Abstract
Rapid, easy and direct DNA extraction protocols are required for use in routine analyses to detect plant pathogenic bacteria. For this purpose, five protocols for extracting bacterial DNA, which do not use toxic chemicals such as phenol and chloroform, were optimized for the detection of plant pathogenic bacteria quickly and directly from plant material using conventional polymerase chain reaction (PCR). The protocols are based on the use of different maceration and extraction buffers to ensure quality bacterial DNA extraction and to remove the PCR inhibitors. Results showed that the protocol #1 based on the use of Tris and TNPEE buffers and protocol #3 based on the use of 0.4% bovine serum albumin and 0.05% Tween 20 were able to detect Erwinia amylovora from apple and pear samples. Protocol #2 that used 2% polyvinyl pyrrolidone in maceration and extraction buffers was found to be efficient for detecting Erwinia amylovora, Agrobacterium tumefaciens, and Allorhizobium vitis directly from plant tissues. However, protocols #4 and #5 based on the use of 1% Triton X-100 and 0.1% Tween 20, respectively, were unable to detect the studied bacteria. Accordingly, the protocol #2 is proposed for efficient, direct and rapid detection of plant pathogenic bacteria in plant materials via PCR.
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Claims 6
Claim · model-written summary
Protocol #1 (Tris and TNPEE buffers) detected Erwinia amylovora from apple and pear samples.
- Taxon
- apple and pear
Malus domestica_Poirier_Fleurs_et_feuilles.jpg?width=60)
Pyrus communise1 - Treatment
- protocol #1 based on the use of Tris and TNPEE bufferse0
- Interactor
- Erwinia amylovora
Erwinia amylovorae1pathogen - Outcome
- able to detect Erwinia amylovorae1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_Poirier_Fleurs_et_feuilles.jpg?width=500)
Verbatim
e0“protocol #1 based on the use of Tris and TNPEE buffers”
e1“were able to detect Erwinia amylovora from apple and pear samples”
Claim · model-written summary
Protocol #3 (bovine serum albumin and Tween 20 buffer) detected Erwinia amylovora from apple and pear samples.
- Taxon
- apple and pearMalus domesticaPyrus communise1
- Treatment
- protocol #3 based on the use of 0.4% bovine serum albumin and 0.05% Tween 20e2
- Interactor
- Erwinia amylovoraErwinia amylovorae1pathogen
- Outcome
- able to detect Erwinia amylovorae1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“were able to detect Erwinia amylovora from apple and pear samples”
e2“protocol #3 based on the use of 0.4% bovine serum albumin and 0.05% Tween 20”
Claim · model-written summary
Protocol #2 (2% polyvinyl pyrrolidone) was efficient for detecting Erwinia amylovora and Agrobacterium tumefaciens, and Allorhizobium vitis, directly from plant tissues.
- Taxon
- apple and pearMalus domesticaPyrus communise1
- Treatment
- Protocol #2 that used 2% polyvinyl pyrrolidone in maceration and extraction bufferse3
- Interactor
- Erwinia amylovoraErwinia amylovorae3pathogen
- Second interactor
- Agrobacterium tumefaciens
Agrobacterium tumefaciense3pathogen - Outcome
- efficient for detectinge3
- Qualifier
- Allorhizobium vitise3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“were able to detect Erwinia amylovora from apple and pear samples”
e3“Protocol #2 that used 2% polyvinyl pyrrolidone in maceration and extraction buffers was found to be efficient for detecting Erwinia amylovora, Agrobacterium tumefaciens, and Allorhizobium vitis directly from plant tissues”
Claim · model-written summary
Protocol #4 (1% Triton X-100) was unable to detect the studied bacteria from apple and pear samples.
- Taxon
- apple and pearMalus domesticaPyrus communise1
- Treatment
- 1% Triton X-100e4
- Outcome
- unable to detect the studied bacteriae4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“were able to detect Erwinia amylovora from apple and pear samples”
e4“protocols #4 and #5 based on the use of 1% Triton X-100 and 0.1% Tween 20, respectively, were unable to detect the studied bacteria”
Claim · model-written summary
Protocol #5 (0.1% Tween 20) was unable to detect the studied bacteria from apple and pear samples.
- Taxon
- apple and pearMalus domesticaPyrus communise1
- Treatment
- 0.1% Tween 20e4
- Outcome
- unable to detect the studied bacteriae4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“were able to detect Erwinia amylovora from apple and pear samples”
e4“protocols #4 and #5 based on the use of 1% Triton X-100 and 0.1% Tween 20, respectively, were unable to detect the studied bacteria”
Claim · model-written summary
Protocol #2 is proposed for efficient, direct and rapid detection of plant pathogenic bacteria in plant materials via PCR.
- Taxon
- apple and pearMalus domesticaPyrus communise1
- Outcome
- efficient, direct and rapid detection of plant pathogenic bacteria in plant materials via PCRe5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“were able to detect Erwinia amylovora from apple and pear samples”
e5“the protocol #2 is proposed for efficient, direct and rapid detection of plant pathogenic bacteria in plant materials via PCR”