Molecular cloning and sequencing for coat protein genes of four important viruses infecting cucurbit and vegetable crops

• Main Authors: Yu-Ling Chen-Hsu, 陳許玉鈴
• Other Authors: Hui-Liang Wang
• Format: Others
• Language: zh-TW
• Published: 2001
• Online Access: http://ndltd.ncl.edu.tw/handle/14138777715378708874

碩士 === 國立高雄師範大學 === 生物科學研究所 === 89 === ZYMV, PRSV-W, TuMV-TW and BlCMV are members of the genus potyvirus and are the most important virueses infecting cucurbit, cruciferous and leguminaceous crops, respectively. In this study, the cDNAs of the 3'' —terminal regions of ZYMV, PRSV-W, TuMV-TW and BlCMV were synthesized by RT-PCR. The nucleotide and amino acid sequences of CP genes and 3'' non-translated regions of ZYMV, PRSV-W, TuMV-TW and BlCMV were analyzed and compared with other reported strains of each virus from different geographical areas. The results showed that the CP gene and 3'' non-translated region of ZYMV-7 had a highest identity with ZYMV-F, and a lowest identity with ZYMV-RU. According to the phylogenetic tree of ZYMV isolates reported in Taiwan showed that CP genes of TW-PT5, TW-TC1, TW-CY2, TW-TN3, TW-TNML1, ZYMV-7 and TW-NT1 share close relationships. No matter cleavage site were at E/S or Q/S between NIb and CP gene, the CP gene of PRSV-W had a higher identity with PRSV-S, and a lower identity with PRSV-India. The 3'' non-translated region of PRSV-W also had a higher identity with PRSV-S, but a lower identity with PRSV-HA. According to the phylogenetic tree of PRSV-W and other reported PRSV isolates in Taiwan showed that PRSV-W had a closer relationship with TW-CI. Comparison of BlCMV with other reported members of bean common mosaic virus subgroup, CP gene of BlCMV had a higher identity with BlCMV-F and CAbMV-TH, and a lower identity with PStV-B and PStV-ChinaG. The 3'' non-translated region of BlCMV has a higher identity with BlCMV-W, and a lower identity with PStV-B. Comparison of the CP gene and 3'' non-translated region of ZYMV, PRSV-W, TuMV-TW and BlCMV indicated that they were distinctly different potyviruses. cDNAs of CP gene of ZYMV, PRSV-W, TuMV-TW and BlCMV used for inserting into pET-21b and pBin-19 vectors were also cloned.