Abstract:

【Objective】 It is of great importance to construct a shareable high-quality crop variety standard DNA fingerprint database for effectively managing the huge number of known varieties.【Method】 Based on fluorescence capillary electrophoresis detection platforms and the plant variety DNA fingerprint database management system, a database containing 3 998 maize authorized accessions was built with 40 SSR primer pairs. Multi-laboratories and multi-detecting platforms were used to conduct the quality evaluation of the database.【Result】 Allele frequency distribution graphs of the 40 corn primers were plotted as characteristic spectrums of each primer, which played the role of the similarity of reference samples in the database construction. A decuplet fluorescent capillary electrophoresis combination was formed and a set of system default PANEL was established in the SSR analyzer. Statistics were conducted on the experimental conditions of the database construction. Of the total samples, 61% of them were subjected to two group independent trials and 33% of them were subjected to three group independent trials. Each sample had 2-5 sets of original experimental data and the corresponding fingerprint maps. The accumulated loss and variable sites of the final built standard fingerprint database accounted for only 0.2%, the data integrity reached 99.8%. The assessment results in different laboratories and different electrophoresis platforms showed that the SSR fingerprint data obtained high agreement on the same electrophoresis fluorescence detection platform, but showed a certain bias in different electrophoretic detection platforms. In order to realize the sharing of SSR fingerprint data in different laboratories, a unified fluorescent primer, analysis software and electrophoresis detection platform were needed. Overall pairwise comparisons were conducted on all the fingerprint data, the results showed that there existed a relative big overall difference among the certification varieties of corn in China. Percentage of different sites among the authorized varieties was mainly concentrated between 80% and 95% (accounted for 78.28%), the percentage of different sites with more than 50% reached 99.21%, and less than 20% of only 0.09%. The average hybrid rate of the authorized varieties reached 64% and mainly concentrated between 50% and 80% (accounted for 89%). By using the corn variety standard fingerprint matching service platform (URL: http: //www.maizedna.org/), a shared fingerprint database is realized. 【Conclusion】 A standard procedure in constructing crop variety SSR fingerprint database was formed in this study and the SSR fingerprint database was constructed with a scale of nearly 4 000 corn authorized varieties. Through joint multi-laboratory comparison tests, the accuracy of database building and the sharing property of database were ensured. A service platform website for corn variety standard fingerprint matching was established in this study, thus achieving sharing of corn authorized variety fingerprint database in national seed identification system, and providing an important reference for other crop species in building high-quality SSR fingerprint database.

Key words: maize, variety identification, DNA fingerprint database, SSR