Cara Preservasi Fitoplasma dari Jaringan Kacang Tanah Bergejala Sapu untuk Deteksi DNA dengan Teknik PCR

Main Authors: Pulogu, Siska Irhamnawati, Mutaqin, Kikin Hamzah, Giyanto, Giyanto
Format: Article info application/pdf Journal
Bahasa: ind
Terbitan: The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia) , 2017
Subjects:
Online Access: https://journal.ipb.ac.id/index.php/jfiti/article/view/17002
https://journal.ipb.ac.id/index.php/jfiti/article/view/17002/12342
ctrlnum article-17002
fullrecord <?xml version="1.0"?> <dc schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd"><title lang="id-ID">Cara Preservasi Fitoplasma dari Jaringan Kacang Tanah Bergejala Sapu untuk Deteksi DNA dengan Teknik PCR</title><creator>Pulogu, Siska Irhamnawati</creator><creator>Mutaqin, Kikin Hamzah</creator><creator>Giyanto, Giyanto</creator><subject lang="id-ID">nested-PCR</subject><subject lang="id-ID">preservation</subject><subject lang="id-ID">peanut witches&#x2019; broom</subject><subject lang="id-ID">standard PCR</subject><description lang="id-ID">Witches&#x2018; broom of peanut caused by phytoplasma is a common disease found in Indonesia. Phytoplasma is able to be detected using polymerase chain reaction (PCR) technique. One of important factor which determine the successful of phytoplasma amplification is the DNA availability from fresh tissues. The research was aimed to evaluate some preservation methods of phytoplasma from infected plant samples. The aspects to be evaluated consisted of time (1, 2, 3, and 4 weeks), temperature (-20 &#xB0;C, 4 &#xB0;C, and 25 &#xB0;C), and preservation medium (1X PGB buffer, 3 M NaCl, CTAB buffer, 70% ethanol, non medium, and FTA-card) for storing the fresh phytoplasma infected samples. Good preservation method will optimize the phytoplasma DNA amplification using PCR standard technique followed by nested-PCR. The results showed that preservation of samples at -20 &#xB0;C, 4 &#xB0;C, and 25 &#xB0;C in CTAB buffer was able to maintain the tissue freshness for 4 weeks and was able to provide the DNA of either quality or quantity sufficiently for PCR detection. PCR standard using a primer pair P1/P7 showed that not all of the preserved DNA of phytoplasma were amplified positively. However, standard PCR followed by nested-PCR using primer pair fU5/rU3 was able to increase the DNA detectability. Preserved samples derived from various medium and stored for 4 weeks gave positive results.&#xA0; This results were in contrary with previous same samples which were detected negatively by standard PCR technique.</description><publisher lang="en-US">The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)</publisher><date>2017-07-05</date><type>Journal:Article</type><type>Other:info:eu-repo/semantics/publishedVersion</type><type>File:application/pdf</type><identifier>https://journal.ipb.ac.id/index.php/jfiti/article/view/17002</identifier><identifier>10.14692/jfi.13.2.43&#x2013;50</identifier><source lang="id-ID">Jurnal Fitopatologi Indonesia; Vol 13 No 2 (2017); 43</source><source lang="en-US">Jurnal Fitopatologi Indonesia; Vol. 13 No. 2 (2017); 43</source><source>2339-2479</source><source>0215-7950</source><language>ind</language><relation>https://journal.ipb.ac.id/index.php/jfiti/article/view/17002/12342</relation><rights lang="en-US">Copyright (c) 2017 Jurnal Fitopatologi Indonesia</rights><recordID>article-17002</recordID></dc>
language ind
format Journal:Article
Journal
Other:info:eu-repo/semantics/publishedVersion
Other
File:application/pdf
File
Journal:Journal
author Pulogu, Siska Irhamnawati
Mutaqin, Kikin Hamzah
Giyanto, Giyanto
title Cara Preservasi Fitoplasma dari Jaringan Kacang Tanah Bergejala Sapu untuk Deteksi DNA dengan Teknik PCR
publisher The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)
publishDate 2017
topic nested-PCR
preservation
peanut witches’ broom
standard PCR
url https://journal.ipb.ac.id/index.php/jfiti/article/view/17002
https://journal.ipb.ac.id/index.php/jfiti/article/view/17002/12342
contents Witches‘ broom of peanut caused by phytoplasma is a common disease found in Indonesia. Phytoplasma is able to be detected using polymerase chain reaction (PCR) technique. One of important factor which determine the successful of phytoplasma amplification is the DNA availability from fresh tissues. The research was aimed to evaluate some preservation methods of phytoplasma from infected plant samples. The aspects to be evaluated consisted of time (1, 2, 3, and 4 weeks), temperature (-20 °C, 4 °C, and 25 °C), and preservation medium (1X PGB buffer, 3 M NaCl, CTAB buffer, 70% ethanol, non medium, and FTA-card) for storing the fresh phytoplasma infected samples. Good preservation method will optimize the phytoplasma DNA amplification using PCR standard technique followed by nested-PCR. The results showed that preservation of samples at -20 °C, 4 °C, and 25 °C in CTAB buffer was able to maintain the tissue freshness for 4 weeks and was able to provide the DNA of either quality or quantity sufficiently for PCR detection. PCR standard using a primer pair P1/P7 showed that not all of the preserved DNA of phytoplasma were amplified positively. However, standard PCR followed by nested-PCR using primer pair fU5/rU3 was able to increase the DNA detectability. Preserved samples derived from various medium and stored for 4 weeks gave positive results. This results were in contrary with previous same samples which were detected negatively by standard PCR technique.
id IOS13793.article-17002
institution Sinarmas Forestry Corporate R&D
institution_id 4603
institution_type library:special
library
library Abdul Gafur Library
library_id 3514
collection Jurnal Fitopatologi Indonesia
repository_id 13793
subject_area Plant Pathology
city Bogor
province JAWA BARAT
repoId IOS13793
first_indexed 2020-04-29T22:43:09Z
last_indexed 2020-12-23T01:24:19Z
recordtype dc
merged_child_boolean 1
_version_ 1800759159139860480
score 16.845257