0 2015 REPORT ON RESEARCH NIMITZ NEMATICIDE FIELD TRIALS Kiwi-1 Ranch, Poplar, CA Kiwi-3 Ranch, Earlimart, CA Abstract Two randomized complete block field trials were conducted at Kiwi-1 Ranch, Poplar, CA and Kiwi-3 Ranch, Earlimart, CA during 2015 to evaluate the efficacy of Nimitz (a.i. fluensulfone) nematicide. The objectives of these trials were to determine if seasonal application of Nimitz nematicide will reduce root-knot nematode (Meloidogyne spp.) soil populations and root infestations in kiwifruit. Nimitz was sprayed onto the top of the berms in 50 gallons per acre spray volume and quickly followed by one inch of irrigation water to move the nematicide into the soil profile. A single application of Nimitz in mid-summer (late July) at 3.5 pints per acre rate provided slight to no control of nematode populations in the soil or within plant roots. A split application of Nimitz (3.5 pts/ac in late May + 3.5 pts/ac in late July) showed trends in reducing soil root knot nematode populations 61 days after the first application and lasting through the summer. The number of nematodes elutriated from fall root samples and the gall severity on roots both trended less for the Split Application compared to the Control and Mid-Summer (3.5 pts/ac in late July) treatments, but high variance among the treatments made it difficult to achieve significance at the 95% confidence level in these trials. No detectable levels of fluensulfone were present in the fruit. John Ojala and Sherod Craig ver. 04/13/16
1 SUMMARY Kiwi-1 Trial Summary - These results indicate that the Mid-Summer (July 27 th ) treatment (3.5 pints Nimitz per acre rate) slightly reduced soil root knot nematode populations thirty-six days after the Nimitz application, but did not sustain this level of control later (94 days after Nimitz application) in the growing season. Fewer root knot nematodes were elutriated from the roots in the Mid-Summer treatment compared to the Control, but not significantly due to high variance between treatments. Gall formation on the roots was not significantly reduced in the first year of Nimitz application. Total fruit yield was significantly greater in the Nimitz Mid-Summer treatment. Fruit grade and fruit quality were not significantly improved by application of Nimitz. This trial demonstrated a weak response to a late July application of 3.5 pints Nimitz per acre for control of root knot nematodes in the soil and roots of kiwifruit. The post-harvest Nimitz treatment will be evaluated in 2016. Kiwi-3 Trial Summary A split application of Nimitz (3.5 pts/ac in late May + 3.5 pts/ac in late July) showed trends in reducing soil root knot nematode populations 61 days after the first application and lasting through the summer. A single application of Nimitz in mid-summer (3.5 pts/ac in late July) did not significantly reduce soil root knot nematode population during the summer months compared to the Control. The less effective control of nematodes in the Mid- Summer compared to the other treatments may be due to the low application rate (3.5 pts/ac) or later application timing, or both. The number of nematodes elutriated from fall root samples and the gall severity on roots both trended less for the Split Application compared to the Control and Mid-Summer (3.5 pts/ac in late July) treatments, but high variance among the treatments made it difficult to achieve significance at the 95% confidence level in this trial. The split application of Nimitz produced significantly fewer large (grade 25) and more medium (grade 36) size fruit compared to the control. Residue Testing No detectable levels of fluensulfone were present in samples of fruit (peel + flesh) from each treatment replication at both trial locations.
2 Nimitz Nematicide Field Trials John Ojala and Sherod Craig Introduction Root knot nematodes are a pervasive problem for kiwifruit growers in California. The most reliable chemical control methods are pre-plant fumigants that work well for the first few years during vineyard establishment. However, nematodes soon re-establish their populations and begin to create galls on roots that impair water uptake, produce sun burning on leaves/fruit, reduce vine vigor and reduce fruit size. There are a few post-plant biological/organic products that may help to suppress nematode populations but they are not generally regarded as reliable or highly effective materials for seasonal nematode control in established vineyards. Nimitz (fluensulfone), distributed by ADAMA Agricultural Solutions, is a new chemistry of nematicides that is currently registered for use on solanaceous and cucurbit crops in California, with additional registration being pursued for other permanent vine and tree crops. Research trials conducted by ADAMA have shown that Nimitz has an efficacy similar to fumigants, yet is much safer and easier to use. This research project evaluates Nimitz application rate and timing on root-knot nematode soil populations and root infestations in kiwifruit, and supports an IR-4 Project whose goal is to add kiwifruit to the Nimitz product label. Two field trails were conducted in 2015, one at the Kiwi-1 Ranch near Poplar, CA and the other at the Kiwi-3 Ranch near Earlimart, CA. Materials & Methods Kiwi-1 Ranch, Poplar The trial was located in a 32 year old stand of Hayward kiwifruit with a 15 ft. by 15 ft. spacing on block 2 of the Sun Pacific Kiwi-1 Ranch near Poplar, CA (Appendix 1). This site was selected because it has a history of root knot galls and nematode soil tests revealed moderate to high (570 to 788 count per 500 cc soil) root knot nematode populations were present in the soil. The trial design was randomized complete block with three treatments and five replications per treatment. The treatments were: 1. 3.5 pts/ac Nimitz applied in mid-summer (July 27th) 2. 3.5 pts/acre Nimitz applied post-harvest (November 13th) 3. Untreated control. Each replication consisted of two female vines (15 ft. row width x 30 ft. length). Berms were approximately 6 ft. wide. A border vine (15ft. of row length) was present between each treatment. The total size of all plots was 0.15 acres (excluding borders) of which 0.10 acres were the Nimitz treatment plots. Sixteen soil subsamples were collected to an eighteen inch depth from each replication on July 17 th, September 1 st and October 29 th. The subsamples from each replication were mixed together to make a single soil sample for each of the fifteen plots. A Pesticide Research Authorization (permit #1507025) to apply restricted materials was obtained from the California Department of Pesticide Regulation. Nimitz nematicide (lot number BO4315002) was hand-sprayed onto the berms of treatment plots with an R&D sprayer fitted with a hand wand containing a flat spray nozzle. The total spray volume applied to the berm of the Nimitz treatments was 50 gallons per acre. The berms of all plots were irrigated with 1.0 inch of water within 48
3 hours of spraying to incorporate Nimitz into the soil profile. Irrigation water was applied by micro sprinklers located about 3 ft. above the berms on 15 ft. spacing (one sprinkler on both sides of the vines). The spray pattern of the sprinklers covered both the berms and the rows. Soil and root samples were collected from each replication post-harvest on November 6 th for nematode analysis. Nematode analyses were conducted by ID Services, McFarland, CA. All samples that were submitted to ID Services were blind and had no Nimitz application identification written on the sample bags, which means they did not know which if any treatment had been applied to any of the samples. Root knot nematode populations were measured in soil samples using a sieving sugar flotation extraction method. Live nematodes were extracted from roots using a Baermann Funnel procedure. Gall injury on root samples were evaluated by visually examining post-harvest root samples. A random sample of fruit from around all areas (high, low, inside, outer) of the vines in each replication was collected on October 20 th and stored under refrigeration (45F) until sent to OMIC USA Inc. (Portland, OR) on October 22 nd for residue analysis of the entire fruit (peel + flesh). Total fruit weight from each replication was measured during harvest on October 20 th and 21 st. Individual weights were also measured during harvest from a random sample of three boxes (85 lbs. to 108 lbs. ranging from 334 to 455 fruit) of kiwifruit from each replication and were used to estimate average fruit size and grade category. A random sample of 60 fruit were collected from each replication during harvest and were measured for total soluble solids content using an Extech portable refractometer and for skin rupture force (maturity) using custom equipment. A small sample of 36 fruit was also collected during harvest and placed into mesh bags for dry matter analysis. The fruit from all plots were treated as crop destruct and after the weight and quality measurements were collected they were discarded into the rows and disked. Trunk circumference was measured for each vine on June 23 rd and again on November 11 th. Kiwi-3 Ranch, Earlimart - The trial was located in a 14 year old stand of Hayward kiwifruit with a 15 ft. by 15 ft. spacing on block 16 of the Sun Pacific Kiwi-3 Ranch near Earlimart, CA (Appendix 1). This site was selected because it has a history of root knot galls and nematode soil tests revealed moderate to high (224 to 982 count per 500 cc soil) root knot nematode populations were present in the soil. The trial design was randomized complete block with three treatments and five replications per treatment. The treatments were: 1) 7.0 pints Nimitz per acre split application (3.5 pints Nimitz per acre applied on May 21 st + 3.5 pints Nimitz per acre applied on July 24 th ) 2) 3.5 pints Nimitz per acre applied in mid-summer (July 24 th ) 3) Untreated control. Each replication consisted of two female vines (15 ft. row width x 30 ft. length). Berms were approximately 6 ft. wide. A border vine (15 ft. of row length) was present between each treatment. The total size of all plots was 0.21 acres (excluding borders) of which 0.15 acres were the Nimitz treatment plots. Sixteen soil subsamples were collected to an eighteen inch depth from each replication on May 20 th, June 25 th, July 21 st, August 26 th and October 30 th. The subsamples from each replication were mixed together to make a single soil sample for each of the fifteen plots. A 12 inch depth soil sample was also collected from each plot on September 10 th. A Pesticide Research Authorization (permit #1505043) to apply restricted materials was obtained from the California Department of Pesticide Regulation. Nimitz nematicide (lot number BO4315002) was hand-sprayed onto the berms of the treatment plots with an R&D sprayer fitted with a hand wand containing a flat spray nozzle. The total spray volume applied to the berms of the Nimitz treatments was 50 gallons per acre. The berms of all plots were irrigated with 1.0 inch of water within 48 hours of spraying to incorporate Nimitz into the soil profile. Irrigation water was applied by micro
4 sprinklers located about 3 ft. above the berms on 15 ft. spacing (one sprinkler on both sides of the vines). The spray pattern of the sprinklers covered primarily the berms with little water application into the rows. Soil and root samples were collected from each replication post-harvest on November 6 th for nematode analysis. Nematode analyses were conducted by ID Services, McFarland, CA. All samples that were submitted to ID Services were blind and had no treatment identification written on the sample bags, which means they did not know which if any treatment had been applied to any of the samples. Root knot nematode populations were measured in soil samples using a sieving sugar flotation extraction method. Live nematodes were extracted from roots using a Baermann Funnel procedure. Galls injury on root samples were evaluated by visually examining roots. A random sample of fruit from around all areas (high, low, inside, outer) of the vines in each replication was collected on October 21 st and stored under refrigeration (45F) until sent to OMIC USA Inc. (Portland, OR) on October 26 th for residue analysis of the entire fruit (peel + flesh). Total fruit weight from each replication was measured during harvest on October 21 st. Individual weights were also measured during harvest from a random sample of three boxes (59 lbs. to 105 lbs. ranging from 233 to 478 fruit) of kiwifruit from each replication and were used to estimate average fruit size and grade category. A random sample of 60 fruit were collected from each replication during harvest and were measured for total soluble solids content using an Extech portable refractometer and for skin rupture force (maturity) using custom equipment. A small sample of 36 fruit was also collected during harvest and placed into mesh bags for dry matter analysis. The fruit from all plots were treated as crop destruct and after harvest measurements were collected they were discarded into the rows and disked. Trunk circumference was measured for each vine on June 24 th and again on November 27 th. Statistical Analyses In both trials, analysis of variance (ANOVA) was conducted on dependent variables containing homogeneous treatment variances. Dependent variables that were non-homogeneous (significant ρ values in a Levene s Test) were transformed (log 10 or square root) and re-tested again for variance homogeneity. If the treatment variances were homogenous then they were analyzed by ANOVA. Any remaining dependent variables that still contained non-homogeneous variances were analyzed with the non-parametric Welch test. Since the post-harvest Nimitz treatment at the Kiwi-1 trial location was applied after all field and lab data was collected in 2015, the post-harvest plots were combined with the untreated plots into a control treatment and statistically compared using a t-test for significance between two means (control and mid-summer treatments). Statistica (StatSoft, Inc., Tulsa, OK) and SPSS (IBM Corporation, New York, NY) software was used to conduct ANOVA, LSD mean separation, Levene s Test, Welch s Test, Brown-Forsythe and other statistical analyses. ANOVA tables were checked for accuracy against hand calculations.
5 Results & Discussion Kiwi-1 Ranch Trial, Poplar The variance and coefficient of variation for soil nematode population within and between treatments was high (Table 1), particularly during the early sampling date for the Control (122% coefficient of variation) and Mid-Summer (63% coefficient of variation) treatments. Treatment variance homogeneity is an important assumption for ANOVA and was found to be true for nearly all measurements (Table A in Appendix 5). Mean separation can be difficult to achieve when the treatment variability is high, as has occurred in this trial. Soil nematode population was significantly different between treatments only on the September 9 th sampling date. However, trends in the data over time should also be examined. Root knot nematode soil populations were lowest in the early samples (July), peaked in late summer (September) and then declined by mid-fall (October) (Fig. 1). The lower nematode population in the Mid-Summer treatment does not indicate nematicide efficacy since the population levels of the Mid-Summer treatment were also lower at the beginning of the season. The Mid-Summer (3.5 pints Nimitz/acre) treatment plots had fewer root knot nematodes than the Untreated on every sampling date, including the pre-treatment samples taken on July 17 th (Fig. 1). Soil root knot nematode populations of the Mid- Summer treatment were significantly less than the Untreated only on the September 1 st sampling date (Table 4 and Fig. 1-B), suggesting some efficacy. For Nimitz to demonstrate nematode control in the soil, the seasonal soil root knot nematode populations in the Mid-Summer treatment should not have a parallel or converging trend when compared to the Untreated. However, the two trend lines are parallel (Fig. 1) during the first two sample dates and ultimately converge on the final sample date, suggesting poor efficacy for the Mid-Summer treatment.
6 The count of root knot nematodes elutriated from after-harvest root samples also trended less in the Mid-Summer treatment (Fig. 3), but not significantly. The gall scores of the after-harvest root samples were not significantly different between treatments (Table 4). Since the post-harvest Nimitz treatment was applied after all field and lab data were collected in 2015, the post-harvest plots were combined with the control plots into one treatment (Untreated). The soil population of Table 1. Root knot nematode soil population, root infestation and galling score in 2015 Nimitz field trial at Kiwi-1 ranch, Poplar, CA. Nimitz Application Plot Soil Sample Date 1 Post-Harvest: 11/06/15 Treatment Replication Rate Number 07/17/15 09/01/15 10/29/15 Roots 2 Galls 3 - - - no. root knot nematodes per 500 cc soil - - - no. per gram root 0 to 10 score Untreated (Control) none applied 1 1 291 601 788 192 5.00 none applied 2 5 442 1,418 357 264 0.00 none applied 3 9 261 1,103 362 409 2.50 none applied 4 11 333 1,430 749 522 3.75 none applied 5 14 2,315 1,624 694 308 6.25 Mean: 728 1,235 590 339 3.50 Std. Deviation: 890 401 213 129 2.40 Variance: 791,362 160,554 45,394 16,690 5.78 Coefficient of Variation: 122.1% 32.4% 36.1% 38.1% 68.7% Post Harvest 4 3.5 pts/acre 1 2-1,152 789 254 5.00 3.5 pts/acre 2 6-788 545 326 5.00 3.5 pts/acre 3 7-1,503 610 267 3.75 3.5 pts/acre 4 12-2,606 704 606 6.25 3.5 pts/acre 5 13-2,279 1,532 344 6.25 Mean: 1,666 836 359 5.25 Std. Deviation: 762 400 143 1.05 Variance: 580,254 159,952 20,379 1.09 Coefficient of Variation: 45.7% 47.8% 39.7% 19.92% Mid-Summer 5 3.5 pts/acre 1 3 267 970 441 100 8.75 3.5 pts/acre 2 4 370 315 550 90 2.50 3.5 pts/acre 3 8 521 491 560 275 5.00 3.5 pts/acre 4 10 39 1,006 367 242 2.50 3.5 pts/acre 5 15 624 1,135 818 502 5.00 Mean: 364 783 547 242 4.75 Std. Deviation: 228 358 171 167 2.56 Variance: 51,830 128,222 29,314 27,956 6.56 Coefficient of Variation: 62.5% 45.7% 31.3% 69.2% 53.9% 1 soil sample depth: 1 to 18 inches; extraction method: sieving sugar flotation. 2 root extraction method: Baermann Funnel procedure. 3 Gall Score: 0 = none (no galls found on roots), 5 = medium (one or more galls), 10 = high (more than 30% of roots have galls). 4 Nimitz application date on the post-harvest application treatment: November 13, 2015. 5 Nimitz application date on the mid-summer application treatment: July 27, 2015.
7 root knot nematodes were significantly (Table 5) less in the Mid-Summer Nimitz treatment (783 counts per 500 cc soil) compared to the untreated Control (1,450 counts per 500 cc soil) in the combined plots (Table 11 in Appendix 2) and diverged away from the Control (Fig. 2). The yield of kiwifruit in the Mid-Summer treatment was significantly greater than the Untreated (Fig. 4). There was no difference between treatments in the average weight of fruit, total soluble solids content of fruit, or fruit maturity as measured by skin pressure (Table 2). There was also no difference between treatments in the grade of fruit (Table 3 and Fig. 5) or in trunk growth (Table 16 in Appendix 2) during the 2015 growing season. Fruit samples from every treatment replication in the trial were collected at harvest and tested for fluensulfone residue by OMIC USA, Inc. Fluensulfone residue in the fruit (peel + flesh) was reported as ND (not detected) in all samples (Appendix 4). Table 2. Yield and fruit quality in the 2015 Nimitz field trial at Kiwi-1 ranch, Poplar, CA. Nimitz Application Total Fruit Treatment Rate Yield Ave. Weight TSS 3 lbs per plot Untreated (Control) none applied 225.2 b 112 ns 6.5 ns 20.1 ns Post-Harvest 1 3.5 pts/acre 214.6 b 116 ns 6.7 ns 19.5 ns Mid-Summer 2 3.5 pts/acre 325.4 a 112 ns 6.4 ns 19.6 ns grams Bx SP 4 psi 1 Nimitz application date on the post-harvest application treatment: November 13, 2015. 2 Nimitz application date on the mid-summer application treatment: July 27, 2015. 3 TSS - the total soluble solids in the fruit juice as estimated by measuring Brix. 4 Skin pressure test - the pressure required to rupture the fruit skin. Numbers in a column followed by the same letters are not significantly (LSD.05 ) different from each other.
8 Table 3. Average fruit grade of each treatment in the 2015 Nimitz field trial at Kiwi-1 ranch, Poplar, CA. Treatment Percent of Total Yield Undersize 45 42 39 36 33 30 27 25 23 20 18 or Larger Untreated (Control) 0.3% ns 0.1% ns 0.7% ns 1.4% ns 2.4% ns 7.5% ns 15.3% ns 17.8% ns 17.7% ns 22.9% ns 8.6% ns 5.3% ns Post-Harvest 0.6% ns 0.2% ns 1.0% ns 2.0% ns 2.5% ns 7.0% ns 14.2% ns 14.3% ns 15.0% ns 19.9% ns 11.0% ns 12.2% ns Mid-Summer 0.5% ns 0.2% ns 1.2% ns 2.2% ns 2.6% ns 8.3% ns 14.9% ns 17.0% ns 14.4% ns 19.3% ns 9.3% ns 10.1% ns Numbers in a column followed by "ns" are not significantly (LSD.05) different from each other. Table 4. Randomized complete block ANOVA probability (ρ) values - Kiwi-1 trial location. Dependent ANOVA Variable ρ Value 1 Nematode Soil & Plant Analyses soil nematode population on 07/17/15 0.34846 soil nematode population on 09/01/15 0.02947 soil nematode population on 10/29/15 0.14558 nematode elutriation from roots on 11/2015 0.29073 gall rating of roots on 11/2015 0.32709 Yield & Fruit Quality total yield from each plot 0.02596 average fruit weight 0.75966 fruit total soluble solids (TSS) 0.50708 fruit skin pressure test (SP) 0.27353 trunk circumference on 06/23/15 0.14891 trunk circumference on 11/11/15 0.13833 delta trunk circumferences 0.69542 Fruit Grade Undersize 0.65620 Grade 45 0.75675 Grade 42 0.77376 Grade 39 0.82033 Grade 36 0.99256 Grade 33 0.93886 Grade 30 0.95465 Grade 27 0.17705 2 Grade 25 0.25525 Grade 23 0.71759 Grade 20 0.76056 Grade 18+ 0.18478 1 probability (ρ) values that are less than 0.05 means that there is a 95% confidence level that a significant difference between treatment means occurred (highlighted in red in the table). 2 Welch's test (used when varainces are heterogeneous).
9 Table 5. T test statistics for comparing all untreated (control + post-harvest) to the mid-summer treatment. T-tests; Grouping: Treatment (Kiwi-1 2015 Nimitz Trial - Combined Controls 2) Group 1: Untreated Group 2: Mid-Summer Mean Mean t-value df p Valid N Valid N Std.Dev. Std.Dev. Variable Untreated Mid-Summer Untreated Mid-Summer Untreated Mid-Summer 07/17/15 728.400 364.2000 0.88687 8 0.401021 5 5 889.5852 227.6614 09/01/15 1450.400 783.4000 2.21223 13 0.045464 10 5 617.0127 358.0814 10/29/15 713.000 547.2000 1.04544 13 0.314872 10 5 328.7468 171.2124 Root Elutr 349.200 241.8000 1.38283 13 0.190009 10 5 128.8563 167.2997 Galls 4.375 4.7500-0.31502 13 0.757747 10 5 1.9764 2.5617 Total Yield 219.890 325.4400-2.35364 13 0.034988 10 5 89.1299 62.5516 Ave. Fruit Wt. 113.752 111.8786 0.41042 13 0.688179 10 5 8.7116 7.4060 TSS 6.583 6.3751 0.93107 13 0.368787 10 5 0.4668 0.2273 SP 19.822 19.5667 0.47640 13 0.641700 10 5 0.9499 1.0363 Grade Under 0.004 0.0051-0.32582 13 0.749744 10 5 0.0048 0.0058 Grade 45 0.001 0.0018-0.45669 13 0.655432 10 5 0.0019 0.0032 Grade 42 0.008 0.0119-0.66819 13 0.515699 10 5 0.0085 0.0122 Grade 39 0.017 0.0221-0.53506 13 0.601643 10 5 0.0169 0.0178 Grade 36 0.025 0.0260-0.11981 13 0.906467 10 5 0.0212 0.0182 Grade 33 0.073 0.0829-0.39166 13 0.701657 10 5 0.0466 0.0471 Grade 30 0.148 0.1488-0.03348 13 0.973799 10 5 0.0506 0.0431 Grade 27 0.161 0.1698-0.48861 13 0.633256 10 5 0.0399 0.0142 Grade 25 0.163 0.1442 1.23101 13 0.240127 10 5 0.0318 0.0194 Grade 23 0.214 0.1934 0.59521 13 0.561925 10 5 0.0622 0.0660 Grade 20 0.098 0.0934 0.18583 13 0.855448 10 5 0.0474 0.0407 Grade 18+ 0.088 0.1007-0.44000 13 0.667164 10 5 0.0599 0.0380 TCircum 6/23/15 39.802 43.1780-1.79137 13 0.096532 10 5 4.0161 1.4786 TCircum 11/11/15 40.036 43.3160-1.85213 13 0.086845 10 5 3.8144 1.1131 Delta Circum 0.234 0.1380 0.31997 13 0.754075 10 5 0.5604 0.5182 Variables (rows) containing probability values (ρ) highlighted in red indicate that there is a significant (95% confidence level) difference between treatment means. Kiwi-1 Trial Summary - These results indicate that the Mid-Summer (July 27 th ) treatment (3.5 pints Nimitz per acre rate) slightly reduced soil root knot nematode populations thirty-six days after the Nimitz application, but did not sustain this level of control later (94 days after Nimitz application) in the growing season. Fewer root knot nematodes were elutriated from the roots in the Mid-Summer treatment compared to the Control, but not significantly due to high variance between treatments. Gall formation on the roots was not significantly reduced in the first year of Nimitz application. Total fruit yield was significantly greater in the Nimitz Mid-Summer treatment. Fruit grade and fruit quality were not significantly improved by application of Nimitz. This trial demonstrated a weak response to a late July application of 3.5 pints Nimitz per acre for control of root knot nematodes in the soil and roots of kiwifruit. The post-harvest Nimitz treatment will be evaluated in 2016.
10 Kiwi-3 Ranch Trial, Earlimart The variability of soil nematode population within treatments was high across all the summer sampling dates in this trial. Large coefficient of variations were present in both soil and root measurements (Table 6). All measured variables were evaluated (and corrected when required) for variance homogeneity (Table B in Appendix 6) before ANOVA tests were conducted. However, the large variances reduced the sensitivity of ANOVA to detect significant (95% confidence level) differences between treatments in soil nematode count means (Table 9). However, trends in the data indicate that Nimitz did have a beneficial effect in reducing root knot nematode population in the soil. Root knot nematode soil populations in the Control and Mid-Summer treatments followed a trend similar to the trial at the Kiwi-1 location. Soil populations were low early in the season, then peaked in late summer and declined after harvest (Fig. 6). The Split Application treatment did not follow this pattern and instead showed consistently low root knot soil populations beginning 61 days after the first Nimitz application and lasting through the summer. The two applications and higher rate (7.0 pints/acre) of Nimitz in the Split Application treatment reduced soil nematode populations better than the Control or the single application (3.5 pts/ac) of Nimitz in the Mid- Summer treatment. However, the nematodes elutriated from the roots (Fig. 7) and galls on the roots (Fig. 8) after harvest were the same in the Mid-Summer and Control treatments. Nematodes presence in the roots and galls appeared to trend lower in the Split Application treatment when compared to either the Control or Mid-Summer treatments.
11 Table 6. Root knot nematode soil population 1, root infestation and galling score in the 2015 Nimitz field trial at Kiwi-3 ranch, Earlimart, CA. Nimitz Application Plot Sample Date Post-Harvest: 11/06/15 Treatment Replication Rate and Timing Number 05/20/15 06/25/15 07/21/15 8/26/2015 10/30/2015 Roots Gall Score 4 - - - - - - - - - - - - no. root knot nematodes per 500 cc soil - - - - - - - - - - - - - no. per gram root 0 to 10 score Untreated (Control) none applied 1 3 672 564 933 824 912 146 3.75 none applied 2 5 509 1,697 1,697 1,782 1,025 279 6.25 none applied 3 7 764 218 909 1,239 416 152 3.75 none applied 4 11 400 230 285 691 521 35 10.00 none applied 5 13 672 721 455 485 774 122 3.75 Mean: 603 686 856 1,004 730 147 5.50 Std. Deviation: 146 605 548 515 257 88 2.74 Variance: 21,372 366,298 300,715 265,060 66,090 7,690 7.50 Coefficient of Variation: 24.2% 88.2% 64.1% 51.3% 35.2% 59.8% 49.8% Split Nimitz Application 2 3.5 pts/acre May + 3.5 pts/ac July 1 2 964 661 800 1,436 1,003 169 6.25 3.5 pts/acre May + 3.5 pts/ac July 2 6 364 570 497 533 851 156 2.50 3.5 pts/acre May + 3.5 pts/ac July 3 8 339 485 448 661 868 13 1.25 3.5 pts/acre May + 3.5 pts/ac July 4 10 473 1,358 885 588 506 137 2.50 3.5 pts/acre May + 3.5 pts/ac July 5 15 679 1,927 1,236 648 451 30 2.50 Mean: 564 1,000 773 773 736 101 3.00 Std. Deviation: 261 623 320 374 243 74 1.90 Variance: 68,033 388,135 102,361 139,891 59,016 5,470 3.59 Coefficient of Variation: 46.3% 62.3% 41.4% 48.4% 33.0% 73.3% 63.2% Mid-Summer 3 3.5 pts/acre July 1 1 - - 1,471 1,127 322 558 10.00 3.5 pts/acre July 2 4 - - 503 800 288 90 0.00 3.5 pts/acre July 3 9 - - 497 1,733 496 68 0.00 3.5 pts/acre July 4 12 - - 1,527 618 505 42 8.75 3.5 pts/acre July 5 14 - - 1,067 1,515 456 92 10.00 Mean: - - 1,013 1,159 413 170 5.75 Std. Deviation: - - 501 469 101 218 5.27 Variance: - - 250,808 219,699 10,277 47,421 27.81 Coefficient of Variation: - - 49.4% 40.5% 24.5% 128.2% 91.7% 1 soil sample depth: 1 to 18 inches; extraction method: sieving sugar flotation. 2 Nimitz application dates on the split application treatment: May 21, 2015 and July 24, 2015. 3 Nimitz application date on the mid-summer application treatment: July 24, 2015. 4 Gall Score: 0 = none (no galls found on roots), 5 = medium (one or more galls), 10 = high (more than 30% of roots have galls).
12 There were no significant differences between treatments in total yield, average fruit weight, fruit total soluble solids content and skin pressure tests for maturity (Fig. 9 and Table 7). Both the Split and Mid-Summer Nimitz applications produced significantly more fruit in the medium 36 grade size and less fruit in the larger 25 grade size compared to the Control (Fig. 10 and Table 8). In general, production of more fruit in the 36 grade size is preferable to the larger 25 grade size. The trunk growth of the Mid-Summer and Split Application treatments were not significantly different from the Control (Table 20 and Fig. Q in Appendix 3) in the 2015 growing season Fruit samples from every treatment replication in the trial were collected at harvest and tested for fluensulfone residue by OMIC USA, Inc. Fluensulfone residue in the fruit (peel + flesh) was reported as ND (not detected) in all samples (Appendix 4). Table 7. Yield and fruit quality in the 2015 Nimitz field trial at Kiwi-3 ranch, Earlimart, CA. Nimitz Application Total Fruit Treatment Rate Yield Ave. Weight TSS 3 lbs per plot grams Bx SP 4 psi Untreated (Control) none applied 233.1 ns 112 ns 5.6 ns 19.8 ns Split Application 1 3.5 pts/acre + 3.5 pts/acre 221.7 ns 106 ns 5.5 ns 19.6 ns Mid-Summer 2 3.5 pts/acre 149.7 ns 112 ns 5.9 ns 19.3 ns 1 Nimitz applications on the split application treatment: 3.5 pts Nimitz/acre on May 21, 2015 and 3.5 pts Nimitz/acre on July 24, 2015. 2 Nimitz application on the mid-summer application treatment: 3.5 pts Nimitz/acre on July 24, 2015. 3 TSS - the total soluble solids in the fruit juice as estimated by measuring Brix. 4 Skin pressure test - the pressure required to rupture the fruit skin. Numbers in a column followed by the same letters are not significantly (LSD.05 ) different from each other.
13 Table 8. Average fruit grade from each treatment in the 2015 Nimitz field trial at Kiwi-3 ranch, Earlimart, CA. Percent of Total Yield Treatment Undersize 45 42 39 36 33 30 27 25 23 20 18 or Larger Untreated (Control) 0.3% ns 0.1% ns 0.6% ns 0.9% ns 1.2% c 5.9% ns 17.1% ns 21.5% ns 18.5% a 20.8% ns 8.5% ns 4.5% ns Split Nimitz Application 1.6% ns 0.5% ns 1.8% ns 2.5% ns 4.4% a 10.6% ns 18.5% ns 18.0% ns 14.3% b 15.9% ns 6.2% ns 7.0% ns Mid-Summer 0.9% ns 0.2% ns 1.4% ns 1.2% ns 2.3% b 6.2% ns 14.8% ns 17.0% ns 15.7% b 22.4% ns 9.9% ns 8.1% ns Numbers in a column followed by the same letters are not significantly (LSD.05) different from each other. Kiwi-3 Trial Summary A split application of Nimitz (3.5 pts/ac in late May + 3.5 pts/ac in late July) showed trends in reducing soil root knot nematode populations 61 days after the first application and lasting through the summer. A single application of Nimitz in mid-summer (3.5 pts/ac in late July) did not significantly reduce soil root knot nematode population during the summer months compared to the Control. The less effective control of nematodes in the Mid- Summer compared to the other treatments may be due to the low application rate (3.5 pts/ac) or later application timing, or both. The number of nematodes elutriated from fall root samples and the gall severity on roots both trended less for the Split Application compared to the Control and Mid-Summer (3.5 pts/ac in late July) treatments, but high variance among the treatments made it difficult to achieve significance at the 95% confidence level in this trial. The split application of Nimitz produced significantly fewer large (grade 25) and more medium (grade 36) size fruit compared to the control.
14 Table 9. ANOVA probability (ρ) values - 2015 Nimitz Trial, Kiwi-3 Ranch, Earlimart, CA. Dependent ANOVA Variable ρ Value 1 Nematode Soil & Root Analyses soil nematode population on 05/20/15 0.75652 soil nematode population on 06/25/15 0.49917 soil nematode population on 07/21/15 0.77846 soil nematode population on 08/26/15 0.48047 soil nematode population on 09/9/15 (12 in.) 0.86895 soil nematode population on 10/30/15 0.09248 2 nematode elutriation from root samples 11/06/15 0.70319 gall rating of root samples 11/06/15 0.27100 4 Yield & Quality Measurements total yield from each plot 0.07563 average fruit weight 0.11237 fruit total soluble solids (TSS) 0.14101 fruit skin pressure test (SP) 0.56900 tree circumference 06/24/15 0.50527 tree circumference 11/27/15 0.59535 delta tree circumference 0.04583 Grade Measurements Undersize 0.32612 Grade 45 0.09876 Grade 42 0.11584 Grade 39 0.08906 Grade 36 0.00217 3 Grade 33 0.05452 Grade 30 0.66002 Grade 27 0.12862 Grade 25 0.00285 Grade 23 0.17942 Grade 20 0.23028 Grade 18+ 0.23481 1 Variables containing probability values (ρ) highlighted in red indicate that there is a significant (95% confidence level) difference between treatment means. 2 log 10 transformed (to provide homogeneous treatment variances). 3 square root transformed (to provide homogeneous treatment variances). 4 Welch's non-parametric test for non-homogeneous variances.
15 Appendix 1 Trial and Plot Maps Map 1: Kiwi-1 Ranch trial site field map Map 2: Kiwi-1 Ranch plot map Map 3: Kiwi-3 Ranch trial site field map Map 4: Kiwi-3 Ranch plot map
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17 Nimitz Field Trial Kiwi-1 Ranch, Block 2 Block # I II III IV V 3 1 2 1 2 1 4 7 10 13 border border border border border E 2 3 1 3 3 N S 2 5 8 11 14 border border border border border W 1 2 3 2 1 3 6 9 12 15 3 buffer 2 buffer 4 buffer 9 buffer 9 buffer plants plants plants plants plants row # 36 35 34 33 32 31 30 Ribbon Color orange yellow red white Treatments 1 = mid-summer application (3.5 pts Nimitz/ac applied in July) 2 = post-harvest application (3.5 pts Nimitz/ac applied in October) 3 = untreated (control) border vines
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19 Nimitz Field Trial Kiwi-3 Ranch, Block 16 Block # I II III IV V 2 2 3 1 3 1 7 10 13 border 4 border border border border W 1 3 1 3 2 S N 2 8 11 14 E border border border border 5 border 3 2 2 1 1 3 9 12 15 6 row # 15 14 13 12 11 10 9 Ribbon Color blue green red white Treatments 1 = split application (3.5 pts Nimitz/ac applied in May + 3.5 pts Nimitz/ac applied in July) 2 = mid-summer application (3.5 pts Nimitz/ac applied in July) 3 = untreated (control) border vines
20 Appendix 2 Nematode & Yield Data: Kiwi-1 Ranch Table 10. Root knot nematode population Fig. A. Root knot nematode soil Fig. B. Root knot nematode roots Fig. C. Root knot nematode gall score Table 11. Root knot nematode population (untreated plots combined) Fig. D. Root knot nematode soil (combined plots) Fig. E. Root knot nematode roots (combined plots) Fig. F. Root knot nematode gall score (combined plots) Table 12. Yield and fruit quality Fig. G. Kiwifruit yield Table 13. Yield and fruit quality (untreated plots combined) Fig. H. Kiwifruit yield (combined plots) Table 14. Fruit grade Fig. I. Fruit grade Table 15. Fruit grade (untreated plots combined) Fig. J. Fruit grade (combined plots) Table 16. Trunk growth Fig. K. Trunk circumference increase in 2015
21 Table 10. Root knot nematode soil population, root infestation and galling score in 2015 Nimitz field trial at Kiwi-1 ranch, Poplar, CA. Nimitz Application Plot Soil Sample Date 1 Post-Harvest: 11/06/15 Treatment Replication Rate Number 07/17/15 09/01/15 10/29/15 Roots 2 Galls 3 - - - no. root knot nematodes per 500 cc soil - - - no. per gram root 0 to 10 score Untreated (Control) none applied 1 1 291 601 788 192 5.00 none applied 2 5 442 1,418 357 264 0.00 none applied 3 9 261 1,103 362 409 2.50 none applied 4 11 333 1,430 749 522 3.75 none applied 5 14 2,315 1,624 694 308 6.25 Mean: 728 1,235 590 339 3.50 Std. Deviation: 890 401 213 129 2.40 Variance: 791,362 160,554 45,394 16,690 5.78 Coefficient of Variation: 122.1% 32.4% 36.1% 38.1% 68.7% Post Harvest 4 3.5 pts/acre 1 2-1,152 789 254 5.00 3.5 pts/acre 2 6-788 545 326 5.00 3.5 pts/acre 3 7-1,503 610 267 3.75 3.5 pts/acre 4 12-2,606 704 606 6.25 3.5 pts/acre 5 13-2,279 1,532 344 6.25 Mean: 1,666 836 359 5.25 Std. Deviation: 762 400 143 1.05 Variance: 580,254 159,952 20,379 1.09 Coefficient of Variation: 45.7% 47.8% 39.7% 19.92% Mid-Summer 5 3.5 pts/acre 1 3 267 970 441 100 8.75 3.5 pts/acre 2 4 370 315 550 90 2.50 3.5 pts/acre 3 8 521 491 560 275 5.00 3.5 pts/acre 4 10 39 1,006 367 242 2.50 3.5 pts/acre 5 15 624 1,135 818 502 5.00 Mean: 364 783 547 242 4.75 Std. Deviation: 228 358 171 167 2.56 Variance: 51,830 128,222 29,314 27,956 6.56 Coefficient of Variation: 62.5% 45.7% 31.3% 69.2% 53.9% 1 soil sample depth: 1 to 18 inches; extraction method: sieving sugar flotation. 2 root extraction method: Baermann Funnel procedure. 3 Gall Score: 0 = none (no galls found on roots), 5 = medium (one or more galls), 10 = high (more than 30% of roots have galls). 4 Nimitz application date on the post-harvest application treatment: November 13, 2015. 5 Nimitz application date on the mid-summer application treatment: July 27, 2015.
22 (the post-harvest treatment is combined with the untreated control) Table 11. Root knot nematode soil population, root infestation and galling score in 2015 Nimitz field trial at Kiwi-1 ranch, Poplar, CA. Nimitz Application Plot Soil Sample Date 1 Post-Harvest: 11/06/15 Treatment Replication Rate Number 07/17/15 09/01/15 10/29/15 Roots 2 Galls 3 - - - no. root knot nematodes per 500 cc soil - - - no. per gram root 0 to 10 score Control (Untreated) none applied 1 1 291 601 788 192 5.00 none applied 2 5 442 1,418 357 264 0.00 none applied 3 9 261 1,103 362 409 2.50 none applied 4 11 333 1,430 749 522 3.75 none applied 5 14 2,315 1,624 694 308 6.25 Control (Post Harvest 4 ) 3.5 pts/acre 1 2-1,152 789 254 5.00 3.5 pts/acre 2 6-788 545 326 5.00 3.5 pts/acre 3 7-1,503 610 267 3.75 3.5 pts/acre 4 12-2,606 704 606 6.25 3.5 pts/acre 5 13-2,279 1,532 344 6.25 Mean: 728 1,450 713 349 4.38 Std. Deviation: 890 617 329 129 1.98 Variance: 791,362 380,705 108,074 16,591 3.91 Coefficient of Variation: 122.1% 42.5% 46.1% 36.9% 45.18% Mid-Summer 5 3.5 pts/acre 1 3 267 970 441 100 8.75 3.5 pts/acre 2 4 370 315 550 90 2.50 3.5 pts/acre 3 8 521 491 560 275 5.00 3.5 pts/acre 4 10 39 1,006 367 242 2.50 3.5 pts/acre 5 15 624 1,135 818 502 5.00 Mean: 364 783 547 242 4.75 Std. Deviation: 228 358 171 167 2.56 Variance: 51,830 128,222 29,314 27,956 6.56 Coefficient of Variation: 62.5% 45.7% 31.3% 69.2% 53.9% 1 soil sample depth: 1 to 18 inches; extraction method: sieving sugar flotation. 2 root extraction method: Baermann Funnel procedure. 3 Gall Score: 0 = none (no galls found on roots), 5 = medium (one or more galls), 10 = high (more than 30% of roots have galls). 4 Nimitz application date on the post-harvest application treatment: November 13, 2015; combined with the control data since Nimitz was applied to this treatment after data collection. 5 Nimitz application date on the mid-summer application treatment: July 27, 2015.
23 Table 12. Yield and fruit quality from each plot in the 2015 Nimitz field trial at Kiwi-1 ranch, Poplar, CA. Nimitz Application Plot Total Fruit Treatment Replication Rate Number Yield Average Weight TSS 1 SP 2 lbs per plot grams Bx psi Untreated (Control) none applied 1 1 419.2 116 6.8 19.9 none applied 2 5 209.8 106 6.6 20.6 none applied 3 9 189.0 101 6.5 20.3 none applied 4 11 184.2 121 6.4 21.4 none applied 5 14 124.0 115 6.1 18.6 Mean: 225.2 112 6.5 20.1 Std. Deviation: 113.0 8 0.3 1.0 Variance: 12,773.6 65 0.1 1.1 Coefficient of Variation: 50.2% 7.2% 3.9% 5.1% Post Harvest 3.5 pts/acre October 1 2 265.3 103 6.1 19.6 3.5 pts/acre October 2 6 112.3 107 7.0 19.3 3.5 pts/acre October 3 7 281.0 121 7.6 20.4 3.5 pts/acre October 4 12 243.0 122 6.1 20.0 3.5 pts/acre October 5 13 171.4 125 6.6 18.2 Mean: 214.6 116 6.7 19.5 Std. Deviation: 70.9 10 0.6 0.8 Variance: 5,030.0 96 0.4 0.7 Coefficient of Variation: 33.05% 8.45% 9.47% 4.33% Mid-Summer 3.5 pts/acre July 1 3 425.1 116 6.5 21.0 3.5 pts/acre July 2 4 256.6 114 6.5 19.3 3.5 pts/acre July 3 8 326.2 117 6.3 19.2 3.5 pts/acre July 4 10 325.2 99 6.0 20.1 3.5 pts/acre July 5 15 294.2 114 6.5 18.3 Mean: 325.4 112 6.4 19.6 Std. Deviation: 62.6 7 0.2 1.0 Variance: 3,912.7 55 0.1 1.1 Coefficient of Variation: 19.22% 6.62% 3.57% 5.30% 1 TSS - total soluble solids. 2 SP - skin pressure test.
24 (the post-harvest treatment is combined with the untreated control) Table 13. Yield and fruit quality from each plot in the 2015 Nimitz field trial at Kiwi-1 ranch. Nimitz Application Plot Total Fruit Treatment Replication Rate Number Yield Average Weight TSS 1 SP 2 lbs per plot grams Bx psi Control (Untreated) none applied 1 1 419.2 116 6.8 19.9 none applied 2 5 209.8 106 6.6 20.6 none applied 3 9 189.0 101 6.5 20.3 none applied 4 11 184.2 121 6.4 21.4 none applied 5 14 124.0 115 6.1 18.6 Control (Post Harvest 3 ) 3.5 pts/acre 1 2 265.3 103 6.1 19.6 3.5 pts/acre 2 6 112.3 107 7.0 19.3 3.5 pts/acre 3 7 281.0 121 7.6 20.4 3.5 pts/acre 4 12 243.0 122 6.1 20.0 3.5 pts/acre 5 13 171.4 125 6.6 18.2 Mean: 219.9 114 6.6 19.8 Std. Deviation: 89.1 9 0.5 0.9 Variance: 7,944.1 76 0.2 0.9 Coefficient of Variation: 40.5% 7.7% 7.1% 4.8% Mid-Summer 3.5 pts/acre July 1 3 425.1 116 6.5 21.0 3.5 pts/acre July 2 4 256.6 114 6.5 19.3 3.5 pts/acre July 3 8 326.2 117 6.3 19.2 3.5 pts/acre July 4 10 325.2 99 6.0 20.1 3.5 pts/acre July 5 15 294.2 114 6.5 18.3 Mean: 325.4 112 6.4 19.6 Std. Deviation: 62.6 7 0.2 1.0 Variance: 3,912.7 55 0.1 1.1 Coefficient of Variation: 19.22% 6.62% 3.57% 5.30% 1 TSS - total soluble solids. 2 SP - skin pressure test. 4 Nimitz application date on the post-harvest application treatment: November 13, 2015; combined with the control data since Nimitz was applied to this treatment after data collection.
25 Table 14. Fruit grade from each plot in the 2015 Nimitz field trial at Kiwi-1 ranch. Nimitz Application Plot Treatment Replication Percent of Total Yield Rate and Timing Number Undersize 45 42 39 36 33 30 27 25 23 20 18 or Larger Untreated (Control) none applied 1 1 0.2% 0.0% 0.0% 0.4% 1.2% 4.6% 14.0% 13.4% 20.2% 27.7% 11.8% 6.4% none applied 2 5 0.3% 0.0% 0.8% 1.6% 3.3% 12.5% 19.3% 22.8% 16.3% 16.0% 3.8% 3.3% none applied 3 9 0.8% 0.3% 2.5% 4.9% 6.6% 15.2% 20.1% 16.8% 12.0% 14.8% 4.3% 1.6% none applied 4 11 0.0% 0.0% 0.0% 0.0% 0.7% 3.1% 8.7% 13.5% 21.5% 26.7% 14.1% 11.6% none applied 5 14 0.0% 0.0% 0.0% 0.2% 0.2% 2.3% 14.7% 22.7% 18.4% 29.2% 8.8% 3.6% Mean: 0.3% 0.1% 0.7% 1.4% 2.4% 7.5% 15.3% 17.8% 17.7% 22.9% 8.6% 5.3% Std. Deviation: 0.3% 0.1% 1.1% 2.0% 2.6% 5.9% 4.6% 4.7% 3.7% 6.9% 4.5% 3.9% Coefficient of Variation: 124.3% 223.6% 165.5% 143.5% 109.0% 78.3% 30.0% 26.2% 21.1% 30.1% 52.8% 73.6% Variance: 0.001% 0.000% 0.012% 0.041% 0.069% 0.348% 0.212% 0.219% 0.139% 0.476% 0.205% 0.153% Post Harvest 3.5 pts/acre October 1 2 0.9% 0.0% 1.3% 3.6% 5.0% 12.3% 23.3% 17.6% 13.6% 12.7% 4.7% 4.9% 3.5 pts/acre October 2 6 1.4% 0.6% 1.7% 3.4% 3.4% 8.9% 16.7% 15.7% 14.6% 16.2% 9.5% 8.0% 3.5 pts/acre October 3 7 0.1% 0.1% 1.0% 1.6% 1.5% 6.5% 12.7% 12.0% 15.6% 20.3% 9.9% 18.8% 3.5 pts/acre October 4 12 0.0% 0.0% 0.0% 0.3% 0.4% 2.7% 8.0% 11.8% 18.3% 27.9% 18.8% 11.8% 3.5 pts/acre October 5 13 0.5% 0.1% 1.0% 1.1% 2.4% 4.8% 10.6% 14.3% 13.0% 22.5% 12.2% 17.5% Mean: 0.6% 0.2% 1.0% 2.0% 2.5% 7.0% 14.2% 14.3% 15.0% 19.9% 11.0% 12.2% Std. Deviation: 0.6% 0.2% 0.6% 1.4% 1.8% 3.7% 6.0% 2.5% 2.1% 5.8% 5.1% 6.0% Coefficient of Variation: 104.0% 140.1% 62.3% 72.0% 70.6% 53.0% 41.9% 17.2% 13.8% 29.2% 46.5% 48.7% Variance: 0.004% 0.001% 0.004% 0.021% 0.032% 0.139% 0.357% 0.061% 0.043% 0.339% 0.263% 0.355% Mid-Summer 3.5 pts/acre July 1 3 0.3% 0.0% 0.4% 1.3% 1.2% 4.4% 13.7% 17.9% 15.6% 23.2% 12.0% 10.0% 3.5 pts/acre July 2 4 0.7% 0.0% 1.1% 2.1% 2.2% 6.4% 12.4% 15.9% 13.9% 21.1% 9.3% 14.9% 3.5 pts/acre July 3 8 0.0% 0.0% 0.0% 0.7% 2.2% 5.4% 12.6% 16.0% 16.8% 22.8% 12.8% 10.7% 3.5 pts/acre July 4 10 1.4% 0.7% 3.1% 5.3% 5.8% 16.1% 22.5% 19.0% 11.6% 7.6% 2.5% 4.3% 3.5 pts/acre July 5 15 0.1% 0.1% 1.3% 1.7% 1.6% 9.1% 13.2% 16.0% 14.2% 22.0% 10.1% 10.5% Mean: 0.5% 0.2% 1.2% 2.2% 2.6% 8.3% 14.9% 17.0% 14.4% 19.3% 9.3% 10.1% Std. Deviation: 0.6% 0.3% 1.2% 1.8% 1.8% 4.7% 4.3% 1.4% 1.9% 6.6% 4.1% 3.8% Coefficient of Variation: 113.8% 183.2% 102.9% 80.5% 69.8% 56.8% 28.9% 8.4% 13.4% 34.1% 43.6% 37.8% Variance: 0.003% 0.001% 0.015% 0.032% 0.033% 0.221% 0.185% 0.020% 0.038% 0.435% 0.166% 0.145%
(the post-harvest treatment is combined with the untreated control) Table 15. Fruit grade from each plot in the 2015 Nimitz field trial at Kiwi-1 ranch. Nimitz Application Plot 26 Rate and Timing Number Undersize 45 42 39 36 33 30 27 25 23 20 18 or Larger 1 Post-harvest plots were treated with 3.5 pints Nimitz per acre on November 13, 2015, after all yield and nematode data had been collected, and therefore were analyzed as control (untreated) plots. Treatment Replication Percent of Total Yield Control (Untreated) none applied 1 1 0.2% 0.0% 0.0% 0.4% 1.2% 4.6% 14.0% 13.4% 20.2% 27.7% 11.8% 6.4% none applied 2 5 0.3% 0.0% 0.8% 1.6% 3.3% 12.5% 19.3% 22.8% 16.3% 16.0% 3.8% 3.3% none applied 3 9 0.8% 0.3% 2.5% 4.9% 6.6% 15.2% 20.1% 16.8% 12.0% 14.8% 4.3% 1.6% none applied 4 11 0.0% 0.0% 0.0% 0.0% 0.7% 3.1% 8.7% 13.5% 21.5% 26.7% 14.1% 11.6% none applied 5 14 0.0% 0.0% 0.0% 0.2% 0.2% 2.3% 14.7% 22.7% 18.4% 29.2% 8.8% 3.6% Control (Post Harvest) 1 3.5 pts/acre 1 2 0.9% 0.0% 1.3% 3.6% 5.0% 12.3% 0.0% 17.6% 13.6% 12.7% 4.7% 4.9% 3.5 pts/acre 2 6 1.4% 0.6% 1.7% 3.4% 3.4% 8.9% 16.7% 15.7% 14.6% 16.2% 9.5% 8.0% 3.5 pts/acre 3 7 0.1% 0.1% 1.0% 1.6% 1.5% 6.5% 12.7% 12.0% 15.6% 20.3% 9.9% 18.8% 3.5 pts/acre 4 12 0.0% 0.0% 0.0% 0.3% 0.4% 2.7% 8.0% 11.8% 18.3% 27.9% 18.8% 11.8% 3.5 pts/acre 5 13 0.5% 0.1% 1.0% 1.1% 2.4% 4.8% 10.6% 14.3% 13.0% 22.5% 12.2% 17.5% Mean: 0.4% 0.1% 0.8% 1.7% 2.5% 7.3% 12.5% 16.1% 16.3% 21.4% 9.8% 8.8% Std. Deviation: 0.5% 0.2% 0.9% 1.7% 2.1% 4.7% 6.0% 4.0% 3.2% 6.2% 4.7% 6.0% Variance: 0.000023 0.000004 0.000072 0.000285 0.000450 0.002173 0.003586 0.001593 0.001011 0.003872 0.002250 0.003585 Coefficient of Variation: 116.0% 166.7% 102.8% 98.8% 85.9% 64.0% 48.0% 24.8% 19.5% 29.1% 48.4% 68.3% Mid-Summer 3.5 pts/acre July 1 3 0.3% 0.0% 0.4% 1.3% 0.0% 4.4% 13.7% 17.9% 15.6% 23.2% 12.0% 10.0% 3.5 pts/acre July 2 4 0.7% 0.0% 1.1% 2.1% 2.2% 6.4% 12.4% 15.9% 13.9% 21.1% 9.3% 14.9% 3.5 pts/acre July 3 8 0.0% 0.0% 0.0% 0.7% 2.2% 5.4% 12.6% 16.0% 16.8% 22.8% 12.8% 10.7% 3.5 pts/acre July 4 10 1.4% 0.7% 3.1% 5.3% 5.8% 16.1% 22.5% 19.0% 11.6% 7.6% 2.5% 4.3% 3.5 pts/acre July 5 15 0.1% 0.1% 1.3% 1.7% 1.6% 9.1% 13.2% 16.0% 14.2% 22.0% 10.1% 10.5% Mean: 0.5% 0.2% 1.2% 2.2% 2.4% 8.3% 14.9% 17.0% 14.4% 19.3% 9.3% 10.1% Std. Deviation: 0.6% 0.3% 1.2% 1.8% 2.1% 4.7% 4.3% 1.4% 1.9% 6.6% 4.1% 3.8% Variance: 0.000033 0.000010 0.000149 0.000317 0.000443 0.002215 0.001855 0.000202 0.000376 0.004354 0.001657 0.001448 Coefficient of Variation: 113.8% 183.2% 102.9% 80.5% 89.1% 56.8% 28.9% 8.4% 13.4% 34.1% 43.6% 37.8%
27 Table 16. Trunk growth measurements - Kiwi-1 trial location. Treatment Nimitz Application Plot Replication Trunk Circumference, cm Rate and Timing Number 6/23/2015 11/11/2015 Increase or Decrease Untreated (Control) none applied 1 1 40.10 39.55-0.55 none applied 2 5 39.03 38.88-0.15 none applied 3 9 35.41 36.11 0.70 none applied 4 11 35.28 36.44 1.16 none applied 5 14 43.45 43.40-0.05 Mean: 38.65 38.88 0.22 Std. Deviation: 3.43 2.94 0.69 Variance: 11.79 8.63 0.48 Coefficient of Variation: 8.9% 7.6% 311.8% Post Harvest 3.5 pts/acre 1 2 42.41 42.22-0.19 3.5 pts/acre 2 6 43.47 43.20-0.27 3.5 pts/acre 3 7 33.34 33.85 0.51 3.5 pts/acre 4 12 45.20 46.06 0.86 3.5 pts/acre 5 13 40.33 40.65 0.32 Mean: 40.95 41.20 0.25 Std. Deviation: 4.61 4.55 0.48 Variance: 21.21 20.75 0.23 Coefficient of Variation: 11.2% 11.1% 193.7% Mid-Summer 3.5 pts/acre 1 3 45.77 45.20-0.57 3.5 pts/acre 2 4 42.71 42.58-0.13 3.5 pts/acre 3 8 42.86 43.11 0.25 3.5 pts/acre 4 10 42.47 43.28 0.81 3.5 pts/acre 5 15 42.08 42.41 0.33 Mean: 43.18 43.32 0.14 Std. Deviation: 1.48 1.11 0.52 Variance: 2.19 1.24 0.27 Coefficient of Variation: 3.4% 2.6% 375.5%
28 Appendix 3 Nematode & Yield Data: Kiwi-3 Ranch Table 17. Root knot nematode population Fig. L. Root knot nematode soil Fig. M. Root knot nematode roots Fig. N. Root knot nematode galls Table 18. Yield and fruit quality Fig. O. Kiwifruit yield Table 19. Fruit Grade Fig. P. Fruit Grade Table 20. Trunk growth Fig. Q. Trunk circumference increase in 2015
29 Table 17. Root knot nematode soil population 1, root infestation and galling score in the 2015 Nimitz field trial at Kiwi-3 ranch, Earlimart, CA. Treatment Nimitz Application Plot Sample Date Post-Harvest: 11/06/15 Replication Rate and Timing Number 05/20/15 06/25/15 07/21/15 8/26/2015 10/30/2015 Roots Gall Score 4 - - - - - - - - - - - - no. root knot nematodes per 500 cc soil - - - - - - - - - - - - - no. per gram root Untreated (Control) none applied 1 3 672 564 933 824 912 146 3.75 0 to 10 score none applied 2 5 509 1,697 1,697 1,782 1,025 279 6.25 none applied 3 7 764 218 909 1,239 416 152 3.75 none applied 4 11 400 230 285 691 521 35 10.00 none applied 5 13 672 721 455 485 774 122 3.75 Mean: 603 686 856 1,004 730 147 5.50 Std. Deviation: 146 605 548 515 257 88 2.74 Variance: 21,372 366,298 300,715 265,060 66,090 7,690 7.50 Coefficient of Variation: 24.2% 88.2% 64.1% 51.3% 35.2% 59.8% 49.8% Split Nimitz Application 2 3.5 pts/acre May + 3.5 pts/ac July 1 2 964 661 800 1,436 1,003 169 6.25 3.5 pts/acre May + 3.5 pts/ac July 2 6 364 570 497 533 851 156 2.50 3.5 pts/acre May + 3.5 pts/ac July 3 8 339 485 448 661 868 13 1.25 3.5 pts/acre May + 3.5 pts/ac July 4 10 473 1,358 885 588 506 137 2.50 3.5 pts/acre May + 3.5 pts/ac July 5 15 679 1,927 1,236 648 451 30 2.50 Mean: 564 1,000 773 773 736 101 3.00 Std. Deviation: 261 623 320 374 243 74 1.90 Variance: 68,033 388,135 102,361 139,891 59,016 5,470 3.59 Coefficient of Variation: 46.3% 62.3% 41.4% 48.4% 33.0% 73.3% 63.2% Mid-Summer 3 3.5 pts/acre July 1 1 - - 1,471 1,127 322 558 10.00 3.5 pts/acre July 2 4 - - 503 800 288 90 0.00 3.5 pts/acre July 3 9 - - 497 1,733 496 68 0.00 3.5 pts/acre July 4 12 - - 1,527 618 505 42 8.75 3.5 pts/acre July 5 14 - - 1,067 1,515 456 92 10.00 Mean: - - 1,013 1,159 413 170 5.75 Std. Deviation: - - 501 469 101 218 5.27 Variance: - - 250,808 219,699 10,277 47,421 27.81 Coefficient of Variation: - - 49.4% 40.5% 24.5% 128.2% 91.7% 1 soil sample depth: 1 to 18 inches; extraction method: sieving sugar flotation. 2 Nimitz application dates on the split application treatment: May 21, 2015 and July 24, 2015. 3 Nimitz application date on the mid-summer application treatment: July 24, 2015. 4 Gall Score: 0 = none (no galls found on roots), 5 = medium (one or more galls), 10 = high (more than 30% of roots have galls).
30 Table 18. Yield and fruit quality from each plot in the 2015 Nimitz field trial at Kiwi-3 ranch, Earlimart, CA. Treatment Nimitz Application Plot Total Fruit Replication Rate and Timing Number Yield Average Weight TSS SP lbs per plot grams Bx psi Untreated (Control) none applied 1 3 335.1 111 5.3 19.4 none applied 2 5 197.0 121 5.6 19.5 none applied 3 7 269.7 108 5.3 18.7 none applied 4 11 213.7 107 6.1 21.6 none applied 5 13 149.8 110 5.9 19.5 Mean: 233.1 112 5.6 19.8 Std. Deviation: 71.4 6 0.3 1.1 Variance: 5,091.4 32 0.1 1.2 Coefficient of Variation: 30.6% 5.1% 6.2% 5.6% Split Nimitz Application 3.5 pts/acre May + 3.5 pts/ac July 1 2 404.6 114 5.1 19.5 3.5 pts/acre May + 3.5 pts/ac July 2 6 63.0 120 6.0 19.4 3.5 pts/acre May + 3.5 pts/ac July 3 8 303.3 97 5.7 20.1 3.5 pts/acre May + 3.5 pts/ac July 4 10 153.2 101 5.3 19.9 3.5 pts/acre May + 3.5 pts/ac July 5 15 184.3 100 5.2 18.9 Mean: 221.7 106 5.5 19.6 Std. Deviation: 133.6 10 0.4 0.5 Variance: 17,847.5 106 0.1 0.2 Coefficient of Variation: 60.3% 9.7% 6.6% 2.4% Mid-Summer 3.5 pts/acre July 1 1 228.2 117 5.6 19.6 3.5 pts/acre July 2 4 116.9 120 6.0 19.4 3.5 pts/acre July 3 9 208.4 100 5.7 18.8 3.5 pts/acre July 4 12 58.7 114 6.1 19.4 3.5 pts/acre July 5 14 136.4 110 6.0 19.2 Mean: 149.7 112 5.9 19.3 Std. Deviation: 69.2 8 0.2 0.3 Variance: 4,785.4 65 0.1 0.1 Coefficient of Variation: 46.2% 7.2% 3.8% 1.6% 2 Nimitz application dates on the split application treatment: May 21, 2015 and July 24, 2015. 3 Nimitz application date on the mid-summer application treatment: July 24, 2015.
31 Table 19. Fruit grade from each plot in the 2015 Nimitz field trial at Kiwi-3 ranch. Nimitz Application Plot Percent of Total Yield Treatment Replication Rate and Timing Number Undersize 45 42 39 36 33 30 27 25 23 20 18 or Larger Untreated (Control) none applied 1 3 0.0% 0.1% 0.3% 0.8% 0.9% 5.6% 17.0% 25.3% 20.1% 20.0% 5.8% 4.0% none applied 2 5 0.0% 0.0% 0.2% 0.0% 0.0% 1.1% 7.2% 15.2% 18.7% 32.7% 17.7% 7.2% none applied 3 7 0.5% 0.1% 0.8% 1.3% 2.3% 8.7% 20.6% 20.1% 15.8% 18.1% 6.7% 4.8% none applied 4 11 0.3% 0.1% 0.5% 1.1% 1.6% 6.8% 23.8% 26.2% 18.3% 15.2% 4.7% 1.4% none applied 5 13 0.4% 0.0% 1.2% 1.4% 1.4% 7.3% 17.0% 20.8% 19.8% 18.0% 7.6% 5.1% Mean: 0.3% 0.1% 0.6% 0.9% 1.2% 5.9% 17.1% 21.5% 18.5% 20.8% 8.5% 4.5% Std. Deviation: 0.2% 0.1% 0.4% 0.6% 0.9% 2.9% 6.2% 4.4% 1.7% 6.9% 5.3% 2.1% Variance: 5.632E-06 6.327E-07 1.726E-05 3.348E-05 7.246E-05 8.421E-04 3.901E-03 1.973E-03 2.849E-04 4.733E-03 2.769E-03 4.387E-04 Coefficient of Variation: 93.1% 91.3% 71.7% 60.9% 69.4% 49.1% 36.5% 20.6% 9.1% 33.0% 61.9% 46.4% Split Nimitz Application pts/acre May + 3.5 pts/ac 1 2 0.1% 0.0% 0.5% 0.3% 1.1% 8.0% 17.0% 16.1% 16.5% 24.2% 10.0% 6.3% 3.5 pts/acre May + 3.5 pts/ac 2 6 0.0% 0.5% 0.5% 0.8% 1.1% 1.6% 7.6% 17.5% 16.0% 27.8% 14.1% 12.5% 3.5 pts/acre May + 3.5 pts/ac 3 8 5.5% 1.5% 4.7% 5.5% 6.8% 13.2% 16.4% 19.9% 11.5% 8.7% 1.2% 5.1% 3.5 pts/acre May + 3.5 pts/ac 4 10 0.5% 0.3% 1.9% 2.1% 5.9% 15.7% 26.9% 21.3% 11.6% 8.3% 2.8% 8.9% 3.5 pts/acre May + 3.5 pts/ac 5 15 1.6% 0.4% 1.7% 3.5% 7.0% 14.7% 24.5% 15.2% 15.7% 10.4% 3.0% 2.3% Mean: 1.6% 0.5% 1.8% 2.5% 4.4% 10.6% 18.5% 18.0% 14.3% 15.9% 6.2% 7.0% Std. Deviation: 2.3% 0.6% 1.7% 2.1% 3.0% 5.9% 7.6% 2.6% 2.5% 9.4% 5.5% 3.9% Variance: 5.363E-04 3.068E-05 2.937E-04 4.478E-04 9.119E-04 3.459E-03 5.802E-03 6.642E-04 6.230E-04 8.763E-03 3.068E-03 1.492E-03 Coefficient of Variation: 148.6% 103.8% 92.9% 86.3% 68.9% 55.4% 41.2% 14.3% 17.5% 59.0% 89.3% 54.9% Mid-Summer 3.5 pts/acre July 1 1 0.0% 0.1% 0.2% 0.0% 0.9% 2.4% 11.6% 19.7% 19.7% 26.8% 9.7% 8.9% 3.5 pts/acre July 2 4 0.2% 0.0% 0.3% 0.3% 0.5% 3.0% 9.7% 13.0% 14.7% 30.3% 15.9% 11.9% 3.5 pts/acre July 3 9 1.0% 0.4% 3.0% 3.7% 4.6% 12.7% 27.8% 20.4% 14.8% 8.9% 2.3% 0.3% 3.5 pts/acre July 4 12 1.8% 0.3% 2.4% 1.4% 3.0% 3.9% 7.1% 14.3% 13.5% 28.1% 13.4% 10.8% 3.5 pts/acre July 5 14 1.3% 0.1% 1.2% 0.8% 2.4% 8.8% 17.7% 17.5% 15.5% 18.0% 8.2% 8.4% Mean: 0.9% 0.2% 1.4% 1.2% 2.3% 6.2% 14.8% 17.0% 15.7% 22.4% 9.9% 8.1% Std. Deviation: 0.8% 0.2% 1.2% 1.5% 1.7% 4.4% 8.3% 3.3% 2.3% 8.9% 5.2% 4.6% Variance: 5.913E-05 2.430E-06 1.561E-04 2.137E-04 2.746E-04 1.959E-03 6.861E-03 1.057E-03 5.519E-04 7.889E-03 2.727E-03 2.081E-03 Coefficient of Variation: 90.2% 81.6% 89.3% 117.3% 72.6% 71.6% 56.0% 19.1% 15.0% 39.6% 52.6% 56.5%
32 Table 20. Trunk growth measurements - Kiwi-3 trial location. Nimitz Application Plot Trunk Circumference, cm Treatment Replication Rate and Timing Number 6/24/2015 11/27/2015 Increase or Decrease Untreated (Control) none applied 1 3 30.29 30.78 0.49 none applied 2 5 31.56 31.72 0.16 none applied 3 7 31.71 31.96 0.25 none applied 4 11 31.12 31.51 0.39 none applied 5 13 30.03 30.38 0.35 Mean: 30.94 31.27 0.33 Std. Deviation: 0.75 0.66 0.13 Variance: 0.57 0.44 0.02 Coefficient of Variation: 2.4% 2.1% 38.8% Split Nimitz Application 3.5 pts/acre May + 3.5 pts/ac July 1 2 33.63 33.97 0.34 3.5 pts/acre May + 3.5 pts/ac July 2 6 30.35 30.31-0.04 3.5 pts/acre May + 3.5 pts/ac July 3 8 33.66 33.93 0.27 3.5 pts/acre May + 3.5 pts/ac July 4 10 29.06 29.06 0.00 3.5 pts/acre May + 3.5 pts/ac July 5 15 35.34 35.73 0.39 Mean: 32.41 32.60 0.19 Std. Deviation: 2.60 2.79 0.20 Variance: 6.78 7.80 0.04 Coefficient of Variation: 8.0% 8.6% 103.5% Mid-Summer 3.5 pts/acre July 1 1 29.77 30.19 0.42 3.5 pts/acre July 2 4 31.37 31.67 0.30 3.5 pts/acre July 3 9 31.75 32.38 0.63 3.5 pts/acre July 4 12 33.08 33.28 0.20 3.5 pts/acre July 5 14 30.14 30.88 0.74 Mean: 31.22 31.68 0.46 Std. Deviation: 1.33 1.22 0.22 Variance: 1.76 1.48 0.05 Coefficient of Variation: 4.2% 3.8% 49.1%