bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular Characterization of the Cucurbit Yellow Stunting Disorder Virus Coat Protein Gene
Authors: Livieratos Ioannis C. , Avgelis Apostolos D. , Coutts Robert H. A.
Published: May 2007

Cucurbit yellow stunting disorder virus (CYSDV) is a partially characterized bipartite closterovirus transmitted by the tobacco whitefly (Bemisia tabaci). CYSDV has emerged as a serious pathogen in southeastern Spain and the Mediterranean Region, causing yellowing disease of cucumber and melon crops. Using a modified reverse-transcription polymerase chain reaction protocol with gel-extracted dsRNA templates, fragments of CYSDV RNA2 were amplified and cloned. Sequence analysis of the cloned fragments revealed open reading frames encoding the heat shock protein 70 homolog, two proteins of unknown function (p58 and p9), and the coat protein (CP) of the virus in a contiguous gene arrangement similar to that of lettuce infectious yellows virus (LIYV) RNA2. The complete CYSDV CP gene is 756 nt long and encodes a protein with a molecular mass of 28.5 kDa. A comparison of the amino acid sequence of the CYSDV CP gene with those of other closteroviruses revealed significant levels of similarity with sweet potato chlorotic stunt virus and LIYV (36 and 27%, respectively), both of which are members of the recently proposed Crinivirus genus of closteroviruses. The complete CYSDV CP gene was cloned into a bacterial expression vector, and the resulting fusion protein was purified and used to produce antiserum. Purified immunoglobulins specifically detected CYSDV in infected plant extracts, both in immunoblot and indirect enzyme-linked immunosorbent assays with a titer exceeding 2,000 times for both assays.
Development of Bacterial Spot on Near-Isogenic Lines of Bell Pepper Carrying Gene Pyramids Composed of Defeated Major Resistance Genes
Authors: Kousik C. S. , Ritchie D. F.
Published: May 2007

Disease severity caused by races 1 through 6 of Xanthomonas campestris pv. vesicatoria on eight near-isogenic lines (isolines) of Early Calwonder (ECW) with three major resistance genes (Bs1, Bs2, and Bs3) in different combinations was evaluated in the greenhouse and field. Strains representing races 1, 3, 4, and 6 caused similar high levels of disease severity, followed by races 2 and 5 on susceptible ECW. Race 3 caused severe disease on all isolines lacking resistance gene Bs2. Race 4, which defeats Bs1 and Bs2, caused less disease on isoline ECW-12R (carries Bs1 + Bs2), than on isolines ECW, ECW-10R (carries Bs1), and ECW-20R (carries Bs2). Similar results were obtained with race 4 strains in field studies conducted during 1997 and 1998. In greenhouse studies, race 6, which defeats all three major genes, caused less disease on isoline ECW-13R (carries Bs1 + Bs3) and ECW-123R (carries Bs1 + Bs2 + Bs3) than on isolines ECW, ECW-10R, ECW-20R, and ECW-30R (carries Bs3), but not on ECW-23R (carries Bs2 + Bs3). In greenhouse studies with commercial hybrids, strains of races 4 and 6 caused less disease on Boynton Bell (carries Bs1 + Bs2) than on Camelot (carries no known resistance genes), King Arthur (carries Bs1), and X3R Camelot (carries Bs2). Race 6 caused less disease on hybrid R6015 (carries Bs1 + Bs2 + Bs3) and Sentinel (carries Bs1 + Bs3) than on Camelot. Residual effects were not as evident in field studies with race 6 strains. Defeated major resistance genes deployed in specific gene combinations (i.e., gene pyramids) were associated with less area under the disease progress curve than when genes were deployed individually in isolines of ECW or commercial hybrids. Successful management of bacterial spot of pepper is achieved incrementally by integrating multiple tactics. Although there is evidence of residual effects from defeated genes, these effects alone likely will not provide acceptable bacterial spot control in commercial production fields. However, when combined with sanitation practices and a judicious spray program, pyramids of defeated resistance genes may aid in reducing the risk of major losses due to bacterial spot.
Sampling for IPM Decision Making: Where Should We Invest Time and Resources?
Authors: Nyrop Jan P. , Binns Michael R. , van der Werf Wopke
Published: May 2007

Guides for making crop protection decisions based on assessments of pest abundance or incidence are cornerstones of many integrated pest management systems. Much research has been devoted to developing sample plans for use in these guides. The development of sampling plans has usually focused on collecting information on the sampling distribution of the pest, describing this sampling distribution with a mathematical model, formulating a sample plan, and sometimes, but not always, evaluating the performance of the proposed sample plan. For crop protection decision making, classification of density or incidence is usually more appropriate than estimation. When classification is done, the average outcome of classification (the operating characteristic) is frequently robust to large changes in the sampling distribution, including estimates of the variance of pest counts, and to sample size. In contrast, the critical density, or critical incidence, about which classifications are made, has a large influence on the operating characteristic. We suggest that rather than investing resources in elaborate descriptions of sampling distributions, or in fine-tuning sample size to achieve desired levels of precision, greater emphasis should be placed on characterizing pest densities that signal the need for management action and on designing decision guides that will be adopted by practitioners.
Molecular Variability Among Isolates of Prunus Necrotic Ringspot Virus from Different Prunus spp.
Authors: Aparicio F. , Myrta A. , Di Terlizzi B. , Pallás V.
Published: May 2007

Viral sequences amplified by polymerase chain reaction from 25 isolates of Prunus necrotic ringspot virus (PNRSV), varying in the symptomatology they cause in six different Prunus spp., were analyzed for restriction fragment polymorphisms. Most of the isolates could be discriminated by using a combination of three different restriction enzymes. The nucleotide sequences of the RNA 4 of 15 of these isolates were determined. Sequence comparisons and phylogenetic analyses of the RNA 4 and coat proteins (CPs) revealed that all of the isolates clustered into three different groups, represented by three previously sequenced PNRSV isolates: PV32, PE5, and PV96. The PE5-type group was characterized by a 5′ untranslated region that was clearly different from that of the other two groups. The PV32-type group was characterized by an extra hexanucleotide consisting of a duplication of the six immediately preceding nucleotides. Although most of the variability was observed in the first third of the CP, the amino acid residues in this region, which were previously thought to be functionally important in the replication cycle of the virus, were strictly conserved. No clear correlation with the type of symptom or host specificity could be observed. The validity of this grouping was confirmed when other isolates recently characterized by other authors were included in these analyses.
Spiroplasma citri Movement into the Intestines and Salivary Glands of Its Leafhopper Vector, Circulifer tenellus
Authors: Kwon Myoung-Ok , Wayadande Astri C. , Fletcher Jacqueline
Published: May 2007

Spiroplasma citri, a helical, wall-less prokaryote in the class Molli-cutes, is transmitted by the beet leafhopper, Circulifer tenellus. Invasion of leafhopper tissues and cytopathological effects by S. citri were investigated by transmission electron microscopy. All eight cell types of the principle salivary glands, as well as the adjacent muscle cells and the cells of the accessory salivary glands, were colonized by the spiroplas-mas. In both midgut epithelia and salivary gland cells, spiroplasmas usually occurred in membrane-bound cytoplasmic vesicles that often were located near the cell periphery. In several salivary gland cells, spiroplas-mas were also observed within membranous pockets apparently formed by invagination of the plasmalemma beneath intact basal lamina. These observations are consistent with spiroplasma entry into the insect cells by receptor-mediated endocytosis. Cytopathological effects of spiroplasma infection in salivary cells included loss of membrane and basal lamina integrity, presence in some cells of irregular inclusion-like structures containing dense matrices of filamentous material that labeled with anti S. citri antibodies, and apparent disorganization of the endoplasmic reticulum. Compared to the tightly aligned fiber bundles in healthy muscle cells, bundles in spiroplasma-containing muscle cells appeared fragmented and loosely arranged. Such symptoms could contribute to the reduction in longevity and fecundity that has been previously reported for S. citri-infected C. tenellus.
Helper Component Mutations in Nonconserved Residues Associated with Aphid Transmission Efficiency of a Pepper Isolate of Potato Virus Y
Authors: Llave César , Martínez Belén , Díaz-Ruíz J. R. , López-Abella D.
Published: May 2007

The aphid transmission properties of a pepper isolate of potato virus Y belonging to the pathotype 1–2 (PVY 1–2) have been characterized. PVY 1–2 was not transmitted in plant-to-plant experiments, although purified virus particles were efficiently transmitted when supplemented with heterologous helper component (HC) of the transmissible isolate PVY 0 AT through membrane acquisition assays, indicating that its coat protein was functional in transmission. Additionally, virions of PVY 1–2 were able to bind to different HCs in in vitro binding assays. Analysis of the sequence of the PVY 1–2 HC gene and comparison with that of PVY 0 AT revealed 19 nucleotide differences, but only 2 resulted in amino acid changes, one of which induced a change of charge. Neither of these two amino acid changes occurred within the cysteine-rich domain, nor did they coincide with conserved motifs of the HC protein known to be involved in aphid transmission and which are present in all known potyvi-ruses. However, both changes are located in positions highly conserved among PVY strains. The possible role of both mutations on the activity of the PVY 1–2 HC in aphid transmission is discussed.
Parasitism of Sedentary Stages of Heterodera glycines by Isolates of a Sterile Nematophagous Fungus
Authors: Timper Patricia , Riggs Robert D. , Crippen Devany L.
Published: May 2007

Isolates of a sterile fungus designated ARF (Arkansas fungus) can be separated into two groups, ARF-C and ARF-L, that differ morphologically and in their ability to suppress numbers of Heterodera glycines on soybean. Our objectives were to determine if the two ARF groups differed in their ability to parasitize juveniles, females, and eggs in the rhizo-sphere of soybean and to proliferate in soil. The experiments were conducted in a greenhouse using soil infested with homogenized ARF mycelium. The ARF-L isolates parasitized more juveniles and young females than did the ARF-C isolates. Suppression of these stages was 67% for ARF-L and 12% for ARF-C isolates 14 days after nematode inoculation. When soybean plants containing gravid females were transplanted into fungus-infested soil, ARF-L isolates parasitized 55 to 98% of nematode eggs, whereas ARF-C isolates parasitized 0 to 22%. In both heat-treated and nonheated soil, the biomass of mycelial mats, a measure of relative proliferation, tended to be greater for ARF-L than for ARF-C isolates. The ability of ARF-L isolates to parasitize a large percentage of both pre-reproductive stages and eggs of H. glycines may contribute to its effectiveness as a biological control agent.
Cytological Investigation of Resistance to Leptosphaeria maculans Conferred to Brassica napus by Introgressions Originating from B. juncea or B. nigra B Genome
Authors: Roussel S. , Nicole M. , Lopez F. , Renard M. , Chèvre A. M. , Brun H.
Published: May 2007

Introgressions into Brassica napus from the B genome, either the B. nigra chromosome B4 or the B. juncea fragment carrying the Jlm1 gene, have given rise to the B. napus-B. nigra addition line (LA4+) and the B. napus-B. juncea recombinant line (MXS), respectively. The resistance of these two lines to Leptosphaeria maculans is characterized by a hypersensitive reaction (HR) on both the cotyledons and leaves, while the collar displays a high degree of resistance. Responses induced in cotyledons of the two lines by L. maculans inoculation were investigated with emphasis on cytological events underlying the HR and on host defense reactions. Features of host cell changes including condensation and lobing of nuclei, fragmentation of chromatin, disruption of the nuclear membranes, and plasma membrane withdrawal were reminiscent of HR cell death in MXS and LA4+ plants. Restriction of pathogen growth to the infection areas in LA4+ was correlated to reinforcement of cell wall barriers, including wall apposition, papillae, and vessel plugging. In MXS, the lower expression of resistance was associated with a delay in plant responses. These results indicate that mechanisms underlying the HR in the B. napus recombinant and addition lines are differently controlled according to the introgressed genes.
Host Range Specificity in Verticillium dahliae
Authors: Bhat R. G. , Subbarao K. V.
Published: May 2007

Verticillium dahliae isolates from artichoke, bell pepper, cabbage, cauliflower, chili pepper, cotton, eggplant, lettuce, mint, potato, strawberry, tomato, and watermelon and V. albo-atrum from alfalfa were evaluated for their pathogenicity on all 14 hosts. One-month-old seedlings were inoculated with a spore suspension of about 107 conidia per ml using a root-dip technique and incubated in the greenhouse. Disease incidence and severity, plant height, and root and shoot dry weights were recorded 6 weeks after inoculation. Bell pepper, cabbage, cauliflower, cotton, eggplant, and mint isolates exhibited host specificity and differential pathogenicity on other hosts, whereas isolates from artichoke, lettuce, potato, strawberry, tomato, and watermelon did not. Bell pepper was resistant to all Verticillium isolates except isolates from bell pepper and eggplant. Thus, host specificity exists in some isolates of V. dahliae. The same isolates were characterized for vegetative compatibility groups (VCGs) through complementation of nitrate nonutilizing (nit) mutants. Cabbage and cauliflower isolates did not produce nit mutants. The isolate from cotton belonged to VCG 1; isolates from bell pepper, eggplant, potato, and tomato, to VCG 4; and the remaining isolates, to VCG 2. These isolates were also analyzed using the random amplified polymorphic DNA (RAPD) method. Forty random primers were screened, and eighteen of them amplified DNA from Verticillium. Based on RAPD banding patterns, cabbage and cauliflower isolates formed a unique group, distinct from other V. dahliae and V. albo-atrum groups. Minor genetic variations were observed among V. dahliae isolates from other hosts, regardless of whether they were host specific or not. There was no correlation among pathogenicity, VCGs, and RAPD banding patterns. Even though the isolates belonged to different VCGs, they shared similar RAPD profiles. These results suggest that management of Verticillium wilt in some crops through crop rotation is a distinct possibility.
Induction of Systemic Resistance to Albugo candida in Brassica juncea by Pre- or Coinoculation with an Incompatible Isolate
Authors: Singh U. S. , Doughty K. J. , Nashaat N. I. , Bennett R. N. , Kolte S. J.
Published: May 2007

In an investigation of the interaction between two isolates of Albugo candida that were compatible (CO) and incompatible (IN) on a Brassica juncea accession, the IN isolate induced both local and systemic protection of cotyledons and true leaves against the CO isolate. The extent of the protection was proportional to the zoosporangia concentration used in the inducing (IN) inoculation. Protection was greatest locally on cotyledons and least on true leaves (the most remote tissue from the point of the inducing inoculation). Protection occurred when the two isolates were inoculated together but was greater when the interval between the IN and CO isolate inoculations was longer. The IN isolate induced only slight protection when it was inoculated after the CO isolate. No induced susceptibility to the IN isolate occurred with any treatment. There was some evidence of competition between CO and IN zoospores for infection sites (stomata). The occurrence of systemic protection and changes detected in phenylalanine ammonia lyase and total soluble peroxidase activities in inoculated cotyledons, particularly after the inducing (IN) inoculation, suggested that host-mediated factors also may be involved in the interaction between the two isolates.
Advances in the Development of Host Resistance in Corn to Aflatoxin Contamination by Aspergillus flavus
Authors: Brown R. L. , Chen Z.-Y. , Cleveland T. E. , Russin J. S.
Published: May 2007

Aflatoxins are toxic, highly carcinogenic secondary metabolites of Aspergillus flavus and A. parasiticus, which when produced during fungal infection of a susceptible crop in the field or after harvest contaminate food and feed and threaten human and animal health. Although there are several management strategies that may reduce aflatoxin contamination of corn, the preeminent strategy for elimination of aflatoxin is to develop preharvest host resistance to aflatoxin accumulation. This strategy has gained even greater prominence due to recent discoveries of natural resistance in corn that can be exploited in plant-breeding strategies. The ability to identify resistant corn genotypes has been enhanced by the development of a laboratory kernel-screening assay and by a strain of A. flavus genetically engineered to produce β-glucuronidase, an enzyme whose activity can be monitored to assess the degree of fungal infection in kernels. Investigations of resistant corn genotypes have associated kernel pericarp wax characteristics with resistance, identified kernel proteins associated with resistance to and inhibition of fungal growth or aflatoxin biosynthesis, and identified chromosome regions associated with resistance to Aspergillus ear rot and aflatoxin production. Such research advances could lead, in the near future, to commercially available, agronomically acceptable corn lines with multiple preharvest resistances to aflatoxin contamination.
A Single Amino Acid Change in Viral Genome-Associated Protein of Potato Virus Y Correlates with Resistance Breaking in ‘Virgin A Mutant’ Tobacco
Authors: Masuta Chikara , Nishimura Mitsuyo , Morishita Hiroshi , Hataya Tatsuji
Published: May 2007

Tobacco cultivar Virgin A Mutant (VAM) is reported to have the recessive potyvirus resistance gene va. Varied levels of resistance were observed in VAM plants inoculated with Japanese potato virus Y (PVY) isolates. VAM was highly resistant to most of the PVY isolates tested and tolerant to three necrotic strain isolates of PVY-T. Based on data obtained from tissue printing and press blotting, the resistance appeared to be mainly at the level of cell-to-cell movement. PVY replicated in VAM proto-plasts, but the replication was 30% lower than in susceptible tobacco, suggesting that impairment of replication also contributes to resistance. To identify the viral gene product or products involved in VAM resistance, we isolated spontaneous resistance-breaking mutants by passing vein-banding (O strain) isolates several times through VAM plants. By comparing the amino acid sequences of the mutants with their original isolates, we identified a single amino acid substitution in the viral genome-associated protein (VPg) domain that is correlated with VAM resistance breaking. Together, these results suggest that, in addition to its role in replication, VPg plays an important role in the cell-to-cell movement of PVY.
Long-Term Biosanitation by Application of Coniothyrium minitans on Sclerotinia sclerotiorum-Infected Crops
Authors: Gerlagh M. , Goossen-van de Geijn H. M. , Fokkema N. J. , Vereijken P. F. G.
Published: May 2007

The effect of the fungal mycoparasite Coniothyrium minitans applied as a spray to crops infected with Sclerotinia sclerotiorum (causal agent of white mold) on contamination of soil with S. sclerotiorum sclerotia was studied in a 5-year field experiment. Sclerotial survival also was monitored during two subsequent years, when the field was returned to commercial agriculture. In a randomized block design, factorial combinations of four crops and three treatments were repeated 10 times. Potato (Solanum tuberosum), bean (Phaseolus vulgaris), carrot (Daucus carota), and chicory (Cichorium intybus), which are all susceptible to S. sclerotiorum, were grown in rotation. Plots were treated with C. minitans or Trichoderma spp. or were nontreated (control). Crops were rotated in each plot, but treatments were applied to the same plot every year. After 3 years during which it showed no effect on sclerotial survival, the Trichoderma spp. treatment was replaced by a single spray with C. minitans during the fourth and fifth years of the trial. The effect of treatments was monitored in subsequent seasons by counting apothecia as a measure of surviving S. sclerotiorum sclerotia and scoring disease incidence. Trichoderma spp. did not suppress S. sclerotiorum, but C. minitans infected at least 90% of S. sclerotiorum sclerotia on treated crops by the end of the each season. C. minitans lowered the number of apothecia compared with the other treatments during the second year after the bean crop. C. minitans reduced the number of apothecia by ≈90% when compared with the control and Trichoderma spp. treatments and reduced disease incidence in the bean crop by 50% during the fifth year of the trial, resulting in a slightly higher yield. In 1993, but not 1994, a single spray with C. minitans was nearly as effective at reducing apothecia as three sprays (monitored in 1995). The final population size of sclerotia in soil at the end of the 7-year period was lower in all C. minitans plots than at the beginning of the trial, even in plots where two highly susceptible bean crops were grown during the period. The results indicate that the mycoparasite C. minitans has the potential to keep contamination of soil with sclerotia low in crop rotations with a high number of crops susceptible to S. sclerotiorum.
Histological Aspects of a Hypersensitive Response in Poplar to Melampsora larici-populina
Authors: Laurans F. , Pilate G.
Published: May 2007

The course of the infection and development of the biotrophic fungus Melampsora larici-populina on leaf tissue from the hybrid poplar Populus deltoides × P. nigra ‘Ogy’ was monitored at the histological level. Leaf disks were inoculated with one of two rust physiological races (E1 and E2), resulting in interactions that were either incompatible (race E1) or compatible (race E2). In the compatible interaction, the fungus rapidly colonized the leaf without inducing any apparent host response. Symptoms appeared on the leaf several days after inoculation just prior to spore dissemination. The incompatible interaction was characterized by the early collapse and disorganization of cytoplasm of infected cells 17 h after inoculation and within 2 h after the appearance of the first haustoria. Resistance to M. larici-populina was mediated through a hypersensitive response, since it was extremely localized and involved only the few cells that were in the immediate vicinity of each infected cell.
Biotic, Molecular, and Phylogenetic Characterization of Bean Calico Mosaic Virus, a Distinct Begomovirus Species with Affiliation in the Squash Leaf Curl Virus Cluster
Authors: Brown J. K. , Ostrow Kristin M. , Idris Ali M. , Stenger Drake C.
Published: May 2007

Bean calico mosaic virus (BCMoV), a whitefly-transmitted geminivirus from Sonora, Mexico, was purified, and the genome components were cloned and sequenced. Purified viral fractions and cloned genome components were infectious by biolistic inoculation to bean, completing Koch's postulates for both. The B biotype of the whitefly Bemisia tabaci efficiently transmitted both native virus and progeny virus derived from cloned DNA inoculum. Host ranges of native virus and of progeny virus derived from cloned DNA were identical based upon whitefly and biolistic mediated transmission, respectively. BCMoV has a relatively wide experimental host range among begomoviruses known to infect bean, encompassing genera and species within the Fabaceae, Malvaceae, and Solanaceae. BCMoV has a bipartite genome, as do other New World begomoviruses. BCMoV DNA-A shared highest nucleotide sequence identities with squash leaf curl virus-E strain (SLCV-E) and cabbage leaf curl virus (CaLCV) at 80.1 and 80.7%, respectively. BCMoV DNA-B shared highest nucleotide sequence identity with SLCV-E at 70.7%. The common region (CR) sequences of BCMoV and SLCV-E are 73 to 76% identical; however, modular cis-acting elements within the CR involved in replication origin function and recognition are 100% conserved. Phy-logenetic analysis indicated that BCMoV DNA-A shares a most recent common ancestor with the DNA-A of two viruses that also occur in the Sonoran Desert, SLCV-E and Texas pepper virus (TPV-TAM), and CaLCV from Florida. In contrast, a phylogenetic analysis indicated that BCMoV DNA-B shares a most recent common ancestor with SLCV-E; whereas DNA-B of CaLCV clustered in a separate clade with pepper hausteco virus. Collectively, biological and molecular characteristics indicate that BCMoV is a distinct begomovirus species with the northernmost distribution of any begomovirus isolated from bean in the Americas. Furthermore, the phylogenetic relationships of begomovirus cognate components are not necessarily identical, suggesting that DNA-A and DNA-B of some begomoviruses may have different evolutionary histories.
Improved Selection with Newly Identified RAPD Markers Linked to Resistance Gene to Four Pathotypes of Colletotrichum lindemuthianum in Common Bean
Authors: Alzate-Marin Ana Lilia , Menarim Henrique , de Carvalho Geraldo Assis , de Paula Trazilbo José , de Barros Everaldo Gonçalves , Moreira Maurilio Alves
Published: May 2007

Three F2 populations derived from crosses between the resistant cultivar AB 136 and the susceptible cultivar Michelite (MiA), and one F2 population derived from a cross between AB 136 and Mexico 222 (MeA), were used to identify markers linked to anthracnose resistance genes present in cultivar AB 136. Primer OPZ04 produced a DNA band (OPZ04560) linked in coupling phase to the resistance gene for pathotype 89 (8.5 ± 0.025 cM) in one population derived from the cross MiA. In the same population, primer OPZ09 produced one band (OPZ09950) linked in repulsion phase (20.4 ± 0.014 cM) to the same resistance gene. The simultaneous use of markers in coupling and in repulsion phases allowed the identification of the three genotypic classes. In the other two populations from cross MiA, OPZ04560 was linked in coupling phase to resistance genes for pathotypes 73 (2.9 ± 0.012 cM) and 81 (2.8 ± 0.017 cM). In population MeA, OPZ04560 was linked in coupling phase (7.5 ± 0.033 cM) to resistance to pathotype 64. These data suggest that a single gene or complex locus of linked resistance genes present in cultivar AB 136 confers resistance to all four pathotypes of C. lindemuthianum.
Behavior of a Virulent Strain Derived from Agrobacterium radiobacter Strain K84 After Spontaneous Ti Plasmid Acquisition
Authors: López-López María José , Vicedo Begonya , Orellana Natividad , Piquer Jaime , López María M.
Published: May 2007

The behavior of the virulent transconjugant K84N6 derived from Agrobacterium radiobacter strain K84 after spontaneous Ti plasmid transfer in crown gall tissue in a biocontrol experiment was studied and compared with the behavior of the wild-type A. tumefaciens donor of the Ti plasmid. The main difference between the strains was a greatly reduced ability of the transconjugant to catabolize nopaline. Host range, ability to induce tumors in several fruit trees, and stability of the pathogenic determinants in isolates from tumors did not differ between the strains. Nevertheless, in a biocontrol experiment, the transconjugant was not controlled by strain K84 or K1026 in peach × almond hybrids and survived in the plant rhizo-sphere for 9 months with larger population densities than the wild strain. The appearance and persistence in soil of strains harboring a Ti plasmid in the K84 chromosomal background could represent a risk in the medium term, if they show good competitive ability.
Molecular Phylogenetic Analyses of Biological Control Strains of Trichoderma harzianum and Other Biotypes of Trichoderma spp. Associated with Mushroom Green Mold
Authors: Ospina-Giraldo M. D. , Royse D. J. , Chen X. , Romaine C. P.
Published: May 2007

A polymerase chain reaction-amplified DNA containing the internal transcribed spacer (ITS)-1, 5.8S, and ITS-2 regions of the nuclear ribosomal DNA transcriptional unit was sequenced for 81 isolates of Trichoderma spp. associated with mushroom culture or used for biological control of plant pathogens. Phylogenetic analyses revealed that the biocontrol isolates were more closely related to an isolate of T. harzianum biotype 1 (Th1) than to the aggressive biotypes 2 and 4. Th1 has been isolated from mushroom compost but is not the cause of widespread green mold epidemics that have occurred during the last 12 years in Europe and North America. Three isolates of T. harzianum obtained from shiitake (Lentinula edodes; Shi1B and S3-96) and maitake (Grifola frondosa; Mai1) substrates were placed within the biocontrol group. We also found evidence suggesting that some isolates of T. harzianum originally identified as Th4 from Pennsylvania are more closely related to Th2 from Europe. Finally, considering the wide range in sequence distribution of our samples, we propose that the consensus sequence found in this investigation be used as the reference sequence for further studies involving the identification and taxonomy of T. harzianum.
Multiphasic Analysis of Xanthomonads Causing Bacterial Spot Disease on Tomato and Pepper in the Caribbean and Central America: Evidence for Common Lineages Within and Between Countries
Authors: Bouzar H. , Jones J. B. , Stall R. E. , Louws F. J. , Schneider M. , Rademaker J. L. W. , de Bruijn F. J. , Jackson L. E.
Published: May 2007

Four hundred thirty-three xanthomonad strains isolated from tomato or pepper plants from 32 different fields in four Caribbean and Central American countries were screened for the ability to hydrolyze starch and sodium polypectate and for resistance to copper and streptomycin. Of these, 95 representative strains were further characterized by various phnetic tests, and 63 of these strains were then analyzed by genomic fingerprinting. Most of the strains (>90%) were tolerant to copper. However, there was much more variability in sensitivity to streptomycin. All strains in Guadeloupe and 93% of the strains in Barbados were sensitive to streptomycin. The majority of strains were typical Xanthomonas campestris pv. vesicatoria group A strains. In Barbados, however, a unique group of strains was identified that was serologically similar to group A strains but was amylolytic. These strains were designated A1. The occurrence of X. campestris pv. vesicatoria group B strains in Central America was found to be limited to two fields in Costa Rica and one in Guatemala. No group B strains were identified in the Caribbean, in contrast to common occurrence in the central United States and in South America. T3 strains were not found in this study, despite the recent increase of such strains in Florida and Mexico. Unique strains from Costa Rica belonging to the X. gardneri group were identified. Little linkage was found among phenotypic and rep-polymerase chain reaction (rep-PCR) genomic fingerprinting profiles of the pathogens except at the species/pathovar level; strains displaying virtually identical fingerprint profiles were found to correspond to distinct races and vice versa. The rep-PCR genomic fingerprinting analyses suggest that certain lineages may have evolved or predominated in specific regions or specific countries.
Amplified Fragment Length Polymorphism Markers Linked to a Major Quantitative Trait Locus Controlling Scab Resistance in Wheat
Authors: Bai Guihua , Kolb Frederic L. , Shaner Gregory , Domier Leslie L.
Published: May 2007

Scab is a destructive disease of wheat. To accelerate development of scab-resistant wheat cultivars, molecular markers linked to scab resistance genes have been identified by using recombinant inbred lines (RILs) derived by single-seed descent from a cross between the resistant wheat cultivar Ning 7840 (resistant to spread of scab within the spike) and the susceptible cultivar Clark. In the greenhouse, F5, F6, F7, and F10 families were evaluated for resistance to spread of scab within a spike by injecting about 1,000 conidiospores of Fusarium graminearum into a central spikelet. Inoculated plants were kept in moist chambers for 3 days to promote initial infection and then transferred to greenhouse benches. Scab symptoms were evaluated four times (3, 9, 15, and 21 days after inoculation). The frequency distribution of scab severity indicated that resistance to spread of scab within a spike was controlled by a few major genes. DNA was isolated from both parents and F9 plants of the 133 RILs. A total of 300 combinations of amplified fragment length polymorphism (AFLP) primers were screened for polymorphisms using bulked segregant analysis. Twenty pairs of primers revealed at least one polymorphic band between the two contrasting bulks. The segregation of each of these bands was evaluated in the 133 RILs. Eleven AFLP markers showed significant association with scab resistance, and an individual marker explained up to 53% of the total variation (R2). The markers with high R2 values mapped to a single linkage group. By interval analysis, one major quantitative trait locus for scab resistance explaining up to 60% of the genetic variation for scab resistance was identified. Some of the AFLP markers may be useful in marker-assisted breeding to improve resistance to scab in wheat.
Bacterial Colonization of Leaves: A Spectrum of Strategies
Authors: Beattie Gwyn A. , Lindow Steven E.
Published: May 2007

Bacteria associated with plant leaves, or phyllobacteria, probably employ a range of colonization strategies. Steps in these colonization strategies include modification of the leaf habitat, aggregation, ingression, and egression. Considerable evidence indicates that bacteria can modify their environment to enhance their colonization of plants, such as by increasing local nutrient concentrations or by producing a layer of extracellular polysaccharides. This local habitat modification may occur on the surface of leaves, as well as in the leaf interior, and may be enhanced by the formation of bacterial aggregates. The conspicuous presence of bacterial aggregates on leaves and the finding that the behavior of bacteria on plants varies in a density-dependent manner indicate the potential importance of cooperative interactions among phyllobacteria. Such cooperative interactions may occur among both homogeneous and heterogeneous populations, thus influencing the development of microbial communities. While the sites commonly colonized by most phyllobacteria have not been unambiguously identified, there is strong circumstantial evidence that a sizable proportion of cells, particularly of phytopathogenic strains, are localized within “protected sites” on plants. The likelihood that these protected sites are located in the interior of leaves indicates that phytopathogenic bacteria have access to more resources and greater protection from stresses associated with the leaf surface than bacteria that are restricted to the leaf surface. The internal and external leaf-associated populations probably form a continuum due to the processes of ingression and egression. For a specific pathogen, however, the extent of egression that occurs prior to disease induction is likely to influence the success of disease predictions based on external population size, i.e., the number of bacteria in leaf washings. In this review, we illustrate the complexity of the ecology of leaf-associated bacteria and propose a model of leaf colonization that emphasizes the common elements in bacterial colonization strategies, as well as allows for distinct behavior of different phyllobacterial species.
Identification, Characterization, and Relatedness of Luteovirus Isolates from Forage Legumes
Authors: Damsteegt V. D. , Stone A. L. , Russo A. J. , Luster D. G. , Gildow F. E. , Smith O. P.
Published: May 2007

Virus isolates from forage legumes collected from eight different states were identified as luteoviruses closely related to soybean dwarf luteovirus dwarfing (SbDV-D) and yellowing (SbDV-Y) described in Japan. All isolates produced reddened leaf margins in subterranean clover and were transmitted in a persistent manner by Acrythosiphon pisum, but not by Aulacorthum solani. Specific monoclonal antibodies raised against SbDV-Y were differentially reactive with endemic isolates. Immunoblots probed with a SbDV-D polyclonal antiserum showed single 26-kDa coat protein bands, confirming close serological relatedness to SbDV. Analyses of genomic and subgenomic double-stranded RNAs and northern blot analyses confirmed genomic relatedness to SbDV. Based on our results, we conclude that the U.S. luteovirus isolates studied comprise a strain or strains of the soybean dwarf virus that have clovers as common hosts and the pea aphid as a common vector.
Enchytraeids and Nematophagous Fungi in Tomato Fields and Vineyards
Authors: Jaffee B. A.
Published: May 2007

I tested the hypothesis that exclusion of enchytraeids and microarthropods in agricultural fields improves establishment of two nematophagous fungi. Soil was collected from three tomato fields and two vineyards and either heat-treated (2 h at 60°C) or not. Alginate pellets containing hyphae of the fungi Hirsutella rhossiliensis or Monacrosporium gephyropagum were added to the soil, which was packed into cages (PVC pipe, 80-cm3volume) sealed with fine (20 μm) or coarse (480 μm) mesh. Cages were buried 22 cm deep in the same fields from which the soil had been collected. After 7 to 50 days, the cages were recovered and fungi and fauna quantified. Fine mesh largely excluded enchytraeids, collembolans, and mites but rarely affected fungus numbers. In contrast, heat treatment of soil rarely affected enchytraeids, collembolans, or mites but frequently increased fungus numbers, regardless of mesh size. The data are inconsistent with the initial hypothesis but are consistent with the idea that organisms narrower than 20 μm interfere with fungal growth from the pellets.
Rapid Detection of the Fusarium oxysporum Lineage Containing the Canary Island Date Palm Wilt Pathogen
Authors: Plyler T. R. , Simone G. W. , Fernandez D. , Kistler H. C.
Published: May 2007

Fusarium oxysporum f. sp. canariensis causes Fusarium wilt disease on the Canary Island date palm (Phoenix canariensis). To facilitate disease management, a polymerase chain reaction diagnostic method has been developed to rapidly detect the pathogen. A partial genomic library of F. oxysporum f. sp. canariensis isolate 95-913 was used to identify a DNA sequence diagnostic for a lineage containing all tested isolates of F. oxysporum f. sp. canariensis. Two oligonucleotide primers were designed and used to amplify a 567-bp fragment with F. oxysporum f. sp. canariensis DNAs. DNA from 61 outgroup isolates did not amplify using these primers. Once the primers were shown to amplify a 0.567-kb fragment from DNA of all the F. oxysporum f. sp. canariensis isolates tested, a rapid DNA extraction procedure was developed that led to the correct identification of 98% of the tested F. oxysporum f. sp. canariensis isolates.
Spatial Pattern Analysis of Strawberry Leaf Blight in Perennial Production Systems
Authors: Turechek W. W. , Madden L. V.
Published: May 2007

Spatial pattern of the incidence of strawberry leaf blight, caused by Phomopsis obscurans, was quantified in commercial strawberry fields in Ohio using statistics for heterogeneity and spatial correlation. For each strawberry planting, two transects were randomly chosen and the proportion of leaflets (out of 15) and leaves (out of five) with leaf blight symptoms was determined from N = 49 to 106 (typically 75) evenly spaced sampling units, thus establishing a natural spatial hierarchy to compare patterns of disease. The beta-binomial distribution fitted the data better than the binomial in 92 and 26% of the 121 data sets over 2 years at the leaflet and leaf levels, respectively, based on a likelihood ratio test. Heterogeneity in individual data sets was measured with the index of dispersion (variance ratio), C(α) test, a standard normal-based test statistic, and estimated θ parameter of the beta-binomial. Using these indices, overdispersion was detected in approximately 94 and 36% of the data sets at the leaflet and leaf levels, respectively. Estimates of the slope from the binary power law were significantly (P < 0.01) greater than 1 and estimates of the intercept were significantly greater than 0 (P < 0.01) at both the leaflet and leaf levels for both years, indicating that degree of heterogeneity was a function of incidence. A covariance analysis indicated that cultivar, time, and commercial farm location of sampling had little influence on the degree of heterogeneity. The measures of heterogeneity indicated that there was a positive correlation of disease status of leaflets (or leaves) within sampling units. Measures of spatial association in disease incidence among sampling units were determined based on autocorrelation coefficients, runs analysis, and a new class of tests known as spatial analysis by distance indices (SADIE). In general, from 9 to 22% of the data sets had a significant nonrandom spatial arrangement of disease incidence among sampling units, depending on which test was used. When significant associations existed, the magnitude of the association was small but was about the same for leaflets and leaves. Comparing test results, SADIE analysis was found to be a viable alternative to spatial autocorrelation analysis and has the advantage of being an extension of heterogeneity analysis rather than a separate approach. Collectively, results showed that incidence of Phomopsis leaf blight was primarily characterized by small, loosely aggregated clusters of diseased leaflets, typically confined within the borders of the sampling units.
Complementation of Coat Protein Mutants of Pepper Huasteco Geminivirus in Transgenic Tobacco Plants
Authors: Guevara-González R. G. , Ramos P. L. , Rivera-Bustamante R. F.
Published: May 2007

The role of the pepper huasteco virus (PHV) coat protein (CP) gene during the infection was investigated in three different hosts by using mutations that produced truncated proteins and by complementation assays in transgenic plants. The infectivity analysis revealed that mutants that express truncated CP (CP7 and CP191) behave like the wild-type virus when inoculated onto pepper and Nicotiana benthamiana plants in terms of symptom expression and viral DNA movement. On the contrary, the CP7 mutant was unable to systemically infect tobacco plants, whereas only 10% of the plants inoculated with the CP191 mutant became infected. The CP7 mutant was complemented by coinoculating it with another geminivirus (taino tomato mottle virus). No complementation was observed in plants from nine transgenic tobacco lines expressing CP under the control of the cauliflower mosaic virus (CaMV) 35S promoter. However, 3 out of 10 lines expressing CP under the control of its own promoter (693 nucleotides) were able to complement the CP7 mutant. Interestingly, upon infection, the levels of CP mRNA in 693CP plants increased dramatically, probably due to transactivation of the CP promoter by the viral protein AC2.
Comparison of Rain Effects on Splash Dispersal of Three Colletotrichum Species Infecting Strawberry
Authors: Ntahimpera N. , Wilson L. L. , Ellis M. A. , Madden L. V.
Published: May 2007

Rain simulation studies were performed to compare splash dispersal of three Colletotrichum species: C. acutatum (C. acutatum-O isolate from Ohio and C. acutatum-M isolate from Mississippi), C. fragariae (isolate from Mississippi), and C. gloeosporioides (isolate from Florida). Conidial dispersal was assessed by counting colonies formed from spore-bearing splash droplets deposited in sheltered petri plates containing a selective medium. Colonies were converted to number of conidia based on germination rates of spores on the media. The interpolated total number of dispersed conidia over a 61 min rain and 72 cm from the point source (Σ) was calculated. For all species, a rain intensity of 30-mm/h resulted in significantly greater dispersal than an intensity of 11-mm/h. C. fragariae had the lowest amount of spore dispersal, and C. acutatum-O had the highest dispersal. C. acutatum-M and C. gloeosporioides were intermediate in magnitude of conidial splash dispersal. However, differences were directly attributed to differences in spore density per fruit at the source. When Σ was corrected for source strength (Σr), the species were very similar, with only C. acutatum-M having a mean Σr significantly less than the others. Proportions and rates of spore removal (per minute) from source fruits were higher for C. acutatum-O and C. gloeosporioides than for other isolates. Wash-off rates of conidia deposited on healthy fruits were the same for all species. Deposition flux density of spores that had been uniformly sprayed over the entire soil surface of the experimental area was affected by species. A significant difference in means was observed between C. acutatum and C. fragariae—the latter had a somewhat lower flux density. This is the first demonstration that closely related species infecting the same plant species are similar in terms of splash dispersal.
Purification and Characterization of Turnip Mosaic Virus Helper Component Protein
Authors: Wang R. Y. , Pirone T. P.
Published: May 2007

The helper component (HC) protein of turnip mosaic virus (TuMV) was concentrated by differential centrifugation followed by ammonium sulfate precipitation. The partially purified HC was then loaded onto a Ni2+-resin column that bound the HC; a histidine tag was not required for binding. The HC eluted from the column migrated as a band of about 50 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In its native state, the HC did not pass through an ultrafiltration membrane with a molecular mass cutoff of 100 kDa, which suggested that the HC is in a multimeric form when it is biologically active. The molecular mass of the multimeric form was determined by gel filtration to be approximately 145 kDa. Purified HC retained its activity for several months at -20°C. Using a protein blotting-overlay protocol, purified HC interacted in vitro with an aphid-transmissible TuMV isolate, but not with a non-aphid-transmissible isolate.