Increasing food self-sufficiency is a critical objective for the State of Hawaii that imports 90% of its food from the global market (Department of Business Economic Development and Tourism DBEDT and Hawaii Department of Agriculture HDOA, State of Hawaii, 2012). Currently, the value of vegetables, melons, potatoes (
Plant-parasitic nematodes are detrimental pests that adversely affect plant health and yields in fruit and vegetable crops. Of particular importance are root-knot (
Fluopyram, first marketed as a fungicide and later as a nematicide, is an inhibitor of the succinate dehydrogenase enzyme (Veloukas and Karaoglanidis, 2012). Exposure of
Azadirachtin is a naturally occurring substance found in seed kernels of neem (
Cultural practices can be implemented to improve soil health and reduce population densities of plant-parasitic nematodes. Planting a sunn hemp (
This study was guided by the hypotheses that the integration of fluopyram or azadirachtin with pre-plant sunn hemp cover crop would not only be suppressive to plant-parasitic nematodes but also mitigate non-target effects of the nematicides on free-living nematodes. The specific objective of this study was to evaluate the effects of fluopyram treated alone and fluopyram or azadirachtin treated in combination with pre-plant sunn hemp cover crop amendment on plant-parasitic nematodes and free-living nematodes.
The hypotheses were tested in three separate field trials, differing to some extent in the number of treatments and the crops grown; the first two with tomato and zucchini and the last one with sweet potato.
The experiments were conducted at Poamoho Experiment Station, Waialua, on the Island of Oahu, HI (21°32'14.8“N and 158°5'20.3“W). The soil type at the experiment site was a Wahiawa silty clay in the Oxisol order with Tropptic Eutrustox, clayey, kaolinitic, isohyperthermic properties, containing 18.6% sand, 37.7% silt, and 43.7% clay, and soil organic matter of approximately 1.08% in the top 25 cm soil.
In 2017, a field trial was conducted to examine the effects of fluopyram (41.5%, Velum® One, Bayer CropScience, Research Triangle Park, NC) and azadirachtin (3%, Molt-X®, BioWorks, Inc., Victor, NY) against root-knot nematodes (
Fluopyram and azadirachtin chemigation schedules for Trials I, II, and III.
Chemigation schedule | |||
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Trial I (2017) | Trial II (2018) | Trial III (2019) | |
Treatments | Zucchini and tomato | Zucchini and tomato | Sweet potato |
Velum I | At planting | At planting | At planting and 3 months post-planting |
Velum II | At planting and 2 weeks post-planting | At 2 weeks post-planting | At 2 weeks and 3 months post-planting |
SH + Velum | At 4 weeks post-planting | At 4 weeks post-planting | At 1 and 3 month(s) post-planting |
SH + MoltX | At monthly interval | At monthly interval | At monthly interval |
Control | Untreated bare ground | Untreated bare ground | Untreated bare ground |
In 2018, a second field trial was superimposed on the first field trial (Trial I) following 3 months of soybean cultivation. All plots were checked for initial population densities of reniform and root-knot nematodes and were found to be not different among plots at the end of the soybean crop prior to soil incorporation. For treatments 3 and 4, sunn hemp cover cropping was planted 2 months before cash crop planting. Soybean or sunn hemp residues were rotor-tilled with a handheld tiller and Trial II was initiated on July 25, 2018 as a repeat of Trial I with a slight modification. While treatments 1, 3, 4, and 5 remained the same as in Trial I, treatment 2 was adjusted to a one-time application of fluopyram at 2 weeks after planting (without the 0-week application as was done in Trial I). Sunn hemp biomass generated in this trial was similar to that in Trial I with an average of 22 Mg/ha.
This trial was conducted in 2019 superimposed on the same field site using sweet potato (
Zucchini canopy width and chlorophyll content from the third matured leaf of each plant was measured bi-weekly. Chlorophyll content was measured using a SPAD-502 Chlorophyll Meter (Konica Minolta, Tokyo, Japan 2003). Due to heavy infestation by melon fly (
Initial soil populations of plant-parasitic nematodes were documented at the time of planting sunn hemp cover crop. These data were used for a background check and were not used in the treatment comparison. For treatment comparison, in Trial I, soil samples from zucchini or tomato were collected at 0, 1, and 2 months after crop planting. In Trial II, soil samples were collected at 0 and 2 months after planting zucchini or 0 and 3 months after planting tomato. In Trial III, soil samples were collected at 0, 2, 3 (prior to fluopyram application), and 6 months (at harvest) after sweet potato planting. At each time of sampling, four soil cores per plot were collected from the top 10 cm using a 7.5 cm diameter GroundShark shovel (Forestry Suppliers Inc. Jackson, Mississippi, USA) and composited in a sampling bag. The soil was sifted using a 4-mm2 mesh screen and homogenized by handshaking prior to collecting a 250-cm3 subsample. Nematodes were extracted from the 250-cm3 soil subsample by elutriation and centrifugal flotation (Byrd et al., 1972; Jenkins, 1964). Individual nematodes present in each sample were identified to genus level except for Rhabditidae which was identified only to the family level under Leica™ Inverted Microscope (Leica Microsystems Co., Wetzlar, Germany) with reference to Goodey (1963) or Smart and Nguyen (1988). All nematodes identified were grouped into one of the five trophic groups: bacterivores, fungivores, herbivores, omnivores, and predators according to Yeates et al. (1993), and the abundance of each trophic group was enumerated.
At the termination of Trials I and II, individual root systems of zucchini and tomato were uprooted as much as possible using a pitchfork. Roots were weighed and the severity of nematode infestation over the whole root system uprooted was rated based on a 0–10 scale, where 0 = no gall and 10 = plant is dead due to severe galling, according to Bridge and Page (1980).
Data from each field trial were checked for normality using Proc Univariate in SAS version 9.4 (SAS Institute Inc., Cary, NC). Wherever necessary, nematode abundance data were normalized using log10 (x + 1). Nematode abundance data were subjected to a repeated-measures analysis of variance using Proc GLM in SAS. If significant interaction between treatment and sampling time was detected, data were analyzed by sampling time. Zucchini, tomato and sweet potato yield, root weight, and root-gall index, were subjected to a one-way ANOVA. Means were separated using the Waller–Duncan
No significant difference was detected among treatments on the abundance of
Abundance of A) reniform and B) root-knot nematodes affected by treatments on zucchini or on tomato (C, D) in Trial I. Means (
Abundance of A) reniform and B) root-knot nematodes on zucchini or on tomato (C, D) affected by treatments in Trial II. Means (
In Trial III, there was no significant difference in population densities of root-knot and reniform nematodes among treatments at sweet potato planting, and no significant interaction between sampling dates and treatments were detected, thus nematode abundance data were combined over the three sampling dates after sweet potato planting (at 2, 3, and 6 months after planting). Similar to results in Trial I and Trial II, SH + MoltX suppressed the soil population of
Abundance of A) reniform and B) root-knot nematodes on sweet potato affected by treatments in Trial III. Means (
In Trial I, zucchini chlorophyll content, canopy width, root weight and fruit numbers were all ranked higher for SH + Velum and SH + MoltX followed by Velum I and II (Table 2). Control treatment resulted in lower chlorophyll content and fruit number/plant than Velum II, SH + Velum and SH + MoltX (
Chlorophyll content, canopy width, fruit number, and
Treatment | Chlorophyll content (SPAD) | Canopy width (cm) | Root weight (g) | Fruit no./plant | Root-gall index (0-10 scale) | |||||
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(Trial) | I | II | I | II | I | II | I | II | I | II |
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Velum I | 36 ± 1bcz | 47 ± 2a | 87 ± 7b | 114 ± 5c | 89 ± 12b | 35 ± 4b | 50 ± 3bc | 7 ± 1ab | 3 ± 1a | 2 ± 0c |
Velum II | 37 ± 1b | 46 ± 1a | 89 ± 8b | 118 ± 6bc | 102 ± 27ab | 31 ± 4b | 40 ± 5c | 5 ± 1bc | 3 ± 1a | 2 ± 1bc |
SH + Velum | 44 ± 1a | 47 ± 1a | 117 ± 4a | 134 ± 3a | 155 ± 12a | 49 ± 4a | 72 ± 3a | 10 ± 1a | 4 ± 1a | 3 ± 1b |
SH + MoltX | 42 ± 2a | 45 ± 1a | 109 ± 6a | 127 ± 6ab | 124 ± 19ab | 39 ± 4ab | 54 ± 5b | 6 ± 1bc | 4 ± 2a | 5 ± 2a |
Control | 34 ± 2c | 44 ± 1a | 69 ± 10c | 98 ± 6d | 71 ± 40b | 35 ± 4b | 20 ± 11d | 3 ± 1c | 7 ± 0a | 5 ± 2a |
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Chlorophyll content (SPAD) | Canopy width (cm) | Root weight (g) | Fruit weight g/plot | Root-gall index (0-10 scale) | ||||||
Velum I | – | 44 ± 1a | – | – | 110 ± 10bc | 53 ± 9a | 3525 ± 717a | 123 ± 50a | 4 ± 1b | 2 ± 1c |
Velum II | – | 43 ± 1ab | – | – | 95 ± 21c | 76 ± 17a | 1074 ± 471c | 101 ± 29a | 3 ± 1b | 4 ± 1b |
SH + Velum | – | 45 ± 1a | – | – | 92 ± 3c | 87 ± 22a | 1995 ± 258b | 86 ± 32a | 2 ± 1b | 6 ± 1ab |
SH + MoltX | – | 40 ± 1b | – | – | 153 ± 11ab | 64 ± 19a | 3460 ± 80a | 105 ± 45a | 6 ± 0ab | 7 ± 1a |
Control | – | 43 ± 2ab | – | – | 164 ± 26a | 77 ± 17a | 1625 ± 604b | 112 ± 65a | 8 ± 0a | 8 ± 0a |
Note: zMeans ± standard error in a column followed by the same letter(s) are not different, according to the Waller–Duncan
In Trial II, similar results were observed on both zucchini and tomato where Velum I resulted in the lowest RGI, followed by lower RGI in Velum II and SH + Velum than in control and SH + MoltX (
In Trial III, where the treatment effects were evaluated over a longer period (6 months) and measurement of RGI or canopy width was difficult on sweet potato, the marketable swollen root number or weight were better indicators of the plant response to the soil treatments. The result showed that all fluopyram treatments yielded higher marketable sweet potato number (
Marketable and unmarketable sweet potato root number and weight affected by treatments in Trial III.
Marketable | Unmarketable | |||
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Treatment | Number | Weight (kg) | Number | Weight (kg) |
Velum I | 2 ± 0az | 0.7 ± 0.2a | 2 ± 1a | 0.4 ± 0.1a |
Velum II | 2 ± 0a | 0.8 ± 0.4a | 2 ± 1a | 0.3 ± 0.1a |
SH + Velum | 3 ± 1a | 1.0 ± 0.3a | 3 ± 1a | 0.7 ± 0.2a |
SH + MoltX | 1 ± 0b | 0.2 ± 0.1a | 2 ± 0a | 0.7 ± 0.3a |
Control | 0 ± 0b | 0.1 ± 0.1b | 2 ± 0a | 0.5 ± 0.1a |
zMeans ± standard error (
In Trial I, population densities of each free-living nematode trophic group prior to crop planting were not different among treatments, thus these data were not presented. This allowed a fair comparison among treatments. There was no interaction between sampling time and treatment for the zucchini trial, thus nematode abundance data were presented from the average of 2 sampling dates (Table 4). Minimal to no predatory nematodes were detected in this field, thus their numbers were not presented. Both Velum I and II resulted in lower abundance (
Abundance (/250 cm3 of soil) of bacterivorous, fungivorous, and omnivorous nematodes on zucchini affected by treatments Trial I.
Treatments | Bacterivores | Fungivores | Omnivores |
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Velum I | 614 ± 178bz | 392 ± 226c | 21 ± 18a |
Velum II | 827 ± 489b | 360 ± 208c | 30 ± 21a |
SH + Velum | 1203 ± 217a | 722 ± 417ab | 23 ± 16a |
SH + MoltX | 1538 ± 682a | 844 ± 488a | 40 ± 28a |
Control | 871 ± 195ab | 416 ± 150bc | 23 ± 13a |
zMeans ± standard error (
Abundance of bacterivorous, fungivorous, and omnivorous nematodes (/250 cm3 of soil) on tomato affected by treatments in Trial I.
Treatments | Bacterivores | Fungivores | Omnivores |
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Velum I | 149 ± 54d | 253 ± 61b | 3 ± 3b |
Velum II | 393 ± 90cd | 410 ± 80b | 7 ± 3b |
SH + Velum | 2193 ± 769a | 900 ± 120a | 40 ± 30ab |
SH + MoltX | 1623 ± 613ab | 1123 ± 185a | 220 ± 157a |
Control | 587 ± 148bc | 293 ± 18b | 10 ± 6b |
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Velum I | 140 ± 36c | 103 ± 60b | 0 ± 0b |
Velum II | 70 ± 25d | 127 ± 15b | 0 ± 0b |
SH + Velum | 320 ± 76b | 407 ± 235a | 0 ± 0b |
SH + MoltX | 817 ± 147a | 407 ± 235a | 57 ± 14a |
Control | 370 ± 35b | 180 ± 104ab | 47 ± 27a |
zMeans ± standard error (
Trial II was superimposed on the field site at Trial I, thus population densities of bacterivorous nematodes at planting were continued to be higher in SH + Velum and SH + MoltX than Velum I, II and the control (
Abundance (/250 cm3 of soil) of bacterivorous, fungivorous, and omnivorous nematodes on zucchini affected by treatments in Trial II.
Bacterivores | Fungivores | Omnivores | ||
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Treatments | 0 week | 8 weeks | ||
Velum I | 40 ± 31bz | 353 ± 175a | 118 ± 41b | 0 ± 0a |
Velum II | 50 ± 25b | 194 ± 95a | 95 ± 37b | 0 ± 0a |
SH + Velum | 547 ± 115a | 510 ± 216a | 450 ± 96a | 8 ± 5a |
SH + MoltX | 470 ± 118a | 547 ± 235a | 553 ± 113a | 13 ± 6a |
Control | 53 ± 15b | 633 ± 111a | 87 ± 47b | 18 ± 13a |
Abundance (/250 cm3 of soil) of bacterivorous, fungivorous, and omnivorous nematodes on tomato affected by treatments in Trial II.
Treatments | Bacterivores | Fungivores | Omnivores |
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Velum I | 68 ± 19cz | 90 ± 16b | 7 ± 7bc |
Velum II | 110 ± 35bc | 77 ± 22b | 5 ± 5c |
SH + Velum | 702 ± 122a | 563 ± 158a | 35 ± 27ab |
SH + MoltX | 1162 ± 457a | 657 ± 194a | 62 ± 30a |
Control | 163 ± 65b | 62 ± 17b | 43 ± 25a |
zMeans ± standard error (
In Trial III where a longer-term impact of soil treatments on free-living nematodes was examined on sweet potato, a similar trend was observed. The impact of Velum I and II on free-living nematodes did not dissipate even after 7 months of crop growth. The abundance of bacterivores was reduced at 2 (
Abundance (/250 cm3 of soil) of bacterivorous nematodes on sweet potato affected by treatments in Trial III.
Bacterivores | ||||||
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Treatments | 0 week | 2 months | 3 months | 7 months | Fungivores | Omnivores |
Velum I | 270 ± 91abz | 313 ± 55c | 137 ± 47bc | 43 ± 20b | 83 ± 29b | 1 ± 1c |
Velum II | 310 ± 95ab | 690 ± 316bc | 117 ± 47c | 70 ± 12b | 113 ± 32b | 4 ± 4c |
SH + Velum | 617 ± 292ab | 723 ± 20ab | 293 ± 48abc | 280 ± 86a | 693 ± 339a | 6 ± 3bc |
SH + MoltX | 820 ± 354a | 1520 ± 525a | 803 ± 158a | 563 ± 206a | 460 ± 207a | 23 ± 8a |
Control | 150 ± 111b | 1223 ± 308a | 350 ± 66ab | 390 ± 83a | 57 ± 12b | 19 ± 8ab |
zMeans ± standard error (
In general, results from all three trials demonstrated that early application of fluopyram at crop planting was key to effective suppression of
The suppression of
The suppressive activity of fluopyram alone on
The current study demonstrated that application of fluopyram at planting is the key to the success of suppressing
On the other hand, planting of sunn hemp cover crop followed by monthly post-plant azadirachtin application (SH + MoltX) suppressed
Regardless of differential effects of azadirachtin and fluopyram on different species of plant-parasitic nematodes, green manure effect of sunn hemp in SH + Velum increased zucchini yield by 2.6 and 2.3 folds in Trials I and II, respectively; and SH + MoltX increased zucchini yield by 1.7 and 2.0 folds in Trials I and II, respectively. No yield improvement was observed on tomato on Trial II, but SH + MoltX and SH + Velum increased tomato yield by 22.8 and 112%, respectively, in Trial I. On the contrary,
Unfortunately, fluopyram imposed some negative effect on the abundance of free-living nematodes in particular on longer-term crops (tomato and sweet potato). Interestingly this negative impact of fluopyram was significantly mitigated by integration with sunn hemp cover cropping. In fact, SH + Velum either increased the abundance of bacterivores or fungivores or both on tomato and sweet potato as compared to the control. Whereas SH + MoltX tended to always increase bacterivorous, fungivorous or omnivorous nematodes. Disturbance of free-living nematodes by other non-fumigant types of nematicides has long been studied (Adams et al., 1979; Simpkin and Coles, 1981). Only limited studies have documented the effects of fluopyram on free-living nematodes. Grabau et al. (2020) reported no effect of fluopyram on any free-living nematodes on peanuts (
This study demonstrated that