Tsai Chi-Chu, Shih Huei-Chuan, Ko Ya-Zhu, Wang Ren-Huang, Li Shu-Ju, Chiang Yu-Chung
Kaohsiung District Agricultural Research and Extension Station, Pingtung 908, Taiwan.
Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
Int J Mol Sci. 2016 Sep 24;17(10):1630. doi: 10.3390/ijms17101630.
Papaya (Carica papaya L.) is an economically important tropical fruit tree with hermaphrodite, male and female sex types. Hermaphroditic plants are the major type used for papaya production because their fruits have more commercial advantages than those of female plants. Sex determination of the seedlings, or during the early growth stages, is very important for the papaya seedling industry. Thus far, the only method for determining the sex type of a papaya at the seedling stage has been DNA analysis. In this study, a molecular technique-based on DNA analysis-was developed for detecting male-hermaphrodite-specific markers to examine the papaya's sex type. This method is based on the loop-mediated isothermal amplification (LAMP) and does not require prior DNA purification. The results show that the method is an easy, efficient, and inexpensive way to determine a papaya's sex. This is the first report on the LAMP assay, using intact plant materials-without DNA purification-as samples for the analysis of sex determination of papaya. We found that using high-efficiency DNA polymerase was essential for successful DNA amplification, using trace intact plant material as a template DNA source.
番木瓜（番木瓜科番木瓜属）是一种具有重要经济价值的热带果树，有雌雄同株、雄株和雌株三种性别类型。雌雄同株植物是番木瓜生产中使用的主要类型，因为它们的果实比雌株的果实具有更多商业优势。在番木瓜幼苗产业中，确定幼苗或早期生长阶段的性别非常重要。到目前为止，在幼苗阶段确定番木瓜性别类型的唯一方法是DNA分析。在本研究中，开发了一种基于DNA分析的分子技术，用于检测雄性-雌雄同株特异性标记，以确定番木瓜的性别类型。该方法基于环介导等温扩增技术（LAMP），无需预先进行DNA纯化。结果表明，该方法是一种简便、高效且廉价的番木瓜性别鉴定方法。这是首次报道使用完整植物材料（无需DNA纯化）作为样本进行番木瓜性别鉴定分析的LAMP检测。我们发现，使用高效DNA聚合酶对于以微量完整植物材料作为模板DNA来源成功进行DNA扩增至关重要。
