Construction Of Molecular Barcode In Crops Based On AFLP-taking The Cruciferae Vegetables In Zhejiang Province As An Example

There are many kinds of crops,and the cultivars are very rich,but with the development of agricultural technology,increasing the effectiveness of breeding,many genetic basis of crops become more and more narrow,identification of applied cultivars and traditional germplasm resources incarnates important Increasingly,on the other hand,more and more difficult.Therefore,looking for simple and effective identification methods will become important especially.Rapid development of Molecular markers theories and methods provides an effective technology,although because it only dependents on a few features to distinguish between tags,not a lot of groups or the individuals,so it is difficult to form a stable,available generally technical system.Here,we will combine the molecular biology and computer technology to make the business bar codes be applied to biometric identification.We proposed molecular barcode and trying to combine modern information technology and bio-technologies,providing a basis for the identification of the biological group and individuals,species identification,and protection of the new cultival rights.In view of Cruciferae vegetables in Zhejiang Province is the largest area of cultivation,the highest total output economic crops,but because of its huge number of cultivars,and new cultivars with narrow genetic diversity increasingly,which makes the identification of species on the morphology more and more difficult. Therefore,this study collected the Zhejiang Cruciferae vegetables main cultivars,using their rich diversity, to explore Construction of the concept of molecular bar code with AFLP marker,and study the construction, standards and feasibility of molecular bar code,On the basis of this technology,established a crop cultivars management platform.The main results are as follows:(1)We identified 10 Chinese cabbage-pak-cho cultivars by 19 morphological characters;identified 4 mustards cultivars by 12 morphological characters;identified 5 radish cultivars by 20 morphological characters.The results indicated that there was rich genetic diversity in Cruciferae cultivars in Zhejiang province.(2)By the research on AFLP system,the results showed that,in 20μL digestion system,the right DNA templet is 250 ng and the pre-amplification is 18 cycles:in 20μL amplification system,dNTPs at 4～8μmol·L^-1,Taq at 0.5～0.8U,Buffer at 2～2.5μL have no significant difference.Compared with two silver staining methods,showed that more diversity had been detected by quick Silver Staining than slow Silver Staining。By using AFLP technique then 251 bands were amplified by eight amplification primers(M5E2,M6E1,M3E2,M5E1,M6E2,M4E7,M1E2,M8E3).Tree dendrograms were constructed based on the AFLP marker data by system cluster analysis of WPGMA,75 cruciferae cultivars were identified except 'Aikanqing','Xinxuanlhao',The results indicated cruciferae vegetable have rich diversity on DNA level, on the side,the results showed that chinese cabbage-pe-tsai have far genetic relationship with turnip chinese cabbage-pak-choi.(3)We posed a molecular barcode concept,and discussed its standar(?)an feasibility.Obtained the molecular barcode of the 75 cruciferae based on PDF417,established a platform for crops management information.