Summary Using a base-specific staining method, three Japanese cultivars of Momordica charantia were investigated. The present method using enzymatic digestion produced a high level of well-spread metaphase chromosome complements. The cultivars investigated showed the same somatic chromosome number 2n=22. In condensation behavior from prophase to metaphase, most chromosomes of all cultivars in this study had early condensing segments at the proximal regions. Moreover, at prophase or earlier stage of metaphase, most chromosomes observed had decondenced segments at the distal regions in both arms. Judging by the determination of centromere positions, the chromosome complements at mitotic metaphases in three cultivars consisted of 22 metacentric chromosomes. Moreover, similar sizes and shapes of the chromosomes indicated that the karyotypes of M. charantia cultivars were symmetrical. In contrast, fluorescent staining showed different sizes and numbers of chromycin A 3 (CMA) positive and 4′,6-diamidino-2-phenylindole (DAPI) negative (CMA + DAPI ) satellites among cultivars. Four CMA + DAPI satellites were observed in M. charantia Abashi-goya and M. charantia Naga-goya, while two satellites were obseved in M. charantia Shiro-goya. All satellites were located at one end of sat-chromosomes. In Shiro-goya, a large size difference of satellites was found between two satchromosomes, which might be homologous chromosomes with polymorphism. Moreover, faintly CMA + DAPI sites were shown at the proximal regions or primary constrictions of most chromosomes in all cultivars.
Abstract:Chalcone synthase (CHS) is a key enzyme in the flavonoid biosynthesis pathway. CHS genes were cloned from genomic DNA and cDNA from the petals of 'Buntharik' white lotus and 'Sattabangkacha' pink lotus by the PCR technique using a specific primer of the CHS gene designed from the GenBank database. Semi-quantitative RT-PCR analysis revealed that the highest CHS gene expression was found in the early budding stage of the pink lotus and was reduced in later stages. Shoot tips from embryos of Buntharik and Rachinee lotus were used to induce shoot clusters by cultivation on a MS medium supplemented with 40 µM NAA and 0.5 µM TDZ for 8 weeks and a MS medium supplemented with 50 µM BA for 8 weeks. An antisense CHS gene (450 bp) from the cDNA of Buntharik lotus was used to construct a plant transformation vector; pCAMBIA1302CHSA. The vector construct was transformed into Buntharik and Rachinee shoot clusters by particle bombardment. After transformant selection and regeneration, two transformants of Buntharik shoot clusters showed GFP green spots and existence of the GFP gene and hptII gene in the genomic DNA amplified by the PCR technique. In the Rachinee transformants, 3 of 5 showed the GFP green spots and the GFP and hptII genes were identified in amplification by PCR. After CHS gene expression analyses by semi-quantitative RT-PCR, two transformed Rachinee shoot clusters had a reduction in CHS gene expression.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.