Application of PCR-RFLP method of chloroplast genome for haplotype evaluation of some tea genotypes in Iran.

Document Type : Research

Authors

1 Tea Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Lahijan, Iran

2 Tea Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization, Lahijan, Iran

Abstract

The PCR-RFLP method was used for the first time in Iran to evaluate and determine haplotype relationships and population structure in 14 tea genotypes from three countries: Iran, India (Assam) and Japan. Three universal chloroplast primers (HK, CS and B1B2) and four restriction enzymes (HinfI, AluI, PstI and BglI) were used in this study and agarose gel was used to separate the bands. The B1B2 primer could not be used in this study due to the production of several bands. Also, the DT primer produced a monomorphic state after cutting, and only the fragments obtained from the HK primer and HinfI enzyme showed a polymorphic state. The mutations identified by the primer-cutting enzyme HK-HinfI were deletion-addition mutations in the range of 20-50 bp, which grouped the samples into five haplotypes (H1, H2, H3, H4 and H5). The distribution of samples in haplotype groups did not correspond to the geographical distribution, and there was no specific haplotype of a sample series. According to the results, it can be said that the PCR-RFLP method is a suitable method for investigating the genetic diversity of tea genotypes at the organ level.

Keywords