Isolation and Amplification of Single Chromosome in Hevea brasiliensis‘RY7-33-97’

doi:10.11924/j.issn.1000-6850.2013-2791

Abstract

Abstract: Hevea brasiliensis was one of the most important sources of natural rubber. The technique of single chromosome microdissection had important significance in genomic study. This experiment took tender leaves of Hevea brasiliensis‘RY-7-33-97’as material. The enzyme and low osmotic method was used to make the chromosom preparation. A technique for single chromosome microdissection by slender glass needles in Hevea brasiliensis was constructed. Eight chromosomes were successfully isolated, and two of them were amplified by linker adaptormediated PCR (LA-PCR) for two rounds, a smear of DNA fragments ranging from 400-1800 bp were generated. Southern blot hybridization results showed that the LA-PCR products were homogeneous with the Hevea brasiliensis genomic DNA, indicating that DNAs from the chromosome have been successfully amplified. This study demonstrated that chromosome microdissection and cloning can be applied in genomic study of Hevea brasiliensis. The results will facilitate for the construction of single chromosomal DNA library of Hevea brasiliensis.

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