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Pak. J. Bot., 47(4): 1333-1340, 2015.

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ESTABLISHMENT OF DNA FINGERPRINTING IN CLONAL TEA IMPROVED CULTIVARS FROM YUNNAN OF CHINA USING ISSR MARKERS

B.Y. LIU*, C.M. ZHAO, X.M. SUN, Y.Y. LI, H.B. JIANG, Y.G. WANG, L. MA AND Z.F. DUAN

Abstract: In this study, DNA fingerprints were constructed by using ISSR markers for 20 clonal improved varieties developed by two breeding institutes in Yunnan province. Seven core ISSR primers were selected from 15 primers. A total of 110 bands were generated by PAGE with seven core primers, 93 of which were polymorphic bands, the percentage of polymorphic band (PPB) was 84.54%, and the mean value of polymorphism information content (PIC) reached 0.417; the genetic similarity coefficient of the cultivars was 0.574-0.854. The two primers, UBC835 and ISSR2, had high PIC values, and could be used to distinguish all cultivars, presenting the most efficient single primers. Among the all of primer combinations from the seven core primers, the three combinations, UBC835/UBC811, UBC835/ISSR2, and UBC835/ISSR3 showed lower similar coefficients, and more efficient in identifying the 20 improved varieties than the other primer combinations. Then these three primer combinations were further scored in 15 traditional cultivars. The results showed that UBC835/ISSR2 was the optimal primer combination, which could be used to distinguish each material among the 20 clonal improved varieties and 15 traditional cultivals. Finally, the DNA fingerprints of the 20 clonal improved varieties were constructed based on country and region code, breeding institute, core primer name and ISSR marker data. The established fingerprints could provide reliable scientific base for the protection of intellectual property right for these clonal improved varieties, and the important molecular information contained in these fingerprints would be useful for the authenticity identification and genetic relationship analysis of tea varieties.

Key words: DNA fingerprinting, Clonal tea improved cultivar, ISSR.

Tea Research Institute of Yunnan Academy of Agricultural Science, Menghai Yunnan 666201, P.R. China

Corresponding author’s e-mail:liusuntao@126.com