Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia cadamba
Flow cytometry (FCM) analysis plays a crucial role in polyploidization studies. It allows for the rapid identification of induced polyploids or mixoploids and plantlets with unchanged ploidy levels compared to the conventional method of chromosome counting. Therefore, the present study aims to optim...
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my.unimas.ir.441732024-01-17T08:17:40Z http://ir.unimas.my/id/eprint/44173/ Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia cadamba Ho, Wei Seng Eng, Wee Hiang Ling, Kwong Hung Q Science (General) S Agriculture (General) SB Plant culture SD Forestry Flow cytometry (FCM) analysis plays a crucial role in polyploidization studies. It allows for the rapid identification of induced polyploids or mixoploids and plantlets with unchanged ploidy levels compared to the conventional method of chromosome counting. Therefore, the present study aims to optimize and develop an efficient FCM analysis procedure to determine the ploidy level and DNA content of Neolamarckia cadamba. This involved investigating various types of leaf tissues of N. cadamba and lysis buffers to identify the best tissue-buffer combination for FCM analysis. The histograms generated by FCM analysis revealed that only fresh leaves combined with LB01 buffer produced clean histograms with sharper peaks, reduced noise, and low coefficient of variation values. FCM analysis effectively classified the nodal explants of N. cadamba treated with colchicine into three distinct groups of polyploid plants: tetraploids, mixoploids, and octoploids. The N. cadamba tetraploids were found to have an estimated DNA content of 2.59 ± 0.09 pg, while octoploids showed an increase of DNA content to 5.35 ± 0.24 pg. These results highlight the effectiveness of FCM as a valuable tool in identifying mixoploids among the colchicine-induced polyploids of N. cadamba, as compared to the conventional method of chromosome counting. Mixoploids, which are characterized by cells with varying ploidy levels, deserve further investigation in future research. Open Science Publishers LLP 2024 Article PeerReviewed text en http://ir.unimas.my/id/eprint/44173/3/Optimizing.pdf Ho, Wei Seng and Eng, Wee Hiang and Ling, Kwong Hung (2024) Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia cadamba. Journal of Applied Biology & Biotechnology, 12 (1). pp. 93-97. ISSN 2347-212X https://jabonline.in/abstract.php?article_id=1060 DOI: 10.7324/JABB.2024.157607 |
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Q Science (General) S Agriculture (General) SB Plant culture SD Forestry Ho, Wei Seng Eng, Wee Hiang Ling, Kwong Hung Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia cadamba |
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Flow cytometry (FCM) analysis plays a crucial role in polyploidization studies. It allows for the rapid identification of induced polyploids or mixoploids and plantlets with unchanged ploidy levels compared to the conventional method of chromosome counting. Therefore, the present study aims to optimize and develop an efficient FCM analysis procedure to determine the ploidy level and DNA content of Neolamarckia cadamba. This involved investigating
various types of leaf tissues of N. cadamba and lysis buffers to identify the best tissue-buffer combination for FCM analysis. The histograms generated by FCM analysis revealed that only fresh leaves combined with LB01 buffer
produced clean histograms with sharper peaks, reduced noise, and low coefficient of variation values. FCM analysis
effectively classified the nodal explants of N. cadamba treated with colchicine into three distinct groups of polyploid plants: tetraploids, mixoploids, and octoploids. The N. cadamba tetraploids were found to have an estimated DNA content of 2.59 ± 0.09 pg, while octoploids showed an increase of DNA content to 5.35 ± 0.24 pg. These results
highlight the effectiveness of FCM as a valuable tool in identifying mixoploids among the colchicine-induced
polyploids of N. cadamba, as compared to the conventional method of chromosome counting. Mixoploids, which are
characterized by cells with varying ploidy levels, deserve further investigation in future research. |
format |
Article |
author |
Ho, Wei Seng Eng, Wee Hiang Ling, Kwong Hung |
author_facet |
Ho, Wei Seng Eng, Wee Hiang Ling, Kwong Hung |
author_sort |
Ho, Wei Seng |
title |
Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia
cadamba |
title_short |
Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia
cadamba |
title_full |
Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia
cadamba |
title_fullStr |
Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia
cadamba |
title_full_unstemmed |
Optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of Neolamarckia
cadamba |
title_sort |
optimizing flow cytometry analysis for determining ploidy level and genome size of colchicine-induced polyploids of neolamarckia
cadamba |
publisher |
Open Science Publishers LLP |
publishDate |
2024 |
url |
http://ir.unimas.my/id/eprint/44173/3/Optimizing.pdf http://ir.unimas.my/id/eprint/44173/ https://jabonline.in/abstract.php?article_id=1060 |
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