Direct LAMP Assay without Prior DNA Purification for Sex Determination of Papaya.
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.
Kaohsiung District Agricultural Research and Extension Station, Pingtung 908, Taiwan. tsaicc@mail.kdais.gov.tw.|Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung 912, Taiwan. tsaicc@mail.kdais.gov.tw.
Chi-Chu
CC
Tsai
Department of Nursing, Meiho University, Pingtung 912, Taiwan. x00002213@meiho.edu.tw.
Huei-Chuan
HC
Shih
Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan. abscl77512@hotmail.com.
Ya-Zhu
YZ
Ko
Kaohsiung District Agricultural Research and Extension Station, Pingtung 908, Taiwan. rhwang@mail.kdais.gov.tw.
Ren-Huang
RH
Wang
Kaohsiung District Agricultural Research and Extension Station, Pingtung 908, Taiwan. gigi7366kimo@yahoo.com.tw.
Shu-Ju
SJ
Li
Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan. yuchung@mail.nsysu.edu.tw.|Department of Biomedical Science and Environment Biology, Kaohsiung Medical University, Kaohsiung 807, Taiwan. yuchung@mail.nsysu.edu.tw.
Yu-Chung
YC
Chiang
D029441:Carica / Q000235:genetics*; D032461:Chromosomes, Plant; D018744:DNA, Plant / Q000032:analysis* / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques; D036226:Seedlings / Q000235:genetics; D017422:Sequence Analysis, DNA; D012732:Sex Determination Analysis / Q000379:methods*
D018744:DNA, Plant
loop-mediated isothermal amplification (LAMP); papaya; sex determination; without DNA purification
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