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This project includes three research parts including the establishment of the mother-stock maintenance technology, purity testing technology for quality management of hybrid seed production, as well as the pathogen detection technology for seed export quarantine. The first part is the establishment of mother-stock gardens of grape, pineapple,and dragon fruit. In the study, growing situation investigation and renew plant of two varieties of pineapple were completed. And specific pathogen of pineapples, grapes and dragon fruit in mother-stock gardens were tested. We established techniques of dragon fruit preservation in tissue culture and mother-stock garden. It could support healthy seedling product industrialization in Taiwan. The second part is the improvement of the molecular testing method for hybrid seed purity. The objective of this study was to develop the specific and co-dominant markers for identification of hybrid seed purity in bitter gourd. A key set of two SNP (AS) markers and another key set of five SSR markers were selected for hybrid identification. The results showed that the kits of SNP marker and SSR marker could be applied to detect the hybrid seed purity of 13 and 16 varieties respectively in bitter gourd and would be a useful tool for early testing the genetic purity in commercial hybrid seed production. The final part is the establishment of inspection system on international important seed-borne diseases. Bacterial fruit blotch (BFB) is one of the important seed-borne diseases on cucurbits. In this study,the PMA-qPCR technique was used to detect viable cells and to avoid the interference from dead cells. The results showed that, in the tests of mixtures of viable and dead cells, PMA-qPCR assay effectively detected live cells and PMA inhibited DNA amplification from dead cells. PMA really made the detection more accurately.
▲Fig1. SNP genotypes of bitter gourd F1 hybrids from both parents by using
allele-specific markers developed by TSIPS. The amplified target products of alleles
of SNP( T/ C) was 548 base pairs.
▲Fig2. The SOP for detecting seed-borne Acit on cucurbit seeds established by TSIPs.