CHAPTER 23: Cucurbit Yellow Vine Disease: A Model for True Bug–Bacteria Relationships

2003 ◽  
Vol 93 (10) ◽  
pp. 1233-1239 ◽  
Author(s):  
J. Rascoe ◽  
M. Berg ◽  
U. Melcher ◽  
F. L. Mitchell ◽  
B. D. Bruton ◽  
...  

A serious vine decline of cucurbits known as cucurbit yellow vine disease (CYVD) is caused by rod-shaped bacteria that colonize the phloem elements. Sequence analysis of a CYVD-specific polymerase chain reaction (PCR)-amplified 16S rDNA product showed the microbe to be a γ-proteobacterium related to the genus Serratia. To identify and characterize the bacteria, one strain each from watermelon and zucchini and several noncucurbit-derived reference strains were subjected to sequence analysis and biological function assays. Taxonomic and phylogenetic placement was investigated by analysis of the groE and 16S rDNA regions, which were amplified by PCR and directly sequenced. For comparison, eight other bacterial strains identified by others as Serratia spp. also were sequenced. These sequences clearly identified the CYVD strains as Serratia marcescens. However, evaluation of metabolic and biochemical features revealed that cucurbit-derived strains of S. marcescens differ substantially from strains of the same species isolated from other environmental niches. Cucurbit strains formed a distinct cluster, separate from other strains, when their fatty acid methyl ester profiles were analyzed. In substrate utilization assays (BIOLOG, Vitek, and API 20E), the CYVD strains lacked a number of metabolic functions characteristic for S. marcescens, failing to catabolize 25 to 30 compounds that were utilized by S. marcescens reference strains. These biological differences may reflect gene loss or repression that occurred as the bacterium adapted to life as an intracellular parasite and plant pathogen.


Plant Disease ◽  
2001 ◽  
Vol 85 (9) ◽  
pp. 1031-1031 ◽  
Author(s):  
R. L. Wick ◽  
J. Lerner ◽  
S. D. Pair ◽  
J. Fletcher ◽  
F. Mitchell ◽  
...  

Commercial plantings of summer squash in Charlemont, Franklin County, MA, were decimated in 1999 by 100% incidence of a yellowing disease resembling cucurbit yellow vine disease (CYVD) (1). Both plantings were established in the same field during the third week of May, one with transplants and the second by direct-seeding. Each planting consisted of four 30-m rows each of yellow zucchini (Cucurbita pepo cv. Gold Rush), summer squash (C. pepo cv. Seneca Prolific), and zucchini (C. pepo cv. Condor). Crops were produced organically and pyrethrum was used to control a high infestation of squash bugs, Anasa tristis (De Geer) (Heteroptera:Coreidae), a putative vector of CYVD (3). Just prior to fruit set, during the first two weeks of June, plants began showing symptoms of foliar chlorosis, plant stunting, or both. All of the plants in the field eventually wilted and collapsed. Cross-sections of the below-ground stem and primary root revealed a honey-brown phloem discoloration and healthy appearing xylem, symptoms characteristic of CYVD. Plants yielded marketable fruit for only about 1 week. When plant samples were tested by polymerase chain reaction (PCR) with CYVD bacterium specific primers (2), a band of the expected size for the CYVD bacterium, identified as Serratia marcescens based on 16s rDNA and groE sequence analyses (4), was amplified in every case. Since all plant samples collected were symptomatic and PCR positive for S. marcescens, asymptomatic greenhouse plants were run simultaneously as a control. All control plants tested negative. A third planting, similar to the two disease-affected plantings and containing the same three squash cultivars from the same seed lot, was established at about the same time approximately 3 km away. No symptoms of CYVD occurred at this site, further evidence that the pathogen is not seed-borne (1). Furthermore, squash bugs were not observed in this field. In 2000, the disease was observed in a planting of ‘Atlantic Giant’ pumpkin in Erving, Franklin County, MA, and confirmed by PCR. Until now, CYVD has been reported only in the states of Oklahoma, Texas, and Tennessee. Confirmation of the disease in Massachusetts significantly increases the known geographical range of CYVD to include the New England area. References: (1) B. D. Bruton et al. Plant Dis. 82:512–520, 1998. (2) U. Melcher et al. Phytopathology 89:S95, 1999. (3) S. D. Pair et al. Pages 145–148 in: Proc. 19th Ann. Hort. Conf., Okla. State Univ. (4) J. Rascoe et al. Phytopathology 90:S63, 2000.


Plant Disease ◽  
2017 ◽  
Vol 101 (1) ◽  
pp. 129-136 ◽  
Author(s):  
K. R. Besler ◽  
E. L. Little

Cucurbit yellow vine disease (CYVD), caused by the squash bug (Anasa tristis)-transmitted bacterium Serratia marcescens, was first identified in Oklahoma and Texas in 1988 and in Georgia in 2012. S. marcescens is a highly diverse species found in many ecological niches. In previous studies, CYVD strains of S. marcescens formed a closely related group separate from non-CYVD strains based on biological and molecular characterization techniques. Multilocus sequence analysis (MLSA) of six housekeeping genes and repetitive elements-based polymerase chain reaction (rep-PCR) using the BOX and ERIC primers were used to assess the genetic diversity of CYVD strains of S. marcescens collected in Georgia together with a strain from Texas and seven non-CYVD strains of S. marcescens. rep-PCR results revealed genetic diversity among CYVD strains while MLSA results showed a 100% similarity across the six loci for all but one of the CYVD strains, which differed at the icd locus by five polymorphisms. For both methods, CYVD strains clustered separately from nonplant-pathogenic S. marcescens strains and were most similar to a rice endophyte strain. One CYVD strain isolated from a squash bug shared genetic similarities with non-CYVD strains, and may be the result of a recombination event between CYVD and non-CYVD strains.


Author(s):  
Kino Emmanuel Maravillas ◽  
Erika Diaz-Almeyda ◽  
Nicole Gerardo

Polycultures, or multicrops, are groupings of plants that grow more prolifically when planted together as compared to when planted alone as monocultures. One of the best known and widely utilized polycultures is the milpa cropping system - the cultivation of maize, beans, and squash together as “the three sisters.” Milpa has been utilized by the indigenous population of Central America for millennia due to its consistent abundant harvests; today it remains a cornerstone of the region’s tradition, diet, and economic growth. Likely contributing to this legacy is the known association of polycultures and heightened resistance to disease, yet the mechanism underlying this relationship in milpa has largely been unexplored. To assess the health of farm soil exposed to milpa monocrops, bicrops, and multicrops, we measured the growth of two bacterial strains: a Burkholderia strain symbiotic of Anasa tristis (the squash bug, an agricultural pest) and a plant pathogenic Serratia strain that is the primary causal agent of cucurbit yellow vine disease (CYVD). We found that after one week in both the polyculture (corn, bean, and squash) soil and the corn monoculture soil, the growth of Burkholderia was significantly inhibited. However, in both corn & bean and bean & squash biculture soils, the growth of the strain was significantly enhanced. The growth of the Serratia strain did not yield any significant increase or decrease after one week in any milpa soil. We conclude that the cultivation of milpa in its polyculture configuration demonstrates antibiotic activity towards the Burkholderia strain SQ4A. Our investigation supports findings that certain multicrop systems are less susceptible to disease than monocultures possibly due to their greater microbial biomass; thus we can infer a higher amount of root exudates present in the soil, of which a substantial amount may be anti-microbials.


Plant Disease ◽  
2003 ◽  
Vol 87 (8) ◽  
pp. 937-944 ◽  
Author(s):  
B. D. Bruton ◽  
F. Mitchell ◽  
J. Fletcher ◽  
S. D. Pair ◽  
A. Wayadande ◽  
...  

Cucurbit yellow vine disease (CYVD), which can inflict heavy losses to watermelon, pumpkin, cantaloupe, and squash in U.S. production areas from the midwest to northeastern states, causes phloem discoloration, foliar yellowing, wilting, and plant decline. Bacteria were cultured from the phloem of crown sections of symptomatic plants of Citrullus lanatas and Cucurbita pepo. Those bacteria testing positive in CYVD-specific polymerase chain reaction (PCR) were all gram negative and appeared morphologically identical, producing creamy white, smooth, entire, convex colonies on Luria-Bertani or nutrient agar. Characterized cucurbit-derived strains of Serratia marcescens were introduced into greenhouse-grown squash plants by puncture inoculation and into field-grown squash plants by enclosure with S. marcescens-fed squash bugs, Anasa tristis. Up to 60% of the bacteria-inoculated plants in the greenhouse and up to 17% of field plants caged with inoculative squash bugs developed phloem discoloration and tested positive for S. marcescens by CYVD-specific PCR. None of the controls developed phloem discoloration or tested positive by PCR. Of the diseased field plants, 12% (2 of 35) also yellowed, wilted, and collapsed, exhibiting full symptom development of CYVD. However, neither plant collapse nor decline was observed in the greenhouse-grown, puncture-inoculated plants. The morphology, growth habit, and PCR reaction of bacteria cultured from crown tissue of a subset of plants in each experimental group were indistinguishable from those of the inoculum bacteria. Evidence presented from our studies confirms that the squash bug can transmit S. marcescens, the CYVD causal bacterium. The S. marcescens-A. tristis relationship described here is the first instance in which the squash bug has been identified as a vector of a plant pathogen. Our experiments represent a completion of the steps of Koch's postulates, demonstrating that S. marcescens is the causal agent of CYVD and that the squash bug, A. tristis, is a vector of the pathogen.


2004 ◽  
Vol 97 (1) ◽  
pp. 74-78 ◽  
Author(s):  
S. D. Pair ◽  
B. D. Bruton ◽  
F. Mitchell ◽  
J. Fletcher ◽  
A. Wayadande ◽  
...  

2003 ◽  
Vol 93 (10) ◽  
pp. 1240-1246 ◽  
Author(s):  
Q. Zhang ◽  
R. Weyant ◽  
A. G. Steigerwalt ◽  
L. A. White ◽  
U. Melcher ◽  
...  

The bacterium that causes cucurbit yellow vine disease (CYVD) has been placed in the species Serratia marcescens based on 16S rDNA and groE sequence analysis. However, phenotypic comparison of the organism with S. marcescens strains isolated from a variety of ecological niches showed significant heterogeneity. In this study, we compared the genomic DNA of S. marcescens strains from different niches as well as type strains of other Serratia spp. through repetitive elements-based polymerase chain reaction (rep-PCR) and DNA-DNA hybridization. With the former, CYVD strains showed identical banding patterns despite the fact that they were from different cucurbit hosts, geographic locations, and years of isolation. In the phylogenetic trees generated from rep-PCR banding patterns, CYVD strains clearly were differentiated from other strains but formed a loosely related group with S. marcescens strains from other niches. The homogeneity of CYVD strains was supported further by the DNA relatedness study, in that labeled DNA from the cantaloupe isolate, C01-A, showed an average relative binding ratio (RBR) of 99%, and 0.33% divergence to other CYVD strains. Used as a representative strain of CYVD, the labeled C01-A had a RBR of 76%, and a 4.5% divergence to the S. marcescens type strain. These data confirm the previous placement of CYVD strains in S. marcescens. Our investigations, including rep-PCR, DNA-DNA hybridization, and previous phenotyping experiments, have demonstrated that CYVD-associated strains of S. marcescens cluster together in a group significantly different from other strains of the species.


2005 ◽  
Vol 71 (12) ◽  
pp. 7716-7723 ◽  
Author(s):  
Q. Zhang ◽  
U. Melcher ◽  
L. Zhou ◽  
F. Z. Najar ◽  
B. A. Roe ◽  
...  

ABSTRACT Cucurbit yellow vine disease (CYVD) is caused by disease-associated Serratia marcescens strains that have phenotypes significantly different from those of nonphytopathogenic strains. To identify the genetic differences responsible for pathogenicity-related phenotypes, we used a suppressive subtractive hybridization (SSH) strategy. S. marcescens strain Z01-A, isolated from CYVD-affected zucchini, was used as the tester, whereas rice endophytic S. marcescens strain R02-A (IRBG 502) was used as the driver. SSH revealed 48 sequences, ranging from 200 to 700 bp, that were present in Z01-A but absent in R02-A. Sequence analysis showed that a large proportion of these sequences resembled genes involved in synthesis of surface structures. By construction of a fosmid library, followed by colony hybridization, selection, and DNA sequencing, a phage gene cluster and a genome island containing a fimbrial-gene cluster were identified. Arrayed dot hybridization showed that the conservation of subtracted sequences among CYVD pathogenic and nonpathogenic S. marcescens strains varied. Thirty-four sequences were present only in pathogenic strains. Primers were designed based on one Z01-A-specific sequence, A79, and used in a multiplex PCR to discriminate between S. marcescens strains causing CYVD and those from other ecological niches.


Sign in / Sign up

Export Citation Format

Share Document