Studies On Virus-free Rapid Propagation And Genetic Variation In Zingiber Officinale Roscoe

Ginger?Zingiber officinale Roscoe?belong to perennial herb.It's traditionally called the small size ginger "xiaohuang ginger".Xiaohuang ginger from Zhexi has the strong market competitive due to high ginger oil and gingerol.Because of the disability to form seed,ginger mainly use rhizomes for asexual reproduction.However asexual reproduction result to deterioration of strains which presented as reduced stress resistant ability and lower quality.Therefore,it is important to studied and established the technology of virus-free seedling of ginger that we can apply in to production.By this,the purpose of expandable production improve quality of ginger will be achieved.To providing a theoretical basis for the large-scale production of xiaohuang ginger from Zhexi,we have studied on the regeneration system and genetic variation of virus-free plants.The main studies are as follows.?1?Sterilization methods of xiaohuang ginger bud explant were identifiedAfter stripping four layers,ginger bud treated with different 0.1% HgCl₂ treatment time?0,3,6,9 mins?combined 75% alcohol for 30 s.Results exhibited that 75% alcohol treatment for 30 s,0.1% HgCl₂ 3min is the best sterilization combination.Stripping different layers of ginger buds?0?9 layers?,then with 75% alcohol for 30 s,0.1% HgCl₂ 3min.results exhibited that after stripping 4 to 6 layers,the explants had the highest disinfection rate and lowest mortality rate.?2?Optimization of culture medium for xiaohuang ginger explantsThe effects of different concentrations of NAA and 6-BA on the growth of ginger tissue culture seedlings were detected using orthogonal method L9?3²?.The data showed that within 3 months' inoculation,the orthogonal group grow multiple shoots at the average of 1.7and adventitious roots 3.1.The differentiation and growth of the multiple shoots and adventitious roots of ginger had a significant advantage,when the medium hormone content was 0.1 mg/L NAA and 4 mg/L 6-BA after analysis,the average of multiple shoots and adventitious roots was at the average of 4.1 and 12,respectively.Thus inferred that this concentration of the hormone can good promote the differentiation and growth of multiple shoots and adventitious roots on tissue culture seedlings.?3?Detoxification tissue culture virus detection of xiaohuang gingerThe TMV content of virus-free tissue culture seedlings was determined by ELISA.The results showed that TMV content of untreated culture seedlings was 3.3174 ?g/L,while the TMV content of the virus-free tissue culture seedlings was lower than that of untreated culture seedlings,and the average concentration of TMV was 0.2995 ?g/L,which indicating that the combination of stem tip toxicity and heat treatment can effectively remove the TMV of xiaohuang ginger.?4?Genetic Variation detection of Virus-free culture seedlingsUse DNA of 8 different cultivars ginger which from different place and PCR technology,5 primers?825,834,846,881,895?with clear bands and good polymorphism were screened out from 100 ISSR primers published the by Columbia University.3 isolates of virus-free seedlings were taken from each generation,using the selected primers to amplify,respectively,the results showed that the 8^th generation of culture seedlings did not occur genetic genetic variation.