DNA分子標誌在番木瓜性型鑑定、親緣關係及品種鑑定之研究

Study on identification of sexual types, phylogenetic relationships and cultivar identification in papaya using DNA molecular markers

柯吟玫(Yin-Mei Ko)

指導教授 ： 張龍生

共同指導教授 ： 洪挺軒 ； 楊雯如(Wen-Ju Yang)

國立臺灣大學/生物資源暨農學院/園藝學研究所/碩士(2005年)

https://doi.org/10.6342/NTU.2005.10306

摘要

無資料

關鍵字

DNA分子標誌 ； 番木瓜 ； 性型鑑定 ； 親緣關係 ； 品種鑑定

並列摘要

Hermaphroditic plants are desired for commercial papaya production in most countries. However, the sex type of papaya plant is difficult to be identified during the seedling stage. To obtain the hermaphroditic plants, papayas used to be cultivated by simultaneously planting 2-3 seedlings in a pit. This cultivated method is not efficient and economic. Thus, the development of a rapid technique for sex identification of papaya and the genetic study of sex determination in papaya are very important to papaya industry. DNA-based markers have been recently applied in sex identification of several plants. This study attempted to use the published papaya sex-specific SCAR primers to identify 39 papaya cultivars or lines in Taiwan. The results showed that SCAR W11 and SCAR T12 existed in all hermaphrodite and male plants, but not in the female plants. The DNA fragments corresponding to W11 SCAR of ‘Sunrise’, ‘TSS No.7’, ‘Thailand’, ‘Florida’, NTU005 and NTU007 were cloned and sequenced. Totally 832 nucleotide sequences from the hermaphrodite and male W11 SCAR were determined, and their sequences were similar. The DNA fragments corresponding to T12 SCAR of ‘Sunrise’, ‘TSS No.7’, ‘Florida’, NTU005 were also cloned and sequenced. Totally 838 nucleotide sequences from the hermaphrodite and male T12 SCAR were determined, and their sequences were similar. Based on the different sequences between hermaphrodite and male T12 SCAR, the primer pair T12-165F and T12-834R was designed to amply PCR products only in male plants of ‘Florida’, II-3-4’91 and VII-7-7’91, but not in hermaphrodite and female plants of ‘Sunrise’ and ‘Thailand’. Nonetheless, it needs strict PCR conditons to make the male-specific band appear steadily and needs more other papaya cultivar samples to certify if it was really a male-specific marker in general. SCAR marker is more reliable and reproducible and is a suitable marker for the precise and rapid identification of sex in papaya.
Estimation the genetic relationships among papaya cultigens of Taiwan and other regions and development of a cultivar identification tool based on random amplified polymorphic DNA (RAPD) were established in this study. Twenty-nine selected primers generated a total of 122 reproducible polymorphic amplification fragments among 44 papaya cultigens and other Vasconcellea species of Caricaceae. The average genetic similarity coefficients of 39 papaya cultigens was 0.662 and ranged from 0.322 for a breeding line of NTU selection and ‘Kapoho’ to 0.942 for K1 and K5. The UPGMA and principal component analysis showed a clear distinction into two clusters reflecting the different geographical origins, Taiwan and other regions. The 30 papaya cultigens of Taiwan clustered into two groups that reflected their origins from different breeding or selection programs. A combination of four primers with five markers was found to be optimum for the discrimination of the 11 papaya cultigens of Taiwan. The marker system can discriminate GMO papaya from other papaya sources of Taiwan. This study showed that RAPD could readily dissect genetic differences between the closely related papaya cultivars or lines and provide a simple, quick and economic method to differentiate the difference among papaya population of Taiwan.

並列關鍵字

sex identification ； phylogenetic relationships ； cultivar identification ； DNA molecular marker ； Carica papaya

參考文獻

陳述、范明仁. 1999. 淺談AFLP分子標幟-種原遺傳歧異分析與品種鑑定的新寵兒. 台灣省農業試驗所技術服務 37: 21-25.

張明聰、呂俊堅. 2000. 番木瓜組織培養兩性株嫁接苗繁殖、生長特性及效益評估. 台南區農業改良場研究彙報第37號：40-55.

楊藹華、林怡廷. 2001. 如何利用分子生物技術鑑定作物品種. 台南區農業專訊 38: 10-14.

翁儷倩. 2001. 桶柑遺傳歧異性之分析. 國立台灣大學園藝學研究所碩士論文.

Ainsworth, C., J. Parker, and V. Buchanan-Wollaston. 1998. Sex determination in plants. Curr. Top. Dev. Biol. 38: 168-223.