Establishment and Optimization of DNA Barcoding in Endangered Orchid Plants

PAN Yingwen; ZHANG Ling; WANG Anshi; LI Jiatong; XIE Tianwei; CHEN Shiming; LIU Fuxiu; LIN Mingguang

doi:10.15886/j.cnki.rdswxb.2019.04.003

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Universal primers were designed for ITS, matK and psbA-trnH sequences of endangered orchids for DNA barcoding and their PCR reaction system optimized. Sixteen species of endangered orchids were amplified by using PCR and sequenced, and their PCR amplification and sequencing efficiencies were compared. The results showed that the primers for the ITS, matK and psbA-trnH sequences had higher amplification success rate and sequencing success rate for species of the endangered orchid plants. The optimum annealing temperatures for the PCR reaction were 54 ℃ for ITS and 50 ℃ for matK and psbA-trnH. The targeted bands obtained under the above reaction systems and conditions were clear and bright, and the ITS, matK and psbA-trnH sequences can be used as a combination of DNA barcodes to identify the species of endangered orchid plants.

Establishment and Optimization of DNA Barcoding in Endangered Orchid Plants

Post-Entry Quarantine Station for Tropical Plants, Haikou Customs, Haikou, Hainan 570311, China

Received Date: 2018-12-05

Rev Recd Date: 2019-07-28

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Abstract: Universal primers were designed for ITS, matK and psbA-trnH sequences of endangered orchids for DNA barcoding and their PCR reaction system optimized. Sixteen species of endangered orchids were amplified by using PCR and sequenced, and their PCR amplification and sequencing efficiencies were compared. The results showed that the primers for the ITS, matK and psbA-trnH sequences had higher amplification success rate and sequencing success rate for species of the endangered orchid plants. The optimum annealing temperatures for the PCR reaction were 54 ℃ for ITS and 50 ℃ for matK and psbA-trnH. The targeted bands obtained under the above reaction systems and conditions were clear and bright, and the ITS, matK and psbA-trnH sequences can be used as a combination of DNA barcodes to identify the species of endangered orchid plants.

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Establishment and Optimization of DNA Barcoding in Endangered Orchid Plants

doi: 10.15886/j.cnki.rdswxb.2019.04.003

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通讯作者: 陈斌, bchen63@163.com

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