Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae
The specific and sensitive nested-PCR method to detect Acidovorax citrulli, a causal agent of bacterial fruit blotch on cucurbitaceae, was developed. PCR primers were designed from the draft genome sequence which was obtained with the Next Generation Sequencing of A. citrulli KACC10651, and the nest...
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doaj-202e2eeecb91475eab3b10bfc23d68d82020-11-24T22:59:08ZengHanrimwon Publishing CompanyResearch in Plant Disease1598-22622233-91912015-06-012127481http://dx.doi.org/10.5423/RPD.2015.21.2.074Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at CucurbitaceaeYoung-Tak Kim0Kyoung-Soo Park1Hye-Seong Kim2Hyok-In Lee3Jae-Soon Cha4 Nongwoo Bio Co., LTD., Yeoju 469-885, Korea Nongwoo Bio Co., LTD., Yeoju 469-885, Korea Nongwoo Bio Co., LTD., Yeoju 469-885, KoreaAnimal and Plant Quarantine Agency, Anyang 430-757, Korea Chungbuk National University, Cheongju 361-763, KoreaThe specific and sensitive nested-PCR method to detect Acidovorax citrulli, a causal agent of bacterial fruit blotch on cucurbitaceae, was developed. PCR primers were designed from the draft genome sequence which was obtained with the Next Generation Sequencing of A. citrulli KACC10651, and the nested-PCR primer set (Ac-ORF 21F/Ac-ORF 21R) were selected by checking of specificity to A. citrulli with PCR assays. The selected nested-PCR primer amplified the 140 bp DNA only from A. citrulli strains, and detection sensitivity of the nested PCR increased 10,000 times of 1st PCR detection limit (10 ng genomic DNA/PCR). The nested PCR detected A. citrulli from the all samples of seed surface wash (external seed detection) of the artificially inoculated watermelon seeds with 101 cfu/ml and above population of A. citrulli while the nested PCR could not detected A. citrulli from the mashed seed suspension (internal seed detection) of the all artificially inoculated watermelon seeds. When the naturally infested watermelon seeds (10% seed infested rate with grow-out test) used, the nested PCR detected A. citrulli from 2 seed samples out of 10 replication samples externally and 5 seed samples out of 10 replication samples internally. We believe that the nested-PCR developed in this study will be useful method to detect A. citrulli from the Cucurbitaceae seeds.http://www.online-rpd.org/journal/view.html?uid=1441&sort=&scale=&key=year&keyword=&s_v=21&s_n=2&pn=vol&year=2015&vmd=FullAcidovorax citrulliBacterial fruit blotchCucurbitaceae seedsNested-PCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Young-Tak Kim Kyoung-Soo Park Hye-Seong Kim Hyok-In Lee Jae-Soon Cha |
spellingShingle |
Young-Tak Kim Kyoung-Soo Park Hye-Seong Kim Hyok-In Lee Jae-Soon Cha Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae Research in Plant Disease Acidovorax citrulli Bacterial fruit blotch Cucurbitaceae seeds Nested-PCR |
author_facet |
Young-Tak Kim Kyoung-Soo Park Hye-Seong Kim Hyok-In Lee Jae-Soon Cha |
author_sort |
Young-Tak Kim |
title |
Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae |
title_short |
Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae |
title_full |
Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae |
title_fullStr |
Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae |
title_full_unstemmed |
Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae |
title_sort |
development of nested-pcr assay to detect acidovorax citrulli, a causal agent of bacterial fruit blotch at cucurbitaceae |
publisher |
Hanrimwon Publishing Company |
series |
Research in Plant Disease |
issn |
1598-2262 2233-9191 |
publishDate |
2015-06-01 |
description |
The specific and sensitive nested-PCR method to detect Acidovorax citrulli, a causal agent of bacterial fruit blotch on cucurbitaceae, was developed. PCR primers were designed from the draft genome sequence which was obtained with the Next Generation Sequencing of A. citrulli KACC10651, and the nested-PCR primer set (Ac-ORF 21F/Ac-ORF 21R) were selected by checking of specificity to A. citrulli with PCR assays. The selected nested-PCR primer amplified the 140 bp DNA only from A. citrulli strains, and detection sensitivity of the nested PCR increased 10,000 times of 1st PCR detection limit (10 ng genomic DNA/PCR). The nested PCR detected A. citrulli from the all samples of seed surface wash (external seed detection) of the artificially inoculated watermelon seeds with 101 cfu/ml and above population of A. citrulli while the nested PCR could not detected A. citrulli from the mashed seed suspension (internal seed detection) of the all artificially inoculated watermelon seeds. When the naturally infested watermelon seeds (10% seed infested rate with grow-out test) used, the nested PCR detected A. citrulli from 2 seed samples out of 10 replication samples externally and 5 seed samples out of 10 replication samples internally. We believe that the nested-PCR developed in this study will be useful method to detect A. citrulli from the Cucurbitaceae seeds. |
topic |
Acidovorax citrulli Bacterial fruit blotch Cucurbitaceae seeds Nested-PCR |
url |
http://www.online-rpd.org/journal/view.html?uid=1441&sort=&scale=&key=year&keyword=&s_v=21&s_n=2&pn=vol&year=2015&vmd=Full |
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