A reliable and inexpensive method of nucleic acid extraction for the PCR-based detection of diverse plant pathogens

被引：212

作者：

Li, R. ^{[1
]}

Mock, R. ^{[1
]}

Huang, Q. ^{[2
]}

Abad, J. ^{[3
]}

Hartung, J. ^{[4
]}

Kinard, G. ^{[1
]}

机构：

[1] USDA ARS, Natl Germplasm Resources Lab, Beltsville, MD 20705 USA

[2] USDA ARS, Floral & Nursery Plants Res Unit, Beltsville, MD 20705 USA

[3] USDA APHIS PPQ PHP, Plant Germplasm Quarantine Program, Beltsville, MD 20705 USA

[4] USDA ARS, Mol Plant Pathol Lab, Beltsville, MD 20705 USA

来源：

JOURNAL OF VIROLOGICAL METHODS | 2008年 / 154卷 / 1-2期

关键词：

CTAB; Extraction; Total nucleic acids; Plant pathogens; PCR-based detection;

D O I：

10.1016/j.jviromet.2008.09.008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A reliable extraction method is described for the preparation of total nucleic acids from at least ten plant genera for subsequent detection of plant pathogens by PCR-based techniques. The method combined a modified CTAB (cetyltrimethylammonium bromide) extraction protocol with a semi-automatic homogenizer (FastPrep (R) instrument) for rapid sample processing and low potential of cross contamination. The method was applied to sample preparation for PCR-based detection of 28 different RNA and DNA viruses, six viroids, two phytoplasmas and two bacterial pathogens from a range of infected host plants including sweet potato, small fruits and fruit trees. The procedure is cost-effective and the qualities of the nucleic acid preparations are comparable to those prepared by commonly used commercial kits. The efficiency of the procedure permits processing of numerous samples and the use of a single nucleic acid preparation for testing both RNA and DNA genomes by PCR, making this an appealing method for testing multiple pathogens in certification and quarantine programs. Published by Elsevier B.V.

引用

页码：48 / 55

页数：8

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