Genetic Diversity and Population Structure of the Xanthomonas campestris pv. campestris Strains Affecting Cabbages in China Revealed by MLST and Rep-PCR Based Genotyping

다국어 초록 (Multilingual Abstract)

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot for cruciferous vegetables worldwide, especially for the cole crops such as cabbage and cauliflower. Due to the lack of resistant cabbage cultivars, black rot has brought about considerable yield losses in recent years in China. Understanding of the pathogen features is a key step for disease prevention, however, the pathogen diversity, population structure, and virulence are largely unknown. In this study, we studied 50 Xcc strains including 39 Xcc isolates collected from cabbage in 20 regions across China, us- ing multilocus sequence genotyping (MLST), repetitive DNA sequence-based PCR (rep-PCR), and pathogenicity tests. For MLST analysis, a total of 12 allelic profiles (AP) were generated, among which the largest AP was AP1 containing 32 strains. Further cluster analysis of rep-PCR divided all strains into 14 DNA groups, with the largest group DNA I comprising of 34 strains, most of which also belonged to AP1. Inoculation tests showed that the representative Xcc strains collected from diverse regions performed differential virulence against three brassica hosts compared with races 1 and 4. Interestingly, these results indicated that AP1/DNA I was not only the main pathotype in China, but also a novel group that differed from the previously reported type races in both genotype and virulence. To our knowledge, this is the first extensive genetic diversity survey for Xcc strains in China, which provides evidence for cabbage resistance breeding and opens the gate for further cabbage-Xcc interaction studies.

목차 (Table of Contents)

• Materials and Methods Results Discussion

참고문헌 (Reference)

1 Achtman, M., "Yersinia pestis, the cause of plague, is a recently emerged clone of Yersinia pseudotuberculosis" 96 : 14043-14048, 1999

2 Zhang, Y., "The occurrence and prevention of bacterial black rot of cruciferous vegetables by Dr. Li Baoju" 17 : 23-25, 2011

3 Lu, Y., "Study on pathotypes of black rot of chinese cabbage in Shaanxi Province" 10 : 132-138, 2008

4 Louws, F. J., "Specific genomic fingerprints of phytopathogenic Xanthomonas and Pseudomonas pathovars and strains generated with repetitive sequences and PCR" 60 : 2286-2295, 1994

5 Muhire, B., "SDT: a virus classification tool based on pairwise sequence alignment and identity calculation" 9 : e108277-, 2014

6 Gao, X., "Rep-PCR analysis of genotypic diversity of Pseudomonas syringae pv. actinidiae strains from Shaanxi province of China" 33 : 340-349, 2016

7 Ah-You, N., "Polyphasic characterization of xanthomonads pathogenic to members of the Anacardiaceae and their relatedness to species of Xanthomonas" 59 : 306-318, 2009

8 Gonçalves, E. R., "Phylogenetic analysis of Xanthomonas species based upon 16S-23S rDNA intergenic spacer sequences" 52 : 355-361, 2002

9 Fargier, E., "Pathogenicity assays restrict the species Xanthomonas campestris into three pathovars and reveal nine races within X. campestris pv. campestris" 56 : 805-818, 2007

10 Rathaur, P. S., "Pathogenic and genetic characterization of Xanthomonas campestris pv. campestris races based on rep-PCR and multilocus sequence analysis" 6 : 317-, 2015

11 Jensen, B. D., "Occurrence and diversity of Xanthomonas campestris pv. campestris in vegetable Brassica fields in Nepal" 94 : 298-305, 2010

12 Bella, P., "Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution" 68 : 278-287, 2019

13 Maiden, M. C. J., "Multilocus sequence typing : a portable approach to the identification of clones within populations of pathogenic microorganisms" 95 : 3140-3145, 1998

14 Urwin, R., "Multi-locus sequence typing : a tool for global epidemiology" 11 : 479-487, 2003

15 Jiang, H., "Microbial diversity in water and sediment of Lake Chaka, an Athalassohaline Lake in Northwestern China" 72 : 3832-3845, 2006

16 Tamura, K., "MEGA6: molecular evolutionary genetics analysis version 6.0" 30 : 2725-2729, 2013

17 Vicente, J. G., "Identification and origin of Xanthomonas campestris pv. campestris races and related pathovars" 91 : 492-499, 2001

18 Ntambo, M. S., "Identification and characterization of Xanthomonas albilineans causing sugarcane leaf scald in China using multilocus sequence analysis" 68 : 269-277, 2019

19 Kong, C., "Germplasm screening and inheritance analysis of resistance to cabbage black rot in a worldwide collection of cabbage (Brassica oleracea var. capitata) resources" 288 : 110234-, 2021

20 Versalovic, J., "Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction" 5 : 25-40, 1994

21 Popović, T., "Genetic diversity and virulence of Xanthomonas campestris pv. campestris isolates from Brassica napus and six Brassica oleracea crops in Serbia" 68 : 1448-1457, 2019

22 Librado, P., "DnaSP v5 : a software for comprehensive analysis of DNA polymorphism data" 11 : 1451-1452, 2009

23 Versalovic, J., "Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes" 19 : 6823-6831, 1991

24 Lema, M., "Discrimination of Xanthomonas campestris pv. campestris races among strains from northwestern Spain by Brassica spp. genotypes and rep-PCR" 133 : 159-169, 2011

25 Garces, F., "Detection and quantification of Xanthomonas albilineans by qPCR and potential characterization of sugarcane resistance to leaf scald" 98 : 121-126, 2014

26 Rademaker, J. L. W., "DNA markers: protocols, applications and overviews" John Wiley & Sons 151-171, 1997

27 Rademaker, J. L., "Comparison of AFLP and rep-PCR genomic fingerprinting with DNADNA homology studies : Xanthomonas as a model system" 50 : 665-677, 2000

28 Hauben, L., "Comparison of 16S ribosomal DNA sequences of all Xanthomonas species" 47 : 328-335, 1997

29 Hyytiä-Trees, E., "Characterisation of ropy slime-producing Lactobacillus sakei using repetitive element sequence-based PCR" 50 : 215-219, 1999

30 Williams, P. H, "Black rot : a continuing threat to world crucifers" 64 : 736-742, 1980

31 Pammel, L. H, "Bacteriosis of Rutabaga (Bacillus campestris n. sp.)" 27 : 130-134, 1895

32 Cruz, J., "Assessment of diversity of Xanthomonas campestris pathovars affecting cruciferous plants in Portugal and disclosure of two novel X. campestris pv. campestris races" 99 : 403-414, 2017

33 Lv, H., "An update on the arsenal : mining resistance genes for disease management of Brassica crops in the genomic era" 7 : 34-, 2020

34 Restrepo, S., "AFLP fingerprinting: an efficient technique for detecting genetic variation of Xanthomonas axonopodis pv. manihotis" 145 : 107-114, 1999

35 Enright, M. C., "A multilocus sequence typing scheme for Streptococcus pneumoniae : identification of clones associated with serious invasive disease" 144 : 3049-3060, 1998

36 Fargier, E., "A multilocus sequence analysis of Xanthomonas campestris reveals a complex structure within crucifer-attacking pathovars of this species" 34 : 156-165, 2011

37 Martin, B., "A highly conserved repeated DNA element located in the chromosome of Streptococcus pneumoniae" 20 : 3479-3483, 1992