Molecular Verification of the UK National Collection of Cultivated Liriope and Ophiopogon Plants
Abstract
:1. Introduction
2. Results
2.1. DNA Barcoding of the rbcLa Region of Liriope and Ophiopogon Accessions
2.2. DNA Barcoding of the rbcLb Region of Liriope and Ophiopogon
2.3. DNA Barcoding of the nrITS Region of Liriope and Ophiopogon
2.4. Deduction of Individual Taxon Identity
2.4.1. Liriope spp.
2.4.2. O. japonicus
2.4.3. O. bodinieri and O. planiscapus
3. Discussion
4. Materials and Methods
4.1. Plant Collections
4.2. Plant Material and Genomic DNA Extraction
4.3. PCR Protocols
- rbcLa PCR: initial denaturation step of 5 min at 95 °C followed by 35 cycles consisting of 30 s at 95 °C, 20 s at 52 °C, and 50 s at 72 °C, with a final extension period of 5 min at 72 °C.
- rbcLb PCR: initial denaturation step of 5 min at 94 °C followed by 35 cycles consisting of 30 s at 94 °C, 30 s at 56 °C, and 30 s at 72 °C, with a final extension period of 5 min at 72 °C.
- ITS PCR: initial denaturation step of 5 min at 95 °C followed by 30 cycles consisting of 30 s at 95 °C, 30 s at 61 °C, and 30 s at 72 °C, with a final extension period of 5 min at 72 °C.
- ITSP PCR: initial denaturation step of 5 min at 95 °C followed by 30 cycles consisting of 30 s at 95 °C, 30 s at 61 °C, and 30 s at 72 °C, with a final extension period of 5 min at 72 °C.
4.4. DNA Sequence Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
nrITS | nuclear ribosomal internal transcribed spacer |
rbcL | large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase gene |
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Primers | References | Sequences 5′–3′ | Annealing Temperature | Amplicon Size (bp) |
---|---|---|---|---|
rbcLaF | [40] | ATGTCACCACAAACAGAGACTAAAGC | 52 °C | ≃ 500/600 |
rbcLaR | [41] | GTAAAATCAAGTCCACCRCG | ||
rbcLbAF | [28] | CGGTGGACTTGATTTTACCA | 56 °C | ≃ 500 |
rbcLbBR | TCATCACGTAATAAATCAAC | |||
ITS1 | [42] | TCCGTAGGTGAACCTGCGG | 61 °C | ≃ 600 |
ITS4 | TCCTCCGCTTATTGATATGC | |||
ITS5P ITS8P | [37] | GGAAGGAGAAGTCGTAACAAGG CACGCTTCTCCAGACTACA | 61 °C | ≃ 600 |
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Masiero, E.; Banik, D.; Abson, J.; Greene, P.; Slater, A.; Sgamma, T. Molecular Verification of the UK National Collection of Cultivated Liriope and Ophiopogon Plants. Plants 2020, 9, 558. https://doi.org/10.3390/plants9050558
Masiero E, Banik D, Abson J, Greene P, Slater A, Sgamma T. Molecular Verification of the UK National Collection of Cultivated Liriope and Ophiopogon Plants. Plants. 2020; 9(5):558. https://doi.org/10.3390/plants9050558
Chicago/Turabian StyleMasiero, Eva, Dipanwita Banik, John Abson, Paul Greene, Adrian Slater, and Tiziana Sgamma. 2020. "Molecular Verification of the UK National Collection of Cultivated Liriope and Ophiopogon Plants" Plants 9, no. 5: 558. https://doi.org/10.3390/plants9050558