2009 Caribbean Division
Meeting Abstracts
(Joint with The Florida Pathological Society)
May 16-19, 2009 - Orlando, Florida
Posted online September 29, 2009
Monitoring resistant populations of Xanthomonas citri subsp.
citri and epiphytic bacteria on young citrus trees treated with copper
or streptomycin
F. BEHLAU (2), J. B. Jones (2), J. H. Graham (1)
(1) Citrus Research and Education Center, University of Florida, Lake
Alfred, FL; (2) Plant Pathology Department, University of Florida,
Gainesville, FL
Since Florida’s citrus canker (Xanthomonas citri subsp. citri,
Xcc) eradication program was halted in 2005 attention has focused on
management strategies that include the use of bactericides such as copper
and streptomycin for disease control. Widespread use of these chemicals in
citrus industries elsewhere in the world has led to development of resistant
strains of Xcc. Cu and Sm resistance were monitored in Xcc and epiphytic
bacterial populations on citrus trees repeatedly sprayed with these
chemicals for control of citrus canker. Copper hydroxide (Cu, Kocide 3000)
or streptomycin sulphate (Sm, Firewall) were sprayed on foliage of young
‘Ray Ruby’ grapefruit every 21 days from March to October 2008. Mature
canker-symptomatic and non-symptomatic leaves were sampled monthly to assay
for resistant Xcc and epiphytic bacteria, respectively. Leaves were washed
with MGY broth + 1 mg/L of CuSO(4) for 2 hrs using 10 mL of liquid medium/g
of leaf and plated on semi-selective medium MGY-KCC + Cu or Sm for isolation
of resistant Xcc or on MGY + Cu or Sm for monitoring resistant population of
epiphytic bacteria. No Cu or Sm resistant strains of Xcc were isolated. No
major differences in total epiphytic bacterial population were observed
among treatments over time in comparison to the check. However, Cu and Sm
sprays increased the ratio of epiphytic bacterial population with resistance
to these chemicals. Overall, the Sm resistant bacterial populations were
proportionally lower than Cu resistant bacterial population.
Survival of Xanthomonas citri subsp. citri on symptomatic
fruit under prolonged ambient and cold storage conditions
G. Bonn (1), E. TAYLOR (3), T. Riley (2), T. Gottwald (3), T. Schubert
(1)
(1) Florida Department of Agriculture & Consumer Services, Division of
Plant Industry, Gainesville, FL; (2) USDA, APHIS, PPQ, Orlando, FL; (3)
USDA, ARS, US Horticultural Research Laboratory, Ft. Pierce, FL
Live cells of Xanthomonas citri subsp. citri (Xcc)
were detected by excising canker lesions from commercial fresh-packed
grapefruit, macerating them in phosphate buffer followed by dilution plating
onto a semi-selective agar medium (KCB). After 4–5 days incubation at 28°C,
separate colonies were counted at 100X using a dissecting microscope.
Confirmation of Xcc was by the use of Agdia’s ImmunoStrip® for suspect plate
colonies and a bioassay by needleless infiltration of leaf lamina of young
‘Duncan’ grapefruit seedlings. Xcc was detected from lesions on fruit
held at both ambient and 5°C for up to 100 days in storage. There was a
general trend for the percentage of Xcc positive lesions and actual
bacterial populations to decrease with storage time. Populations of Xcc
decreased faster at ambient temperature than at 5°C, possibly due to the
higher metabolic activity of cells or microbial competition at the elevated
ambient temperature. The low numbers of viable canker bacteria associated
with peel lesions in grapefruit, especially over time, suggest that the risk
of canker transmission from them is extremely low in comparison to active
lesions on the leaves, stems and fruit during the growing season.
Distribution of Candidatus Liberibacter asiaticus in huanglongbing
infected citrus trees
R. H. BRLANSKY (1), K. S. Pelz-Stelinski (1)
(1) University of Florida, CREC
The distribution of the huanglongbing (HLB) associated Candidatus
Liberibacter asiaticus (Las) bacterium in mature field infected citrus trees
was evaluated. The number of tissue samples collected per tree ranged from
16–32 and included: fruit peduncles, bark phloem, and symptomatic leaf
midribs and petioles taken from throughout the tree canopy. Overall, the
percentage of Las-positive plant tissue samples obtained ranged from 23–44%
based on real-time PCR results. Although samples taken from bark phloem
varied, phloem from one-year old bark consistently tested positive for Las.
Similar variation in the detection of Las occurred in samples obtained from
leaf petioles and fruit peduncles. The percentage of positive leaf petioles
ranged from 0–100%, while fruit peduncles ranged from 17–56%. Additional
replicates continue to be collected in order to firmly establish whether
one-year old bark phloem is consistently positive when another part of the
tree has tested positive for Las via PCR. These results indicate that
distribution of the HLB-associated pathogen varies widely within
symptomatic, PCR-positive citrus trees and thus illustrate the importance of
obtaining multiple samples from trees where an infection is suspected.
Infectious clones and characterization of a previously unreported
bean-infecting begomovirus from Rynchosia minima (L.), an endemic
legume species from Puerto Rico
J. K. BROWN (2), M. Rehman (1), A. M. Idris (2)
(1) Department of Microbiology, Hazara University, Mansehra, Pakista; (2)
Department of Plant Sciences, The University of Arizona, Tucson, AZ, USA
R. minima plants exhibiting mild mosaic symptoms that were
reminiscent of begomovirus infection were observed in Puerto Rico during the
summer, 1997. Total nucleic acids were extracted from symptomatic R.
minima leaves using the CTAB method. The viral single-stranded DNA was
subjected to rolling circle amplification. The SacI-linearized,
multimeric DNA band was ligated into SacI-digested pGEM7Zf+ and
cloned. Eight clones bearing a ~2.6 kbp fragment were sequenced using primer
walking. Analysis of the resultant sequences indicated that five and three
of the clones were DNA-A and DNA-B components, respectively. The genome
organization and number of open reading frames (six) was typical of other
bipartite begomoviral genomes from the Western Hemisphere. Comparative
analysis of the DNA-A (n = 5) and DNA-B (n = 3) component sequences shared
98–99% and 99% nucleotide (nt) identity, respectively. Inspection of the
common region (CR) revealed that they were cognate components, and shared an
identical iteron. The DNA-A component shared 80% nt identity with its
closest relatives, Macroptilium mosaic Puerto Rico virus and
Rhynchosia golden mosaic virus. The DNA-B component shared 64% and 62%
nt identity with RhGMV and Cabbage leaf curl virus, respectively.
This previously undescribed begomovirus species is herein named Rhynchosia
mild mosaic virus (RhMMV). Clones of the RhGMV DNA-A and DNA-B component
were released with SacI and biolistically inoculated to R. minima and
bean (Phaselous vulgaris) seedlings. R. minima seedlings
developed mild mosaic symptoms like those observed in field-infected bean
plants, thereby fulfilling Koch’s postulates, and the bean seedlings
developed severe green-yellow mosaic symptoms, confirming that the virus
also infects bean. This previously undescribed virus could pose a serious
threat to bean crops in the Caribbean region.
Developing an effective international education program for management of
Ralstonia solanacearum Race 3 biovar 2
P. G. CHAMPOISEAU (1), J. B. Jones (1), C. Allen (3), T. M. Momol (2)
(1) University of Florida, Department of Plant Pathology, Gainesville,
FL, USA; (2) University of Florida, District Directors Office, Gainesville,
FL, USA; (3) University of Wisconsin-Madison, Department of Plant Pathology,
Madison, WI, USA
Because it threatens both potato and ornamental production, Ralstonia
solanacearum Race 3 biovar 2 (R3bv2) is considered a serious quarantine
pest in Canada and Europe and is listed as a Select Agent plant pathogen in
the United States, where it is subject to the strictest biosecurity
regulations. Although this pathogen is not known to be established in the
US, import of infected geranium cuttings from off-shore production sites has
already proved to be a possible pathway for introduction. Previous
accidental introductions resulted in multi-million dollar losses due to
quarantine responses. Therefore it is critical to prevent further
re-introduction and possible spread of R3bv2 in the US. This involves
exclusion, early detection, and unambiguous identification of the pathogen
at both national and international levels. This can be achieved by use of
reliable diagnostic tools for the pathogen and effective phytosanitary
measures; however, this is not enough. It is also essential to ensure
preparedness and effective training of official regulators, diagnosticians
and other individuals responsible for first detection and response to a
possible R3bv2 discovery in the US. We have therefore developed an
integrated education and outreach program, as part of a USDA-founded project
for a better management of R3bv2. This program involves development of
educational and training content by a team of experts, delivery of
educational materials to target audiences by diverse means including current
web-based technologies, as well as use of various evaluation tools to assess
program effectiveness. Monitored access of our Ralstonia/bacterial
wilt-dedicated website shows that stakeholders from diverse organizations
both within and outside the US regularly use this resource to obtain updated
and accurate information on R. solanacearum R3bv2 and bacterial wilt
disease management.
Nitrocellulose membranes as a solid matrix for Cucumber mosaic virus
immuno-detection and subgroup identification by RT-PCR
P. S. CHANG (1), S. A. Tolin (1)
(1) Virginia Polytechnic Institute and State University, Blacksburg, VA,
USA
Cucumber mosaic virus (CMV) is an important and widespread plant
virus. The strains and isolates of CMV are highly diverse and assigned to
either subgroup 1A, 1B or 2. Here we report the application of tissue blot
immunoassay (TBIA) followed by reverse transcription-polymerase chain
reaction (RT-PCR) for the immuno-detection and subgroup identification of
CMV from various hosts and locations. Freshly torn leaves were blotted onto
nitrocellulose membranes (NCM), which were used as sources of viral RNA
after processing by TBIA. CMV positive samples show a purple precipitate at
the blot site. A 3 mm disc was removed from the positive sites and cleaned
by rinsing with Triton X-100, followed by TE buffer, then dried and added
directly to RT reactions that included a reverse primer to the CMV coat
protein (CP) gene. The resulting cDNA was added to PCR reactions containing
forward and reverse primers for the CMV CP gene. Agarose gel electrophoresis
revealed amplicons of the expected size. The subgrouping of CMV samples was
predicted from sequences of PCR products and confirmed by monoclonal
antibodies specific to CMV subgroups 1 and 2. Successful amplification was
possible from NCM blotted and TBIA processed up to 15 months previously, but
amplification levels from older blots were lower. The PCR protocol was
adjusted by increasing the number of cycles for consistent results from
blots older than 8 months. This method eliminates the need for leaf tissue
storage or costly RNA extraction when sampling for CMV diversity or
monitoring virus prevalence and incidence. NCM are thus a suitable matrix
for obtaining viral RNA for RT-PCR and archival storage of viral nucleic
acids, similar to Whatman’s FTA® Plant Cards.
Sugarcane orange rust, an emerging disease in the western hemisphere
J. C. COMSTOCK (1), N. C. Glynn (1), L. A. Castlebury (2)
(1) USDA-ARS Sugarcane Field Station, Canal Point, FL; (2) Yystematic
Mycology and Microbiology Laboratory, ARS, USDA, Beltsville, MD
Symptoms consistent with sugarcane orange rust were first observed in
Florida in June 2007, these were subsequently confirmed morphologically and
molecularly as being caused by Puccinia kuehnii, the causal agent of
orange rust. This was the first documented occurrence of sugarcane orange
rust in the Western Hemisphere. Since then it has been reported in
Guatemala, Costa Rica and Nicaragua and has been confirmed in several other
Central American and Caribbean Countries. A comparison of brown rust and its
causal agent, P. melanocephala and P. kuehnii, will be
presented. Orange rust has impacted both the commercial production and the
cultivar development program in Florida. One major difference in the
epidemiology of the two pathogens is that P. kuehnii tolerates warmer
temperatures and orange rust severity continues throughout the summer and
early fall lasting much longer than brown rust. This is significant as it
means that commercial cultivars susceptible to both pathogens are impacted
by either one or both pathogens depending on the month of the growing
season. A cultivar that occupies 25% of the acreage in Florida, CP 80-1743,
is susceptible to the disease and has had reduced cane yields. It is being
withdrawn from production. Results from a comprehensive approach towards
developing sugarcane cultivars resistant to orange rust that is being
adopted in the Canal Point breeding program will be presented. This involves
identifying and discarding susceptible sugarcane clones as early in the
breeding program as possible, the development of novel screening methods and
the identification of sources of resistance for breeding.
‘Candidatus Liberibacter solanacearum’ on tomato and potential
losses in field production
R. D. FRENCH-MONAR (3), R. W. Wallace (2), J. A. Abad (1), T. A. Wheeler
(4)
(1) APHIS-PPQ-PGQP, Beltsville, MD, USA; (2) Dept. of Horticultural
Sciences, AgriLife Extension-Texas A&M System, Lubbock, TX, USA; (3) Dept.
of Plant Pathology and Microbiology, AgriLife Extension-Texas A&M System,
Amarillo, TX, USA; (4) Dept. of Plant Pathology and Microbiology, AgriLife
Research-Texas A&M System, Lubbock, TX, USA
In August 2008, tomato (Solanum lycopersicum) plots in Lubbock
County, TX that were utilized for a chemical test aimed at management of
root knot nematode became infected with ‘Candidatus Liberibacter
solanacearum’. Overall symptoms on tomato cv. Spitfire included leaf
yellowing, lateral stem dieback, upward leaf curling, enlargement of stems,
adventitious roots, and swollen nodes. PCR amplification was done using 16S
rDNA OA2 and OI2c primers for ‘Ca. L. solanacearum’ used for potato,
tomato, and other solanaceous crops in New Zealand, which amplifies a 1.1 kb
fragment of the 16S rRNA gene of this new species. A 1.1 kb fragment was
obtained, sequenced, and found to be 99.9% identical in sequence to a ‘Ca.
L. solanacearum’ obtained last year from a potato production field in Texas.
In the tomato field, a total of 32 plots (one-row wide, 7.7 m long)
comprised of 24 plants per plot were evaluated for disease symptoms and
galling by root-knot nematode. Foliar disease incidence in plots ranged from
one (4.2%) to 19 (79.2%) plants showing symptoms by the last harvest date.
Regression analysis was used to determine losses in yield associated with
the bacterium and with root-knot nematode. Percent galling by root-knot
nematode only explained 14% of the variability in yield, and 100% galling
was predicted to cause a 1.5% loss in yield, based on the regression model.
In contrast, for each 1% incidence in plants with disease symptoms, there
was a 0.9% loss in yield. In essence, there was no yield contribution if a
plant developed symptoms (R(^2) = 0.41). The potential exists for ‘Ca.
L. solanacearum’ to be a detriment in tomato production and a source for
survival of this bacterium that has been found to be associated with the
Zebra Chip disease in potato, tomato, pepper (Capsicum annuum), and
other solanaceous crops.
Diverse tomato cropping systems affect arbuscular mycorrhizal fungal
community diversity and structure
E. G. JOHNSON (3), D. O. Chellemi (2), T. Wu (1), J. H. Graham (3)
(1) Division of Cell Biology, Microbiology, and Molecular Biology,
University of South Florida, Tampa, FL, USA; (2) USDA-ARS, US Horticulture
Research Laboratory, Fort Pierce, FL, USA; (3) University of Florida, Citrus
Research and Education Center, Lake Alfred, FL, USA
In conventional agricultural systems, AMF are greatly affected by factors
including soil disruption, intermittent lack of host root tissue, and plant
inhibition of AMF colonization due to high soil fertilization. To determine
the effect of diverse agricultural land management and crop production
practices on the AMF community structure and diversity, five tomato crop
production systems consisting of bahiagrass pasture cover, conventional,
continuous removal of vegetation (disk fallow), organic, and undisturbed
(weed fallow) were initiated. The plots were adjusted to the new management
regime, except for conventional, for three or four years followed by one or
two years of tomato cropping. Soil DNA samples were taken in the off season,
at planting, and after harvest. Phylogenetic analysis of AMF 18S rDNA
sequence combined with multivariate statistical analysis using PRIMER-E was
used to compare community structure and diversity. Initial analysis shows
that bahiagrass, weed fallow, and organic land management practices support
different, diverse AMF communities, while disk fallow and conventional
practices greatly reduced detection of AMF sequences. Tomato cropping caused
the emergence of common sequences for the Glomus mosseae group, in
all cropping systems. Bahiagrass and weed fallow diversity were unaffected
by the emergence of the G. mosseae group, while organic, conventional
and disk fallow all converge on a low diversity community dominated by the
G. mosseae group. Current analyses will determine if the shift in AMF
community caused by tomato cropping in organic and bahiagrass plots is
seasonal or persistent, and if other factors such as soil fertility and
disease incidence correlate to these community changes.
Identification and characterization of powdery mildew caused by
Golovinomyces cichoracearum on sunn hemp
S. A. JORDAN (1), G. S. Maia (1), A. J. Gevens (1)
(1) University of Florida, Gainesville, FL, USA
Crotalaria juncea, or sunn hemp, is a warm season legume grown in FL
as a cover crop. In 2008, powdery mildew was observed on sunn hemp in a
research field in Hastings, FL. This disease is important because it has the
potential to impact the quality of sunn hemp and this powdery mildew can
infect cucurbits which are grown in north FL in late summer. Fungal growth
appeared first on lower, more mature leaves as white, powdery mildew
colonies initially seen on upper leaf surfaces and later moving to
undersides; petioles and floral parts were disease-free. As disease
progressed, colonies enlarged, coalesced, and covered entire leaf surfaces;
heavily infected leaves senesced and abscised. Mycelia produced white
accumulations of conidiophores and conidia. Hyphae were superficial with
papillate appressoria and produced conidiopohores with cylindrical foot
cells that measured 48.5 × 10.0 µm. Conidia were hyaline,
short-cylindrical-ovoid, lacked fibrosin bodies, borne in short chains, had
sinuate edge lines with other immature conidia, and measured 22.5-40.0 ×
12.5-20.0 µm. The teleomorph was not observed. The nuclear rDNA internal
transcribed spacer (ITS) regions were amplified by PCR and sequenced. On the
basis of morphological characteristics of the asexual state and ITS sequence
data, the pathogen was identified as G. cichoracearum. Pathogenicity
was confirmed on healthy plants. This is the first report of G.
cichoracearum causing powdery mildew on C. juncea.
Phytophthora cactorum a serious problem on prefinished Cattleya
orchid liners from Thailand
R. T. MCMILLAN (1), A. J. Palmateer (2), R. A. Cating (2)
(1) Kerrys, Homestead, FL; (2) University of Florida, Homestead, FL
The major pathogen on Cattleya orchids in Florida and in the New and Old
World countries is Phytophthora cactorum (Lebert & Cohn) J. Schroet.,
which causes Black Rot during the wet months of the year. All species of
Cattleya and their interspecific and intergeneric hybrids are susceptible.
Phytophthora cactorum infects leaves, pseudobulbs, rhizomes, and
flower buds. Shipments of prefinished Cattleya orchid liners from Thailand
during the monsoon season often infected with P. cactorum. Orchid
plants with visual symptoms of P. cactorum were removed from the
shipment and drenched with fungicides such as Banrot, Natriphene, Shield
Brite, Truban, and Phyton 27, Heritage, Shield Brite, Stature, Truban,
Pentathlon, Aliette, Subdue Maxx, and Insignia, in an effort to salvage some
of the plants. In spite of the effort to save the orchids, the level of
Phytophthora over rode the attempt to control the fungus resulting in
destroying the shipments. Cattleya liner shipments during the dry season are
found to be P. cactorum free.
Controlling angular leaf spot in Florida annual strawberry
J. C. MERTELY (1), N. A. Peres (1)
(1) University of Florida-GCREC, Wimauma, FL, USA
Angular leaf spot (ALS) is a bacterial disease caused by Xanthomonas
fragariae that produces unsightly lesions on strawberry leaves and
sepals. Leaves with numerous spots and/or vein-following lesions become
blighted and die prematurely. During the 2008–2009 growing season, an
epidemic of ALS occurred in the principal Florida strawberry production area
west of Tampa. During the season, a replicated trial evaluating products for
ALS control was conducted at the Gulf Coast Research and Education Center in
Wimauma, FL. Treatments included the plant defense activator
acibenzolar-s-methyl (Actigard); copper fungicides Badge, Cuprofix, IRF070,
Kentan, Kocide, and Quint; hydrogen peroxide (Oxidate); and Streptomyces
lydicus (Actinovate) applied weekly throughout the season to the foliage
with a CO(2) back pack sprayer. A late-season evaluation of foliar symptoms
showed that Badge, IRF070, Kentan, Kocide, and Actigard significantly
reduced the proportion of leaves killed and partially blighted by X.
fragariae. Alternating applications of Actinovate and Cuprofix also
showed this effect. However, only Badge and the low rate of Actigard
significantly increased marketable yield during a season with markedly high
disease pressure. Future experiments may target applications to periods
favorable to infection and disease spread, such as rain events associated
with approaching cold fronts, and periods of prolonged overhead irrigation
for freeze protection.
Cultivar susceptibility, temperature and leaf wetness durations required
for lesion production by Alternaria alternata on tangerine and
tangerine hybrids
S. N. Mondal (1), L. Timmer (1), M. M. DEWDNEY (1)
(1) University of Florida
Alternaria brown spot, caused by A. alternata, is an important
disease of tangerines and their hybrids in many citrus-producing regions. A
prediction model for fungicide applications, the Alter-Rater, was developed
previously, but it was unknown whether the relationship between temperature
and leaf wetness duration (LWD) would be consistent across all tangerine
hybrids. We tested the LWD and temperature relationships on 5 tangerine and
tangerine hybrids: Dancy, Minneola, Murcott, Nova and Sunburst. The LWDs
were 2, 4, 8, 16, 24 and 30h at temperatures of 20, 24, 28 and 32°C. The
rating scale for the number of lesions/leaf was: 0 = 0; 1 = 1-2; 2 = 3-5; 3
= 6-10; 4 = 11-15; 5 => 15 lesions/leaf and the data were taken from 15
leaves. The experiment was an incomplete block design. Cultivar differences
were observed; lesions formed on Minneola and Dancy with as little as 2h of
leaf wetness at all temperatures. Lesions were observed on Murcott, Nova and
Sunburst with 4h of leaf wetness. The optimal temperature range for lesion
production was 24 and 28°C for all LWDs. On the more susceptible Minneola
and Dancy, 24h LWD was required to reach the max. lesion rating, but that
level was never reached on Murcott, Nova and Sunburst even with 30h of leaf
wetness. The results should be incorporated into the Alter-Rater model so
that unnecessary sprays are not applied to less susceptible tangerine and
tangerine hybrids.
Identification of the Florida torreya canker pathogen
L. L. MOUNT (1), J. A. Smith (1)
(1) University of Florida
The Florida torreya (Torreya taxifolia Arn.) is a conifer of the
Taxaceae. The native population is restricted to within 20 miles of the
Apalachicola River in northern Florida and southern Georgia. Torreya wood is
resistant to decomposition. For this reason, it was lumbered for railroad
ties in the 19th century. Surveys conducted prior to 1970 detail a dramatic
reduction of population numbers, size and health. Historically, trees
reached 18 m at maturity. Of the 58 trees surveyed in November 2008, no
individual surpassed a height of one meter. Disease symptoms associated with
this decline are leaf spot, shoot tip dieback, and cankers. Fungal cultures
were isolated from symptomatic tissue and from the initial 58 trees tested,
cultures of Fusarium spp. and Botryosphaeria spp. were each
isolated 20 times. The same tree sample often produced both genera. Products
were amplified by polymerase chain reaction using internal transcribed
spacer region rDNA (ITS-rDNA) specific primers and the sequences obtained
were compared to those deposited in the GenBank database. Four unique
sequences of Fusarium spp. and Botryosphaeria obtusa were
identified. Fusarium solani and F. lateritium matched GenBank
sequences to the species level. Fusarium lateritium was previously
identified as the causal agent of the leaf spot, but was cultured during
this study directly from cankered tissue. Two species of Phomopsis,
as well as Diaporthe, Lasiodiplodia, and Hypoxylon were also
infrequently isolated and identified in the same manner from dying shoots
and cankers sampled in November 2008 and January 2009. Inoculations of as
many of these genera as possible were conducted on torreyas grown using
sterile tissue culture at Atlanta Botanical Gardens in April 2009.
The impact of silicon soil amendments on cucumber anthracnose in the
greenhouse
J. PALENCHAR (1), S. Taber (1), L. E. Datnoff (2), and A. J. Gevens (1)
(1) Department of Plant Pathology, University of Florida, Institute of
Food and Agricultural Sciences, Gainesville, FL; (2) Current address: Dept.
of Plant Pathology and Crop Physiology, Louisiana State University
Cucumber is an economically important crop in Florida and in other parts
of the U.S. In Florida, one of the most common diseases on cucumber is
anthracnose, caused by the ascomycetous fungus Colletotrichum orbiculare.
Anthracnose can cause serious yield and quality losses, and produces
symptoms on all aboveground plant parts at any stage of growth. Organic
producers have limited options for anthracnose control. The use of silicon
(Si) as a tool for disease control has been established in other crop
systems. In the greenhouse, the control of anthracnose was demonstrated on
2-week-old ‘Straight Eight’ cucumber seedlings by amending soil (organic
Fafard FOF 30) with a high rate of Si (Vansil W50). Treatments included
cucumber seeds planted into 1) Si-amended soil (600 kg Si/ha) + no C.
orbiculare inoculation 2) Si-amended soil (600 kg Si/ha) with C.
orbiculare inoculation, 3) non-amended soil with no C. orbiculare
inoculation, and 4) non-amended soil with C. orbiculare inoculation.
Each treatment included 5 replications and the experiment was repeated 4
times. Disease evaluations (Horsfall-Barrett scale) were recorded for leaves
at 3, 7, and 14 days post inoculation (dpi) and Si levels in plant tissues
were determined. Significant differences between treatments were observed at
7 and 14 dpi. Si treatment reduced disease severity on leaves by 20–60% when
compared to the inoculated control. This is the first study demonstrating
the efficacy of soil-applied Si for the control of cucumber anthracnose.
Diagnostics and emerging plant pathogens
M. E. PALM-HERNANDEZ (1)
(1) USDA/APHIS/PPQ, Molecular Diagnostic Laboratory, Beltsville, MD, USA
Accurate identification of plant pathogens is essential for making
management decisions and determining appropriate regulatory actions. An
accurate identification is facilitated when a group is well understood
taxonomically based on robust systematics studies. Such studies provide
information on which morphological and molecular characters are
taxonomically useful and tools can then be developed to use in detection and
identification. Identification of emerging plant pathogens poses a
particular challenge in that they are often understudied and poorly
characterized. Phytophthora ramorum, the causal agent of sudden oak
death and ramorum blight, exemplifies how regulatory diagnostics evolve as
scientific knowledge about the disease and its causal pathogen is gathered
and evaluated. The discovery of new and related taxa, including P.
foliorum that cross-reacted in the P. ramorum nested assay, led
to the development of additional assays. The use of new markers to compare
large numbers of P. ramorum isolates has provided a clearer picture
of the genetic diversity in the pathogen and a means of tracing origins of
newly found isolates. This case study is one of many examples of the
importance of a strong understanding of the systematics of a group as the
basis for accurate identification and the development of diagnostic tools.
Basil downy mildew in Florida: A disease of new importance
R. N. RAID (1), P. D. Roberts (4), P. F. Harmon (2), A. J. Palmateer (3),
S. A. Jordan (2)
(1) University of Florida, Belle Glade, FL, USA; (2) University of
Florida, Gainesville, FL, USA; (3) University of Florida, Homestead, FL,
USA; (4) University of Florida, Immokalee, FL, USA
Sweet basil (Ocimum basilicum L.) is one of the most important
herbs currently grown in Florida, with both commercial field and greenhouse
production. In addition, it is one of the most commonly propagated herbs in
home gardens. Fortunately, it has had very few foliar disease problems and
has, for that reason, required little or no disease management. During fall
2007, a new disease was first reported on field-grown basil in south
Florida. Symptoms initially appeared as a yellowing of the lower canopy,
with chlorotic areas frequently delineated by leaf veins. Gray, fuzzy fungal
growth was apparent on the abaxial leaf surface. The disease was
subsequently reported to be incited by a species of Peronospora.
Yield losses during this initial outbreak were near total, since
preventative control measures were formerly unnecessary, and therefore,
non-existent. Since the initial outbreak, basil downy mildew has become
firmly established in Florida. It has been observed from all regions within
the state, as well as in numerous other states. Although there is ample
evidence that the disease may have been introduced on infested seed,
alternative sources (i.e. from closely related hosts) have not been totally
ruled out. With widely-scattered, year-round greenhouse and/or field
production providing a host continuum, it is very likely that basil downy
mildew will be a disease to contend with on a permanent basis. Nearly all
basil varieties or types appear susceptible at this time. Management
programs are currently under development. Aside from cultural practices to
limit leaf wetness and hence fungal infection, preventive foliar
applications of phosphonates and strobilurin fungicides have proven useful.
Used together in a program, these have provided economic but not total
control.
Orange rust of sugarcane: Prospects for fungicidal control
R. N. RAID (2), J. C. Comstock (1), N. C. Glynn (1)
(1) USDA Sugar Cane Field Station, Canal Point, FL, USA; (2) University
of Florida, Belle Glade, FL, USA
Orange rust of sugarcane, incited by Puccinia kuehnii, was first
observed in Florida during June 2007 on one of the industry’s most important
commercial cultivars, CP80-1743. This was the first report of this disease
in the Western Hemisphere. It has since been reported in several other
Central American and Caribbean Countries. With host-plant resistance being a
worthy long-term goal, studies were initiated to investigate the feasibility
of fungicides serving as an interim or supplementary management strategy.
Thirteen different fungicide treatments were examined for their efficacy in
controlling orange rust during the 2008/2009 growing season. Experimental
units consisted of two rows of cane 15m in length replicated four times in a
randomized complete block design. Fungicide treatments consisted of select
candidates from two major classes of fungicides, the strobilurins (FRAC
group 11) and triazoles (FRAC group 3), alone, and in combination or
alternation. Fungicide applications were made using a CO(2) backpack sprayer
and were initiated following canopy closure (approx. 1.5-m ht) at 21 day
intervals. Rust severity in the trial area was moderately severe, with
severities in excess of 30% on the distal third of the fourth leaf beneath
the top-visible-dewlap leaf in the untreated check. Results indicate that
the strobilurin fungicides provided the highest level of control, followed
by strobilurin/triazole combinations, and finally, the triazole fungicides
alone. In separate trials using the strobilurin fungicide pyraclostrobin,
fungicide treatments were demonstrated capable of reducing orange rust to
levels sufficient to significantly reduce yield losses by as much as 40%.
While economic factors will ultimately be an important consideration, levels
of orange rust control obtained in these studies show promise regarding
prospects for fungicides as a potential management tool.
Fungicidal control of basil downy mildew
R. N. RAID (1)
(1) University of Florida, Belle Glade, FL, USA
Sweet basil (Ocimum basilicum L.) is one of the most common herbs
grown by home gardeners in Florida. Commercially, basil ranks as Florida’s
most important potted herb and the state ranks second nationally in field
production, shipping to the entire eastern seaboard. Since 2007, basil downy
mildew, incited by a species of Peronospora, has caused considerable
losses for commercial basil growers in the U.S. In the absence of control,
total crop failure is common. Two fungicide field trials were conducted to
determine the efficacy of various foliar applications for the control of
this disease. The tests included both registered and non-registered
compounds. Experimental units consisted of four rows 2m in length separated
on the ends by alleyways and replicated four times in a randomized complete
block design. All experimental compounds were topically applied on
approximately a weekly basis using a CO(2) backpack sprayer equipped with 3
flat-fan nozzles mounted on a hand-held boom. Treatment commenced at the 4-6
leaf stage, with mildew present in the area at time of initial application.
Disease severity was considered extreme. Products tested in these trials
included: acibenzolar, azoxystrobin, Bacillus subtilis,
chlorothalonil, copper hydroxide, cyazofamid, cymoxanil/famoxadone,
dimethomorph, fenamidone, mandipropamid, mefenoxam, potassium phosphanates,
propamocarb, pyraclostrobin/boscalid, and Streptomyces lydicus. All
products provided for significant suppression of downy mildew early in the
trials, but only a few provided for significant control by the end of the
tests. Dimethomorph, mandipropamid, cyazofamid, fenamidone, and mefenoxam
provided the highest levels of control, but are not currently labeled for
use on basil. Of currently registered products, only azoxystrobin and the
potassium phosphonates provided control levels that could be considered
acceptable from a marketing perspective. No compounds licensed for use in
organic herb production provided acceptable levels of mildew control when
sprayed on a weekly basis in these trials.
Current distribution of Texas Phoenix palm decline in Florida
D. A. RESTOM GASKILL (2), N. A. Harrison (3), M. L. Elliot (3), T. R.
Smith (1)
(1) Florida Dept. of Agriculture and Consumer Services, Gainesville, FL;
(2) USDA-APHIS-PPQ, Sarasota, FL; (3) University of Florida IFAS, Fort
Lauderdale, FL
Texas Phoenix palm decline (TPPD) is a fatal disease of date (Phoenix
dactylifera, P. sylvestris, P. canariensis, P.
reclinata), queen (Syagrus romanzoffiana) and cabbage (Sabal
palmetto) palms caused by a ‘Candidatus Phytoplasma
palmae’-related strain belonging to subgroup 16SrIV-D. In addition to the
economic costs of disease management in nurseries and landscapes, the
potential ecological impact due to reduction in S. palmetto
populations is incalculable. TPPD was first reported and characterized in
2002 from samples obtained in Corpus Christi, Texas, and was detected in the
Tampa Bay area of Florida in 2006. Surveys were conducted by the Cooperative
Agricultural Pest Survey (CAPS) in 2008 and 2009 to determine the current
distribution in Florida. Palms displaying characteristic symptoms of TPPD
were sampled and analyzed by polymerase chain reaction assay. Phytoplasma
positive samples from new locations were sequenced. Samples were also
submitted by personnel from the University of Florida, Institute of Food and
Agricultural Sciences (UF-IFAS) and from private landscape companies. TPPD
was determined to occur at a high incidence in a three county area
(Hillsborough, Manatee, Sarasota), while it was found to be of limited
distribution in seven additional counties (Pinellas, Polk, Hardee, Desoto,
Highlands, Lake, Duval). Spread of TPPD is likely occurring locally through
an unknown insect vector and over long distances through the transportation
of infected palms.
Biosafety regulation and biotechnology: How it affects public research in
Latin America and the Caribbean
M. M. ROCA (1)
(1) Zamorano University, Tegucigalpa, Honduras
While poverty in developing countries is usually linked with low
agricultural output, pest and plant diseases are major factors that
contribute significantly to this low productivity. Genetic engineering and
transgenic’s have great potential to improve crop production. However, the
application and use of this biotechnology has not materialized for the
public sector because of the politics associated with the regulatory
process. As a result, the time, effort and expense required for
commercialization of transgenic crops are way beyond what public-sector
investigators can muster leaving only the private sector to accomplish this
task. Restrictive regulations were established when the commercial use of
transgenic crops was just beginning, and have not taken into account the
more than 12 years of extensive experience gained on crops tested on more
than 100 million hectares in 23 countries. This information has
scientifically demonstrated that crops obtained through biotechnologies do
not have risk profiles that are any different from those developed through
more traditional plant breeding methods. The potential health and
environmental risks originally foreseen have not materialized. Furthermore,
it has been demonstrated that this biotechnology provides environmental and
economic benefits. As it now stands, the current biosafety regulatory
standards in place continue to delay the development and use of transgenic
technology. The time has come where there is a great need to consider both
the benefits and the risks of this technology, and analyze them relative to
those of the present agricultural production systems especially in Latin
America and the Caribbean.
Low molecular variability of Potato yellow vein virus (PYVV) isolates of
Solanum phureja and Solanum tuberosum from Colombia
P. Rodriguez-Burgos (2), G. Chaves (2), L. Franco-Lara (1), M.
GUZMAN-BARNEY (2)
(1) Militar University Nueva Granada, Bogotá-Colombia; (2) National
University of Colombia, Bogotá-Colombia
PYVV Closteroviridae-Crinivirus, is a quarentine, phloem-limited
potato virus, with a tripartite ssRNA(+). It causes yellowing of foliage
with reduction of yield. It is found in Colombia, Peru, Venezuela and
Ecuador and is transmitted by white fly Trialeurodes vaporariorum and
tubers. To study variability, CPg of 75 isolates of PYVV from Solanum
phureja and 50 from S. tuberosum, from 5 Colombian regions, was
amplified by RT-PCR. Amplicons were analyzed by SSCP and 32 were sequenced
directly. Ten SSCP patterns were observed (P1 to P10); P1 represented 78% of
the isolates, P9 9.6% and P6 4.8%. Phylogenetic analysis of 70% of CPg
produced two groups: Group I (29 isolates) and Group II (3 isolates). In
group I, isolates 1084 of S. phureja and 1114 of S. tuberosum,
showed evidence of possible recombination within CPg. No direct
correspondence between the number of SSCP patterns and the sequence clusters
was found, but P1 was present in Group 1 and P6 was found in all 3 isolates
of Group II. The aa relationship dN/dS = 0.214 indicated negative selection,
suggesting that PYVV has a tendency for low mutation fixation. This might be
related to a selection pressure coming from the insect vector. According to
variability of the CPg there are at least 3 virus variants circulating in
the Country, although variability among them is low.
Asparagus as host of Phythophtora species prevalent in Michigan
and its importance as rotational crop
L. RODRIGUEZ-SALAMANCA (1), M. Hausbeck (1)
(1) Michigan State University, East Lansing, MI, USA
Michigan ranks third in U.S. asparagus production, after Washington and
California, with 11.200 acres that produced 12 million kg of asparagus
spears in 2007. Phytophthora spear and crown rot has been recently
identified in Michigan fields as a major limiting disease of asparagus.
Although different species of Phytophthora have been reported in
other production areas as causing disease in asparagus, only one species has
been found in Michigan and has identified as P. asparagi.
Phytophthora sp. isolated from vegetables and ornamentals in Michigan
were tested for their ability to infect asparagus spears and cause lesions.
A series of growth chambers studies were conducted to determine; (i)
an optimum inoculation point for detached asparagus spears using three P.
asparagi isolates when incubated at 15, 20 and 25°C, (ii) the
ability of different Phythophthora sp. present in Michigan
agriculture to infect asparagus spears. All the studies were conducted three
times. When detached spears were wounded and inoculated at 2, 9 or 16 cm
from asparagus tip and incubated at 20°C, all the inoculation points
developed similar-sized lesions. However, when inoculated spears were
incubated at either 15 or 25°C, the size of the resulting lesion differed
significantly among the inoculation points. Among the select
Phythophthora sp. used to inoculate asparagus spears, P. capsici
was able to cause small lesions. Further studies that investigate
pathogenicity of commonly encountered Phythophthora sp. is important
in determining appropriate crop rotation strategies.
Fungal diversity associated with rambutan (Nephelium lappaceum L.)
in Puerto Rico
L. M. SERRATO (2), L. I. Rivera (2), R. J. Goenaga (1)
(1) USDA TARS, Mayaguez, Puerto Rico; (2) University of Puerto
Rico-Mayaguez Campus, Mayaguez, Puerto Rico
Rambutan (Nephelium lappaceum L.) is an exotic tropical fruit of
increasing importance in international markets, and that has awakened great
interest from farmers in Puerto Rico. During 2008 and 2009, fruit rot and
lesions on leaves, branches, and flowers were observed in rambutan orchards
through the island. To examine fungal diversity associated with rambutan,
samples from different organs were collected in symptomatic and asymptomatic
trees. Plant tissue was superficially disinfected and transferred to
acidified potato dextrose agar to promote the development of fungi. A total
of 311 fungal isolates were obtained, which include 19 genera. Based on
morphology, the following species have been identified: Beltrania
rhombica, Botryodiplodia theobromae, Botryosphaeria spp.,
Colletotrichum gloeosporioides, Colletotrichum spp.,
Curvularia spp., Cylindrocladium spp., Dolabra nepheliae,
Fusarium spp., Gliocephalotrichum bulbilium, Lasmenia
spp., Phomopsis spp., and Septoria spp. Pathogenicity tests
are in progress under laboratory and greenhouse conditions, using seedlings
and detached fruit. PCR amplification of the rDNA ITS region and the
beta-tubulin gene will complement morphological identification of fungi.
Use of bio-enhanced organic mulches for integrated management of nutsedge
in tomato
Y. SHABANA (2), E. Rosskopf (5), R. Charudattan (2), A. Abou Tabl (1), W.
Klassen (3), J. Morales-Payan (4)
(1) Mansoura University, El-Mansoura, Egypt; (2) University of Florida,
Gainesville, FL, USA; (3) University of Florida, Homestead, FL; (4)
University of Puerto Rico-Mayaguez, Mayaguez, PR; (5) USDA, ARS, USHRL, Fort
Pierce, FL, USA
Purple and yellow nutsedges (Cyperus rotundus and C. esculentus,
respectively) are among the world’s most problematic weeds that impact
virtually every horticultural crop grown in Florida and the Caribbean. As an
alternative to conventional methods of control that includes the use of soil
fumigation with methyl bromide, we tested nine hay mulches (shoot straw of
bahiagrass, cogongrass, cowpea, millet, yellow nutsedge, sorghum Sudangrass,
sunnhemp, and rye) and three green mulches (shoot biomass of cowpea, millet,
and sorghum Sudangrass) as a means to suppress nutsedge growth in a
raised-bed tomato (cv. Tygress) field. In addition, two fungus-infested
cogongrass hays (infested with the nutsedge pathogen Dactylaria higginsii
[Dh] or the saprophytic fungus Trichoderma sp. [Tri]), and two
plastic mulches (black and infra-red transmissible [IRT]) were tested. The
black plastic mulch and the Dh-infested cogongrass mulch consistently
reduced nutsedge emergence and growth more than the other organic mulches
and the IRT plastic mulch. Among the organic mulches, cogongrass infested
with Dh or Tri and cowpea, sunnhemp, Bahiagrass, and cogongrass provided the
highest levels of nutsedge suppression. No disease symptoms developed on
nutsedge plants when Dh- or Tri-infested cogongrass was used as the mulch.
Both plastic mulches (black and IRT) and Tri-infested cogongrass enhanced
tomato yield and the proportion of larger fruits. The highest yield of extra
large tomatoes per plant was obtained when these mulches were applied.
Laurel wilt of avocado: Management and mitigation research in Florida
J. A. SMITH (1), R. C. Ploetz (2), T. J. Dreaden (1)
(1) School of Forest Resources and Conservation, University of Florida,
Gainesville, FL, USA; (2) Tropical Research and Education Center, University
of Florida, Homestead, FL, USA
Laurel wilt, caused by the fungus Raffaelea lauricola and
transmitted by the exotic redbay ambrosia beetle, Xyleborus glabratus,
threatens the U.S. avocado industry with elimination if drastic measures are
not taken in the near future. Since its introduction in Georgia
approximately 6 years ago, the disease has spread on several hosts in the
Lauraceae on the southeastern coastal plain and now looms only 100 miles
from commercial avocado groves (7,500 acres worth $34 million/yr) in
Miami-Dade County, FL. Within the last 2 years, door-yard avocadoes have
been rapidly killed and serve as a source of inoculum for the epidemic.
Current research efforts include: examining extant avocado germplasm for
resistance; using a taxon-specific real-time PCR technique to diagnose the
pathogen and identify it in screening and epidemiology studies; and
fungicide efficacy trials. The results from these studies will be presented
and future work will be discussed. Laurel wilt threatens avocado production
worldwide. Thus, we will address its potential impact and preventing its
movement to new areas.
Combating the loss of red bay and other native species to laurel wilt
J. A. SMITH (3), M. A. Hughes (1), C. Anderson (3), R. C. Ploetz (4), A.
E. Mayfield (2)
(1) Department of Plant Pathology, University of Florida, Gainesville,
FL, USA; (2) Florida DACS Division of Forestry, Gainesville, FL, USA; (3)
School of Forest Resources and Conservation, University of Florida,
Gainesville, FL, USA; (4) Tropical Research and Education Center, University
of Florida, Gainesville, FL, USA
Laurel wilt, caused by Raffaelea lauricola, currently threatens
all native and some exotic species in the Lauraceae in the United States.
Since 2003, the disease has devastated native stands of redbay, Persea
borbonia, and threatens several other taxa in the family, including
avocado. Two natives, pondspice (Litsea aestivalis) and pondberry (Lindera
melissifolia), which are on state endangered and federal critically
endangered lists, respectively, face extinction. Despite sanitation and
other efforts to slow the movement of laurel wilt, it continues to move to
new areas every year, largely due to the efficiency of the disease vector,
the exotic redbay ambrosia beetle (Xyleborus glabratus). There are
many gaps in our current state of knowledge about the biology of the
disease, and several studies are underway. Current research has focused on:
protecting existing trees via fungicides; identifying and utilizing putative
resistance in redbay and avocado; elucidating the disease’s epidemiology and
host range; and determining to what extent genetic and pathogenic variation
exist in the R. lauricola population. An update on the results from
these studies will be given and future research needs will be discussed.
Effect of acibenzolar-S-methyl on bacterial leaf spot of shrub roses
caused by a Xanthomonas sp.
G. E. VALLAD (1)
(1) Gulf Coast REC, University of Florida, Wimauma, FL, USA
A bacterial leaf spot was recently identified on shrub rose varieties
‘Knockout’ and ‘Double Knockout’ caused by a Xanthomonas sp. and can
be problematic during vegetative propagation and nursery production.
Acibenzolar-S-methyl, the active ingredient of Actigard (Syngenta,
Greensboro, NC), is an elicitor of plant defenses that has demonstrated
efficacy in the control of several bacterial diseases of vegetable crops.
Greenhouse and nursery trials were established to test the effect of
Actigard on the severity of bacterial leaf spot on ‘Knockout’ and ‘Double
knockout’ roses. While lower rates of 0.25 to 0.5 oz of Actigard per 100
gallons were effective at reducing disease severity on potted roses and
liners, higher rates of 0.75 to 1.0 oz gave the best results. Multiple
applications of Actigard (1.0 oz/100 gal) prior to disease development
improved bacterial leaf spot control over single applications. Results
demonstrate the potential to use Actigard for disease management on
ornamental and nursery species.
Effect of acibenzolar-S-methyl on the management of early blight and
target spot of tomato
G. E. VALLAD (1)
(1) Gulf Coast REC, University of Florida, Wimauma, FL, USA
Acibenzolar-S-methyl, the active ingredient of Actigard (Syngenta,
Greensboro, NC), is an elicitor of plant defenses. While labeled for tomato,
usage is currently limited to the control of bacterial leaf spot (Xanthomonas
spp.) and bacterial speck (Pseudomonas syringae pv. tomato).
In 2008, two field trials assessed the performance of Actigard (8 weekly
applications at 0.75 oz per acre) when integrated into a standard spray
program that included weekly applications of copper sulfate (2.1 lbs a.i.
per acre) mixed with either mancozeb (1.5 lbs a.i. per acre) or
chlorothalonil (1.5 lbs a.i. per acre). The addition of Actigard reduced the
severity of early blight (Alternaria solani) and target spot (Corynespora
cassiicola) by 22 to 44% over the standard spray program alone, and by
31 to 60% compared to the non-treated plots. In the spring trial, plots
treated with Actigard yielded 336 more cartons (25 lbs) of marketable
tomatoes per an acre than those receiving the standard alone, and 1,179
cartons more per an acre than the non-treated plots. No yield improvement
was observed in the fall trial, due to the development of disease in the
late season. Results demonstrate the benefit of including Actigard as part
of an overall spray program to manage common foliar diseases caused by
bacterial and fungal pathogens of tomato.
Salmonella outbreaks associated with vegetables: How high is the
risk?
A. VAN BRUGGEN (1)
(1) Emerging Pathogens Institute and Plant Pathology Department, IFAS,
University of Florida, Gainesville, FL, USA
Gastro-enteritis outbreaks increased in the 1990s and have remained
steady since then. Most outbreaks have been associated with seafood, but
most individual cases with vegetables and fruits (38% of all cases).
Salmonella enterica is the most common pathogen involved in outbreaks
associated with vegetables. Several Salmonella outbreaks were traced
back to contaminated tomatoes. Salmonella enterica is very versatile:
there are more than 2500 serovars, which occur in various environments,
including many plant and animal species. The main reservoirs are the
intestines of birds, pigs, cattle, wild mammals and reptiles, but they are
also harbored by protozoa, earthworms, nematodes and snails. They can
multiply in the rhizosphere of various plants and occur on plant surfaces as
well as in the endosphere. Because of the human as well as economic costs
associated with Salmonella outbreaks, it is important to study the
risk of an outbreak to occur. However, there are different kinds of risk:
calculated probabilities as well as perceived risks. These last risks are
concerns voiced by consumers on a comparative scale. Among various safety
concerns, microbiological risks are ranked high, due to some knowledge and
experience and the feeling of not being able to control exposure. Perceived
risks do not necessarily coincide with calculated probabilities, but may be
more influential in terms of the response to an outbreak. Quantitative
microbial risk assessment consists of several steps: hazard identification
and characterization, exposure assessment and risk characterization. In a
project on risk assessment of enteric pathogens in the vegetable production
chain, we limited ourselves to exposure assessment through lettuce
contaminated from manure and soil. The occurrence and survival of
enteropathogens in cattle manure were primarily determined by the feed given
to the cattle: low-fiber feed resulted in more shedding and longer survival
in the low-fiber and low-pH manure. Other risk factors were low numbers of
nonpathogenic coliform bacteria and high dissolved organic carbon contents.
Constant temperatures and low oxygen levels also contributed to long
survival times in manure. Survival times in soil were negatively correlated
to microbial diversity and positively to dissolved organic carbon contents.
A probabilistic exposure model for E. coli O157:H7 resulted in a
relatively low probability of about 1 contaminated head in 10,000 lettuce
heads. The risk can best be reduced at the beginning of the production
chain, the cattle farm. There are several quantitative risk assessment
models for Salmonella, but most of them are for animal products,
except for one model for almond contamination and on for vegetable
contamination from irrigation water. No risk model was found for tomato
production and processing. In a tomato safety research workshop, research
needs were identified, but control at the source of the chain was not
mentioned while this was the crucial factor in our lettuce risk model. Risk
models based on calculated probabilities could be used to influence
perceived risks by the general public.
STAR-D: The NPDN accreditation program for diagnostic laboratories
A. B. VITORELI (1), C. L. Harmon (1)
(1) University of Florida, Gainesville, FL, USA
The Food and Agriculture Defense Initiative was established in 2002 to
enable the United States Department of Agriculture to develop a network
linking plant and animal disease diagnostic facilities across the USA. The
National Plant Diagnostic Network (NPDN) is the plant disease component of
this network. The mission of the NPDN requires quick and accurate diagnosis
of high consequence plant pathogens, weeds and insect pests that threaten
national security; communication of such information response authorities;
the ability to scale up and manage sample surge as needed; and diagnostic
data security. To accomplish these objectives, the NPDN relies on diagnostic
data generated by laboratories in Land-Grant Universities, State Departments
of Agriculture, and USDA-APHIS. Traditionally, these laboratories have
provided diagnostic services at the State or regional levels at a high level
of competence. However, to accomplish the national objectives listed above,
a standardized approach to diagnosis is required, particularly if the
diagnosis has regulatory implications. The NPDN System for True, Accurate,
and Reliable Diagnostics (STAR-D) has been developed to enable participating
laboratories to meet standards of quality for laboratory management,
facilities, equipment, and trained personnel. The ISO 17025 Quality Standard
can be adapted to testing done by NPDN diagnostic laboratories by providing
a basis for a quality system to meet the needs of the NPDN STAR-D.
Levels of P in Areca catechu leaves following phosphorous acid
application through adventitious roots
G. C. WALL (1)
(1) University of Guam, Mangilao, GU 96923, USA
Bud rot disease of betel nut (Areca catechu L) has been shown to be
caused by Phytophthora palmivora. Fosphite, or phosphorous acid, is
recommended for the control of Phytophthora, applied by injection to the
trunk. After finding evidence of damage associated with injection sites in
the trunks of betel nut trees, a decision was made to look for other ways of
applying the fungicide in order to avoid damaging the trees. A paired t test
was devised to study the effect of applying phosphorous acid solution via
adventitious roots of betel nut trees. A group of mature trees was sampled
pre- and post-application. Levels of P were determined from leaf samples
collected from each frond per tree. There were 12 pairs of trees in the
study; one set of trees was treated with the recommended rate (applied by
absorption through an adventitious root) and half were controls, treated
only with sterile distilled water also via one adventitious root. After the
appropriate statistical analysis (NCSS, Kaysville, UT), differences were
found in the level of P in the leaf samples according to treatment. Control
trees had higher levels of P in their leaf tissue compared to trees given
phosphorous acid. The underlying hypothesis was that application of the
fungicide via adventitious roots of trees would result in a systemic
distribution of the fungicide throughout the tree. It was expected that all
leaf samples from treated trees would show higher levels of P compared to
untreated controls. Surprisingly, P levels were lower in treated trees, yet
there was no difference between fronds, suggesting an even effect throughout
treated trees. No explanation is known at this time for the reduced P levels
observed after treatment; however, results were consistent enough to yield
highly significant differences statistically.
Epidemiology of soybean rust (Phakopsora pachyrhizi) in soybean (Glycine
max) sentinel plots in Florida
H. M. YOUNG (2), J. J. Marois (2), D. L. Wright (2), D. F. Narvaez (1),
G. K. O’Brien (2)
(1) Monsanto, St. Louis, MO, USA; (2) University of Florida, NFREC,
Quincy, FL, USA
The overwintering of soybean rust (SBR) in the Southeastern United States
has been variable due to weather conditions which may influence disease
incidence and severity in the major soybean producing regions of the
Midwest, making it important to understand the epidemiology of the pathogen
in Florida. This study examined the incidence and severity of SBR in
relation to prevailing weather data, growth stage, and maturity group
(MGIII, MGV, MGVII) in 15 m square soybean plots across the Panhandle of
Florida from 2005 through 2008. Of the three maturity groups, the MGIII
soybean became infected first the least often. Plots became infected first
at growth stage R4 (full pod) or later. On average, plots became infected 40
days earlier in 2008 than 2005. Precipitation was the principle factor
affecting disease progress, where disease increased rapidly after rain
events and was suppressed during dry periods. The area under the disease
progress curves (AUDPC) for incidence and severity was the lowest in 2007,
most likely due to dry conditions. In 2008, there was a significant increase
in disease incidence and severity as reflected in the AUDPC. This can be
attributed in part to the occurrence of Tropical Storm Fay, which deposited
up to 290 mm of water in the plot locations during the third week of August.
Results from this study may lead to a better understanding of the impact of
weather on the epidemiology of this pathogen.
Effect of rhizobacteria, acibenzolar and silicon on bacterial spot of
tomato
S. ZHANG (1), T. L. White (1), W. Klassen (1), M. C. Martinez (1)
(1) Tropical REC, University of Florida, IFAS, Homestead, FL, USA
Bacterial spot, caused by Xanthomonas perforans, is one of the
most economically important diseases of tomato in Florida and other tomato
grown areas worldwide. Chemical controls have been only partially effective
due to the wet and warm climate in Florida and the development of resistance
in populations of this bacterial pathogen. It is imperative that practical
alternative strategies be developed to sustain the production of tomatoes.
Greenhouse and field trials have been conducted to investigate the effect of
plant growth-promoting rhizobacteria (PGPR), acibenzolar-S-methyl (ASM) and
silicon nutrient on bacterial spot of tomato. In the greenhouse, eight
bacilli PGPR strains were evaluated on two cultivars of tomato (FL47R and
Tygress). Tomato seeds were sown into pro-mix in 128-cell Styrofoam flats
and grown for 1–2 weeks when solutions of PGPR, Actigard 50WG (ASM) and
silicic acid were applied weekly as soil drenches. Tomato seedlings were
transplanted into 4-inch pots containing potting mix after 3–4 soil
drenches, and inoculated by foliar spray with suspensions of X. perforans
at 1 × 10(^8) CFU/ml. Results indicated that PGPR strain SE76 and INR7
significantly (P < 0.05) reduced disease severity of bacterial spot on both
tomato cultivars compared to the nontreated control. SE52 on cv. FL47R and
SE34, IN937a and IN937b on cv. Tygress each had a significant effect on
disease reduction. In the first field trial on tomato cv. FL47R, Actigard 50
WG at 30 mg/l significantly suppressed bacterial spot rated at 8, 9, 10 and
11 weeks after transplanting, whereas silicic acid at 0.15 and 1.5 mM did so
only at 8 weeks. In another field experiment, the same eight PGPR strains
and Actigard 50WG at 30 and 3 mg/l were tested on tomato cv. Tygress.
Significant disease reduction was observed on tomato plants treated with
PGPR strains IN937a and IN937b 3 weeks after transplanting.