Edward J. Sikora

Application of rhizobacteria for induced resistance

This article provides a review of experiments conducted over a six-year period to develop a biological control system for insect-transmitted diseases in vegetables based on induced systemic resistance (ISR) mediated by plant growth-promoting rhizobacteria (PGPR). Initial experiments investigated the factors involved in treatment with PGPR led to ISR to bacterial wilt disease in cucumber caused by Erwinia tracheiphila. Results demonstrated that PGPR-ISR against bacterial wilt and feeding by the cucumber beetle vectors of E. trachiphiela were associated with reduced concentrations of cucurbitacin, a secondary plant metabolite and powerful beetle feeding stimulant. In other experiments, PGPR induced resistance against bacterial wilt in the absence of the beetle vectors, suggesting that PGPR-ISR protects cucumber against bacterial wilt not only by reducing beetle feeding and transmission of the pathogen, but also through the induction of other plant defense mechanisms after the pathogen has been introduced into the plant. Additional greenhouse and field experiments are described in which PGPR strains were selected for ISR against cucumber mosaic virus (CMV) and tomato mottle virus (ToMoV). Although results varied from year to year, field-grown tomatoes treated with PGPR demonstrated a reduction in the development of disease symptoms, and often a reduction in the incidence of viral infection and an increase in tomato yield. Recent efforts on commercial development of PGPR are described in which biological preparations containing industrial formulated spores of PGPR plus chitosan were formulated and evaluated for use in a transplant soil mix system for developing plants that can withstand disease attack after transplanting in the field.

Plant Growth-Promoting Rhizobacterial Mediated Protection in Tomato Against Tomato mottle virus

Tomato plants treated with plant growth-promoting rhizobacteria (PGPR), applied as an industrially formulated seed treatment, a spore preparation mixed with potting medium (referred to as powder), or a combined seed-powder treatment, were evaluated under field conditions for induced resistance to Tomato mottle virus (ToMoV). The PGPR strains used, based on their ability to induce resistance in previous experiments, included Bacillus amyloliquefaciens 937a, B. subtilis 937b, and B. pumilus SE34. Experiments were conducted in the fall of 1997 and the spring and fall of 1998 at the University of Floridas Gulf Coast Research & Education Center, Bradenton. All plants were rated for symptoms and analyzed for the presence of ToMoV DNA at 40 days after transplant (dat). Whitefly densities were determined on individual plants in each trial, and marketable fruit yields were determined at least two times during each trial. The highest level of protection occurred in the fall 1997 trial when, at 40 dat, ToMoV disease severity ratings were significantly less in all PGPR powder-based treatments than in either of the seed or control treatments. Detection of viral DNA using Southern dot blot analyses correlated with symptom severity ratings, as did fruit yields. A reduction in ToMoV symptom severity ratings and incidence of viral DNA were also observed for some PGPR treatments in the spring 1998 trial, although corresponding yield responses were not apparent. Little or no resistance was observed in the fall 1998 trial. No differences in disease severity, detection of ToMoV DNA, or yield occurred among treatments in any of the trials at 80 dat. These data show that up to 40 dat under natural conditions of high levels of vector-virus pressure, some PGPR treatments resulted in reduced ToMoV incidence and disease severity and, in some cases, a corresponding increase in fruit yield. The use of PGPR could become a component of an integrated program for management of this virus in tomato.

A Multivirus Epidemic of Tomatoes in Alabama

During 1992, a multivirus epidemic reduced tomato production by as much as 25% in the major tomato-growing region of Alabama. Estimated yield losses of 100% resulted from the epidemic in over 250 ha in two counties of North Alabama. Cucumber mosaic cucumovirus (CMV), alone or in combination with potato potyvirus Y (PVY) and/or tobacco etch potyvirus (TEV), was responsible for the crop failure. CMV was detected alone or in combination with PVY and/or TEV in over 70% of the samples tested and was present in 90% of the fields surveyed. In 1993, 21 tomato fields were monitored weekly from transplanting through harvest for CMV, PVY, TEV, tobacco mosaic tobamovirus, and tomato spotted wilt tospovirus. All 5 viruses were detected, with CMV occurring most frequently. Incidence of CMV at the 61% level or higher was found in 16 of the 21 fields surveyed. Tomatoes transplanted in April and May were least effected and had relatively low virus incidence until late in their development. Tomatoes transplanted in June and July were infected at an earlier age, had the highest virus incidence, were the most severely affected, and suffered the greatest loss in yield. In 1994,Aphis gossypii, the cotton aphid, was the most common virus vector found during an aphid monitoring/virus spread study. Populations of A. gossypii peaked in late June, immediately preceding a period of rapid CMV incidence and spread.

Confirmed Distribution and Occurrence of Megacopta cribraria (F.) (Hemiptera: Heteroptera: Plataspidae) in the Southeastern United States

Abstract Megacopta cribraria (F) (Hemiptera: Heteroptera: Plataspidae) was first discovered in North America in 9 counties in northeastern Georgia (USA) in October 2009. By the end of 2012, surveys conducted in Georgia and neighboring states confirmed that the insect had spread into 383 additional counties in the southeastern U.S., including the states of Alabama, Florida, Georgia, Mississippi, North Carolina, South Carolina, Tennessee and Virginia. It was reported from 33 species of plants representing 15 taxonomic families in these surveys, with 17 of those from the family Fabaceae (legumes). Kudzu (Pueraria montana Lour. [Merr.] variety lobata [Willd.] Maesen & S. Almeida) was the most frequently reported host. All life stages of the insect were observed only on kudzu and soybean (Glycine max [L.] Merrill) which, to date, are the only confirmed reproductive host plants of M. cribraria in its expanded North American range.

A Coordinated Effort to Manage Soybean Rust in North America: A Success Story in Soybean Disease Monitoring

Existing crop monitoring programs determine the incidence and distribution of plant diseases and pathogens and assess the damage caused within a crop production region. These programs have traditionally used observed or predicted disease and pathogen data and environmental information to prescribe management practices that minimize crop loss. Monitoring programs are especially important for crops with broad geographic distribution or for diseases that can cause rapid and great economic losses. Successful monitoring programs have been developed for several plant diseases, including downy mildew of cucurbits, Fusarium head blight of wheat, potato late blight, and rusts of cereal crops. A recent example of a successful disease-monitoring program for an economically important crop is the soybean rust (SBR) monitoring effort within North America. SBR, caused by the fungus Phakopsora pachyrhizi, was first identified in the continental United States in November 2004. SBR causes moderate to severe yield losses globally. The fungus produces foliar lesions on soybean (Glycine max) and other legume hosts. P. pachyrhizi diverts nutrients from the host to its own growth and reproduction. The lesions also reduce photosynthetic area. Uredinia rupture the host epidermis and diminish stomatal regulation of transpiration to cause tissue desiccation and premature defoliation. Severe soybean yield losses can occur if plants defoliate during the mid-reproductive growth stages. The rapid response to the threat of SBR in North America resulted in an unprecedented amount of information dissemination and the development of a real-time, publicly available monitoring and prediction system known as the Soybean Rust-Pest Information Platform for Extension and Education (SBR-PIPE). The objectives of this article are (i) to highlight the successful response effort to SBR in North America, and (ii) to introduce researchers to the quantity and type of data generated by SBR-PIPE. Data from this system may now be used to answer questions about the biology, ecology, and epidemiology of an important pathogen and disease of soybean.

Increased Occurrence of Target Spot of Soybean Caused by Corynespora cassiicola in the Southeastern United States

Target spot of soybean (Glycine max (L.) Merr.) caused by Corynespora cassiicola (Berk. & Curt.), although found in most soybean-growing countries, is considered to be a disease of limited importance (1) and has never been reported to cause soybean yield loss in the southeastern United States (2,3). Soybean plants submitted to the North Carolina Plant Disease and Insect Clinic (NCPDIC) in August 2004 from Beaufort, Robeson, Wilson, and Johnston counties, NC had symptoms consistent with target spot. Symptoms consisted of roughly circular, necrotic leaf lesions from minute to 11 mm in diameter, though typically approximately 4 to 5 mm in diameter, and with a yellow margin. Large lesions occasionally exhibited a zonate pattern often associated with this disease. Microscopic examination of the lesions revealed the presence of spores (conidia) typical of C. cassiicola (1). Conidia were mostly three to five septate with a central hilum at the base and ranged in size from 7 to 22 wide × 39 to 520 μm long. Three commercial soybean fields near Blackville, SC (Barnwell County) were severely affected by this disease and it caused premature defoliation. Nineteen of twenty-seven maturity group VII and VIII genotypes in the 2004 Clemson University soybean variety trial near Blackville, SC had visible symptoms of target spot. Heavy rainfall associated with hurricanes during September 2004 probably enhanced the incidence of this disease, and yield suppression due to target spot was estimated at 20 to 40% in some fields. In 2005, 20 of 161 soybean samples submitted to the NCPDIC or collected in surveys from 16 counties were positive for target spot on the basis of microscopic examination. Target spot also was diagnosed in six counties (Baldwin, DeKalb, Elmore, Fayette, Macon, and Pickens) in Alabama and in four additional counties (Bamberg, Hampton, Orange-burg, and Calhoun) in South Carolina in 2005. Records from the NCPDIC indicate that target spot had not been diagnosed on soybean in North Carolina since 1981. The large increase in incidence of target spot in the southeast may be related to changes in weather patterns, changes in pathogen virulence, and/or the introduction of more susceptible host genotypes. References: (1) J. B. Sinclair. Target spot. Page 27 in: Compendium of Soybean Diseases. G. L. Hartman et al. eds. The American Phytopathological Society, St. Paul, MN, 1999. (2) J. A. Wrather et al. Plant Dis. 79:1076. 1995. (3) J. A. Wrather et al. On-line publication. doi:10.1094/PHP-2003-0325-01-RV. Plant Health Progress, 2003.

Performance of Virus Resistant Transgenic Yellow Summer Squash in Alabama

Abstract Production of summer squash in Alabama and the southeastern United States is generally limited to spring and early summer due to the abundance of aphid transmitted viruses during the late summer and fall. Cucumber mosaic virus (CMV), Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV), and Papaya ring spot virus (PRSV) are the most common viruses affecting Cucurbits in the southeastern United States. The objective of this study was to evaluate virus-resistant squash cultivars with naturally occurring virus disease pressure to determine if they are cost effective. Experiments were conducted in 2000 and 2002 at the E. V. Smith Research Center in Shorter, AL. Cultivars evaluated included ‘Destiny III’ a semi crookneck type with transgenic resistance to CMV, WMV and one or more strains of ZYMV; ‘Liberator III’, a yellow straight neck with transgenic resistance to CMV, WMV and ZYMV; and ‘Conqueror IIP’, a yellow greenstem straight neck with transgenic resistance to CMV, WMV and ZYMV, and intermediate resistance to PRSV. Results indicated that the transgenic cultivars produced greater marketable yields than the susceptible controls (‘Dixie’ and ‘Lemon Drip’ in 2000; ‘Dixie’ in 2002) during both years of the study under high virus pressure. Growers considering production of a yellow squash cultivar in the late summer or fall in Alabama should strongly consider using a virus-resistant yellow squash cultivar when field history and conditions suggest the likelihood of virus disease. The economic impact of growing a virus-resistant cultivar will depend on multiple factors that include seed cultivar/cost, types of viruses present during a particular growing season and their relative severity levels, and the market price of squash at the time of harvest.

Delayed Senescence in Soybean: Terminology, Research Update, and Survey Results from Growers

The terms used to describe symptoms of delayed senescence in soybean often are used inconsistently or interchangeably and do not adequately distinguish the observed symptoms in the field. Various causes have been proposed to explain the development of delayed senescence symptoms. In this article, we review published reports on delayed senescence symptoms in soybean, summarize current research findings, provide examples of terms related to specific symptoms, and present an overview of the results of a multi-state survey directed to soybean growers to understand their concerns about delayed soybean senescence. Some of these terms, such as green bean syndrome and green stem syndrome, describe symptoms induced by biotic factors, while other terms describe symptoms associated with abiotic factors. Some delayed senescence terms involve the whole plant remaining green while other terms include just the stem and other plant parts such as pods. In the grower survey, 77% reported observing soybean plants or plant parts that remained green after most plants in the field were fully mature with ripe seed. Most respondents attributed these symptoms to changes in breeding and choice of cultivars. At the end of this article, we standardized the terms used to describe delayed senescence in soybean. Disciplines Agricultural Science | Agronomy and Crop Sciences | Plant Biology | Plant Pathology Comments This article is published as Harbach, C. J., Allen, T. W., Bowen, C. R., Davis, J. A., Hill, C. B., Leitman, M., Leonard, B. R., Mueller, D. S., Padgett, G. B., Phillips, X. A., Schneider, R. W., Sikora, E. J., Singh, A. K., and Hartman, G. L. 2016. Delayed senescence in soybean: Terminology, research update, and survey results from growers. Plant Health Prog. 17:76-83. doi: 10.1094/PHP-RV-16-0008. Posted with permission. Rights Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted. Authors C. J. Harbach, T. W. Allen, C. R. Bowen, J. A. Davis, C. B. Hill, M. Leitman, B. R. Leonard, D. S. Mueller, G. B. Padgett, X. A. Phillips, R. W. Schneider, E. J. Sikora, A. K. Singh, and G. L. Hartman This article is available at Iowa State University Digital Repository: http://lib.dr.iastate.edu/agron_pubs/341

First report of soybean rust (Phakopsora pachyrhizi) on Florida Beggarweed (Desmodium tortuosum) in Alabama.

Soybean rust (SBR), caused by the fungus Phakopsora pachyrhizi, was detected on Florida Beggarweed (Desmodium tortuosum) for the first time in Alabama in November, 2009. The pathogen was not observed in 2010 or 2011, probably because of the exceptionally dry, hot weather in the region. The pathogen was observed on multiple mature leaves of plants, evenly distributed through a field at the Wiregrass Research and Extension Center in Headland, Alabama, located in the southeast region of the state. Florida Beggarweed can serve as an overwintering host for SBR. Symptoms on leaves were consistent with SBR symptoms previously described on soybeans (1). Sori in multiple pustules were observed on the undersurface of the leaves. Urediniospores and paraphyses were observed microscopically and identified as P. pachyrhizi. Symptomatic leaves from 20 plants were analyzed using an Envirologix monoclonal antibody strip test kit at the Auburn University Plant Diagnostic Laboratory. A subsample of 20 plants were positive for the pathogen. Representative symptomatic leaves were sent to the USDA Molecular Diagnostic Laboratory in Beltsville, Maryland, for confirmation. DNA was extracted from sori aseptically removed from leaves using a Qiagen DNeasy Plant Mini Kit, and amplified with primers Ppa1 and NL4. The resulting partial ITS2 and 28S ribosomal RNA sequences were 100% identical to GenBank entry DQ354537. Voucher specimens were deposited in the USDA Agricultural Research Service, National Fungus Collection (BPI). To our knowledge, this is the first report of the disease on Florida Beggarweed in Alabama. References: (1) A. Carcamo Rodriguez et al. Plant Dis. 90:1260, 2006. (2) R. D. Frederick et al. Phytopathology 92:217, 2002.

Detection of cucumber mosaic cucumovirus in weed species: a cautionary report on nonspecific reactions in ELISA

The reliability of enzyme-linked immunosorbent assay (ELISA) to detect cucumber mosaic cucumovirus (CMV) coat protein (CP) in three weed species (Taraxacum officinale, Gnaphalium obtusifolium, and Conyza canadensis) was compared with other detection methods, including Western blot analysis and a bioassay. Analysis of extracts prepared from these species by double antibody sandwich (DAS) ELISA yielded high absorbance values (A4O5nm) that were indicative of CMV concentrations of purified CMV standards of greater than 50 mg/mL. However, results from an ELISA procedure designed such that no viral antigen should be detected suggested that the strong ELISA absorbance values were nonspecific reactions with material(s) in the weed extracts. These findings were substantiated when no CMV CP was detected by Western blot analysis or infectious CMV was detected in bioassays involving inoculation of extracts onto an indicator host (Cucurbita pepo ‘Crookneck’ (squash)). In both DAS ELISA and indirect ELISA procedures, t...