Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks
Rice Research and Development Institute (RRDI) at Bathalagoda and its’ substations are solely conducting rice breeding in Sri Lanka. Recently, RRDI has identified five exportable rice varieties/lines. As rice export in Sri Lanka is undertaken by the private sector, the RRDI does not receive any fore...
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| Format: | Article |
| Language: | English |
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Faculty of Science, University of Peradeniya, Sri Lanka
2018-12-01
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| Series: | Ceylon Journal of Science |
| Subjects: | |
| Online Access: | https://cjs.sljol.info/articles/7548 |
| _version_ | 1811193686945431552 |
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| author | G.K.S. Ananda W.W.M.U.K. Wijesundara U.A.K.S. Udawela S.D.S.S. Sooriyapathirana |
| author_facet | G.K.S. Ananda W.W.M.U.K. Wijesundara U.A.K.S. Udawela S.D.S.S. Sooriyapathirana |
| author_sort | G.K.S. Ananda |
| collection | DOAJ |
| description | Rice Research and Development Institute (RRDI) at Bathalagoda and its’ substations are solely conducting rice breeding in Sri Lanka. Recently, RRDI has identified five exportable rice varieties/lines. As rice export in Sri Lanka is undertaken by the private sector, the RRDI does not receive any foreign revenues or credit for breeding rice genotypes. Therefore, RRDI currently is in need of a reliable protocol to claim the Plant Breeders’ Rights (PBR) of these exportable genotypes. The first step of establishing such a protocol based on DNA fingerprinting is depending on the ability to extract good quality DNA from rice caryopses. Therefore, the present study compared the suitability of commonly available DNA extraction methods to purify DNA from rice caryopsis. Three rice genotypes; Bg 250 and At 353 (varieties for local consumption) and Bw-Bs-1-2-31 (with export potential) were selected. Three parboiled rice samples each from long- and short-grain groups were also selected. The DNA extraction, quantification and purity assessment were carried out using four methods viz., CTAB, modified CTAB, Promega kit and Qiagen kit. The extracted DNA was subjected to simplex and multiplex PCR using four rice specific DNA markers. Although positive PCR bands were obtained for all four methods, the modified CTAB method yielded the significantly highest DNA yield (6196.67μg/ml) compared to other methods. Thus, RRDI can employ modified CTAB method in the protocol to claim its PBR and rice varietal ownerships. |
| first_indexed | 2024-04-12T00:13:45Z |
| format | Article |
| id | doaj.art-6246415393cd46f99204ca9a656174cb |
| institution | Directory Open Access Journal |
| issn | 2513-2814 2513-230X |
| language | English |
| last_indexed | 2024-04-12T00:13:45Z |
| publishDate | 2018-12-01 |
| publisher | Faculty of Science, University of Peradeniya, Sri Lanka |
| record_format | Article |
| series | Ceylon Journal of Science |
| spelling | doaj.art-6246415393cd46f99204ca9a656174cb2022-12-22T03:55:54ZengFaculty of Science, University of Peradeniya, Sri LankaCeylon Journal of Science2513-28142513-230X2018-12-0147431131710.4038/cjs.v47i4.75485726Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulksG.K.S. Ananda0W.W.M.U.K. Wijesundara1U.A.K.S. Udawela2S.D.S.S. Sooriyapathirana3University of PeradeniyaUniversity of PeradeniyaRice Research and Development InstituteUniversity of PeradeniyaRice Research and Development Institute (RRDI) at Bathalagoda and its’ substations are solely conducting rice breeding in Sri Lanka. Recently, RRDI has identified five exportable rice varieties/lines. As rice export in Sri Lanka is undertaken by the private sector, the RRDI does not receive any foreign revenues or credit for breeding rice genotypes. Therefore, RRDI currently is in need of a reliable protocol to claim the Plant Breeders’ Rights (PBR) of these exportable genotypes. The first step of establishing such a protocol based on DNA fingerprinting is depending on the ability to extract good quality DNA from rice caryopses. Therefore, the present study compared the suitability of commonly available DNA extraction methods to purify DNA from rice caryopsis. Three rice genotypes; Bg 250 and At 353 (varieties for local consumption) and Bw-Bs-1-2-31 (with export potential) were selected. Three parboiled rice samples each from long- and short-grain groups were also selected. The DNA extraction, quantification and purity assessment were carried out using four methods viz., CTAB, modified CTAB, Promega kit and Qiagen kit. The extracted DNA was subjected to simplex and multiplex PCR using four rice specific DNA markers. Although positive PCR bands were obtained for all four methods, the modified CTAB method yielded the significantly highest DNA yield (6196.67μg/ml) compared to other methods. Thus, RRDI can employ modified CTAB method in the protocol to claim its PBR and rice varietal ownerships.https://cjs.sljol.info/articles/7548plant breeders’ rights (pbr)rice varietal authenticitydna extraction from rice grainsmodified ctab method for rice grainsexporting rice in sri lanka |
| spellingShingle | G.K.S. Ananda W.W.M.U.K. Wijesundara U.A.K.S. Udawela S.D.S.S. Sooriyapathirana Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks Ceylon Journal of Science plant breeders’ rights (pbr) rice varietal authenticity dna extraction from rice grains modified ctab method for rice grains exporting rice in sri lanka |
| title | Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks |
| title_full | Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks |
| title_fullStr | Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks |
| title_full_unstemmed | Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks |
| title_short | Identification of a robust protocol for isolating PCR amenable DNA from caryopses to employ DNA fingerprinting to claim the plant breeders’ rights and varietal ownership of the exporting rice bulks |
| title_sort | identification of a robust protocol for isolating pcr amenable dna from caryopses to employ dna fingerprinting to claim the plant breeders rights and varietal ownership of the exporting rice bulks |
| topic | plant breeders’ rights (pbr) rice varietal authenticity dna extraction from rice grains modified ctab method for rice grains exporting rice in sri lanka |
| url | https://cjs.sljol.info/articles/7548 |
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