Genetic Transformation Of Drought- And Salt-tolerance Gene Avp1、FNR-Fld、OsSiz1 Into Barley Commercial Varieties By Agrobacterium Tumefaciens Infection

The problems of drought and salinization are very serious environmental stresses while bringing about severe adverse effects on the agricultural production, which were recognized throughout the world. Recently, with the gradual deterioration of ecological environment, the scope of the drought and salinization land is continuing to expand internationally. Using transgenic technology to introduce droughtand salt-tolerance genes into barley for improving drought- and-salt resistance and cultivating new varieties, plant breeders could solve the problem of drought and salt effectively. In this study, we optimized the regeneration system of barley immature embryo, on the basis of which, the regeneration system of barley mature embryo was established. Meanwhile, we make a variety of barley varieties as recipient materials and use the Agrobacterium-mediated genetic transformation system to transform the droughtand –salt tolerant genes Avp1、FNR-Fld and Os Siz1 into them for obtaining new transgenic barley varieties with the trait of drought- and-salt resistance. The results were as follows:(1) In the process of barley callus induction, adding 0.2mg/L ABA is helpful to barley immature callus induction, and adding 0.4mg/L ABA is helpful to barley mature callus induction.(2) For the callus induction of barley mature embryo, the method of removing the embryonic axis is superior to the method of adding 0.4mg/L ABA to the whole embryo.(3) The genotypes of barley have a great influence on callus induction. In terms of callus induction, the two-rowed barley varieties are better than the six-rowed barley varieties, the hulled barley varieties are better than the naked barley varieties.(4) In the Agrobacterium-mediated genetic transformation of barley immature embryo callus, drought- and salt-tolerance genes Avp1, FNR-Fld and Os Siz1 were transformed into five different barley varieties, including Amagi Nijo, Kanto Nijo 22, Misato Golden, Zhenongda 3, Huadamai 6, and Supi 1. We identified a total of 5 transgenic plants, including 3 lines of Amagi Nijo, 1 line of Misato Golden, and 1 line of Zhenongda 3 with PCR analysis from 36 regeneration plants.(5) In barley mature embryo callus genetic transformation mediated by Agrobacterium, drought- and salt-tolerance gene Avp1, FNR-Fld and Os Siz1 were co-transformation into four different barley varieties, including Amagi Nijo, Misato Golden, Zhenongda 3, and Supi 1. We got 13 regenerated plants in all. Nevertheless, only 1 transgenic line of Amagi Nijo was identified as the positive plant by PCR analysis, and all the regenerated plants from the other barley varieties were negative.(6)The varieties of barley have a great influence on genetic transformation. In Agrobacterium-mediated genetic transformation the two-rowed barley varieties Amagi Nijo is the best recipient material.