1 FLUENSULFONE EFFICACY TO MANAGE ROOT-KNOT NEMATODES ON DRIP- IRRIGATED FRESH-MARKET TOMATOES AND SPATIAL DISTRIBUTION IN SANDY SOILS USING SEEPAGE IRRIGATION By GILMA XIOMARA CASTILLO LICONA A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2017
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FLUENSULFONE EFFICACY TO MANAGE ROOT-KNOT NEMATODES ON DRIP-IRRIGATED FRESH-MARKET TOMATOES AND SPATIAL DISTRIBUTION IN SANDY
SOILS USING SEEPAGE IRRIGATION
By
GILMA XIOMARA CASTILLO LICONA
A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF
Literature Review .................................................................................................... 18 Root-Knot Nematodes: Disease Cycle of a Global Plant Pathogen ................. 18 Alternatives for Root-Knot Nematode Management in Vegetable Production .. 19
Cultural practices ....................................................................................... 19 Chemical management .............................................................................. 20
Fluensulfone, a Recent Introduction of New Chemistry for Management of Root-Knot Nematodes ................................................................................... 22
Modeling Pre-Plant-Incorporated Fluensulfone with HYDRUS 2D/3D Using Seepage Irrigation ......................................................................................... 23
2 EFFECTS OF FLUENSULFONE AND SOIL FUMIGATION ON ROOT-KNOT NEMATODES AND FRUIT YIELD OF DRIP-IRRIGATED FRESH-MARKET TOMATO ................................................................................................................ 26
Introduction ............................................................................................................. 26 Materials and Methods............................................................................................ 29
Field Preparation and Treatment Application ................................................... 29
Data Collection ................................................................................................. 31
Plant Vigor, Root-Knot Nematode Soil Density, and Root Galling .................... 33 Tomato Fruit Yield and Grade Distribution ....................................................... 34
3 EVALUATION OF FLUENSULFONE ON SOIL SPATIAL DISTRIBUTION AND MOVEMENT, PLANT GROWTH, FRUIT YIELD, AND POSTHARVEST QUALITY OF TOMATO USING SEEPAGE IRRIGATION ...................................... 41
Introduction ............................................................................................................. 41 Materials and Methods............................................................................................ 43
Field Preparation and Treatment Application ................................................... 43 Data Collection ................................................................................................. 45 Statistical Analysis ............................................................................................ 47
Results .................................................................................................................... 48 Weather Conditions, Water Table Depth, and Soil Water Matric Potential ....... 48 Fluensulfone Concentration and HYDRUS 2D/3D Modeling ............................ 49 Plant Growth, Fruit Yield, and Postharvest Quality ........................................... 50
Table page 1-1 Current fumigant and non-fumigant nematicides available in Florida to
manage plant-parasitic nematodes on vegetables. ............................................ 25
2-1 Summary of minimum, mean, and maximum daily average air and soil temperatures and total rainfall accumulation during the fall of 2014, the spring of 2016, and 10-year fall and spring averages for Myakka City, FL. ........ 38
2-2 Effect of pre-plant drip-injected fluensulfone on plant vigor, root-knot nematode soil population density, and root galling index in tomato grown during the fall of 2014 and the spring of 2016 in Myakka City, FL. ..................... 39
2-3 First, second, third, and total season marketable and unmarketable tomato fruit yield in response to pre-plant drip-injected fluensulfone during the fall of 2014 and the spring of 2016 in Myakka City, FL. ................................................ 40
3-1 Summary of minimum, mean, and maximum daily average air temperatures, solar radiation, total rainfall, and evapotranspiration during the spring and fall of 2016, and 10-year spring and fall averages for Immokalee, FL. ..................... 56
3-2 Effect of pre-plant application of fluensulfone on plant dry biomass on seepage-irrigated fresh-market tomato crops grown during the spring and fall seasons of 2016 in Immokalee, FL. .................................................................... 62
3-3 Effect of pre-plant application of fluensulfone on first, first and second harvests combined, and total harvest marketable and unmarketable tomato fruit yield during the spring and fall seasons of 2016 in Immokalee, FL. ............ 63
3-4 Fluensulfone treatment effects on tomato fruit firmness (expressed as fruit deformation), exterior fruit color, pH, and total soluble solids at first harvest during the spring and fall seasons of 2016 in Immokalee, FL. ............................ 64
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LIST OF FIGURES
Figure page 3-1 Water table level (centimeters from the top of the bed) observed in seepage-
irrigated fresh-market tomato crops grown during the spring and fall seasons of 2016 in Immokalee, FL.. ................................................................................. 57
3-2 Soil water matric potential observed in seepage-irrigated tomato crops grown during the spring and fall seasons of 2016 in Immokalee, FL. ............................ 58
3-3 Fluensulfone concentration in the soil profile at 0-10 and 10-20 cm deep from the top of the bed observed in seepage-irrigated fresh market tomatoes during the spring and fall seasons of 2016 in Immokalee, FL. ............................ 59
3-4 HYDRUS 2D/3D simulation describing water flow and fate of fluensulfone treatments at 6, 20, 30 days after treatment application (DAA) under seepage irrigation conditions during the spring of 2016 in Immokalee, FL. ....................... 60
3-5 HYDRUS 2D/3D simulation describing water flow and fate of fluensulfone treatments at 8, 21, 30 days after treatment application (DAA) under seepage irrigation conditions during the fall of 2016 in Immokalee, FL. ............................ 61
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LIST OF ABBREVIATIONS
ANOVA Analysis of Variance
AZ Arizona
CA California
CEC Cation exchange capacity
CO2 Carbon Dioxide
CSB Chemical Safety Board
DAA Days after application
DAFH Days after first harvest
DAT Days after transplanting
DW Dry weight
EC Electrical conductivity
EPA Environmental Protection Agency
ET Evapotranspiration
FAO United Nations Food and Agriculture Organization
FAWN Florida Automated Weather Network
FIFRA Federal Insecticide, Fungicide, and Rodenticide Act
FL Florida
GA Georgia
ID Idaho
IL Illinois
IN Indiana
J2 Second-stage juveniles
K Potassium
LC/MS-MS Liquid chromatography tandem mass spectrometry
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K2SO4 Potassium sulfate
MA Massachusetts
Max Maximum
MeBr Methyl bromide
Min Minimum
N Nitrogen
NH4NO3 Ammonium nitrate
NC North Carolina
OM Organic matter
P Phosphorus
Pic-Clor 60 1,3-dichloropropene plus chloropicrin (40:60, w/w)
PVC Polyvinyl chloride
RHU Representative harvest unit
RKN Root-knot nematode
SWFREC Southwest Florida Research and Education Center
TSS Total soluble solids
UF/IFAS University of Florida/Institute of Food and Agricultural Sciences
UF/SWFREC University of Florida/Southwest Florida Research and Education Center
U.S. United States
USDA United States Department of Agriculture
WI Wisconsin
1D One dimensional
2D/3D Two- and three-dimensional
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Abstract of Thesis Presented to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science
FLUENSULFONE EFFICACY TO MANAGE ROOT-KNOT NEMATODES ON DRIP-
IRRIGATED FRESH-MARKET TOMATOES AND SPATIAL DISTRIBUTION IN SANDY SOILS USING SEEPAGE IRRIGATION
CA) grown in 128-cell styrofoam trays (Mobley Plant World, LaBelle, FL), were
transplanted 61 cm apart in a single row for each bed establishing 20 plants per plot
and a population of 8,970 plants·ha-1. ‘HM 1823’ is an early season tomato with 70-74
days to maturity, which required tying and staking and had no resistance to RKNs.
Fertigation was used to supplement the pre-plant fertilizer with 48–3–40 kg·ha-1 N–P–K
on a weekly basis. Both trials were irrigated using drip irrigation. Irrigation run times
were 57 min·ha-1 for 6 days per week with a flow rate of 0.30 m3·ha-1·min-1 for a total
water discharge of 17 m3·ha-1·day-1. Pest management tactics were applied based on
weekly scouting reports and UF/IFAS recommendations (Santos et al., 2013).
Data Collection
Average minimum, mean, maximum daily air and soil temperatures, and total
rainfall accumulation were recorded by the Florida Automated Weather Network
(FAWN) for Balm, FL. Prior to treatment application, at midseason, and at final harvest,
six soil cores per plot were randomly collected at 20 cm deep using a soil probe for
sandy soils (Oakfield Apparatus, Inc., Oakfield, WI). Soil cores from each plot were
mixed thoroughly to create one composite sample of 250 cm3 of soil. Samples were
placed in a cooler for preservation. Soil samples were then sent to LLH Ag and
Research Services, LLC, Tifton, GA for nematode quantification and identification. The
levels of soil population densities of RKN second-stage juveniles (J2) were categorized
as low, medium, or high according to Barker et al., 1976. A representative harvest unit
(RHU) of 10 plants was marked at the center of each plot. At midseason, on 31 Oct.
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2014 and 8 Mar. 2016 [49 and 48 days after planting (DAP)], and final harvest, on 23
Dec. 2014 (102 DAP) and 10 May 2016 (111 DAP), three plants at the edges of the
RHU and six plants within RHU were selected for RKN root galling evaluation,
respectively. Root-knot nematode root galling index was visually assessed according to
Hussey and Janssen (2002) rating system using a 0 to 5 continuous scale, where zero
= no traces of root galling, 1 = infection with few small galls, 2 = less than 25% of roots
galled, 3 = between 25 to 50%, 4 = between 51 and 74%, and 5 = greater than 75 % of
roots galled. Plant vigor was visually assessed at 25 and 20 DAP in the fall of 2014 and
spring of 2016, respectively, based on a 1-10 continuous rating scale, where 1 = poor
overall plant growth and 10 = optimal uniform plant growth. Tomato fruit within RHU
were manually harvested and weighed three times at mature-green stage at 73, 88, and
102 DAP in 2014 and at 92, 103, and 111 DAP in 2016. Fruit yield was classified into
marketable and unmarketable. Marketable fruit yield was graded according to USDA
size category specifications—extra-large (diameter > 7.00 cm), large (6.35 to 7.00 cm),
and medium (5.72 to 6.43 cm) (USDA, 1997). Tomato fruit were considered
unmarketable based upon size less than 5.72 cm and the presence of defects such as
sunscald, scratch, off-shape, catfaced, and graywall (Jones et al., 1991; Ozores-
Hampton et al., 2010).
Statistical Analysis
Plant vigor, RKN soil population density, root galling index, and fruit yield were
subjected to analysis of variance (ANOVA) using the GLM procedure and means were
separated according to Duncan’s multiple range test at 5% confidence level using SAS
(SAS 9.3, SAS Institute Inc., Cary, NC, 2012). Root-knot nematode densities were
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transformed by square-root transformation prior to analysis to obtain a normal
distribution.
Results
Weather Conditions
Average minimum, maximum, and mean air and soil temperatures were within
the range of average temperatures recorded in the previous 10 years for both fall 2014
and spring 2016 seasons (Table 2-1). There were no freezing events reported during
both seasons. Total rainfall accumulation for fall 2014 and spring 2016 were 276.1 and
104.9 mm greater than the previous 10-year average (Table 2-1).
Plant Vigor, Root-Knot Nematode Soil Density, and Root Galling
Application of fluensulfone to fumigated soil (Pic-Clor 60) did not affect plant
vigor in 2014 and 2016. Tomato plants exhibited optimal uniform growth with all
treatments during both years (Table 2-2). Initial population densities of RKN J2 before
treatments were applied were low in 2014 and 2016 (10/100 and 30/100 cm3 of soil,
respectively). In 2014, at final harvest, Pic-Clor 60 followed by 2.0 and 2.8 kg a.i·ha-1
fluensulfone decreased population densities of RKN J2 as compared to Pic-Clor 60
alone by approximately 96 and 81%, respectively (P ≤ 0.001). Similarly, in 2016, at final
harvest, Pic-Clor 60 followed by 2.0 and 2.8 kg a.i·ha-1 fluensulfone decreased
population densities as compared to Pic-Clor 60 alone by approximately 85 and 94%,
respectively (P < 0.05). However, there were no significant differences among
fluensulfone rates at midseason and final harvest for both years [P > 0.05 (Table 2-2)].
In 2014 and 2016, at final harvest, combining fluensulfone at 2.0 kg a.i·ha-1 with Pic-
Clor 60 reduced root galling index as compared to Pic-Clor 60 alone by 57 and 90%,
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respectively (P = 0.0001). There were no significant differences among fluensulfone
rates in either year.
Tomato Fruit Yield and Grade Distribution
In 2014, neither Pic-Clor 60 alone nor Pic-Clor 60 followed by fluensulfone had
an effect on any tomato fruit size categories or total marketable and nonmarketable
yield at first and second harvest, separately [P > 0.05 (Table 2-3)]. However, when first
and second harvests were combined, differences among treatments were only found in
the extra-large fruit size category (data not shown). Pic-Clor 60 alone and Pic-Clor 60
followed by fluensulfone at 2.8 kg a.i·ha-1 accounted for the greatest extra-large fruit
yield (P = 0.04) (data not shown).
At the third harvest in 2014, both 2.0 and 2.8 kg a.i·ha-1 fluensulfone produced
highest fruit yield for all tomato size categories and total marketable yield [P < 0.05
(Table 2-3)], except for the unmarketable yield where no differences were found among
treatments (P > 0.05) (data not shown). Pic-Clor 60 alone and Pic-Clor 60 followed by
fluensulfone at 2.8 kg a.i·ha-1 accounted for the greatest total season extra-large fruit
yield (P = 0.05). There were no differences for the remaining tomato size categories and
for the total season marketable and unmarketable yields in 2014 (P > 0.05). In 2016,
inclusion of fluensulfone at 2.0 and 2.8 kg a.i·ha-1 in the fumigated soil did not have a
significant effect on yield in any harvest or tomato fruit size category [P > 0.05 (Table 2-
3)].
Discussion
This study demonstrated that fluensulfone in combination with Pic-Clor 60 can be
an effective tool to manage RKNs in drip-irrigated fresh-market tomato grown in sandy
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soils with high RKN infestation. In the fall of 2014 when RKN infestation was high, the
weather conditions may have contributed in facilitating RKN reproduction and survival
(Table 2-1). Optimum air and soil temperatures for M. hapla and related species range
from 15 to 25 °C and 15 to 20 °C, respectively; and corresponding temperatures for M.
javanica and related species range about 5 °C higher (Taylor and Sasser, 1978;
Wallace, 1964). The cumulative fall season rainfall was 276.1 mm higher than the
previous 10-year average, which offered nematodes ideal conditions to complete their
life cycles since high moisture content enhances nematode movement (Djian-
Caporalino et al., 2009) and egg hatching (Van Gundy, 1985). In the spring of 2016
when RKN infestation was low, the cool air and soil temperatures at the beginning of the
growing season (e.g. 0.9 and 13.6 °C on 25 Jan. 2016, respectively) may have played a
role in increasing the duration of the RKN life cycle and decreasing reproduction and
hatching (Collange et al., 2011).
During the fall of 2014 and the spring of 2016, injection of fluensulfone through
the drip tape after soil fumigation with Pic-Clor 60 did not affect plant vigor observed at
25 and 20 DAT, respectively. Similarly, in a tomato-cucumber double-cropping system,
application of fluensulfone did not affect tomato plant vigor; nonetheless, fluensulfone
improved cucumber plant vigor as compared to a non-treated control as well as reduced
root galling (Morris et al., 2015).
Although initial RKN population densities before treatment application were low
for both seasons, the effect of fluensulfone was observed when RKN population
densities reached higher levels throughout the season, especially at the third harvest.
Both fluensulfone rates reduced RKN populations and root galling during the fall of 2014
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and the spring of 2016. However, the level of suppression was greater in the fall of 2014
when RKN infestation was high, which may be explained by the fact that nematode
damage and dynamics are population density dependent (Seinhorst, 1965). In the fall of
2014, inclusion of drip-injected fluensulfone to the nematode management program
provided a more effective level of RKN control compared to Pic-Clor 60 alone. In the
presence of high RKN infestation, the combination of fluensulfone and Pic-Clor 60
improved tomato yield at the third harvest. In contrast during the spring of 2016, tomato
yield did not reflect the effect of the nematicide due to the low RKN infestation. These
findings are consistent with the results of Morris et al. (2015) who showed that tomato
yield did not respond to fluensulfone in fields with low RKN infestation. Norshie et al.
(2016) indicated that fluensulfone treatments reduced potato cyst nematode (Globodera
pallida) root infection by 43% and increased yield by 61.5% at greater root infection
(1125.8 RKN J2·g-1 root) whereas potato yield did not respond when there was lower
infection (444.2 RKN J2·g-1 root).
Summary
In 2014 and 2016, at final harvest, Pic-Clor 60 followed by 2.0 and 2.8 kg a.i·ha-1
fluensulfone decreased population densities of RKN J2 as compared to Pic-Clor 60
alone by 96 and 81% and 85 and 94%, respectively. Combining fluensulfone at 2.0 kg
a.i·ha-1 with Pic-Clor 60 reduced root galling index as compared to Pic-Clor 60 alone by
57 and 90% at final harvest in 2014 and 2016, respectively. However, an increase of
total marketable fruit yield was only observed at the third harvest of the fall of 2014
when there was a high level of RKN infestation (3265 RKNs·cm-3 soil). Inclusion of
fluensulfone at 2.0 and 2.8 kg a.i·ha-1 after Pic-Clor 60 did not have an effect on yield in
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any harvest or tomato fruit size category when RKN infestation was low in 2016 (800
RKNs·cm-3). Further research is needed to support the results presented in this study,
particularly considering other integrated management practices, commercial crops, and
different soil types.
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Table 2-1. Summary of minimum (Min.), mean, and maximum (Max.) daily average air and soil temperatures and total rainfall accumulation during the fall of 2014, the spring of 2016, and 10-year fall and spring averages for Myakka City, FL.
Period
Temperature (°C) Total
rainfall Air Soil (-10 cm depth)
Min. Mean Max. Min. Mean Max. (mm)
Fall 2014 Septembera 21.1 24.7 30.9 25.4 26.3 27.2 228.9 October 16.4 22.4 29.2 23.5 24.3 25.2 54.9 November 10.9 16.8 23.7 18.8 19.6 20.4 182.9 December 10.5 16.1 23.0 16.8 17.6 18.4 4.1 Average/total 14.7 20.0 26.7 21.1 21.9 22.8 470.7 Fall 10-year average 15.4 20.9 27.6 23.0 23.7 24.5 194.6 Spring 2016 January 7.9 13.7 20.0 15.5 16.1 16.9 66.5 February 9.1 15.5 22.3 16.4 17.2 18.2 42.4 March 14.7 20.5 27.3 19.7 20.6 21.6 49.3 April 15.5 21.9 28.6 22.1 23.2 24.3 56.6 May 15.5 22.2 28.7 22.6 23.8 25.2 84.1 Average/total 12.6 18.8 25.4 19.3 20.2 21.2 299.0 Spring 10-year average 11.9 18.9 26.2 19.6 20.7 21.8 194.1
a The temperature averages and rainfall totals were recorded daily from 12 Sept. through 23 Dec. 2014 and from 20 Jan. through 10 May 2016. Data source: Florida Automated Weather Network station located in Balm, FL, at 64.4 km of distance from the field (http://fawn.ifas.ufl.edu/).
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Table 2-2. Effect of pre-plant drip-injected fluensulfone on plant vigor, root-knot nematode [Meloidogyne spp. (RKN)] soil population density, and root galling index in tomato grown during the fall of 2014 and the spring of 2016 in Myakka City, FL.
a Plant vigor was visually assessed based on a 1-10 scale, where 1 = poor overall plant growth and 10 = optimal uniform plant growth. b 0 = no root galling, 1 = trace infection with a few small root galls, 2 ≤ 25% root galls, 3 = 25-50%, 4 = 51-74%, and 5 ≥ 75 % of root galls (Hussey and Janssen, 2002). c Second-stage juveniles (J2) count data were transformed using the square-root function before statistical analysis. d DAT = days after transplanting e Pic-Clor 60 = 1,3-dichloropropene plus chloropicrin (40:60, w/w) at 280 kg·ha-1. f Within season means followed by different letters are significantly different according to Duncan’s multiple range test at 5%. NS *, **, *** Nonsignificant or significant at P ≤ 0.05, 0.01, or 0.001, respectively.
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Table 2-3. First, second, third, and total season marketable and unmarketable tomato fruit yield by size categories in response to pre-plant drip-injected fluensulfone during the fall of 2014 and the spring of 2016 in Myakka City, FL.
a XL = extra-large (greater than 7.00 cm); L = large (6.35 to 7.00 cm); M = medium (5.72 to 6.43 cm); ); TM = total marketable, UM = unmarketable [fruit with defects such as sunscald, scratch, off-shape, catfaced, and graywall (Jones et al., 1991; Ozores-Hampton et al., 2010b)]. b Pic-Clor 60 = 1,3-dichloropropene plus chloropicrin (40:60, w/w) at 280 kg·ha-1. c Within season means followed by different letters are significantly different according to Duncan’s multiple range test at 5%. NS *, **, *** Nonsignificant or significant at P ≤ 0.05, 0.01, or 0.001, respectively.
Treatment First harvest Second harvest Third harvest Total season harvest
128-cell styrofoam trays (Mobley Plant World, LaBelle, FL), were transplanted 61 cm
apart in a single row for each bed establishing 60 plants per plot or a population of
8,966 plants·ha-1. The crop was irrigated using seepage irrigation with a water furrow
located every six beds. Management of pests and foliar pathogens was accomplished
based on weekly scouting reports and UF/IFAS recommendations (Freeman et al.,
2015).
Data Collection
Averages for minimum, mean, maximum daily air temperatures, and for total
rainfall accumulation, daily solar radiation, and evapotranspiration (ET) were recorded
by the Florida Automated Weather Network (FAWN) located at the UF/IFAS/SWFREC
in Immokalee, FL. Irrigation was managed by installing monitoring wells made from a
1.2-m-long, 10-cm-diameter polyvinyl chloride (PVC) pipe (Smajstrla and Muñoz
Carpena, 2011). Monitoring wells were installed at the center of each replication and
water table depth was monitored weekly throughout the growing season. To indicate
water levels, a float was attached to the bottom end of the PVC pipe, and marks were
made every 2.54 cm to show water table depth below the polyethylene mulch.
Tensiometers (Irrometer Company, Inc., Riverside, CA) were located at the center of
each replication to monitor soil moisture by measuring the soil water matric potential
weekly throughout the growing season. Fluensulfone concentration in the soil and
gravimetric water content were measured immediately after treatment application, and
subsequently at 2, 7, 14, and 21 DAA. Five soil cores were randomly collected to the
depths of 0-10 cm and 10-20 cm at the center bed of each plot and mixed to create one
composite sample. Soil cores were collected using a 25.4-cm-diameter recovery probe
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with individual acetate liners (AMS, Inc., American Falls, ID). Subsamples of 100 g were
oven dried at 105 oC for 24 h and weighed to determine soil moisture. Then, frozen
subsamples were sent to EAG Laboratories, Hercules, CA, where soil concentration of
fluensulfone was determined by liquid chromatography tandem mass spectrometry
(LC/MS-MS). To simulate fluensulfone movement in seepage irrigation at 6, 20, and 30
DAT for the spring and at 8, 21, and 30 DAT for the fall, HYDRUS 2D/3D software (PC-
Progress, Prague, Czech Republic) was used by ADAMA Agricultural Solutions Ltd.,
Airport City, Israel.
Roots, stems, leaves, and fruits of two plants per plot were randomly collected
and oven dried at 65 oC until constant weight at 30, 60, and 90 DAT to determine dry
weight (DW) (Mills and Jones, 1996). Total plant DW was calculated by adding the DW
of the roots, stems, leaves, and fruits. Tomato fruit of 10 representative plants at the
center bed for each plot were manually harvested and weighed three times at mature-
green stage at 85, 92, and 98 DAT during the spring and at 81, 95, and 102 DAT during
the fall of 2016. Tomato fruit yield was classified into marketable and unmarketable.
Marketable fruit yield was graded according to USDA size category specifications—
extra-large (diameter > 7.00 cm), large (6.35 to 7.00 cm), and medium (5.72 to 6.43 cm)
(USDA, 1997). Tomato fruit were considered unmarketable according to the presence of
defects such as sunscald, scratch, off-shape, catface, and graywall (Jones et al., 1991;
Ozores-Hampton et al., 2010b). At first harvest, 20 mature-green tomato fruit per plot
were collected, placed in paper bags, and exposed to ethylene at 20 °C and 85 to 90%
relative humidity until they achieved stage two of ripeness (Sargent et al., 2014) at
Gargiulo, Inc. packing house in Immokalee, FL. After achieving stage two, tomatoes
47
were transported to UF/IFAS/SWFREC, Vegetable Horticulture Laboratory in
Immokalee, FL, where they were allowed to achieve table-ripe stage at room
temperature (23 to 24 °C) for postharvest quality assessments (Ozores-Hampton et al.,
2010b). Four fruit from each plot were then selected to determine their firmness, which
is based on the amount of fruit deformation recorded by an 11-mm probe when a 1-kg
force is applied to the fruit “equatorial” area for 5 s by a portable digital firmness tester
(Model C125EB, Mitutoyo Co., Aurora, IL). Exterior color for each of the same four fruit
was measured using a one to six scale where one = green and six = red (USDA, 1997).
One-fourth of each of the four fruit was selected, and the fourths were then combined to
measure total soluble solids [(TSS), °Brix] and pH. The samples were homogenized and
centrifuged (Model Sorval ST16; Thermo Scientific, Waltham, MA) at 7177 gn for 20
min. After that, the supernatant was filtered using cheesecloth. The filtered was used to
measure TSS with a portable refractometer (Model Eclipse 45-02; Bellingham + Stanley
Inc., Suwanee, GA) and pH with a pH meter (Orion 4 star benchtop; Thermo Electron
Co., West Palm Beach, FL).
Statistical Analysis
Because soil chemical and physical properties varied between seasons, all data
parameters were analyzed separately by season. Using the TTEST procedure in SAS
software package (SAS 9.3, SAS Institute Inc., Cary, NC, 2012), fluensulfone soil
concentrations among the two rates were subjected to Student’s t-distribution at 5%
confidence level. Plant DW (roots, stems, leaves and fruits), yield, and postharvest fruit
quality parameters were subjected to analysis of variance (ANOVA) using the GLM
48
procedure and means were separated according to Duncan’s Multiple Range Test at
5% confidence level.
Results
Weather Conditions, Water Table Depth, and Soil Water Matric Potential
Averages for minimum, mean, and maximum daily air temperatures during the
growing season (planting to third harvest) were 14.5, 21.6, and 29.4 oC in the spring
and 19.2, 24.4, and 31.5 oC in the fall, respectively (Table 3-1). Average daily solar
radiation was 219.3 and 174.0 W·m-2 in the spring and fall of 2016, respectively. Total
rainfall accumulation and ET were 186.4 and 350.8 mm in the spring and 300.0 and
318.8 mm in the fall, respectively. Daily air temperatures were within the range of
average temperatures recorded in the previous 10 years (2006-2015) and there were no
freezing events reported for both seasons. Daily solar radiation, however, was 14.8 and
12.7 W·m-2 lower than the historical 10-year average. Total rainfall accumulation and ET
for the spring and fall seasons were 13.1 and 12.3 mm lower and 12.7 and 37.9 mm
lower than the previous 10-year average, respectively.
Water table depth in the monitoring wells fluctuated between 53 and 82 cm and
46 and 87 cm in the spring and fall seasons, respectively (Figure 3-1). Water table
levels were closer to the bed top during the first five weeks of the growing season. At
planting, on 9 Feb. (spring) and 25 Aug. (fall), water table levels were raised closer to
the bed top to facilitate the establishment of the transplants and ensure adequate
nutrient uptake. As a result of rainfall events recorded on 13 and 14 Aug. (24 mm)
during the fall, water table reached the highest level of 46 cm from bed top. Soil water
matric potential fluctuated between 2.3 and 11.8 kPa and 2.8 and 11.8 kPa in the spring
49
and fall seasons, respectively (Figure 3-2). At planting, soil tension was low for the
spring (2.3 kPa) and fall (2.8 kPa) seasons to facilitate plant lower energy requirements
to extract water from the soil.
Fluensulfone Concentration and HYDRUS 2D/3D Modeling
During the spring of 2016, fluensulfone soil residues measured at 0-10 cm deep
from the top of the bed were not statistically different among the two treatments at 0, 2,
14, 21 DAA [P > 0.05 (Figure 3-3)]. However, at seven DAA, soil residue of fluensulfone
at 4.0 kg a.i·ha-1 (1.12 mg·kg-1) was higher than at 2.0 kg a.i·ha-1 (0.55 mg·kg-1) (P =
0.05). Fluensulfone soil residues measured at 10-20 cm deep were not statistical
different at 0, 2, 7, 14 DAA (P > 0.05), except at 21 DAA in which fluensulfone at 4.0 kg
a.i·ha-1 was higher (2.22 mg·kg-1) than at 2.0 kg a.i·ha-1 (0.99 mg·kg-1) (P = 0.03).
During the fall of 2016, fluensulfone soil residues measured at 0-10 cm deep at 0, 2, 14,
21 DAA, were not statistically different among the two treatments (P > 0.05). However,
at seven DAA, soil residue of fluensulfone at 4.0 kg a.i·ha-1 (2.38 mg·kg-1) was higher
than at 2.0 kg a.i·ha-1 (0.47 mg·kg-1) (P = 0.04). Fluensulfone soil residues measured at
10-20 cm deep were not statistical different at 0, 2, and 21 DAA (P > 0.05). Soil residue
of fluensulfone at 4.0 kg a.i·ha-1 was higher than the lower treatment at 7 and 14 DAA
with 0.99 and 0.92 mg·kg-1 at 10-20 cm deep, respectively (P = 0.001 and 0.01).
During the spring conditions, at six DAA, simulation of fluensulfone at 2.0 kg
a.i·ha-1 with HYDRUS 2D/3D showed the highest soil residue concentration (1.23
mg·kg-1) at 0 cm deep (bed surface) (Figure 3-4). Similarly, during the fall, at eight DAA,
the highest soil residue concentration (1.01 mg·kg-1) occurred at 0 cm deep (Figure 3-
5). In the spring, fluensulfone at 2.0 kg a.i·ha-1 appeared to undergo a rapid dissipation
50
to the bed shoulders and upward movement, with the highest soil concentration of 0.36
mg·kg-1 found at 0 cm deep at 20 DAA. In the fall, at 21 DAA, the highest residual
concentration of 0.35 mg·kg-1 was found at 11 cm deep. HYDRUS 2D/3D predicted that
at 30 DAA, the highest residual concentrations of 0.22 mg·kg-1 and 0.29 mg·kg-1 were
found at 0 and 9 cm for the spring and fall seasons, respectively.
In the spring, at six DAA, simulation of fluensulfone at 4.0 kg a.i·ha-1 accounted
for the highest soil residue concentrations of 1.40 mg·kg-1 allocated at 12 cm deep from
bed top. A rapid dissipation occurred at 20 DAA to bed shoulders with a concentration
of 0.50 mg·kg-1 at 0 cm deep. Lastly, at 30 DAA, HYDRUS 2D/3D predicted a
concentration decrease to 0.32 mg·kg-1 found at 0 cm. In the fall, at eight DAA,
simulation of fluensulfone at 4.0 kg a.i·ha-1 accounted for the highest soil residue
concentrations of 1.73 mg·kg-1 allocated at 4 cm deep from bed top. A rapid dissipation
occurred at 21 DAA to bed shoulders with a concentration of 0.61 mg·kg-1 at 10 cm
deep. Finally, at 30 DAA, HYDRUS 2D/3D predicted a concentration decline to 0.5
mg·kg-1 found at 9 cm.
Plant Growth, Fruit Yield, and Postharvest Quality
During the spring, total plant and fruit DW at 90 DAT in the non-treated control
were on average 29 and 34% higher than both fluensulfone treatments, respectively
(Table 3-2). Similarly, during the fall, at 60 DAT, fruit DW in the non-treated control was
on average 18% higher than both fluensulfone treatments whereas there was no effect
of fluensulfone on total plant DW at 60 DAT (P > 0.05). However, in the spring and fall
seasons, DW of roots, stems, and leaves were not affected by fluensulfone treatments
on any sampling dates (P > 0.05). Total plant and fruit DW were not affected by
51
fluensulfone rates at 30 and 60 DAT and at 30 and 90 DAT in the spring and fall
seasons, respectively P > 0.05).
In the spring, at third harvest, fluensulfone at 4.0 kg a.i·ha-1 accounted for the
highest large and medium size categories and total marketable and unmarketable yield
with an increase of 81, 70, 64, and 121%, respectively (Table 3-3). Total season large
and medium fruit were on average 43 and 61% higher with fluensulfone at 4.0 kg a.i·ha-
1. However, there was no effect on the extra-large fruit size category (P > 0.05). Total
season extra-large, total marketable and unmarketable yields were not affected by any
fluensulfone treatments (P > 0.05). Application of fluensulfone did not have an effect on
any tomato fruit size categories or total marketable and unmarketable yield at first,
second, and first and second harvests combined (P > 0.05). In the fall, at first harvest,
fluensulfone at 2.0 kg a.i·ha-1 and the non-treated control accounted for the highest
extra-large fruit size category and total marketable yield (Table 3-3). However,
fluensulfone at 4.0 kg a.i·ha-1 and the non-treated control accounted for equal yields. At
third harvest, fluensulfone at 4.0 kg a.i·ha-1 accounted for the highest large and medium
size categories and total marketable yield with an increase of 34, 52, and 39%,
respectively, whereas there were no significant differences for the unmarketable yield (P
> 0.05). Fluensulfone treatments did not have an effect on any tomato fruit size
categories or total marketable and unmarketable yield at second, first and second
harvests combined and total season yield (P > 0.05).
Postharvest evaluation as fruit deformation, exterior color, TSS, and pH were not
influenced by any fluensulfone treatment during the spring or fall season [P > 0.05,
(Table 3-4)].
52
Discussion
Simulation by HYDRUS 2D/3D suggested that fluensulfone concentrated in the
shallower soil layers based on soil properties and weather conditions. However, soil
fluensulfone concentrations were below the threshold to cause plant phytotoxicity during
the spring and fall seasons. During the spring, which had cooler temperatures and lower
rainfall, fluensulfone concentrated in the upper layer, showing a dissipation over time at
six DAA when water table levels reached 53 cm from bed top. Rubin et al., 2011
demonstrated that when fluensulfone was applied through drip irrigation, the chemical
migrated to deeper layers (30 cm), which can be attributed to the downward movement
of water by gravity. In contrast, using seepage irrigation, HYDRUS 2D/3D modeling
suggests that fluensulfone residues concentrated in the soil upper layers, which can be
ascribed by the upward movement of the water table.
During the fall, which had higher temperatures, rainfall, and OM, fluensulfone
residues were located at 5-11 cm whereas in the spring the residues migrated to
surface of the soil (0 cm). Previous studies with clay, Hamra sandy, and loamy sand
soils have shown that fluensulfone exhibits an affinity to OM, potentially limiting the
movement in the soil through binding by sorption [Van der Waal's forces, hydrogen
bonding, hydrophobic bonding (Oka et al., 2013; Morris et al., 2015; Moss et al., 1975;
Leistra and Smelt, 1981; Bollag et al., 1992; Whitehead, 1988)]. Therefore, high OM
content in the fall (2.5%) could possibly explain retention of fluensulfone, but may not be
considered as the only factor since during both seasons, water upward flow using
seepage irrigation appeared to be the prevailing factor on the fate of fluensulfone.
These findings were expected since fluensulfone, as a non-fumigant, moves through the
53
soil water, which suggests that during the first weeks of the growing seasons,
fluensulfone transport and fate were governed by water flow and less by chemical
reactions (adsorption) or biological degradation (E. Segal, personal communication).
Although symptoms of phytotoxicity did not appear in either season, each
fluensuflone treatment yielded the lowest total plant and fruit DW in the spring at 90
DAT and the lowest total fruit DW in the fall at 60 DAT. However, fluensulfone
treatments did not decrease root, stem, leaf DW, or fruit yield. Reduction in plant and
fruit DW could be attributed to fluensulfone systemic activity (Oka et al., 2013). Recent
studies by Morris et al., 2016 have presented that foliar applications of fluensulfone
caused a reduction on plant DW in eggplant (Solanum melongena) and tomato. In
addition, high rates of fluensulfone (12 g a.i·l-1) were phytotoxic to both crops at 12 and
28 DAA. Several studies have verified that some systemic chemicals have an influence
on plant growth and production (Pless et al., 1971; Lee, 1977; Olofinboba and
Kozlowski, 1982; Reddy et al., 1986; Araya et al., 1988; Cranshaw and Thorton, 1988).
However, there is no information on fluensulfone mode of absorption, translocation, and
detoxification within the plant, translocatable form, or the concentration of molecules in
the plant cells, especially in the fruit. Further studies are needed to assess the ability of
fluensulfone to penetrate plant cells and the implications on DW.
Following label recommendation, when crop transplanting needs to be at least
seven DAA, residual concentrations of fluensulfone at 2.0 and 4.0 kg a.i·ha-1 were 0.55
and 1.12 mg·kg-1 and 0.47 and 2.38 mg·kg-1 at 0-10 cm deep for the spring and fall
seasons, respectively. Twenty-one DAA, which is the planting interval for some
available pre-plant fumigants used extensively by the vegetable industry, the residual
54
concentrations of 2.0 and 4.0 kg a.i·ha-1 at 0-10 cm deep decreased to 0.46 and 1.42
mg·kg-1 and 0.37 and 1.38 for the spring and fall seasons, respectively. Therefore,
delayed transplanting after seven DAA of fluensulfone application could be a possible
strategy to avoid a decrease in DW.
Pre-plant incorporation of fluensulfone in seepage irrigation did not reduce fruit
yield or negatively influence tomato fruit postharvest quality. Overall, total marketable
yield obtained during both seasons were in accordance with those observed by Ozores-
Hampton et al. (2012, 2015) in Immokalee, FL. In addition, fluensulfone did not
decrease extra-large fruit yield which will be the grower’s preferred size category
because they are valued at premium prices (Bierlen and Grunewald, 1995). These
results were not expected since fluensulfone residual concentration of approximately
1.0 mg·kg-1 caused phytotoxicity when applied through foliar spray or drip injection (E.
Segal, personal communication). However, our results suggest that fluensulfone
residual concentrations greater than 1.0 mg·kg-1 did not decrease fruit yield in seepage
irrigation. However, further research needs to be conducted with replicated years and
different weather conditions to corroborate the results of these studies. In addition,
studies are needed to assess fluensulfone nematicidal efficacy in seepage irrigation.
Summary
Using seepage irrigation, fluensulfone concentrated in the upper bed profile as
predicted by HYDRUS 2D/3D software with soil residues of 2.0 and 1.01 mg·kg-1 at six
DAA and 0.36 and 0.35 mg·kg-1 at 21 DAA for the spring and fall seasons, respectively.
Fluensulfone treatments showed lower total plant and fruit dry weight at 90 DAT during
the spring and lower fruit DW at 60 DAT during the fall. However, fluensulfone
55
treatments below 2.0 mg·kg-1 did not show phytotoxicity or decrease root, stem, leaf
DW, fruit yield, or impact postharvest fruit quality. These results suggest that the pre-
plant incorporation of fluensulfone did not negatively influence fresh-market tomato
production grown in sandy soils using seepage irrigation. However, the movement and
distribution of fluensulfone and the effects on plant growth and yield need to be further
evaluated for different climates, soil types, and crops.
56
Table 3-1. Summary of minimum (Min.), mean, and maximum (Max.) daily average air temperatures, solar radiation, total rainfall accumulation, and evapotranspiration (ET) during the spring and fall of 2016, and 10-year spring and fall averages for Immokalee, FL.
a ET = Evapotranspiration is presented as the United Nations Food and Agriculture Organization (FAO) Penman-Monteith. bThe temperature and solar radiation averages, and rainfall and evapotranspiration totals were recorded daily from 2 Feb. through 18 May and from 18 Aug. through 5 Dec. 2016. Data source: Florida Automated Weather Network station located at University of Florida/Institute of Food and Agricultural Science/Southwest Florida Research and Education Center in Immokalee, FL (http://fawn.ifas.ufl.edu/).
Period Temperature (°C)
Solar radiation
Total rainfall
ETa
Min. Mean Max. (W·m-2) (mm) (mm)
Spring
Februarya 10.6 17.2 24.5 176.3 62.0 63.5
March 15.4 21.8 29.3 200.3 17.3 94.7
April 15.2 22.9 31.2 248.6 34.8 117.6
May 16.9 24.4 32.5 252.0 72.4 74.9
Average/total 14.5 21.6 29.4 219.3 186.4 350.8
Spring 10-year average 13.5 20.9 29.2 234.1 199.5 363.1
Fall
August 24.3 28.0 34.2 203.0 68.6 55.6
September 23.4 27.3 34.0 194.7 184.7 109.5
October 19.7 24.4 30.7 171.5 45.5 86.1 November 12.6 20.0 28.9 159.4 0.8 58.2 December 16.0 22.0 29.8 141.3 0.5 9.4
Average/total 19.2 24.4 31.5 174.0 300.0 318.8
Fall 10-year average 18.7 23.9 31.3 186.7 337.9 323.3
57
Figure 3-1. Water table level (from the top of the bed) in seepage-irrigated fresh-market tomato crops grown during the
spring and fall seasons of 2016 in Immokalee, FL. Means of four replications. Vertical bars represent ± SE.
58
Figure 3-2. Soil water matric potential in seepage-irrigated fresh-market tomato crops grown during the spring and fall
seasons of 2016 in Immokalee, FL. Means of four replications. Vertical bars represent ± SE. Readings of 0–5 kPa: soils are saturated or nearly saturated; 10–15 kPa: crops should be irrigated as soon as possible; 25 kPa and higher: plants probably present symptoms of water stress (Migliaccio et al., 2012).
59
Figure 3-3. Fluensulfone concentration in the soil profile at 0-10 and 10-20 cm deep from the top of the bed in seepage-
irrigated fresh market tomatoes during the spring and fall seasons of 2016 in Immokalee, FL. Vertical lines represent planting dates.
60
Figure 3-4. HYDRUS 2D/3D simulation describing water flow and fate of fluensulfone treatments at 6, 20, 30 days after
treatment application (DAA) in seepage irrigation conditions during the spring of 2016 in Immokalee, FL.
61
Figure 3-5. HYDRUS 2D/3D simulation describing water flow and fate of fluensulfone treatments at 8, 21, 30 days after
treatment application (DAA) in seepage irrigation conditions during the fall of 2016 in Immokalee, FL.
62
Table 3-2. Effect of pre-plant application of fluensulfone on plant dry biomass on seepage-irrigated fresh-market tomato crops grown during the spring and fall seasons of 2016 in Immokalee, FL.
aDAT = days after transplanting bTotal represents roots, stems, and leaves. cWithin columns, means followed by different letters are significantly different according to Duncan’s multiple range test at 5%. NS, *, **, ***, Non-significant or significant at P ≤ 0.05, 0.01, 0.001, respectively.
Treatment
Dry biomass (g·plant-1)
Spring (DATa)
30 60 90
Root Stem Leaves Totalb Root Stem Leaves Fruits Total Root Stem Leaves Fruits Total
Table 3-3. Effect of pre-plant application of fluensulfone on first, first and second harvests combined, and total season harvest (three harvests combined) marketable and unmarketable tomato fruit yield graded by size categories during the spring and fall seasons of 2016 in Immokalee, FL.
a XL = extra-large (diameter greater than 7.00 cm); L = large (6.35 to 7.00 cm); M = medium (5.72 to 6.43 cm); ); TM = total marketable, UM = unmarketable [fruit with defects such as sunscald, scratch, off-shape, catface, and graywall (Jones et al., 1991; Ozores-Hampton et al., 2010b)]. b Within columns, means followed by different letters are significantly different according to Duncan’s multiple range test at 5%.
NS, *, **, ***, Non-significant or significant at P ≤ 0.05, 0.01, 0.001, respectively.
Treatment First harvest First and second harvests Total season harvest
Table 3-4. Fluensulfone treatment effects on tomato fruit firmness (expressed as fruit deformation), exterior fruit color, pH, and total soluble solids at first harvest during the spring and fall seasons of 2016 in Immokalee, FL.
Treatment Deformation
(mm) (8 DAFHa)
Color stage (1–6 scale)b
pH Total soluble solids (°Brix)
Spring Non-treated control 2.83 5.38 4.00 3.95 Fluensulfone 2.0 kg a.i·ha-1 2.68 5.25 3.90 3.96 Fluensulfone 4.0 kg a.i·ha-1 2.79 5.63 3.90 3.89 P-value 0.49 0.11 0.07 0.90 Significancec NS NS NS NS Fall Non-treated control 2.20 5.44 3.90 4.50 Fluensulfone 2.0 kg a.i·ha-1 2.52 5.69 3.80 4.53 Fluensulfone 4.0 kg a.i·ha-1 2.21 5.56 3.95 4.54 P-value 0.19 0.51 0.24 0.56 Significance NS NS NS NS
aDAFH = Days after first harvest b1 = green and 6 = red (USDA, 1997) cNS = Non-significant at P > 0.05.
65
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BIOGRAPHICAL SKETCH
Gilma Castillo was born in Yoro, Honduras in 1992. She graduated from Ave
Maria University, FL in 2013 with a B.A. in biology and a minor in chemistry. In 2014,
she joined the Vegetable Horticulture Program in Immokalee, FL as part of her optional
practical training (OPT). In August 2016, she started her graduate studies at University
of Florida to pursue a master’s degree in horticultural sciences. Her research focuses
on the evaluation of the efficacy of fluensulfone to manage root-knot nematodes on drip-
irrigated fresh-market tomato and on the study of the distribution and movement of pre-
plant-incorporated fluensulfone in sandy soils irrigated via a seepage method.