[논문]Analysis of intraspecific genetic diversity in Acidovorax citrulli causing bacterial fruit blotch on cucurbits in Korea

Song, Jeong Young; Oo, May Moe; Park, Su Yeon; Seo, Mun Won; Lee, Seong-Chan; Jeon, Nak Beom; Nam, Myeong Hyeon; Lee, Youn Su; Kim, Hong Gi; Oh, Sang-Keun

doi:10.7744/kjoas.20180086

Analysis of intraspecific genetic diversity in Acidovorax citrulli causing bacterial fruit blotch on cucurbits in Korea 원문보기

Korean journal of agricultural science, v.45 no.4, 2018년, pp.575 - 582

Song, Jeong Young (Department of Applied Biology, Chungnam National University) , Oo, May Moe (Department of Applied Biology, Chungnam National University) , Park, Su Yeon (Department of Applied Biology, Chungnam National University) , Seo, Mun Won (Department of Applied Biology, Chungnam National University) , Lee, Seong-Chan (Protected Horticulture Research Institute, National Institute of Horticultural and Herbal Science) , Jeon, Nak Beom (Fruit and Vegetable Research Center, Chungnam ARES) , Nam, Myeong Hyeon (Fruit and Vegetable Research Center, Chungnam ARES) , Lee, Youn Su (Division of Bio-resource Sciences, Kangwon National University) , Kim, Hong Gi (Department of Applied Biology, Chungnam National University) , Oh, Sang-Keun (Department of Applied Biology, Chungnam National University)

Abstract ▼ AI-Helper

Bacterial fruit blotch (BFB) caused by Acidovorax citrulli is a devastating disease found in many cucurbits cultivation fields. The genetic diversity for 29 strains of A. citrulli collected from various cucurbits in South Korea was determined by DNA fingerprinting with a pathogenicity test, multi locus analysis, Rep-PCR (repetitive sequence polymerase chain reaction), and URP (universal rice primers) PCR bands. Two distinct groups (Korean Clonal Complex, KCC1 and KCC2) in the population were identified based on group specific genetic variation in the multi locus phylogeny using six conserved loci and showed a very high similarity with DNA sequences for representative foreign groups [the group I (CC1-1 type) and the group II (CC2-5 type)] widely distributed worldwide, respectively. Additionally, in the case of phaC, a new genotype was found within each Korean group. The KCC1 was more heterogeneous compared to the KCC2. The KCC1 recovered mainly from melons and watermelons (ratio of 6 : 3) and 15 of the 20 KCC2 strains recovered from watermelons were dominant in the pathogen population. Accordingly, this study found that two distinct groups of differentiated A. citrulli exist in South Korea, genetically very similar to representative foreign groups, with a new genotype in each group resulting in their genetic diversity.

주제어

표/그림 (5)

표 Table 1. Host, geographic origin and multi-locus genotypes of Acidovorax citrulli strains isolated from cucurbit cultivation felds in Korea.
그림 Fig. 1. Maximum likelihood phylogeny of 29 Korean strains of Acidovorax citrulli with two foreign strains; A. citrulli CC1 and CC2 types (Feng et al., 2009) and two Acidovorax spp. as the outgroup. The tree was constructed from 1,823 bp using four genes: pilT, gltA, ugpB and phaC. Bootstrap values with 1,000 replicates for the major divisions of A. citrulli from the outgroups. C, cucumber; M, melon; W, watermelon; P, pumpkin.
표 Table 2. Comparison of DNA variation sites for six genes of Korean KCC1 and KCC2 groups with two foreign groups of Acidovorax citrulli.
그림 Fig. 2. Cluster analysis of Acidovorax citrulli strains and A. valerianella 12066 based on DNA fingerprint profiles generated by polymerase chain reaction (PCR) amplification of repetitive elements and universal rice primers (URP) primers. Distance matrix data was generated using Dice’s (1945) coefficient of similarity and the dendrogram was constructed using the unweighted pairwise group method with arithmetic mean (UPGMA) algorithm. C, cucumber; M, melon; W, watermelon; P, pumpkin.
그림 Supplementary Fig 1. Representative DNA fingerprinting by four universal rice primers-polymerase chain reaction (URP-PCR) and two repetitive elements amplification of Acidovorax citrulli strains. M; 100 bp plus DNA ladder, Lane 1; 11246, 2; 11247, 3; 11248, 4; 14194, 5; 14201, 6; 14202, 7; 12091, 8; 12158, 9; 14222, 10; 11147, 11; 11162, 12; 11163, 13; 11164, 14; 11165, 15; 11201, 16; 11251, 17; 11259, 18; 12089, 19; 12090, 20; 13034, 21; 13211, 22; 17005, 23; 17912, 24; 13255, 25; 11171, 26; 12170, 27; 13217, 28; 14236, 29; 17913 of A. citrulli and 30; A. valerianella 12066.

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제안 방법

For the multi-locus analysis, DNA sequences of six housekeeping genes; adk (437 bp), glyA (563 bp), gltA (487 bp), pilT (405 bp) obtained from our previous work (Song et al., 2015) and ugpB (452 bp) and phaC (479 bp) were selected and PCR products were performed using Primers of ugpB (ugpB1 & ugpB2) and phaC (phaCF & phaCR).
However, these studies couldn’t give enough information to understand genetic characteristics of Korean population. Therefore, the current study was tried to clarify the intraspecific genetic diversity within Korean A. citrulli population through multi-locus, repetitive-sequence-polymerase chain reaction (Rep-PCR) and universal rice primers (URP)-PCR analysis in order to obtain reliable data for developing resistance cultivars and the integrated control of bacterial fruit blotch in Korea.

이론/모형

Distance matrix data was generated using Dice’s (1945) coefficient of similarity and the dendrogram was constructed using the unweighted pairwise group method with arithmetic mean (UPGMA) algorithm.
RAPD bands were scored from the agarose gel and recorded as present (1) or absent (0) and assembled into a data matrix. The combined six results were analyzed with NTSYS (Exter Biological Software), and dendrograms were generated using the unweighted pair group method with average (UPGMA).

성능/효과

In this study, the allelic profile analysis using the base sequence of the genes including adk, gltA, glyA, pilT, ugpB except phaC presented the existence of species-specific and group-specific base mutation in 7 sites of KCC1 and KCC2 groups in 29 Korean A. citrulli strains with foreign isolates (Table 2). The 30th bases were cytosine (C), thymine (T) in adk; 439th, 442th, and 451th bases were C, guanine (G), adenine (A) and G, A, C in gltA; 452th were A and G in glyA; 94th were C and T in pilT; and 280th were G and C in ugpB in the strains of KCC1 and KCC2 groups, respectively (Table 2).

후속연구

citrulli existed in Korea, very similar to two foreign groups widely distributed worldwide, with a variety of genotypes in each group resulting in the genetic diversity and this is very meaningful. Upon further comparison analysis on the pathogenicity differences using various hosts of these strains and on the features of the responses against various control agents, it is considered to be the very useful data in the development of effective resistance cultivars and for the establishment of the disease control system against BFB due to A. citrulli.

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