| Strawberry(Fragaria×ananassa Duch.),belongs to Rosaceae Fragaria,is one of the most important fruit products in daily life.But strawberry cultivars are mostly octoploid,and due to difficult breeding associated with high heterozygosity,it has a limited potential using traditional breeding methods.Moreover,the strawberry virus disease will gradually accumulate with the continuation of asexual reproduction in the growth process,which affect the yield of strawberry and fruit quality seriously.Therefore,strawberry seedlings generally need 2 to 3 years for a change seedlings.In order to improve the differentiation rate of strawberry seedling and Proliferation coefficient to shorten the regeneration period.Strawberry seed,leaf,petiole and the base of the test-tube plantlet were used as explants,by changing the hormone ratio and concentration,establish the efficient regeneration system using the base of the test-tube seedlings as explant.In order to further improve the differentiation efficiency,the base of the test-tube seedlings was divided into two parts of the stalk of the stem of stipules and the base of the test-tube seedlings as explant,and established a more rapid and efficient regeneration system.The genetic stability of strawberry plants obtained by different routes were detected by SRAP molecular marker technique,by exploring the similarity between the offspring and the female parent,which provided the technical basis for the preservation of in vitro strawberry germplasm resources.Strawberry will be affected by the external environment esaily in the growth process.Saline soil is widely distributed in China,salt stress can inhibit the growth and development of plants tissues and organs.According to statistics,salt damage will lead to global crop annual reduce more than20% in production.How to improve the salt resistance of strawberry is the urgent problem to be solved at present.Wu Keying has shown that resistance of the tobacco which introduced into Cjhppd gene is significantly increased.In this study,the transformation system of Agrobacterium tumefaciens-mediated strawberry was established.The Cjhppd gene was introduced into strawberry plants to obtain transgenic plants,then study transgenic characters,which laid the foundation for the improvement of strawberry quality by using geneticengineering technology.The main findings include:1.Establishe The efficient regeneration system of strawberry: using the stem of stipules and the base of the test-tube seedlings as explant were culture on MS medium supplemented with IBA 0.2 mg·L-1+6-BA1.5mg·L-1.The explants can directly differentiate adventitious buds without producing obvious "callus",and the regeneration period was short.The differentiation rate is 100% in the fastest one week,and the average value is 6 and 6.73 buds,up to 13.Buds transferred to MS medium,can form a normal plant,rooting rate of 90% or more.2.The strawberry genomic DNA was extracted by CTAB method,and the SRAP-PCR appropriate reaction system of strawberry was established.Seventeen pairs Random primer combinations can amplify clear and stable bands,a total of 199 bands,the polymorphic band ratio of 58.79%,the confidence probability of 99.99999%,can be used to identify any kind of sample.And it was proved that the genetic stability of strawberry basement was higher,which provided a basis for the rapid propagation of strawberry germplasm resources.3.Establish Agrobacterium tumefaciens-mediated transformation system of strawberry test-tube plantlets using gfp as marker gene,The highest transformation rate is 22.73%.4.Using the above system,the Cjhppd gene was introduced into strawberry to obtain the transformed plants. |
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