| The experiment had been conducted in Shandong Agriculture University .Shandong Grape Brewing Research Institute and the Seedling Nursery of Dongying Forest Bureau. Grapes (vitis. vinifera. var. ruby seedless and v. vinifera. var. red globe) transferred with BADH gene were used as the experimental material. The result of transformation identified by antibiotic filtrating , GUS gene dyeing and PCR testing.Salt and alkali tolerance study of transgenic grapes were conducted through tissue culture , pot cuture and water culture. Biological characteristics of transgenic grapes were also studied. The main results were as follow: l.The micropropagation system of grape transfered with BADH gene was established. Through lots of experiments, a series of micropropagation medium were obtained,such as differentiation medium, bud elongating medium and rooting medium. The optimum bud differentiation was the medium with B5 + 6-BAO. 4mg. 1-1 , which induced many buds, high bud density and green leaf. The medium of B5 + KT3. 0mg. 1-1+NAAO. Img. 1-1 was the best bud-elongating medium, on which bud elongated obviously and had a protein growth. The medium of B5 + IAAO. 4mg. 1-1was the best seedling rooting medium, on which induced roots through 7 days , induced many roots and had a protein growth. The result of simplified rooting method indicated that IAA had good rooting effect. And roots had protein growth, lots ofsubordinated roots and high survival retio.2. The identification of transgenic grape.(l)The transgenic grape grew well on the differentiation medium containing 50mg. 1"' kan, but the control died on the same condition.(2)Using GUS gene dyeing identified that the transgenic grapes became blue, but the contol had no changing.(3) Using PCR cloning special gene examined the transgenic gropes (v. vinifera. var. ruby seedless and v. vinifera. var. red globe) that were filtrated by kan. Ruby seedless were examined 6 individual plants. The result indicated that 4 individual plants had the special strips about 1.5kb. Red globe were examined 5 individual plants. The result indicated that 4 individual plants had the special strips about 1.5kb. The identified plants were.examined again. The result indicated that 6 individual plants entirely had the special strips. Enlarging the 3' end of BADH gene in transgenic red globe grapes , the result indicated that 2 individual plants had the special strips about 300bp in 5 individual plants. One identified transgenic red globe grape was enlarged BADH gene and 3' end.The results obtained 2 special strips.3. The results of salt and alkali tolerance tests of transgenic grape were obtained by tissue culture,pot culture and water culture. The main results were as the following:(1)In tissue culture condition, transgenic grape delayed the time of beginning differntiation,decreased their differentiation rate and bud density, weakened their growth status and enhanced the death rate with salt content increased. At 5%o salt content, the transgenic grapes grew well, but the control grew well only 3% salt content. The transgenic grapes delayed the time of beginning rootage, decreased the average length daily changing of one grade roots and the average root number, weakened their growthstatus, enhanced the death rate with salt content increased. Throgh 40 days rooting culture, the average height of one grade seedlings and the average length of one grade roots entirly decreased, and chlorophyl content gradully decreased. Below 5% salt content,the transgenic grape grew well.(2)In tissue culture condition, grape transfered with BADH gene compared with the control that the time of beginning differentiation, bud density , death rate, rooting rate, the average length daily changing of one grade roots, the average of one grade seedlings height and biological characteristics had no obviously different. The transgenic grape had no obviously fastness to alkalinity. The diversified targets of transgenic grape had no obviously different in all alkalinity. The result indicated that the...
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