| In this vineyard Some Chinese Wild Vitis specis including Shangnan-24 (in whichresveratrol content is very high) and Baihe-35-1 (which is resistant to Powdery Mildew) andsome Vitis vinifera specis (the primary material is Fenghuang-51) were studied. By thetechnology of RT-PCR ,we explored some aspects in this direction of modern breeding, andsome results of our research are as follows: 1. By the new strategy for cloning disease resistance gene and the new efficienttechnology, we got three gene fragments from baihe-35-1 which is one resistant stain toPowdery Mildew in Chinese wild Vitis species, These three gene fragments are named 9-2,7#and15#,with the length of 200bp,565bp,211bp respectively.and the accession number ofthem in GeneBank are: AY656730, AY656725, AY656731. 9-2 gene fragment shows highhomlogous to those ESTs from abiotic stressed leaves of Vitis vinifera var. Another anylysismethod shows that it is homlogous to Vitis vinifera VvmybA1 gene for myb-relatedtranscription factor, this indicates this gene fragment may be a part of the gene of a transcriptfact, and it may play an important role in the resistance to disease .The homlogous anylysisindicated that 7# gene fragment may be a part of some gene which take part in the regulationof the fiber synthesis, which result in the induced resistance of structure. By this way, thisgene fragment shows high homlogous to those ESTs from abiotic stress expressed sequencetag database for abiotic stressed leaves of Vitis vinifera var. too.15# gene fragment has nohomlogous sequence in any bioinfomatic database in the internet, this indicated that it is anew gene, which may have some special function. 2. By the technology of RT-PCR, we cloned three members of stilbene synthase genefamily from the total RNA of Chinese wild Vitis species Shangnan-24, which can producemore Resveratrol in the same condition. with the length of772bp,720bp,283bp respectively,and the accession number of them in GeneBank are: The accession number of them inGeneBank are:AY656724, AY656725, AY656726. Besides these gene fragments, we gotanother four gene fragments by the same method, they are parts of the genes as follows:chalcone synthase gene, polyphenol oxidase gene, and phenylalanine ammonia-lyase gene. Inthese gene fragments, two fragments of chalcone synthase gene are gotten from leaves andfruit peel of shangnan-24 respectively, and the fragment of phenylalanine ammonia-lyasegene is special because it is amplified from DNA of shangnan-24.The length are 460bp,340bp,267bp respectively, and the accession number of them in GeneBank are:AY656727,AY656728, AY656729,AY656733。 3. The research of the mechanism of the different display in the different strain isinteresting. Now, we have gotten two stilbene synthase gene promoter similar sequences,considered that the fragment we have cloned and some results of RT-PCR, we have gottensome new solution about the mechanism of the different display in different strain. Forexample, the different activity of different members in gene family and SNP in different strainmay contribute a lot to the different display in different strain. 4.In the field of mechanism of disease resistance, we researched the activity of stilbenesynthase gene after the different time of inoculation of Powdery Mildew, whichcontributed to the research of cloning of resistance gene, and it helps us know more about themechanism of different strains. 6.A more efficient technology to isolate total RNA has been developed ,what's more, wedeveloped a new method to ascertain the period in which the disease resistance genetranscript, which is a basis to clone new disease resistance gene. How to clone diseaseresistance gene is a difficult problem in this research field. After many research in this field,we decided to use a new technology, which was proved to be very efficient. 7. In our research, we applied the gene engineering of secondary... |
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