Potential Impact of Regalia Treatments on Tree Health and Vigor

POTENTIAL IMPACT OF REGALIA® TREATMENTS ON TREE HEALTH AND VIGOR

V. 1.5

Alan L. Yelvington

Major Project/Report submitted to the faculty of the Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of

Online Master of Agricultural and Life Sciences In Plant Science and Pest Management

Dr. Keith S. Yoder, Research and Extension Tree Fruit Pathologist, Virginia Tech AREC Dr. Sherif Sherif, Research and Extension Tree Fruit Pomologist, Virginia Tech AREC Dr. Kari A. Peter, Tree Fruit Pathologist, Penn State FREC

1/3/2019

Keywords: Regalia, Reynoutria sachalinensis, ISR, russet, vigor, apple, Malus ×domestica

Version History

Version Date Comments 1.0 Working draft 1.1 Updated bar graphs in PDF 1.2 25 Nov 2018 Incorporated Dr. Yoder comments 1.3 3 Dec 2018 Incorporated Dr. Peter comments Removed rough log Added hardware appendix 1.4 21 Dec 2018 Conversion to Vancouver style 1.5 3 Jan 2019 Committee comments and edits

POTENTIAL IMPACT OF REGALIA® TREATMENTS ON TREE HEALTH AND VIGOR

Alan L. Yelvington

Abstract

Regalia®, an extract from giant knotweed (Reynoutria sachalinensis (REYSA)), is an organic immune system response (ISR) treatment that can mitigate fungal and bacterial infections by stimulating the genetic immune response system of a plant. The mode-of-action of Regalia® is to penetrate the plant tissue and stimulate a biodynamic response to resist pathogens. The primary response to Regalia® is the production of reactive oxygen species (ROS) that disable pathogens, and a thickening of cell walls to inhibit pathogen penetration. Regalia® is an uncommon fungicide since the protection is provided by the plant itself and not directly by the agent, and because the activity is internal rather than external. This process provides protection from pathogens that get past surface protectants. Because ISR agents require plant resources to be effective, there is a risk that they can disrupt the metabolic processes of the plant, or there can be an allocation cost that could diminish the plant vigor. This study examines if the ISR activity of Regalia® presents any phytoxicity or impact to vigor in four varieties of cider apples (Malus ×domestica cvs. Dabinett, Medaille d’Or, Chisel Jersey, and Redfield). This quantitative study, based on measured metrics of plant growth during the study period, showed that Regalia, applied with Cueva or Serenade, had no effect on primary or secondary shoot length or the number of shoot leaves on any of the four test varieties.

Table of Contents Version History ...... i Abstract ...... i List of Figures ...... iv List of Tables ...... iv Introduction ...... 1 Background and Setting ...... 1 Problem Statement ...... 2 Purpose of the Project ...... 2 Project Objectives ...... 2 Definition of Terms ...... 2 Significance of the Problem ...... 2 Review of Literature ...... 3 Summary ...... 3 Project Methodology ...... 4 Target Population ...... 4 Participating Audience ...... 4 Materials and Methods: ...... 4 Treatments: ...... 5 Treatment period: ...... 5 Preparation: ...... 6 Applications ...... 7 Data Collection ...... 8 Project Results, Discussion, and Recommendations...... 9 Results ...... 9 Discussion ...... 13 Regalia® and nutritional status...... 13 Russeting ...... 13 Recommendations ...... 13 References ...... 15 Appendix A MaryBlyt™ report ...... A-1 Appendix B Lab reports ...... B-1 Soil Samples ...... B-1

Tissue Samples ...... B-1 Appendix C Trial block plan ...... C-1 Appendix D Spray record ...... D-1 Appendix E Variety descriptions ...... E-1 Appendix F Spray application equipment ...... F-1 Appendix G First order data calculations ...... G-1 Appendix H Anecdotal Observations ...... H-1 Disease ...... H-1 Deer damage ...... H-1 Leaf size and turgor ...... H-1 Early drops ...... H-1 Russeting ...... H-1 Appendix I Lessons Learned ...... I-1

iii

List of Figures Figure 1: Trial flagging tape ...... 4 Figure 2: Measuring tape ...... 8 Figure 3: Tally counter ...... 8 Figure 4: Summary (LS means) – Treatments ...... 12 Figure 6: Fimco sprayer mounted on rear of BX-24 ...... F-1 Figure 7: Solo wand with nozzle holder ...... F-1 Figure 8: TeeJet fan and spot nozzles ...... F-1 Figure 9: Dabinett russet (Row 1) ...... F-2 Figure 10: Dabinett russet (Row 2) ...... H-2 Figure 11: Medaille d’Or (Row 3) ...... H-2 Figure 12: Medaille d’Or (Row 4) ...... H-2 Figure 13: Chisel Jersey Control (Row 5) ...... H-3 Figure 14: Chisel Jersey Replicate (Row 6) ...... H-3 Figure 15: Redfield Control (Row 7) ...... H-3 Figure 16: Redfield Replicate (Row 7) ...... H-3 Figure 17: Redfield Replicate (Row 8) ...... H-4

List of Tables Table 1: Trial block varieties and rootstocks ...... 5 Table 2: Application dates ...... 7 Table 3: Summary (LS means) – Treatments ...... 10 Table 4: Summary statistics (Quantitative data): ...... 11 Table 5: Standard russet values for trial varieties ...... H-1

Introduction Cider varieties are a specialty apple (Malus ×domestica) commodity since they have limited use outside of hard cider production. Traditional cider varieties (bitter sharps and bitter sweets) that were imported from Europe typically lack resistance to N. American pathogens, notably apple scab (Venturia inaequalis) and cedar apple rust (Gymnosporangium juniperi-virginianae).

Russetting and cosmetic blemishes (e.g. sooty blotch and fly speck) are not concerns on cider fruit since none of it is used for fresh market sales and much is promptly pressed and stored as juice. Although the use of copper is endorsed, rotation with other fungicides is strongly recommended to reduce selection pressure for resistant strains.

It’s not uncommon for hard cider producers to take advantage of “wild ferments” to create a signature terroir in their products. The use of broad spectrum fungicides can disrupt native yeast colonies and prevent that terroir from developing (1). Terroir has been cultivated in old world vineyards and orchards for generations (https://www.calvados-dupont.com/en/cidre-reserve.htm), and is considered a valuable marketing tool in new world vineyards and orchards. Reducing the amount of fungicides applied to an orchard might help to foster more wild yeast and a local terroir.

Background and Setting Organic options to prevent or reduce fungal infections in apple trees have previously been limited to products that provide protective barriers to infection; they either coat the surface with a biocide (e.g. copper) or populate the surface with benign organisms (e.g. Bacillus subtillis) or byproducts that limit the ability for pathogens to become established. Induced Resistance (IR) solutions trigger genetic response mechanisms within the plant to create a hostile environment for pathogens that works within the plant itself. Keeping the tree response mechanism at an active state reduces the risk that a pathogen would invade and infect by defeating the genetic response.

Regalia® is a bioprotectant/biopesticide (FRAC group P 05 Biological Plant Activator) extracted from giant knotweed (Reynoutria sachalinensis (REYSA)) . Its principle modes of action include accumulation of reactive oxygen species (ROS) and lignification and papilla formation in cell walls (2). Regalia® can be applied to apples as a drip (3), injection, or foliar spray (4). Soil drench applications or root dips are intended for pre-planting treatment of small plants or seeds (5).

A comparative trial of two Regalia® treatments was applied during the 2018 season on four different varieties of cider varieties (Redfield, Dabinett, Medaille d’Or, and Chisel Jersey) at Happy Dog Farm in NW Pennsylvania (https://goo.gl/maps/af9T1qpgDN62). The 2018 season was marked by periods of high disease pressure caused by temperature and extended wetting periods. A MaryBlyt™ report of orchard weather is located in Appendix A.

Applications were made on June 8th, 12th, 21st, and July 3rd. The period was shortly after full leaf, and blossom treatments for fire blight had ended. Previous treatments included Cueva® (copper) that resulted in all of the fruit being russeted. This study did not determine if the Regalia/Serenade™ treatment causes russeting. Applications were made as weather permitted.

The trial block indicated signs of nutrient deficiency in past seasons. Foliar applications of calcium and potassium were applied as covers before and during the trial period to address those deficiencies. Tissue analysis performed by the Penn State Agricultural Analytical Services Laboratory (University Park, PA) indicated that calcium levels remained low, but overall levels improved with nutritional treatment. Leaf size and turgor improved after the foliar feeding began. Soil and tissue lab reports are located in Appendix B.

Problem Statement Authors have speculated that maintaining a heightened level of induced resistance could have an allocation cost to the vigor of the plant (6). The tradeoff is that the use of broad spectrum fungicides could diminish the terroir that results from fermentation using wild yeasts.

If the treatments do not impact the vigor of the trees, then Regalia® has potential as an effective fungicide solution while trying to establish wild yeast colonies.

Purpose of the Project This project fulfills the project and report requirement of the Virginia Tech OMALS program.

Project Objectives The project examines if the application of an IR bioprotectant (Regalia®) has a measurable impact on the vigor of four cider varieties compared to their controls.

Definition of Terms Russet: Russet is a discoloration of the fruit surface that can also have an impact on the surface texture. It can vary from a light shadow to cracking (7). Russeting is considered a discouraging trait for fresh market fruit since it’s misunderstood by consumers. Conversely, russeted fruit are suitable for hard cider production (8).

Terroir: Terroir is a French term used to describe a unique flavor profile unique to a given place. That flavor profile is a reflection of production processes, plant varieties, and abiotic qualities of the place (9).

Induced Resistance (IR) – Induced resistance is when the genetic response mechanism of a plant is activated by a non-pathogenic agent, but provides resistance to pathogenic agents. It is typically short-lived and must be periodically provoked to provide continuous protection. IR is typically only a localized benefit where the stimulating agent has contacted the plant. Regalia® is a SAR agent.

Systemic Acquired Resistance (SAR) - SAR is a whole-plant response to an IR agent. Regalia® is not a SAR agent.

Significance of the Problem Finding solutions that enable wild yeast colonies could promote the practice and enable market differentiation. Also, it’s important to measure the impact of IR/SAR agents on plant vigor and productivity.

Review of Literature Optimizing induced resistance is a literature review report done by another Virginia Tech OMALS graduate student (10). Although it does not include apples, it is an excellent introduction to the concept of induced resistance (IR) and much of the literature up to that time. Critical elements include:

• Differentiating IR products from conventional products. IR agents do not provide immediate protect and require time to elicit an immune response from the host plant. Also, the level of response depends on the plant more than the agent. Some species may demonstrate no benefit, and some more than others. • Allocative costs and impact of abiotic stress. IR agents depend on plant resources to thicken cell walls and create protective agents such as reactive oxygen species (ROS) (2). Although stimulating all available defense mechanisms has the highest opportunity to fend off infection, this continuous state of “battle stations” is energy intensive and has the potential to impact plant vigor and yield (11). • Application timing. Light generated ROS can be used as a defense mechanism, and consume less nicotinamide adenine dinucleotide phosphate (NADPH) than production in the absence of light. Consequently, applying an IR agent at the beginning of a sunny day enables the host to exploit photosynthesis for the production of ROS. (It also ensures faster leaf drying to further hinder fungal infection.) • Potential impacts on yield. Although authors predict negative impact on yields, this is out of the scope of this project since this planting is pre-productive with no yield history.

There is an abundance of literature regarding trials using Regalia® as a plant defense agent that are out of scope for this project. However, Regalia® has the added feature of serving as a mild blossom thinning agent when applied during bloom (12). Growers should be aware of this behavior if Regalia® is used outside of a conventional thinning program.

Summary Regalia® represents a novel approach to disease prevention in apple orchards. Reducing the use of conventional antifungal agents has the potential to improve soil health, foster mycorrhizal colonies, and improve the quality and yield of organically produced cider fruit. The demonstrated success of Regalia® with cedar apple rust and other pathogens merits closer examination and more trials (13).

Project Methodology This is a quantitative trial based on measured metrics of plant growth during the trial period.

Target Population The target audience is growers looking for organic disease control solutions. Growers for hard cider production are the most likely to benefit because cider varieties tend not to be inherently disease resistant, and russeting is not a concern.

Hard cider producers wanting to develop an orchard terroir also stand to benefit since biocides and broad spectrum fungicides are likely to reduce wild yeast types that would develop in an orchard. Although out of scope for this project, Regalia® might enable wild yeast populations to become established that other products would suppress.

Participating Audience This project was sponsored by Happy Dog Farm (HDF) in Russell PA. HDF is a cider orchard that grows fruit for local hard cider makers.

Materials and Methods: The trial trees are in eight parallel rows oriented north to south in the southern section of the orchard. A map of the trial block is located in Appendix C. Each variety is grouped in pairs of adjacent rows (See Table 1). The trees were planted in 2015 and all were inoculated with mycorrhizal fungi (BioOrganics LLC). Lime and potash were applied during the spring in response to soil tests from the previous year. Treatments alternated between rows with all rows having untreated control trees. Rows were marked with distinct flagging tape indicating which treatment was to be applied. Control and sample trees were also uniquely flagged (See Figure 1) to ensure that they were appropriately treated or isolated. Foliar treatments were made as weather permitted along with nutritional and insecticidal applications. The orchard spray record is located in Appendix D.

Treatment row flags Replicate flags Control flags Figure 1: Trial flagging tape

Table 1 lists the locations and varieties used in the trial. Detailed cultivar descriptions from the U.S. National Plant Germplasm System (https://npgsweb.ars-grin.gov/gringlobal/search.aspx?) for each variety are located in Appendix E.

Table 1: Trial block varieties and rootstocks

Rows Variety Rootstock Reference 1 & 2 Dabinett B.9 (14) 3 & 4 Medaille d’Or G.16 (15) 5 & 6 Chisel Jersey B.9 (16) 7 & 8 Redfield B.9 (17)

A 25 gallon Fimco wand sprayer (Tractor Supply Co.) with an electric pump was used for maintenance (insecticides, foliar fertilizers, etc.) and treatment in the trial block. The stock Fimco wand was replaced with a Solo 4900170N wand (Amazon) using a TeeJet 6504 nozzle (North East Fruit Growers) to ensure a constant volume and spray pattern (at 40 psi) when wetting the trees.

A graduated bucket (e-Encore Mix’n Measure 5 gallon, Lowes) and pesticide measuring cup (Accu Pour 1 liter, Tractor Supply Co.) were used to prepare the treatment solutions. The standard protocol of filling the tank half full with water, mixing in the agents, and topping off the tank to the desired volume was used. The spray tank was agitated with the pressure pump in full bypass.

Images of the application equipment are located in Appendix F.

Treatments: Two treatments were compared using Regalia® (Marrone Bio Innovations, EPA reg. 84059-3) as the common ingredient. Both Serenade™ (Bayer Crop Science, EPA reg. 264-1152) and Cueva® (Certis USA, EPA reg. 67702-2-70051) are established protectants with dissimilar modes of action.

Serenade™ (Specifically Serenade-ASO) is a biological protectant used to cover the surface of the treated area with the fermentation byproduct of Bacillus subtilis to deny the pathogen a suitable environment. Serenade™ does contain some live B. subtilis that are benign and compete with pathogenic agents. Serenade™ is not an IR agent, and there is no reported synergy between Serenade™ and Regalia®.

Cueva® is a copper-based protectant that acts as a slow-release topical biocide. Copper is an elemental biocide that can be phytotoxic if used off-label or as part of an acidic mix.

Both products are listed for use on apple (Malus pumila). The treatments were compared on parallel rows of each variety.

Treatment period: Fire blight infection periods were predicted with Maryblyt™ 7.1 using data from a Kestrel D-3 data logger and rainfall data. A MaryBlyt™ report for the season is located in Appendix A.

The trial treatment period was after petal fall until early July with data collected on July 26-27. This was a period of vigorous growth and high disease pressure. Applications were made during weather breaks

and alternated with foliar feedings and insecticide treatments. A complete list of trial applications and dates can be found in Table 2 on page 7. The spray log is found in Appendix D.

Preparation: Pre-trial fire blight and scab controls included oil (TriTek), copper (Cueva®), streptomycin (Agrimycin-17), Bacillus subtilis (Serenade-ASO™), giant knotweed extract (Reynoutria sachalinensis (REYSA)) (Regalia®), and oxytetracycline (Mycoshield).

Soil and tissue samples were taken at the end of the 2017 season to provide a comparative baseline for the 2018 trial. Lab results are located in Appendix B.

Foliar nutrients were applied prior to and concurrently during the trial based on 2017 soil and tissue samples. The following products were used for foliar fertilizers: Nutri-K (CSI Chemical Corporation) , Nutri-Cal (CSI Chemical Corporation), and Solubor (U.S. Borax, Inc.). Multiple Nutri-K and Nutri-CAL applications were made at the manufacturer’s recommended rate. A single application of Solubor was made at half the manufacturer’s recommend rate.

Application rates are as concentrations rather than gallons/acre because of the small volumes used. Two treatments were used:

• Treatment A (rows 1, 3, 5, 7): 1% Regalia® + 1% Serenade™ • Treatment B (rows 2, 4, 6, 8): 1% Regalia® + 1% Cueva® • Untreated control trees were located in all rows

Regalia® was the common IR agent in both treatments, and Serenade™ and Cueva® were the complementary protective agents.

Applications Applications were made with calm wind to minimize drift between rows and onto control trees. All applications were made until runoff. Applications were made on the following dates:

Table 2: Application dates

Date Agent(s) Rows Comments April 30 Cueva® All FB Silver tip May 7 Cueva® All FB Tight cluster May 9 Agri-mycin All FB Bloom May 13 Serenade™ ASO 7, 8 (Redfield) FB Bloom May 15 Serenade™ + All FB Bloom Agri-mycin May 17 Serenade™ + Regalia® All FB Bloom May 19 Serenade™ + Cueva® All FB Bloom May 24 Agri-mycin All FB Bloom May 26 Agri-mycin All FB Bloom May 30 Mycoshield All FB Bloom June 1 Cueva® All FB Bloom / Post bloom June 8 Regalia® + Serenade™ 1, 3, 5, 7 Treatment spray June 8 Regalia® + Cueva® 2, 4, 6, 8 Treatment spray June 12 Regalia® + Serenade™ 1, 3, 5, 7 Treatment spray June 12 Regalia® + Cueva® 2, 4, 6, 8 Treatment spray June 21 Regalia® + Serenade™ 1, 3, 5, 7 Treatment spray June 21 Regalia® + Cueva® 2, 4, 6, 8 Treatment spray July 3 Regalia® + Serenade™ 1, 3, 5, 7 Treatment spray *Potassium bicarbonate added July 3 Regalia® + Cueva® 2, 4, 6, 8 Treatment spray *Potassium bicarbonate added *Potassium bicarbonate was added as a foliar feeding and anti-fungal treatment.

Foliar feeding and insecticidal cover sprays were applied to trial and control trees during the period. The cover sprays consisted of insecticides and foliar feedings of Ca, B, and K.

Data Collection The following measurements were taken as metrics of plant vigor:

• Annual growth of 20 vertical shoots (cm) • Annual growth of a random sample of 20 lateral limbs (cm) • Leaf count on annual growth of the 20 vertical shoot • Leaf count on annual growth of the 20 sampled lateral limbs

Annual growth data was collected on July 26 - 27, 2018. Apical and lateral annual growth was measured using a tailor’s tape measure (Figure 2). Leaf counts were tallied using a mechanical hand counter (See Figure 3).

First order calculations (medians) are located in Appendix G.

Figure 2: Measuring tape

Figure 3: Tally counter

Although weather data was not a part of the study, it reveals the fire blight infection pressure that was present prior to the beginning of the trial. A Kestrel D3 (Kestrel Meters, Boothwyn, PA) data logger recorded weather data at 30 minute intervals. Rainfall was measured using a Stratus Rain Gauge (Productive Alternatives, Inc, Fergus Falls, MN) and recorded each morning. There was no capability to record light intensity or the duration of wetting periods. A MaryBlyt™ weather data and fire blight pressure report is located in Appendix A.

Project Results, Discussion, and Recommendations The trial evaluated the treatments on four different varieties of trees. Differences in morphology between varieties made it inappropriate to compare the data between varieties.

Results The data reflected no difference between treatment and untreated controls indicating that Regalia® is benign in terms of apple vigor. Untreated controls and replicates were not yet at terminal bud set when measurements were taken, indicating continued tree growth.

The data indicate that Regalia®, applied with Cueva® or Serenade™, does not have a detrimental or beneficial impact on the vigor of the four varieties tested. Although the lack of a detrimental impact could be presumed since Regalia is listed for pome fruit, there was the possibility that one or more of the test varieties could have had a phytotoxic sensitivity.

Dr. Sherif Sherif (Virginia Tech) was kind enough to provide a template for submitting the median data, and then run the statistical analysis for me using XLStat (https://www.xlstat.com/en/).

Table 3: Summary (LS means) – Treatments

Rows 1&2: Dabinett Primary shoot Secondary shoot # of apical # of lateral (cm) (cm) leaves leaves Regalia + Serenade™ 41.750 a 39.781 a 33.750 a 29.750 a Regalia+ Cueva® 38.625 a 35.719 a 30.000 a 26.719 a Control 38.000 a 38.750 a 33.500 a 29.625 a Pr > F(Model) 0.585 0.069 0.795 0.134 Significant No No No No

Rows 3&4: Medaille d’Or Primary shoot Secondary shoot # of apical # of lateral (cm) (cm) leaves leaves Regalia + Serenade™ 46.000 a 45.208 a 33.833 a 29.917 a Regalia+ Cueva® 49.500 a 40.500 a 34.000 a 27.063 a Control 47.000 a 37.500 a 32.667 a 28.917 a Pr > F(Model) 0.848 0.289 0.979 0.563 Significant No No No No

Rows 5&6: Chisel Jersey Primary shoot Secondary shoot # of apical # of lateral (cm) (cm) leaves leaves Regalia + Serenade™ 54.000 a 44.781 a 27.750 a 24.719 a Regalia+ Cueva® 51.875 a 46.063 a 29.625 a 31.063 a Control 54.500 a 45.813 a 27.000 a 28.313 a Pr > F(Model) 0.881 0.803 0.846 0.335 Significant No No No No

Rows 7&8: Redfield Primary shoot Secondary shoot # of apical # of lateral (cm) (cm) leaves leaves Regalia + Serenade™ 54.000 a 48.906 a 23.625 a 22.750 a Regalia+ Cueva® 50.286 a 53.357 a 20.143 a 22.214 a Control 61.500 a 61.063 a 26.250 a 28.500 a Pr > F(Model) 0.611 0.355 0.470 0.237 Significant No No No No

Table 4: Summary statistics (Quantitative data):

Rows 1&2: Dabinett Obs. Obs. w/data w/o data Std. Variable Observations missing missing Min. Max. Mean deviation Primary shoot (cm) 20 0 20 26.000 51.000 39.750 6.843 Secondary shoot (cm) 20 0 20 32.750 45.500 37.950 3.675 # of apical leaves 20 0 20 22.000 63.000 32.200 11.316 # of lateral leaves 20 0 20 23.750 36.000 28.513 3.276

Rows 3&4: Medaille d’Or Obs. Obs. w/data w/o data Std. Variable Observations missing missing Min. Max. Mean deviation Primary shoot (cm) 20 7 13 29.000 65.000 47.308 8.760 Secondary shoot (cm) 20 7 13 30.250 55.250 41.981 7.063 # of apical leaves 20 7 13 15.000 46.000 33.615 8.471 # of lateral leaves 20 7 13 20.500 35.500 28.808 3.881

Rows 5&6: Chisel Jersey Obs. Obs. w/data w/o data Std. Variable Observations missing missing Min. Max. Mean deviation Primary shoot (cm) 20 0 20 35.000 71.000 53.250 9.585 Secondary shoot (cm) 20 0 20 38.750 52.000 45.500 3.813 # of apical leaves 20 0 20 14.000 47.000 28.350 7.903 # of lateral leaves 20 0 20 21.000 61.250 27.975 8.397

Rows 7&8: Redfield Obs. Obs. w/data w/o data Std. Variable Observations missing missing Min. Max. Mean deviation Primary shoot (cm) 20 1 19 21.000 81.000 54.211 17.265 Secondary shoot (cm) 20 1 19 24.500 83.000 53.105 13.438 # of apical leaves 20 1 19 11.000 38.000 22.895 7.944 # of lateral leaves 20 1 19 10.000 37.250 23.763 6.218

Sunday, November 25, 2018

Figure 4: Summary (LS means) - Treatments

Rows 1&2: Dabinette Rows 3&4: Medaille d’Or

Summary (LS means) - Treatments Summary (LS means) - Treatments

45 60 A A A A A A 40 A A A A 50 A A 35 A A A A

A 30 A 40 A A A 25 A A 30 A 20 LS means 15 LS means 20 10 10 5 0 0 Primary shoot Secondary # of apical # of lateral Primary shoot Secondary # of apical # of lateral (cm) shoot (cm) leaves leaves (cm) shoot (cm) leaves leaves Dependent variables Dependent variables

Regalia + Serenade control Regalia+ Cueva Regalia + Serenade control Regalia+ Cueva

Rows 5&6: Chisel Jersey Rows 7&8: Redfield

Summary (LS means) - Treatments Summary (LS means) - Treatments

60 70 A A A A A 60 50 A A A A A A A 50

40 A A 40 A A A 30 A A 30 A A A

LS means LS means A 20 A 20

10 10

0 0 Primary shoot Secondary # of apical # of lateral Primary shoot Secondary # of apical # of lateral (cm) shoot (cm) leaves leaves (cm) shoot (cm) leaves leaves Dependent variables Dependent variables

Regalia + Serenade control Regalia+ Cueva Regalia + Serenade control Regalia+ Cueva

Discussion The manufacturer of Regalia® claims that Serenade™ only has an additive value and is not synergistic (2). The conditional approval of Serenade™ for the State of New York mentions a report that Serenade™ induces systemic acquired resistance (SAR) but did not provide a reference (18). Consequently, I would not expect this treatment to have had a higher metabolic load on the plant than treatment of Regalia® and Cueva®. Although the study was performed at the maximum recommended rate of Regalia® (1% v/v), I recommend testing on personal plantings at the lower recommended rates to reduce the risk of phytotoxicity.

Serenade™ is a biological disease control product. Manufacturer marketing literature claims systemic qualities (19), but independent sources attribute the foliar benefits to fermentation byproducts that are antibacterial and antifungal. Controlled field trials verify that Serenade™ does in fact not have IR qualities (K. Peter, personal communication, December 5, 2018). Although spores are present in the product, they are not considered instrumental in the applied product (20). Consequently, Serenade™ would not present any metabolic benefit or load to the host plant.

Cueva® (10% copper octanoate) is a general non-selective biocide. Copper can be phytotoxic on leaves if used in an inappropriate concentration, or mixed with acidic water. Cueva® has a relatively low concentration of metallic copper (1.8%) with small particle sizes for good coverage (21). The risk of copper is that it is phytotoxic when used in excess or in an acidic solution. Cueva® was applied according to label using hard (>425 mg/l) well water.

Regalia® and nutritional status. Induced resistance requires plant resources to be effective, so plants under abiotic nutritional stress may not be able to fully exploit the affect for a sustained period. This was a concern before starting the trial based on previous soil and tissue tests. Appendix B includes test results from 2017. Fertilizer and lime were applied as a side dress and foliar applications before and during the 2018 season. Nutritional cover sprays of Ca, K, and B appear to have improved leaf turgor on both treated and untreated control trees and the rest of orchard, so leaf turgor was not attributed to test treatments. The tissue sample from 2018 reflects changes in Medaille d’Or trees that appear to have benefited from the nutritional treatments.

Russeting Russeting on fruit from both treated replicates and untreated controls might be from copper treatments during bloom. Copper can promote russeting in pome fruit, but there are also other factors including weather, disease (powdery mildew), and natural yeasts (Aureobasidium pullalans). It’s not possible to attribute any of the observed russeting to any of the treatment products.

Recommendations Regalia® can be used on apple for disease treatments without risk of phytoxicity when applied according to the label. However, it should not be applied with the expectation of improving plant vigor. Regalia appeared to be benign to tree vigor when applied after fruit set. Literature indicates that Regalia® has a gentle thinning effect when applied during bloom (12). Growers need to be aware of the thinning effect when incorporating it as part of a disease treatment program during blossom.

Future research to determine the impact of Regalia® on native yeast colonies may be appropriate for growers attempting to develop orchard terroir.

Label notes and anecdotal information indicate that the IR benefits of Regalia® are short-lived, localized, and frequent applications are necessary. (The application only benefits the wetted surfaces.) There is no evidence of the IR benefits of Regalia® being carried from season to season. Testing for benefits carried over from season to season would require more resources and planning.

Regalia® offers a potential option to organic cider apple growers looking for disease treatment solutions that may enable building wild yeast populations for terroir in their cider products.

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10. Sirois J. Optimizing induced resistance (IR). 2011 May [cited 2018 Oct 12]; Available from: https://vtechworks.lib.vt.edu/handle/10919/51519

11. Bolton MD. Primary Metabolism and plant defense-fuel for the fire. Mol Plant Microbe Interact. 2009 May;22(5):487–97.

12. Peck GM, Combs LD, DeLong C, Yoder KS. Precision apple flower thinning using organically approved chemicals. Acta Hortic. 2016 Aug;(1137):47–52.

13. DeLong CN, Yoder KS, Cochran AE, Kilmer SW, Royston WS, Combs LD, et al. Apple disease control and bloom-thinning effects by lime sulfur, Regalia, and JMS Stylet-Oil. Plant Health Prog. 2018 Jan;19(2):143–52.

14. Nat’l Plant Germplasm System. Dabinett, PI 589073 [Internet]. GRIN-Global Web v 1.10.3.6. [cited 2018 Oct 20]. Available from: https://npgsweb.ars- grin.gov/gringlobal/AccessionObservation.aspx?id=1008346

15. Nat’l Plant Germplasm System. Medaille d’Or, PI 594108 [Internet]. GRIN-Global Web v 1.10.3.6. [cited 2018 Oct 20]. Available from: https://npgsweb.ars- grin.gov/gringlobal/AccessionObservation.aspx?id=1018993

16. Nat’l Plant Germplasm System. Chisel Jersey, PI 588806 [Internet]. GRIN-Global Web v 1.10.3.6. [cited 2018 Oct 20]. Available from: https://npgsweb.ars- grin.gov/gringlobal/AccessionObservation.aspx?id=1002937

17. Nat’l Plant Germplasm System. Redfield, PI 589211 [Internet]. GRIN-Global Web v 1.10.3.6. [cited 2018 Oct 20]. Available from: https://npgsweb.ars- grin.gov/gringlobal/AccessionObservation.aspx?id=1011987

18. Riggs R. bacillus subtilis NYSDEC-Conditional Registration 4/01 [Internet]. Bacillus subtilis NYSDEC-Conditional Registration 4/01. 2001 [cited 2018 Oct 21]. Available from: http://pmep.cce.cornell.edu/profiles/fung-nemat/aceticacid- etridiazole/bacillus_subtilis/bacillus_label_401.html

19. Merckling T, Manker D, Ricci M. Development of Serenade as a biopesticide against plant bacterial diseases. In: Annual COST873 Meeting—Management Committee Meeting. 2009. p. 1–35. Available from: https://www.researchgate.net/publication/312086009_Development_of_Serenade_as_a_biopesticide _against_plant_bacterial_diseases

20. Caldwell B, Brown Rosen E, Shelton A, Smart C. Bacillus subtilis material fact sheet | Horticulture International [Internet]. Resource guide for organic insect and disease management. 2004 [cited 2018 Oct 11]. Available from: https://hortintl.cals.ncsu.edu/articles/bacillus-subtilis-material-fact- sheet

21. Peter K. Demystifying copper for disease management. Power Point presented at: 2016 Winter Meeting; 2016 Mar 5.

22. Marschner H, Marschner P, editors. Marschner’s mineral nutrition of higher plants. 3rd ed. London ; Waltham, MA: Elsevier/Academic Press; 2012. 651 p.

23. Nat’l Plant Germplasm System. Fruit russet intensity [Internet]. GRIN-Global Web v 1.10.3.6. [cited 2018 Oct 20]. Available from: https://training.ars- grin.gov/gringlobal/descriptordetail.aspx?id=115054

24. Nat’l Plant Germplasm System. Fruit russet type [Internet]. GRIN-Global Web v 1.10.3.6. [cited 2018 Oct 20]. Available from: https://training.ars- grin.gov/gringlobal/descriptordetail.aspx?id=115056

Appendix A MaryBlyt™ report Pre-trial weather and disease pressure were recorded using MaryBlyt™. MaryBlyt™ does not support data entry for an entire season.

A-1

Intentionally blank Page 1 Happy Dog Farm - Ciders Date Phenology Max Temp (F) Min Temp (F) Wetness (in) Trauma Spray Notes Avg Temp (F) EIP BHWTR BBS CBS SBS TBS 4/1/2018 D 42.4 20.6 0.00 Frost 31.5 ------4/2/2018 D 51.9 29.6 0.00 40.8 ------4/3/2018 D 57.3 37.3 0.00 47.3 ------4/4/2018 D 41.3 20.8 0.00 Frost 31.0 ------4/5/2018 D 36.1 22.6 0.00 Frost 29.4 ------4/6/2018 D 44.0 10.9 0.00 Frost 27.4 ------4/7/2018 D 39.7 17.8 0.00 Frost 28.8 ------4/8/2018 D 35.6 15.9 0.00 Frost 25.8 ------4/9/2018 ST 41.6 18.3 0.00 Frost 30.0 ------4/10/2018 ST 43.9 23.4 0.00 Frost 33.6 ------4/11/2018 ST 46.4 35.1 0.15 40.8 ------4/12/2018 ST 70.3 42.0 0.00 56.2 ------4/13/2018 ST 80.4 40.3 0.02 Heavy dew and 2% Dormant Oil 60.4 ------4/14/2018 ST 66.0 33.6 0.02 49.8 ------4/15/2018 ST 53.7 39.3 1.35 46.5 ------4/16/2018 ST 42.3 28.6 0.01 35.4 ------4/17/2018 ST 34.5 28.5 0.30 31.5 ------4/18/2018 ST 39.7 29.7 0.00 34.7 ------4/19/2018 ST 34.8 24.9 0.00 Frost 29.8 ------4/20/2018 ST 47.8 20.7 0.01 Frost 34.2 ------4/21/2018 ST 59.1 20.9 0.01 Frost 40.0 ------4/22/2018 ST 68.5 27.4 0.00 Frost 48.0 ------4/23/2018 ST 75.2 23.6 0.08 Rain Frost 49.4 ------4/24/2018 ST 58.9 50.0 0.27 54.4 ------4/25/2018 ST 57.3 35.8 0.07 46.6 ------4/26/2018 ST 62.0 31.1 0.00 Dry 46.6 ------4/27/2018 ST 71.6 42.8 0.43 Redfield and Wickson Crab 57.2 ------4/28/2018 ST 47.6 30.9 0.14 39.2 ------4/29/2018 ST 50.9 20.3 0.00 Frost 35.6 ------4/30/2018 ST 66.4 29.8 0.00 Yes 1% oil and 1% copper 48.1 ------5/1/2018 GT 84.6 39.8 0.00 62.2 - - - 6 - - 5/2/2018 GT 88.1 60.7 0.00 74.4 --- 15 -- 5/3/2018 GT 83.3 60.5 0.00 71.9 --- 24 -- 5/4/2018 TC 81.9 38.7 0.05 60.3 --- 29 -- 5/5/2018 TC 82.0 48.8 0.00 65.4 --- 35 -- 5/6/2018 TC 72.7 48.2 0.10 60.4 --- 39 -- 5/7/2018 TC 74.5 37.8 0.00 Yes 1% copper 56.2 --- 42 -- 5/8/2018 TC 80.4 43.8 0.00 62.1 --- 47 -- 5/9/2018 B 87.5 56.8 0.05 Yes 100 ppm streptomycin 72.2 - + - + + H - 56 -- 5/10/2018 B 74.0 35.3 0.20 54.6 24 + - + - M - 59 -- 5/11/2018 B 59.0 45.1 0.75 52.0 16 + - + - M - 59 -- 5/12/2018 B 54.8 47.1 1.58 51.0 8 + - + - M - 59 -- 5/13/2018 B 62.3 50.6 0.00 Yes 1% Serenade on blossoms 56.4 - + - + - M - 61 -- 5/14/2018 B 79.9 55.3 0.09 67.6 61 + - + + H - 67 -- Page 2 Happy Dog Farm - Ciders Date Phenology Max Temp (F) Min Temp (F) Wetness (in) Trauma Spray Notes Avg Temp (F) EIP BHWTR BBS CBS SBS TBS 5/15/2018 B 80.3 45.3 0.04 Yes 100ppm Stres with 1% Serenade 62.8 - + - + + H - 72 -- 5/16/2018 B 81.6 54.3 0.00 dry day 67.9 73 + - - + M - 79 -- 5/17/2018 B 87.6 49.8 0.10 Yes 100ppm Strep Regalia Nutri-K 68.7 - + - + + H - 87 -- 5/18/2018 B 85.4 54.1 0.00 69.8 97 + - + + H - 94 -- 5/19/2018 B 83.6 55.9 0.16 Yes 1% serenade 1% Cueva 4oz sticker 69.8 - + - + + H - CMS -- 5/20/2018 B 66.1 45.2 0.36 55.6 - + - + - M - 4 - - 5/21/2018 B 86.4 56.3 0.01 71.4 97 + - + + H - 19 -- 5/22/2018 B 78.9 54.0 0.35 Rain 66.4 158 + + + + I - 31 -- 5/23/2018 B 80.9 44.4 0.01 Heavy dew 62.6 218 + + + + I 10 a 41 -- 5/24/2018 B 87.5 46.8 0.00 Yes Agrimycin 100 ppm 67.2 - + - + + H 23 a 54 -- 5/25/2018 B 91.7 50.8 Dew Dew 71.2 121 + + + + I 40 a 71 -- 5/26/2018 B 90.7 60.7 0.00 Yes Agrimycin 200 ppm 75.7 - + - + + H 60 a 91 -- 5/27/2018 B 91.0 56.8 0.47 73.9 133 + + + + I 79 a CBS -- 5/28/2018 B 92.5 56.8 0.00 74.6 267 + + + + I 97 a --- 5/29/2018 B 80.0 50.0 0.00 65.0 327 + + - + H 108 a - 11 - 5/30/2018 B 88.6 68.1 Dew Yes Oxytetracyclin Dew 78.4 - + - + + H 120 b - 33 - 5/31/2018 B 87.7 65.9 0.08 76.8 145 + + + + I 112 c - 54 - 6/1/2018 B 85.8 62.5 0.22 Yes 1% Cueva on all 74.2 - + - + + H 91 d - 73 - 6/2/2018 B 73.0 54.0 0.02 Damp 63.5 24 + - + + H 100 d - 82 - 6/3/2018 B 69.7 52.1 0.21 60.9 36 + - + + H 87 e - 87 - 6/4/2018 B 65.9 45.0 0.01 55.4 36 + - + - M 91 e - 91 - 6/5/2018 PF 71.2 47.7 0.02 59.4 -- 97 e - 97 - 6/6/2018 PF 62.2 42.0 Dew Dew 52.1 -- 99 e - 99 - 6/7/2018 PF 84.0 51.6 Dew Dew 67.8 -- 112 e - 112 - 6/8/2018 PF 86.5 44.6 0.00 Project trial START 65.6 -- 70 f --- 6/9/2018 PF 82.9 52.6 0.00 67.8 -- 83 f --- 6/10/2018 PF 68.2 52.6 0.48 60.4 -- 88 f --- 6/11/2018 PF 89.6 51.7 0.00 70.6 -- 104 f --- 6/12/2018 PF 85.9 43.4 0.59 Wind Wind 64.6 ------6/13/2018 PF 86.3 43.4 0.03 Wind Wind 64.8 ----- 12 a 6/14/2018 PF 80.0 40.8 0.00 60.4 ----- 20 a 6/15/2018 PF 80.8 45.8 0.00 63.3 ----- 30 a 6/16/2018 87.6 54.6 0.00 71.1 ----- 46 a 6/17/2018 96.9 65.4 0.00 81.2 ----- 70 a 6/18/2018 92.8 64.2 1.03 Thunderstorms 78.5 ----- 92 a 6/19/2018 82.5 57.2 0.06 69.8 ----- 107 a 6/20/2018 74.5 54.1 0.00 64.3 ----- 104 b 6/21/2018 87.7 51.7 0.00 69.7 ------6/22/2018 74.9 60.6 0.30 67.8 ------6/23/2018 84.2 62.2 0.50 73.2 ------6/24/2018 78.1 56.1 0.14 67.1 ------6/25/2018 77.7 40.6 0.00 59.2 ------6/26/2018 86.5 63.9 0.00 75.2 ------6/27/2018 75.0 62.6 0.55 Rain overnight 68.8 ------Page 3 Happy Dog Farm - Ciders Date Phenology Max Temp (F) Min Temp (F) Wetness (in) Trauma Spray Notes Avg Temp (F) EIP BHWTR BBS CBS SBS TBS 6/28/2018 83.3 57.3 0.00 70.3 ------6/29/2018 95.4 59.2 0.00 77.3 ------6/30/2018 96.9 59.7 0.00 78.3 ------7/1/2018 96.2 66.3 0.00 Trapped first JB 81.2 ------7/2/2018 93.6 63.8 0.61 Heavy rain 78.7 ------7/3/2018 95.1 66.5 0.00 80.8 ------7/4/2018 94.7 68.1 0.00 81.4 ------7/5/2018 95.9 65.5 0.00 80.7 ------7/6/2018 78.5 42.5 0.00 60.5 ------7/7/2018 86.4 45.8 0.00 66.1 ------7/8/2018 95.1 48.9 0.00 72.0 ------7/9/2018 92.3 55.9 0.00 74.1 ------7/10/2018 89.4 51.5 0.00 70.4 ------7/11/2018 83.3 44.6 0.00 64.0 ------7/12/2018 90.1 57.3 0.00 73.7 ------7/13/2018 92.9 61.5 0.00 77.2 ------7/14/2018 93.5 66.0 0.00 79.8 ------7/15/2018 93.3 63.2 0.00 78.2 ------7/16/2018 96.0 66.8 0.82 81.4 ------7/17/2018 85.2 49.0 0.00 67.1 ------7/18/2018 82.6 47.6 0.00 65.1 ------7/19/2018 90.3 53.7 0.00 72.0 ------7/20/2018 88.8 64.7 0.00 76.8 ------7/21/2018 80.2 57.6 1.58 68.9 ------7/22/2018 81.0 60.0 0.00 70.5 ------7/23/2018 84.0 66.5 0.00 75.2 ------7/24/2018 90.0 67.4 0.00 78.7 ------7/25/2018 86.6 58.2 0.18 72.4 ------7/26/2018 90.6 61.1 0.00 75.8 ------Collected Data 7/27/2018 85.9 60.8 0.73 73.4 ------Collected Data 7/28/2018 79.6 53.1 0.00 66.4 ------7/29/2018 82.4 50.8 0.00 66.6 ------7/30/2018 74.0 56.0 0.43 65.0 ------7/31/2018 80.5 61.2 0.40 70.8 ------8/1/2018 81.9 64.0 0.00 73.0 ------8/2/2018 88.0 62.5 0.00 75.2 ------8/3/2018 86.0 61.9 0.00 74.0 ------8/4/2018 90.7 60.5 0.00 75.6 ------8/5/2018 93.7 59.0 0.00 76.4 ------8/6/2018 96.5 64.2 0.00 80.4 ------8/7/2018 82.5 66.1 0.00 74.3 ------8/8/2018 89.8 63.5 0.00 76.6 ------8/9/2018 89.2 59.7 0.23 74.4 ------8/10/2018 87.7 61.6 0.00 74.6 ------Page 4 Happy Dog Farm - Ciders Date Phenology Max Temp (F) Min Temp (F) Wetness (in) Trauma Spray Notes Avg Temp (F) EIP BHWTR BBS CBS SBS TBS 8/11/2018 86.8 55.3 0.00 71.0 ------8/12/2018 88.7 50.2 0.00 69.4 ------8/13/2018 82.2 53.9 0.00 68.0 ------8/14/2018 84.8 58.5 0.00 71.6 ------8/15/2018 88.1 56.4 0.00 72.2 ------8/16/2018 90.2 60.0 0.00 75.1 ------8/17/2018 84.8 67.5 0.00 76.2 ------8/18/2018 82.6 55.2 0.25 68.9 ------8/19/2018 80.1 52.2 0.00 66.2 ------8/20/2018 83.2 56.8 0.00 70.0 ------8/21/2018 76.5 65.2 0.00 70.8 ------8/22/2018 71.2 50.4 3.50 60.8 ------8/23/2018 80.2 46.2 0.00 63.2 ------8/24/2018 84.3 49.4 0.00 66.8 ------8/25/2018 83.7 58.3 0.00 71.0 ------8/26/2018 88.5 63.2 0.00 75.8 ------8/27/2018 92.4 66.3 0.00 79.4 ------8/28/2018 93.7 68.8 0.00 81.2 ------8/29/2018 93.6 66.2 0.00 79.9 ------8/30/2018 77.8 53.2 0.00 65.5 ------8/31/2018 86.5 52.7 0.56 69.6 ------9/1/2018 91.4 63.5 0.00 77.4 ------9/2/2018 92.5 67.8 0.00 80.2 ------9/3/2018 92.0 64.8 0.00 78.4 ------9/4/2018 97.1 64.3 0.00 80.7 ------9/5/2018 96.0 66.6 0.00 81.3 ------9/6/2018 0.0 0.0 0.20 Frost Frost 0.0 ------

Appendix B Lab reports

Soil Samples Soils samples were taken at the end of the 2017 season and submitted to the Penn State Agricultural Analytical Services laboratory in University Park, PA for analysis. The results reflected pH and nutrient imbalances that were addressed by liming, side-dressing, and foliar feeding. 2018 soil samples were not available at the time of this report.

Tissue Samples Tissue samples were taken from the control trees for each variety and submitted to the Penn State Agricultural Analytical Services laboratory in University Park, PA for analysis. The results were compared to a single sample taking the previous year from Medaille d’Or (rows 3 & 4). The control trees continue to show a deficiency in calcium despite foliar feeding. This is a reflection of a nutrient deficiency should not be attributed to SAR activity. I was concerned about calcium levels since it appears to be a critical nutrient for signaling pathways (22). It appears that the calcium level was adequate.

B-1

Intentionally blank SOIL TEST REPORT FOR: ADDITIONAL COPY TO: ALAN YELVINGTON RUSSELL 373 TOWLINE RD RUSSELL PA 16345

DATE LAB # SERIAL # COUNTY ACRES FIELD ID SOIL 10/26/2017 S17-41719 Warren 1-2

SOIL NUTRIENT LEVELS Deficient Optimum Exceeds Crop Needs Soil pH 5.4

Phosphate (P2O5) 147 lb/A

Potash (K2O) 221 lb/A Magnesium (MgO) 704 lb/A Calcium (CaO) 4055 lb/A

RECOMMENDATIONS FOR: Apples-To Plant Limestone: 6000 lb/A

Magnesium (Mg): NONE

Phosphate (P2O5): 150 lb/A

Potash (K2O): 120 lb/A

MESSAGES

Nitrogen should be applied based on leaf analysis and shoot growth. In absence of a current season's leaf analysis, shoot growth on bearing trees should be 12 to 18 inches. Another general guideline is to apply 0.02 lb of actual N per tree per year of tree age. If following this guideline, do not exceed 0.30 lb of actual N per tree per year.

If terminal growth was excessive, fruit color was inadequate or major renovative pruning was performed, a reduction in the rate of N application is in order.

LABORATORY RESULTS: Optional Tests: 1pH 2P lb/A Exchangeable Cations (meq/100g) % Saturation of the CEC Organic Nitrate-N Soluble salts 3Acidity 2K 2Mg 2Ca 4CEC K Mg Ca Matter % ppm mmhos/cm 5.4 64 6.9 0.2 1.8 7.3 16.2 1.5 10.9 45.0 1 2 3 4

Test Methods: 1:1 soil:water pH, Mehlich 3 (ICP), Mehlich Buffer pH, Summation of Cations Tree Fruit-1

5420 SOIL TEST REPORT FOR: ADDITIONAL COPY TO: ALAN YELVINGTON RUSSELL 373 TOWLINE RD RUSSELL PA 16345

DATE LAB # SERIAL # COUNTY ACRES FIELD ID SOIL 10/26/2017 S17-41720 Warren 3-4

SOIL NUTRIENT LEVELS Deficient Optimum Exceeds Crop Needs Soil pH 5.8

Phosphate (P2O5) 96 lb/A

Potash (K2O) 242 lb/A Magnesium (MgO) 797 lb/A Calcium (CaO) 4518 lb/A

RECOMMENDATIONS FOR: Apples-To Plant Limestone: 4000 lb/A

Magnesium (Mg): NONE

Phosphate (P2O5): 175 lb/A

Potash (K2O): 80 lb/A

MESSAGES

If terminal growth was excessive, fruit color was inadequate or major renovative pruning was performed, a reduction in the rate of N application is in order.

Test Methods: 1:1 soil:water pH, Mehlich 3 (ICP), Mehlich Buffer pH, Summation of Cations Tree Fruit-1

5421 SOIL TEST REPORT FOR: ADDITIONAL COPY TO: ALAN YELVINGTON RUSSELL 373 TOWLINE RD RUSSELL PA 16345

DATE LAB # SERIAL # COUNTY ACRES FIELD ID SOIL 10/26/2017 S17-41721 Warren 5-6

SOIL NUTRIENT LEVELS Deficient Optimum Exceeds Crop Needs Soil pH 5.6

Phosphate (P2O5) 110 lb/A

Potash (K2O) 175 lb/A Magnesium (MgO) 740 lb/A Calcium (CaO) 4321 lb/A

RECOMMENDATIONS FOR: Apples-To Plant Limestone: 7000 lb/A

Magnesium (Mg): NONE

Phosphate (P2O5): 175 lb/A

Potash (K2O): 120 lb/A

MESSAGES

If terminal growth was excessive, fruit color was inadequate or major renovative pruning was performed, a reduction in the rate of N application is in order.

Test Methods: 1:1 soil:water pH, Mehlich 3 (ICP), Mehlich Buffer pH, Summation of Cations Tree Fruit-1

5422 SOIL TEST REPORT FOR: ADDITIONAL COPY TO: ALAN YELVINGTON RUSSELL 373 TOWLINE RD RUSSELL PA 16345

DATE LAB # SERIAL # COUNTY ACRES FIELD ID SOIL 10/26/2017 S17-41722 Warren 7-8

SOIL NUTRIENT LEVELS Deficient Optimum Exceeds Crop Needs Soil pH 5.7

Phosphate (P2O5) 55 lb/A

Potash (K2O) 149 lb/A Magnesium (MgO) 677 lb/A Calcium (CaO) 3780 lb/A

RECOMMENDATIONS FOR: Apples-To Plant Limestone: 5000 lb/A

Magnesium (Mg): NONE

Phosphate (P2O5): 175 lb/A

Potash (K2O): 120 lb/A

MESSAGES

If terminal growth was excessive, fruit color was inadequate or major renovative pruning was performed, a reduction in the rate of N application is in order.

Test Methods: 1:1 soil:water pH, Mehlich 3 (ICP), Mehlich Buffer pH, Summation of Cations Tree Fruit-1

5423 SOIL TEST REPORT FOR: ADDITIONAL COPY TO: ALAN YELVINGTON RUSSELL 373 TOWLINE RD RUSSELL PA 16345

DATE COUNTY 10/26/2017 Warren

Soil Test Report1

Lab ID Field ID Copper Iron Manganese Zinc

mg/kg

S17-41719 1-2 1.83 235.58 17.64 2.05 S17-41720 3-4 1.81 227.46 23.91 1.68 S17-41721 5-6 2.16 226.66 31.62 1.89 S17-41722 7-8 1.87 220.79 35.73 1.47

1Mehlich 3 Soil Test PLANT TISSUE ANALYSIS FOR: ADDITIONAL COPY TO: Alan Yelvington Happy Dog Farm LLC 373 Townline Road Russell PA 16345