Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles

Abstract. Junaid M, Purwantara A, Guest G. 2021. Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles. Biodiversitas 22: 2838-2843. Understanding the biology of the fastidious Basidiomycete Ceratobasidium theobromae occupying host-tissue is essential for plant dis...

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Autores principales:	Muhammad Junaid, David Guest, AGUS PURWANTARA
Formato:	article
Lenguaje:	EN
Publicado:	MBI & UNS Solo 2021
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/e666df53a0fc44e0b6a09719d7eff9b7
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spelling	oai:doaj.org-article:e666df53a0fc44e0b6a09719d7eff9b72021-11-22T12:16:55ZFungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles1412-033X2085-472210.13057/biodiv/d220734https://doaj.org/article/e666df53a0fc44e0b6a09719d7eff9b72021-06-01T00:00:00Zhttps://smujo.id/biodiv/article/view/8394https://doaj.org/toc/1412-033Xhttps://doaj.org/toc/2085-4722Abstract. Junaid M, Purwantara A, Guest G. 2021. Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles. Biodiversitas 22: 2838-2843. Understanding the biology of the fastidious Basidiomycete Ceratobasidium theobromae occupying host-tissue is essential for plant disease management. Direct pathogen DNA extraction from infected plant tissue avoids the need for isolation in artificial media. We report a modified DNA isolation protocol to obtain total plant DNA designed to overcome DNA extraction and isolation problems caused by infected petioles rich in polysaccharides and phenolic substances as a primary source of gummosis. This study examined and compared total plant DNA isolated from petioles high in polysaccharides and polyphenolic compounds using two methods: conventional CTAB lysis buffer and Kits (the standard method), and a new modified CTAB protocol to address these PCR inhibitors. The modified method resulted in higher quality and quantity of C. theobromae crude DNA and amplified PCR product. The modified method produced large quantities of clear, transparent, aqueous DNA-containing lysate (crude DNA) with a clear separation between the upper crude DNA and organic waste layers. Mean DNA absorbance was 1.80, and the lowest DNA yield was 836.6 ng/µl. With the standard method, the blurred, viscous lysate obtained showed signs of gummosis, with poor separation between layers of crude DNA, polysaccharides, protein, and organic waste layers and low yield. Gel electrophoresis indicated poor quality DNA extract. We conclude that this modified method will be valuable for genetic diversity and disease studies in a range of previously challenging plant tissues and their pathogens.Muhammad JunaidDavid GuestAGUS PURWANTARAMBI & UNS SoloarticleBiology (General)QH301-705.5ENBiodiversitas, Vol 22, Iss 7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Muhammad Junaid David Guest AGUS PURWANTARA Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles
description	Abstract. Junaid M, Purwantara A, Guest G. 2021. Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles. Biodiversitas 22: 2838-2843. Understanding the biology of the fastidious Basidiomycete Ceratobasidium theobromae occupying host-tissue is essential for plant disease management. Direct pathogen DNA extraction from infected plant tissue avoids the need for isolation in artificial media. We report a modified DNA isolation protocol to obtain total plant DNA designed to overcome DNA extraction and isolation problems caused by infected petioles rich in polysaccharides and phenolic substances as a primary source of gummosis. This study examined and compared total plant DNA isolated from petioles high in polysaccharides and polyphenolic compounds using two methods: conventional CTAB lysis buffer and Kits (the standard method), and a new modified CTAB protocol to address these PCR inhibitors. The modified method resulted in higher quality and quantity of C. theobromae crude DNA and amplified PCR product. The modified method produced large quantities of clear, transparent, aqueous DNA-containing lysate (crude DNA) with a clear separation between the upper crude DNA and organic waste layers. Mean DNA absorbance was 1.80, and the lowest DNA yield was 836.6 ng/µl. With the standard method, the blurred, viscous lysate obtained showed signs of gummosis, with poor separation between layers of crude DNA, polysaccharides, protein, and organic waste layers and low yield. Gel electrophoresis indicated poor quality DNA extract. We conclude that this modified method will be valuable for genetic diversity and disease studies in a range of previously challenging plant tissues and their pathogens.
format	article
author	Muhammad Junaid David Guest AGUS PURWANTARA
author_facet	Muhammad Junaid David Guest AGUS PURWANTARA
author_sort	Muhammad Junaid
title	Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles
title_short	Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles
title_full	Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles
title_fullStr	Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles
title_full_unstemmed	Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles
title_sort	fungal basidiomycete ceratobasidium theobromae dna obtained directly from cocoa petioles
publisher	MBI & UNS Solo
publishDate	2021
url	https://doaj.org/article/e666df53a0fc44e0b6a09719d7eff9b7
work_keys_str_mv	AT muhammadjunaid fungalbasidiomyceteceratobasidiumtheobromaednaobtaineddirectlyfromcocoapetioles AT davidguest fungalbasidiomyceteceratobasidiumtheobromaednaobtaineddirectlyfromcocoapetioles AT aguspurwantara fungalbasidiomyceteceratobasidiumtheobromaednaobtaineddirectlyfromcocoapetioles
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Fungal Basidiomycete Ceratobasidium theobromae DNA obtained directly from cocoa petioles

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