Showing papers in "Plant Disease in 2005"

Epidemics of Soybean Rust (Phakopsora pachyrhizi) in Brazil and Paraguay from 2001 to 2003.

Abstract: In 5 March 2001, a severe rust outbreak was recorded at Pitapo, Paraguay, and the causal organism was determined to be Phakopsora pachyrhizi using polymerase chain reaction (PCR) and DNA s...

First Report of Soybean Rust Caused by Phakopsora pachyrhizi in the Continental United States.

Abstract: Asian soybean rust, caused by Phakopsora pachyrhizi Sydow, has been known to occur in the eastern hemisphere for nearly a century. More recently, it was reported from Hawaii in 1994, eastern and so...

First Report of a Huanglongbing-Like Disease of Citrus in Sao Paulo State, Brazil and Association of a New Liberibacter Species, “Candidatus Liberibacter americanus”, with the Disease

Abstract: Huanglongbing (HLB) (ex-greening) is one of the most serious diseases of citrus. The causal agent is a noncultured, sieve tube-restricted α-proteobacterium, "Candidatus Liberibacter africanus" in Africa and "Candidatus Liberibacter asiaticus" in Asia (2). The disease has never been reported from the American continent. However, Diaphorina citri, the Asian psyllid vector of HLB, is found in South, Central, and North America (Florida and Texas). Early in 2004, leaf and fruit symptoms resembling those of HLB were observed in several sweet orange orchards near the city of Araraquara, Sao Paulo State. Leaf mottling on small and large leaves was the major symptom. Shoots with affected leaves were yellowish. Fruits were small and lopsided, contained many aborted seeds, and appeared more severely affected than were plants infected with classic HLB. Forty-three symptomatic samples and twenty-five samples of symptomless sweet orange leaves from five farms were analyzed for the presence of the HLB-liberibacters using polymerase chain reaction (PCR) with two sets of HLB-specific primers for amplification of 16S rDNA (2,3) and ribosomal protein genes (1). None of the 43 symptomatic leaf samples gave a positive PCR amplification, while HLB-affected leaves from the Bordeaux HLB collection produced the characteristic amplicons with both sets of primers. The 43 symptomatic and the 25 symptomless leaf samples were then analyzed using PCR with universal primers for amplification of bacterial 16S rDNA (4). All symptomatic leaf samples, but none of the symptomless leaf samples, yielded the same 16S rDNA amplification product, indicating the presence of a bacterium in the symptomatic leaves. This was confirmed using the observation of a sieve tube restricted bacterium by electron microscopy. The 16S rDNA product was cloned, sequenced, and compared with those of "Ca. L. africanus" and "Ca. L. asiaticus". While the 16S rDNAs of these two liberibacter species have 97.5% sequence identity, the 16S rDNA sequence of the new bacterium shared only 93.7% identity with that of "Ca. L. asiaticus" and 93.9% with that of "Ca. L. africanus". The 16S rDNA sequence of the new bacterium had a secondary loop structure characteristic of the α subdivision of the proteobacteria and possessed all the oligonucleotide signatures characteristic of the liberibacters. For these reasons, the new bacterium is a liberibacter and is sufficiently different phylogenetically from known liberibacters to warrant a new species, "Candidatus Liberibacter americanus". Specific PCR primers for amplification of the 16S rDNA of the new species have been developed. They were able to detect "Ca. L. americanus" in 214 symptomatic leaf samples from 47 citrus farms in 35 municipalities, while the "old" species, "Ca. L. asiaticus", has been found only four times within the 47 farms. References: (1) A. Hocquellet et al. Mol. Cell. Probes, 13:373, 1999. (2) S. Jagoueix et al. Int. J. Syst. Bacteriol. 44:379, 1994. (3) S. Jagoueix et al. Mol. Cell. Probes 10:43, 1996. (4) W. G. Weisburg et al. J. Bacteriol. 173:697, 1991.

Crop Damage Estimates for Crown Rot of Wheat and Barley in the Pacific Northwest.

Abstract: Crown rot of wheat and barley in the Pacific Northwest is caused by a complex of Fusarium pseudograminearum, F. culmorum, F. avenaceum, Bipolaris sorokiniana, and Microdochium nivale. Yield-loss estimates were made by evaluating yield components on tillers collected from commercial fields and sorted by disease severity classes, and by comparing yields for field plots inoculated with F. pseudograminearum with yields in naturally infested soil. Increasing crown rot severity caused an increase in grain protein content and reduction in grain yield, kernels per head, kernel weight, test weight, tiller height, and straw weight. Crown rot reduced winter wheat yield as much as 1,550 kg/ha (35%, $219/ha) in commercial fields, with a 13-field mean of 9.5% ($51/ha). Inoculation reduced yields as much as 2,630 kg/ha (61%, $372/ha) over that caused by the native pathogen flora. Rain-induced crusting of the soil surface greatly amplified preemergence damping-off caused by F. pseudograminearum. Crown rot caused the greatest losses during seasons of lowest precipitation but also damaged crops under wet conditions. Aboveground symptoms were not always apparent under conditions of moderate infection and yield constraint. Damage from crown rot in the Pacific Northwest is more widespread and damaging than previously recognized.

Breeding for Resistance to Soybean Rust.

Abstract: Soybean rust occurs in all major soybean-growing regions of the world including the North American mainland. Soybean rust, caused by Phakopsora pachyrhizi, is the most destructive foliar disease of soybean, and yield losses of over 50% are common when environmental conditions are conducive for disease development. Heavily infected plants defoliate and mature more rapidly than plants not infected with rust. P. pachyrhizi has a broad host range and can infect many other legumes including some native to Australia. A number of physiological races of the fungus have been reported on these native legumes from Australia and on soybean. In addition, four single genes for rust resistance were previously identified in four different soybean plant introductions. These sources of resistance also have been reported to be susceptible in some field locations and when challenged with certain isolates of P. pachyrhizi. Partial resistance, expressed as reduced pustule number and increased length of latent period, has also been reported but has not been widely used in breeding programs. Yield stability has been used in the past and compares percentage of yields in fungicide and nonfungicide plots. Cultivars or lines with a higher percentage of yield have greater yield stability in the presence of rust. Although soybean rust only recently was found in the continental United States, a proactive project to evaluate the USDA soybean germ plasm collection for rust resistance was initiated in 2002 at the Fort Detrick plant biocontainment facility and at six international locations. Part of this project is to discover soybean lines with greater yield stability, and additional single and partial resistance. To help minimize the impact of soybean rust, the first line of defense will be fungicides, with host resistance and yield stability augmenting the long-term management of soybean rust.

Effect of the F129L Mutation in Alternaria solani on Fungicides Affecting Mitochondrial Respiration

Abstract: Pasche, J. S., Piche, L. M., and Gudmestad, N. C. 2005. Effect of the F129L mutation in Alternaria solani on fungicides affecting mitochondrial respiration. Plant Dis. 89:269-278. Isolates of Alternaria solani previously collected from throughout the Midwestern United States and characterized as being azoxystrobin sensitive or reduced sensitive were tested for sensitivity to the Quinone outside inhibitor (QoI) fungicides famoxadone and fenamidone and the carboxamide fungicide boscalid. All three fungicides affect mitochondrial respiration: famoxadone and fenamidone at complex III, and boscalid at complex II. A. solani isolates possessing reducedsensitivity to azoxystrobin also were less sensitive in vitro to famoxadone and fenamidone compared with azoxystrobin-sensitive isolates, but the shift in sensitivity was of lower magnitude, approximately 2- to 3-fold versus approximately 12-fold for azoxystrobin. The in vitro EC50 values, the concentration that effectively reduces germination by 50% relative to the untreated control, for sensitive A. solani isolates were significantly lower for famoxadone and azoxystrobin than for fenamidone and boscalid; whereas, for reduced-sensitive isolates, famoxadone EC50 values were significantly lower than all other fungicides. Isolates of A. solani with reducedsensitivity to azoxystrobin were twofold more sensitive in vitro to boscalid than were azoxystrobin-sensitive wild-type isolates, displaying negative cross-sensitivity. All isolates determined to have reduced-sensitivity to azoxystrobin also were determined to possess the amino acid substitution of phenylalanine with leucine at position 129 (F129L mutation) using real-time polymerase chain reaction. In vivo studies were performed to determine the effects of in vitro sensitivity shifts on early blight disease control provided by each fungicide over a range of concentrations. Reduced-sensitivity to azoxystrobin did not significantly affect disease control provided by famoxadone, regardless of the wide range of in vitro famoxadone EC50 values. Efficacy of fenamidone was affected by some azoxystrobin reduced-sensitive A. solani isolates, but not others. Boscalid controlled azoxystrobin-sensitive and reduced-sensitive isolates with equal effectiveness. These results suggest that the F129L mutation present in A. solani does not convey cross-sensitivity in vivo among all QoI or related fungicides, and that two- to threefold shifts in in vitro sensitivity among A. solani isolates does not appreciably affect disease control. Additional keywords: Solanum tuberosum

Susceptibility of Oregon Forest Trees and Shrubs to Phytophthora ramorum: A Comparison of Artificial Inoculation and Natural Infection

Abstract: Phytophthora ramorum is an invasive pathogen in some mixed-hardwood forests in California and southwestern Oregon, where it causes sudden oak death (SOD) on some members of Fagaceae, ramorum shoot dieback on some members of Ericaceae and conifers, and ramorum leaf blight on diverse hosts. We compared symptoms of P. ramorum infection resulting from four different artificial inoculation techniques with the symptoms of natural infection on 49 western forest trees and shrubs; 80% proved susceptible to one degree or another. No single inoculation method predicted the full range of symptoms observed in the field, but whole plant dip came closest. Detached-leaf-dip inoculation provided a rapid assay and permitted a reasonable assessment of susceptibility to leaf blight. Both leaf age and inoculum dose affected detached-leaf assays. SOD and dieback hosts often developed limited leaf symptoms, although the pattern of midrib and petiole necrosis was distinctive. Stem-wound inoculation of seedlings correlated with field symptoms for several hosts. The results suggested that additional conifer species may be damaged in the field. Log inoculation provided a realistic test of susceptibility to SOD, but was cumbersome and subject to seasonal variability. Pacific rhododendron, salmonberry, cascara, and poison oak were confirmed as hosts by completing Koch's postulates. Douglas-fir was most susceptible to shoot dieback shortly after budburst, with infection occurring at the bud.

Soybean Cyst Nematode Management Reconsidered.

Abstract: A well-known nematologist once said (in public) that soybean cyst nematode (SCN, Heterodera glycines Ichinohe) was easy to control: “Don’t grow soybeans.” Shortly thereafter, following a barrage of good-natured criticism for this bit of unhelpful advice, he amended the statement to: “Don’t grow soybeans more than once every 5 years or so.” (He made a similar statement in print [22]). From a nematological standpoint, he was quite right in that rotation to a nonhost is the most effective way to reduce SCN population densities and thereby avoid the soybean yield losses associated with moderate to heavy infestations. However, reality often trumps theory, and the reality is that most soybean producers in the U.S. Soybean Belt states (Illinois, Indiana, Iowa, Minnesota, Ohio, Missouri, Wisconsin, and adjacent states) have been unwilling to intersperse their soybean crops with more than a year or two of a nonhost, usually corn. In the South, soybean production has declined dramatically (9). For this reason, most of what follows will emphasize SCN in the Midwestern and North Central states, identified in the preceding sentence as the Soybean Belt. Information on a wide range of issues related to SCN management may be found in the recently published Biology and Management of the Soybean Cyst Nematode, Second Edition (29). As the bulk of soybean production in the United States has shifted northward and westward, so has the known distribution of SCN (Figs. 1 and 2). The most recent discovery of SCN was in North Dakota in 2004 (3). For about the past decade, SCN has been considered the most economically important pathogen of soybean in the United States (7,37,38). Since the last time a feature article appeared in Plant Disease about SCN (36), some changes have occurred in our thinking about this pathogen and in approaches to managing soybean in infested fields. These changes include recommendations for scouting, the meaning and use of damage thresholds, and the means of classifying field populations according to virulence phenotypes (“races”). Some of what is included in this article is still very much a matter of debate, and readers are invited to check with their favorite nematologists regarding varying opinions, or with recent reviews (21,29).

Reverse Transcription Loop-Mediated Isothermal Amplification of DNA for Detection of Potato virus Y.

Abstract: A reverse transcription loop-mediated isothermal amplification of DNA (RT-LAMP) for detection of Potato virus Y (PVY) was developed. In this procedure, a set of four primers matching a total of six sequences of the coat protein (CP) gene of PVY were designed in such a way that a loop could be formed and elongated during DNA amplification. Using PVY CP complementary DNA clones as templates, the LAMP reaction was optimized by adjusting the concentrations of MgSO4, dNTPs, and Bst DNA polymerase. The effects of fragment length of target DNA on LAMP also were investigated. Two-step and one-step RT-LAMPs were performed using RNA extracts of various PVY cultures, and the results were correlated with two-step reverse transcription polymerase chain reaction (RT-PCR) for detection of PVY. Further, the turbidity caused by precipitation of magnesium pyrophosphate formed in positive RT-LAMP reactions was used to measure the amplification by utilizing a time-saving spectrophotometric method. The one-step RT-LAMP-turbidity method gave results comparable with the two-step RT-PCR method for detection of PVY from potato leaf and tuber samples. Of the total 240 samples, 234 were diagnosed similarly by both methods.

Characterization of California Isolates of Fusarium oxysporum f. sp. vasinfectum

Abstract: Thirty isolates of Fusarium oxysporum f. sp. vasinfectum from California, Australia, China, and the American Type Culture Collection were characterized by partial sequences of translational elongation factor (EF-1α), phosphate permase (PHO), and beta-tubulin (BT) genes, restriction digests of the intergenic spacer (IGS) region of nuclear rDNA, and pathogenicity tests. Based on phylogenetic analysis of combined sequences of EF-1α, PHO, and BT genes, California isolates represented four lineages. Lineage I contained race 3, lineage II contained races 1, 2, and 6, lineage III contained race 8, and lineage IV contained race 4. The Australian isolates formed a strongly supported independent clade. There were nine haplotypes based on restriction digests of the IGS region. In greenhouse pathogenicity tests with California isolates, those from the race 4 lineage were highly aggressive on certain Pima cotton (Gossypium barbadense) cultivars and less aggressive on Upland cotton (Gossypium hirsutum) cultiva...

Evaluation of Thymol as Biofumigant for Control of Bacterial Wilt of Tomato Under Field Conditions

Abstract: Volatile plant essential oils thymol and palmarosa oil, used at a concentration of 0.7%, were evaluated under field conditions for control of bacterial wilt of tomato caused by Ralstonia solanacearum. The experimental fields were artificially infested with the bacterial pathogen. Two hours after infestation, the plant essential oils were applied, then the plots were sealed with plastic mulch for 3 or 6 days. Tomato seedlings were transplanted into the field 7 days later. In fall of 2002, 92.5% of tomato plants (cv. Equinox) wilted in the untreated control plots. Both thymol and palmarosa oil soil treatments reduced bacterial wilt incidence significantly. Thymol was more effective than palmarosa oil based on the final assessment, when 33.1 and 48.1% of the plants had wilted in plots treated with thymol and palmarosa oil, respectively. Soil treatment with either thymol or palmarosa oil produced significantly higher yield of tomato than the untreated control. In 2003, only thymol was evaluated. Thymol application significantly reduced bacterial wilt incidence on the susceptible cultivar Solar Set. Disease incidence in untreated plots reached 65.5%, while in thymol treated plots only 12% of plants wilted. Thymol treatment also increased yield of Solar Set significantly compared with the untreated control. This is the first report on the use of thymol for controlling a plant disease under field conditions, which indicated that this compound provided effective control of bacterial wilt on susceptible tomato cultivars when used as preplant treatment of soils. Because of its volatile property and broad-spectrum functions, thymol shows potential to be used as a soil biofumigant for the management of various plant pathogens.

Epidemiology and Chemical Control of Soybean Rust in Southern Africa.

Abstract: Phakopsora pachyrhizi was discovered on soybeans in Uganda in 1996. This was the initial confirmation of the pathogen on soybeans in Africa, although there had been earlier unsubstantiated listings on other legumes. Thereafter, it was wind-dispersed southward to Rwanda, Zimbabwe, and Zambia in February 1998, where it severely damaged commercial plantings. It also devastated small-scale fields in eastern Nigeria at about this time. Rust continued its southward movement to southern Mozambique in early 2000, and into eastern South Africa in March 2001. By early 2003, substantial losses were being reported from western Cameroon. Scientists in Zimbabwe and South Africa have coordinated their research to combat the pathogen and have developed a strategy based on the effective, economical use of fungicides and the development of resistant germ plasm. The chemical and spraying recommendations resulting from field studies are discussed in relation to their practicalities, and a preliminary analysis of the meteorological data recorded will show the fundamental factors that influence the development of an epidemic.

Genetics of Chickpea Resistance to Five Races of Fusarium Wilt and a Concise Set of Race Differentials for Fusarium oxysporum f. sp. ciceris

Abstract: Sharma, K. D., Chen, W., and Muehlbauer, F. J. 2005. Genetics of chickpea resistance to five races of Fusarium wilt and a concise set of race differentials for Fusarium oxysporum f. sp. ciceris. Plant Dis. 89:385-390. Genetics of resistance in chickpea accession WR-315 to Fusarium wilt was investigated, and a concise set of differentials was developed to identify races of Fusarium oxysporum f. sp. ciceris. A population of 100 F7 recombinant inbred lines (RILs) from a cross of WR-315 (resistant) and C-104 (susceptible) was used to study genetics of resistance to races 1A, 2, 3, 4, and 5 of F. oxysporum f. sp. ciceris, and a population of 26 F2 plants from a cross between the same two parents was used to study inheritance of resistance to race 2. Segregations of the RILs for resistance to each of the five races suggest that single genes in WR-315 govern resistance to each of the five races. A 1:3 resistant to susceptible ratio in the F2 population indicated that resistance in WR-315 to race 2 was governed by a single recessive gene. A race-specific slow disease progress reaction was observed in chickpea line FLIP84-92C(3) to infection by race 2, a phenomenon termed as slow wilting, that is different from previously reported late wilting with respect to latent period, disease progress rate, and final disease rating. Twenty-nine germ plasm lines (27 Cicer arietinum and two C. reticulatum) including previously used differentials were evaluated for their reactions to infection by the five races. Only eight of the 29 germ plasm lines differentiated at least one of the five races based on either resistant or susceptible reactions, whereas the remaining germ plasm lines were either susceptible or resistant to all five races or differentiated them by intermediate reactions. A concise set of eight chickpea lines comprised of four genotypes and four F7 RILs with vertical resistance was developed as differentials for race identification in F. oxysporum f. sp. ciceris. These differential lines were characterized by early appearance of wilt symptoms, and clear and consistent disease phenotypes based on no wilt or 100% wilt incidence, which offers important improvements over previously available differential sets and provides more precise and unambiguous identification of the races.

Influence of pH and NaHCO3 on Effectiveness of Imazalil to Inhibit Germination of Penicillium digitatum and to Control Postharvest Green Mold on Citrus Fruit

Abstract: In vitro, spores of Penicillium digitatum germinated without inhibition between pH 4 and 7, but were inhibited at higher pH. Estimated concentrations of imazalil (IMZ) in potato-dextrose broth-Tris that caused 50% reduction in the germination of spores (ED50) of an IMZ-sensitive isolate M6R at pH 4, 5, 6, and 7 were 0.16, 0.11, 0.015, and 0.006 μg/ml, respectively. ED50 IMZ concentrations of an IMZ-resistant isolate D201 at pH 4, 5, 6, and 7 were 5.9, 1.4, 0.26, and 0.07 μg/ml, respectively. The natural pH within 2-mm-deep wounds on lemon was 5.6 to 5.1 and decreased with fruit age. IMZ effectiveness to control green mold and its residues increased with pH. The pH in wounds on lemon fruit 24 h after immersion in 1, 2, or 3% NaHCO3 increased from pH 5.3 to 6.0, 6.3, and 6.7, respectively. NaHCO3 dramatically improved IMZ performance. Green mold incidence among lemon fruit inoculated with M6R and treated 24 h later with IMZ at 10 μg/ml, 1% NaHCO3, or their combination was 92, 55, and 22%, respectiv...

Integration of Biological Control Agents and Systemic Acquired Resistance Inducers Against Bacterial Spot on Tomato.

Abstract: Two strains of plant growth-promoting rhizobacteria, two systemic acquired resistance inducers (harpin and acibenzolar-S-methyl), host-specific unformulated bacteriophages, and two antagonistic bacteria were evaluated for control of tomato bacterial spot incited by Xanthomonas campestris pv. vesicatoria in greenhouse experiments. Untreated plants and plants treated with copper hydroxide were used as controls. The plant growth-promoting rhizobacteria or a tap water control were applied as a drench to the potting mix containing the seedlings, while the other treatments were applied to the foliage using a handheld sprayer. The plant growth-promoting rhizobacteria strains, when applied alone or in combination with other treatments, had no significant effect on bacterial spot intensity. Messenger and the antagonistic bacterial strains, when applied alone, had negligible effects on disease intensity. Unformulated phage or copper bactericide applications were inconsistent in performance under greenhouse conditions against bacterial spot. Although acibenzolar-S-methyl completely prevented occurrence of typical symptoms of the disease, necrotic spots typical of a hypersensitive reaction (HR) were observed on plants treated with acibenzolar-S-methyl alone. Electrolyte leakage and population dynamics experiments confirmed that acibenzolar-S-methyl-treated plants responded to inoculation by eliciting an HR. Application of bacteriophages in combination with acibenzolar-S-methyl suppressed a visible HR and provided excellent disease control. Although we were unable to quantify populations of the bacterium on the leaf surface, indirectly we determined that bacteriophages specific to the target bacterium reduced populations of a tomato race 3 strain of the pathogen on the leaf surface of acibenzolar-S-methyl-treated plants to levels that did not induce a visible HR. Integrated use of acibenzolar-S-methyl and phages may complement each other as an alternative management strategy against bacterial spot on tomato.

Seedling and Adult Plant Resistance to Powdery Mildew in Chinese Bread Wheat Cultivars and Lines

Abstract: Powdery mildew, caused by Blumeria graminis f sp tritici, is a widespread wheat disease in China Identification of race-specific genes and adult plant resistance (APR) is of major importance in breeding for an efficient genetic control strategy The objectives of this study were to (i) identify genes that confer seedling resistance to powdery mildew in Chinese bread wheat cultivars and introductions used by breeding programs in China and (ii) evaluate their APR in the field The results showed that (i) 98 of 192 tested wheat cultivars and lines appear to have one or more resistance genes to powdery mildew; (ii) Pm8 and Pm4b are the most common resistance genes in Chinese wheat cultivars, whereas Pm8 and Pm3d are present most frequently in wheat cultivars introduced from CIMMYT, the United States, and European countries; (iii) genotypes carrying Pm1, Pm3e, Pm5, and Pm7 were susceptible, whereas those carrying Pm12, Pm16, and Pm20 were highly resistant to almost all isolates of B graminis f sp tritici tested; and (iv) 22 genotypes expressed APR Our data showed that the area under the disease progress curve, maximum disease severity on the penultimate leaf, and the disease index are good indicators of the degree of APR in the field It may be a good choice to combine major resistance genes and APR genes in wheat breeding to obtain effective resistance to powdery mildew

Outbreak of Pitch Canker Caused by Fusarium circinatum on Pinus spp. in Northern Spain.

Abstract: During the winter of 2003-2004, dieback symptoms were observed on Pinus radiata and P. pinaster in pine nurseries in Asturias (northern Spain). Small groups of affected seedlings appeared randomly distributed throughout the nurseries. The seedlings died rapidly, showing basal needle dieback, stem lesions, resin exudations, and wilting. Isolations from infected material onto potato dextrose agar (PDA) supplemented with 0.5 mg/ml of streptomycin sulfate and Komada's medium consistently yielded Fusarium sp. cultures. The isolates were transferred to PDA and Spezieller Nahrstoffarmer agar and incubated at 25°C for 10 days with a 12-h photoperiod. The cultures were identified as Fusarium circinatum Nirenberg & O'Donnell (= Fusarium subglutinans Wollenweb. & Reinking), causal agent of pitch canker disease, on basis of the presence of polyphialides and characteristic sterile, coiled, hyphae (2). To further confirm their identity, a polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFL...

Genetic Structure of Botrytis cinerea Populations from Different Host Plants in California

Abstract: The population structure of Botrytis cinerea was investigated by using transposable elements, DNA fingerprinting generated by microsatellite primed-polymerase chain reaction (MP-PCR), and sensitivity to the hydroxyanilide fungicide, fenhexamid, for 234 isolates collected from fig, grape, kiwifruit, pea, and squash in California. Among 234 isolates tested, 195 had two transposable elements, Boty and Flipper (transposa type), 38 had only the Boty element (Boty type), and one had neither of these elements (vacuma type). Four of these 234 isolates, which belonged to the Boty type, were resistant to fenhexamid. A phenogram generated based on MP-PCR markers showed that the isolates were not clustered based on their source hosts or the presence of transposable elements. Analysis of molecular variance (AMOVA) showed that there were no significant genetic differentiations among isolates collected from grape, kiwifruit, pea, and squash at the Kearney Agricultural Center. A more detailed analysis based on AMOVA partition of the total genetic variance indicated that 96% of the variation occurred within populations. The parsimony tree length permutation (PTLPT) and index of association ( IA) analyses of MP-PCR data set were consistent with absence of sexual recombination in sampled populations of this pathogen.

Application of Acibenzolar-S-Methyl Enhances Host Resistance in Tomato Against Ralstonia solanacearum

Abstract: The chemical elicitor acibenzolar-S-methyl (ASM; Actigard 50 WG), which induces systemic acquired resistance (SAR), was investigated to determine the effect on bacterial wilt of tomato caused by Ralstonia solanacearum on moderately resistant cultivars under greenhouse and field conditions. In greenhouse experiments, ASM was applied as foliar spray and/or soil drench (3 μg/ml) before and as foliar spray (30 μg/ml) after transplanting. The chemical elicitor was ineffective in reducing bacterial wilt incidence on susceptible tomato cultivars Equinox and FL 47 when plants were inoculated with R. solanacearum. However, greenhouse studies indicated that ASM significantly enhanced resistance in cultivars with moderate resistance to bacterial wilt such as Neptune and BHN 466. It appeared that ASM-mediated resistance was partially due to prevention of internal spread of R. solanacearum toward upper stem tissues of tomato plants. The effect of ASM on moderately resistant cultivars was consistent in field e...

Distribution and Characterization of Rice yellow mottle virus: A Threat to African Farmers.

Abstract: Africa produces only 2.7% of the world’s rice and is the second-largest riceimporting region in the world (6.5 Mt in 2003). This amount represents about 25% of the world rice importation (40). With an average of 2 t/ha, excluding Egypt, rice production in Africa remains significantly below the world average (Asia 3.8 t/ha, Latin America 3.0 t/ha, United States 7.0 t/ha; Food and Agriculture Organization of the United Nations [FAO], published online). Insects and diseases are the two major constraints limiting rice production in Africa and Asia. Of all the rice diseases, the one caused by the Rice yellow mottle virus (RYMV), discovered in Kenya in 1970, is one of the most damaging in Africa. RYMV has been reported in many countries in East and West Africa, where in some cases whole fields have been devastated. RYMV has only been reported from the African continent and is endemic in every country where it has been reported. RYMV is a member of the genus Sobemovirus and possesses all the characteristic biophysical and biological properties of the members of the genus. The major insect vectors are chrysomelids, which play an essential role in primary infection, while secondary infection is due mainly to plant-to-plant contact. The virus particles are stable in infected dried leaves. Limitations to the spread of the virus include restricted mobility of the insect vectors, a limited host range, and absence of seed transmission. Partially resistant and highly resistant varieties have been identified, but currently the disease is not controlled adequately, and its incidence is increasing significantly in Africa. Due to the importance of rice as a staple food, the scientific community is active in studying the virus and the resistant mechanisms in rice. At the Institut de Recherche pour le Developpement (IRD) and the International Laboratory for Tropical Agricultural Biotechnology (ILTAB), different tools, such as RYMV polyclonal and monoclonal antibodies, have been produced for RYMV detection; and a series of studies has been initiated for a better understanding of the virus–host interactions. The synthesis of an infectious clone, the sequencing of many isolates of the virus, and structural studies of virus particles permitted a study of the function of the different viral proteins and an increased understanding of the virus cycle. African virologists also have initiated studies on the epidemiology of the RYMV, mainly the virulence and variability of virus strains collected throughout Africa (36,45). The relationships between genetic variability of the virus and geographical distances have been studied (19). Currently, phytopathologists are working in the field to better assess the agronomic impact of the virus (36,61,62). Breeders and geneticists at IRD and West African Rice Development Association (WARDA) in Cote d’Ivoire are working together to identify and isolate natural resistance genes (6,41). The International Institute for Tropical Agriculture (IITA) in Nigeria undertook breeding programs in Africa. Molecular biologists are offering new ways of controlling the disease through production of transgenic rice plants (37,52). This article is intended to provide a review of the disease and give an update on recent research results. The disease’s impact on rice production in Africa, its distribution, symptomatology, and epidemiology, the physical and genetic characteristics of the virus, and resistance mechanisms and breeding efforts for resistance will be discussed.

Characterization of the tufB-secE-nusG-rplKAJL-rpoB Gene Cluster of the Citrus Greening Organism and Detection by Loop-Mediated Isothermal Amplification

Abstract: Okuda, M., Matsumoto, M., Tanaka, Y., Subandiyah, S., and Iwanami, T. 2005. Characterization of the tufB-secE-nusG-rplKAJL-rpoB gene cluster of the citrus greening organism and detection by loop-mediated isothermal amplification. Plant Dis. 89:705-711. Thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) was performed to amplify the uncharacterized regions adjacent to the nusG-rplKAJL-rpoB gene cluster of citrus greening organism (GO) isolates from different locations in Japan and Indonesia. Conventional PCR was used to amplify the internal nusG-rplKAJL-rpoB gene cluster of these isolates, and the complete sequence of this 6.1-kb fragment was determined. Comparisons with other bacterial sequences showed that the fragment is the tufB-secE-nusG-rplKAJL-rpoB gene cluster. The organization of this gene cluster is similar to that of the homologous cluster found in Escherichia coli. Except for three nucleotide changes, the sequence was identical among Japanese and Indonesian isolates. A loop-mediated isothermal amplification (LAMP) assay based on the conserved sequence of the nusG-rplKAJL-rpoB gene cluster was developed for the detection of the GO. The LAMP product was rapidly detected on nylon membranes by staining with AzurB. LAMP could detect as low as about 300 copies of the nusG-rplKAJL-rpoB fragment of the Japanese and Indonesian isolates of GO. The LAMP-based detection method, which does not depend upon a thermal cycler and electrophoresis apparatus, will be useful for under-equipped laboratories, including those found in extension centers and quarantine offices. Additional keywords: Diaphorina citri, quantitative PCR

Pathogenicity of Fungi Associated with the Wheat Crown Rot Complex in Oregon and Washington.

Abstract: Crown rot of wheat in the Pacific Northwest is caused by a complex including Bipolaris sorokiniana, Fusarium avenaceum, F. culmorum, F. pseudograminearum, and Microdochium nivale. Relative pathogenicity was examined under greenhouse conditions for 178 isolates of the five species, and under field conditions for 24 isolates of B. sorokiniana, F. culmorum, and F. pseudograminearum. In the greenhouse, all five species reduced (P < 0.05) plant height relative to noninoculated controls. Disease severity was inversely correlated with plant height for the three Fusarium spp. In one or more of four experiments with spring wheat in the field, all three species reduced stand establishment and density of mature heads and increased the incidence and severity of crown rot. F. culmorum and F. pseudograminearum caused the greatest disease severity and plant damage and were the only pathogens that reduced grain yield. Virulence ratings were variable among isolates for each species in all greenhouse and field experiments. Isolate variability was especially high for the location and year variables in field experiments. Mixtures of multiple isolates are required for future research.

Pathogenicity of Fusarium Species on Different Plant Parts of Spring Wheat Under Controlled Conditions.

Abstract: Most of the Fusarium species responsible for Fusarium head blight of wheat in Saskatchewan, Canada, have also been isolated from discolored subcrown internodes/crowns of wheat. It was therefore of interest to compare the susceptibility of heads and ground/underground tissue of wheat to isolates of Fusarium species from different sources. Controlled-environment pathogenicity tests were conducted on heads, seeds, and seedlings of spring wheat. Overall, F. culmorum and F. graminearum were the most pathogenic species, although the former was more pathogenic than the latter. F. equiseti and F. poae were the least pathogenic species, whereas F. avenaceum had intermediate pathogenicity in the head and seed tests, but low pathogenicity in the seedling test. There was a similar pathogenicity among isolates of each Fusarium species from different sources to heads and ground/underground plant parts, indicating a lack of adaptation of these isolates. Our observations suggest that Fusarium inoculum on or in infected seed or plant debris might infect plants at or below soil level, which could then become a source of inoculum for infection of heads in the following season(s). Survival of fungal inoculum in underground plant parts might be important during dry conditions.

A New Method for the Quantification of Rhizoctonia solani and R. oryzae from Soil

Abstract: Paulitz, T. C., and Schroeder, K. L. 2005. A new method for the quantification of Rhizoctonia solani and R. oryzae from soil. Plant Dis. 89:767-772. Rhizoctonia solani anastomosis group (AG) 8 and R. oryzae are important root pathogens on wheat and barley in the dryland production areas of the inland Pacific Northwest. R. solani AG-8 is difficult to isolate from root systems and quantify in soil because of slow growth and low population densities. However, both pathogens form extensive hyphal networks in the soil and can grow a considerable distance from a food base. A quantitative assay of active hyphae was developed, using wooden toothpicks as baits inserted into sample soils. After 2 days in soil, toothpicks were placed on a selective medium, and the numbers of colonies that grew after 24 h were counted under a dissecting microscope. R. solani and R. oryzae could be distinguished from other fungi based on hyphal morphology. This method was tested in natural soils amended with known inoculum densities of R. solani AG-8 and R. oryzae. Regressions were used to compare the inoculum density or toothpick colonization curves to a predicted curve based on the volume of the toothpicks. The slopes and y intercept of log-log transformed regressions did not differ from the predicted curves in most cases. This technique was used to assess the hyphal activity of R. solani AG-8 and R. oryzae from soil cores taken from various positions in and around Rhizoctonia bare patches at two locations. Activity of R. solani was highest in the center and inside edge of the patch, but there was no effect of patch position on R. oryzae. This simple and inexpensive technique can be used for detection and diagnosis in grower fields and to study the ecology and epidemiology of Rhizoctonia spp.

Molecular, Physiological, and Host-Range Characterization of Acidovorax avenae subsp. citrulli Isolates from Watermelon and Melon in Israel.

Abstract: Burdman, S., Kots, N., Kritzman, G., and Kopelowitz, J. 2005. Molecular, physiological, and host-range characterization of Acidovorax avenae subsp. citrulli isolates from watermelon and melon in Israel. Plant Dis. 89:1339-1347. Bacterial fruit blotch (BFB), caused by Acidovorax avenae subsp. citrulli, is a serious disease of cucurbit plants. The first important occurrence of BFB in Israel was during 2000 to 2003 on watermelon and melon. Twelve bacterial isolates associated with these outbreaks were confirmed as A. avenae subsp. citrulli by pathogenicity assays, gas chromatography of fatty-acid methyl esters, and substrate-utilization profiles. The isolates were characterized in terms of their aggressiveness in different hosts by seed, seedling, and fruit inoculations, and according to their DNA fingerprinting profiles using pulse-field gel electrophoresis (PFGE) and repetitive-PCR approaches. Results from the present work agree with previous studies supporting the existence of two differentiated groups within A. avenae subsp. citrulli, one including strains that are more associated with watermelon (group II), the other consisting of strains that are usually associated with nonwatermelon cucurbits (group I). This study indicates that isolates from both groups have been introduced to Israel. PFGE analysis revealed that the 12 analyzed isolates can be divided into five different haplotypes, of which four were previously unreported. Additional differentiating features between group I and II strains are presented. Additional keywords: Citrullus lanatus, Cucumis melo, Cucurbitaceae

Use of Boron for the Control of Eutypa Dieback of Grapevines.

Abstract: Eutypa dieback is a perennial canker disease of grapevine (Vitis vinifera) caused by Eutypa lata. The fungus produces ascospores, which infect grapevines through pruning wounds during the dormant season. Management of the disease has been achieved with fungicide applications during the dormant period. However, no effective fungicide was available for this purpose after Benlate was withdrawn from the market. Boric acid (17.5% a.i. boron), a potential alternative to Benlate, was tested in the present study against E. lata. The EC50 values for inhibition of mycelial growth and ascospore germination were 125 and 475 μg of boric acid per ml (22 and 83 μg a.i./ml), respectively. Two boron-based treatments were developed and tested in vitro and in four field trials during 2001 to 2003. One product, biopaste, contained 5% boric acid (8.75 mg a.i./ml) in a commercial paste. The second product, bioshield, contained 5% boric acid in a spore suspension of Cladosporium herbarum. Both products significantly re...

Physiologic Specialization of Puccinia triticina on Wheat in the United States in 2003.

Abstract: Collections of Puccinia triticina were obtained from rust infected wheat leaves by cooperators throughout the United States and from surveys of wheat fields and nurseries in the Great Plains, Ohio Valley, Southeast, California, and the Pacific Northwest, in order to determine the virulence of the wheat leaf rust fungus in 2003. Single uredinial isolates (580 in total) were derived from the wheat leaf rust collections and tested for virulence phenotype on lines of Thatcher wheat that are near-isogenic for leaf rust resistance genes Lr1, Lr2a, Lr2c, Lr3, Lr9, Lr16, Lr24, Lr26, Lr3ka, Lr11, Lr17, Lr30, LrB, Lr10, Lr14a, and Lr18. In the United States in 2003, 52 virulence phenotypes of P. triticina were found. Virulence phenotype MBDS, which has been selected by virulence to resistance gene Lr17, was the most common phenotype in the United States. MBDS was found in the Southeast, Great Plains, the Ohio Valley, and California. Virulence phenotype THBJ, which has been selected by virulence to genes Lr...

Comparison of Visual and Multispectral Radiometric Disease Evaluations of Cercospora Leaf Spot of Sugar Beet

Abstract: Visual assessments of disease severity are currently the industry standard for quantification of the necrosis caused by Cercospora beticola on sugar beet (Beta vulgaris) leaves. We compared the precision, reproducibility, and sensitivity of a multispectral radiometer to visual disease assessments. Individual wavebands from the radiometer, as well as vegetative indices calculated from the individual wavebands, were compared with visual disease estimates from two raters at each of two research sites. Visual assessments and radiometric measurements were partially repeated immediately after the first assessment at each site. Precision, as measured by reduced coefficients of variation, was improved for all vegetative indices and individual waveband radiometric measures compared with visual assessments. Visual assessments, near-infrared single-waveband reflectance values, and four of the six vegetative indices had high treatment F values, suggesting greater sensitivity at discriminating disease severity levels. Reproducibility, as measured by a test-retest method, was high for visual assessments, single-waveband reflectance at 810 nm, and several of the vegetative indices. The use of radiometric methods has the potential to increase the precision of assessments of Cercospora leaf spot foliar symptoms of sugar beet while eliminating potential bias. We recommend this method be used in conjunction with visual disease assessments to improve precision of assessments and guard against potential bias in evaluations.

Occurrence of resistance-breaking Beet necrotic yellow vein virus of sugar beet

Abstract: Rhizomania is an important virus disease of sugar beet and is caused by Beet necrotic yellow vein virus (BNYVV). During 2002-03, several sugar beet fields with cultivars partially resistant to BNYVV grown in the Imperial Valley of California were observed with severe rhizomania symptoms, suggesting that resistance conditioned by Rz1 had been compromised. Soil testing with sugar beet baiting plants followed by enzyme-linked immunosorbent assay (ELISA) was used to diagnose virus infection. Resistant varieties grown in BNYVV-infested soil from Salinas, CA, were ELISA-negative. In contrast, when grown in BNYVV-infested soil collected from the Imperial Valley, CA, all resistant varieties became infected and tested positive by ELISA. Based on host reaction, eight distinct BNYVV isolates have been identified from Imperial Valley soil (IV-BNYVV) by single local lesion isolation. Reverse transcription-polymerase chain reaction (RT-PCR) assays showed that the eight IV-BNYVV isolates did not contain RNA-5. Single-strand conformation polymorphism banding patterns for the IV-BNYVV isolates were identical to A-type and different from P-type. Sequence alignments of PCR products from BNYVV RNA-1 near the 3' end of IV-BNYVV isolates revealed that both IV-BNYVV and Salinas BNYVV isolates were similar to A-type and different from B-type. Our results suggest that the resistance-breaking BNYVV isolates from Imperial Valley likely evolved from existing A-type isolates.