| Apricot department rosaceae apricot genus, Apricot is one own fruit trees of China, it distributing east-north of our country. Because of poignant chance of the temperature in early spring, the bud, flower and even young fruit will be hurt by the low temperature, and the production of the apricot fruit will be lower.So it is very important to breeding frost-resistant strong varieties .Last ten years it has made rapid development in transgenic technology.It has been widely used in fruit breeding at home and abroad. Transgenic has become one of the most promising methods to improve fruit varieties.Now to import the DNA by pollen tube it has been successfully used in ccrops, but rarely in fruit, which is reason to the long breeding cycle in fruit.So it is very meaningful to find a effective methods in breeding. My study use Daiyu apricot transfer the cold hardiness gene (AmEBP1) from Sand Holly by pollen tube and then determin the physiological indexes of the seedling cultivation. We have gain results followed.1.After Haidai jade apricot♀cross Kate♂and transferred with outer DNA, fruit-bearing rate reached 57%, horizontal diameter,vertical diameter and side diameter got 4.081 cm, can reach 4.403 cm, 4.412 cm,which are higher than Tarzan haidai jade apricot in the natural pollination., There heterosis performance appears.We gained the highest rate of conversion of DNA by cutting the 1/2 tube of flower,But there were certain influence to fruit-bearing rate.So take the method of cutting the 1/2 instead. Fruit-bearing rate by pollen gene treatment were less down 9.5% than normal pollen fruit-bearing rate;2.Using different concentrations of DNA conversion rate have certain effect, when DNA concentration for 200μg/mL respectively/mL, 500μg/mL, 800μg/mL, fruit-bearing rate for targeting were 21.5%,26.4%, 12.7%. Fruit-bearing rate was not as exogenous DNA concentration levels and gradually raise or lower, but the most with 500μg/mL and cutting the 1/2 tube of flower.which can achieve 75% in the first seven days, 30% 60 days later. But there were still that gene lost trend still exists after gene transgenic. 3.Take the modified WPM media for the different embryos period, The results show that it can form callus only when PF>0.3,and the callus induction get the most by PF=0.7 and 6-BA=0.6 mg/ml. Mature embryos can budding on the the modified WPM and MS medias.But there appears little leaves, curly, deformity, or vitrification, stressing the situation. poor growth situations.4. The results show that low processing time and the germination of 4 species apricot was positively correlated. With the extension of treatment time, signs of germination about the 4 varieties of apricot are more obvious, seedling rate has increase. In dealing with Daguo apricot 60 days, the field emergence almost can reach 100%,which is the the most vulnerable to seedling specie.Taishan Daiyu has a high rate of embryo rot either under 4℃or 0℃,and it has the low rate of seedling. That shows embryo of Taishan Daiyu is dysplasia and perishable.5. We get some transgenic seedlings of Taishan Daiyu apricot in this study ,16 tested positive for PCR.And Southern hybridization with digoingenin labeled DNA probe results showed 11 tissue culture had 1-3 respectively.It showed that exogenous AmEBPI gene were successfully transformed into T0 tissue culture of Taishan Daiyu.6. By the cold resistance contrast detection between transgenic plants and non-transgenic plants,it showed that LT50 of the transgenic plants was -4.37℃ 0.05), which did not cause disease to animals. |
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