "S" curve, and the MS medium for callus proliferation was better than N6 medium. Calluses growed slowly after 30 days, then calluses needed to replace the same composition of medium for continuing to grow. Sucrose concentration changes the physiology of leaf callus and directly affect the maturation of somatic embryos, the study found that concentrations of sucrose increased in the medium is conducive to callus to the transformation of somatic embryogenesis. Therefore, the culture medium for the proliferation of embryogenic callus was:MS+0.2 mg/L NAA+5.0 mg/L 6-BA+30 mg/L sucrose+mg/L agar; and the culture medium for embryogenic callus into somatic embryogenesis was:MS+0.2 mg/L NAA+5.0 mg/L 6-BA+5.0% sugar+0.6% agar.3. Lower concertrations of NAA and higher concertrations of 6-BA was conductive to callus and plant regeneration, and 6-BA which promoted organ differentiation was better than KT. Mediun for "Laiwu" ginger callus and plant regeneration was MS supplemented with 6-BA(10 mg/L) and NAA(0.2 mg/L), and the index of plantlet regeneration was 8.5; Callus differentiation index of "Shannong No.1" ginger could although to achieve 34.1, but the differentiation rate was lower 11.42% than "Laiwu" ginger, and the index of plantlet regeneration was 5.9. The effect of activated carbon added to promote callus differentiation was not obvious, and which added 0.5% activated carbon easily lead to a galass vitro regeneration ginger plants, detrimental to their growth.4. Different colchicine concentration and treatment time, can significantly influence the ginger plantlet survival rate, mutation rate and the doubling rate of shoot regeneration. We calculated that the doubling rate of ginger plantlets was expected to reach more than 2% by regression analysis, when the colchicine concentration and treatment time were 0.118% 0.215% and 9.04 d～14.48 d, and when colchicine concentration and treatment time were 0.034%～0.187%,4.91～15.62 d, the survival rate of ginger plantlets could achieve more than 80%; and colchicine concentration and treatment time were 0.116%～0.197%,10.63 d～16.29 d, the mutation rate could expected to get more than 5%. The tetraploid plants exibited some morphological, including larger stomata and fewer stomata per unit area. There were no plantlets from callues after colchicine concentration treated by morphological trait and flow cytometry techniques.5. In order to improve the efficiency of micropropagation of ginger, the effects of different light quality on the growth and micro-rhizome induction of in vitro cultured ginger were studied. The results showed that compared with white light, green and red. lights increased plant height of in vitro ginger, and green light also reduced its propagation coefficients, significantly, but that of the blue light just was opposite, and yellow light had a little effect on the growth of in vitro ginger. The time of micro-rhizome formation of of in vitro ginger cultured under green, blue and red lights was earlier than that of white light, but later under yellow light. Moreover, the micro-rhizome fresh weight of in vitro ginger under green light was higher, which increased by over 60% compared with that of white light, but there was no significant difference among the other light treatments. Otherwise the dry mass content, starch and crude cellulose of micro-rhizome formated under green light were lowest, but highest under blue light. Soluble sugar content was higher under red and yellow light, while lower under green and blue light, but contrary result appeared with soluble protein content. The Gs and transpiration rate were higher under blue and yellow light, but lower under red and green light.