Source: UNIV OF MARYLAND submitted to 
CHARACTERIZING THE EFFECTS OF CULTURAL MANAGEMENT ON PHYTOCHEMICAL CONCENTRATIONS IN THE FRUIT OF THE ALTERNATIVE CROP ARONIA
Sponsoring Institution
State Agricultural Experiment Station
Project Status
NEW
Funding Source
Reporting Frequency
Annual
Accession No.
1000894
Grant No.
(N/A)
Project No.
MD-UME-7390.
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Aug 14, 2013
Project End Date
Jun 30, 2014
Grant Year
(N/A)
Project Director
RISTVEY, AN, .
RISTVEY, AN, .
Performing Department
Cooperative Extension Service
Cooperative Extension Service
Non Technical Summary
Aronia is a new specialty crop fruit, which has recently garnered interest by a number of groups including growers (farmers and other land owners), producers of foods and juices, pharmaceutical companies, amongst others. Aronia is a pome-fruit bearing shrub in the Rose family which has been shown to possess very high concentrations of anthocyanins in the fruit. These polyphenolic phytochemicals have been implicated to benefit and promote cardio vascular, gastrointestinal, urological, pulmonary, and musculoskeletal health. This makes the fruit a very marketable commodity which could bring profit to those invested in culture, processing and sales of Aronia products. Presently, there is much to understand about Aronia production especially with the effects cultural management on the health of the plant and quality of the fruit. This study proposes to investigate the effects of fertility, and other cultural practices including plant spacing on plant health and fruit quality, including phytochemical content. The results of this research program will be disseminated through an already nationally recognized University of Maryland Extension program, extension publications and peer-reviewed journal manuscripts. This grant may serve to assist investigators in applying for a USDA/NIFA SCRI Planning Grant for 2014, if available, a 2014 North East SARE Grant .
Aronia is a new specialty crop fruit, which has recently garnered interest by a number of groups including growers (farmers and other land owners), producers of foods and juices, pharmaceutical companies, amongst others. Aronia is a pome-fruit bearing shrub in the Rose family which has been shown to possess very high concentrations of anthocyanins in the fruit. These polyphenolic phytochemicals have been implicated to benefit and promote cardio vascular, gastrointestinal, urological, pulmonary, and musculoskeletal health. This makes the fruit a very marketable commodity which could bring profit to those invested in culture, processing and sales of Aronia products. Presently, there is much to understand about Aronia production especially with the effects cultural management on the health of the plant and quality of the fruit. This study proposes to investigate the effects of fertility, and other cultural practices including plant spacing on plant health and fruit quality, including phytochemical content. The results of this research program will be disseminated through an already nationally recognized University of Maryland Extension program, extension publications and peer-reviewed journal manuscripts. This grant may serve to assist investigators in applying for a USDA/NIFA SCRI Planning Grant for 2014, if available, a 2014 North East SARE Grant .
Animal Health Component
75%
Research Effort Categories
Basic
(N/A)
Applied
75%
Developmental
25%
Classification
| Knowledge Area (KA) | Subject of Investigation (SOI) | Field of Science (FOS) | Percent |
|---|---|---|---|
| 701 | 1199 | 2000 | 100% |
Knowledge Area
701 – Nutrient Composition of Food;
701 – Nutrient Composition of Food;
Subject Of Investigation
1199 – Deciduous and small fruits, general/other;
Field Of Science
2000 – Chemistry;
Goals / Objectives
Objectives – Using the presently established orchard with the two cultivars Aronia mitschurinii 'Viking' and Aronia mitschurinii 'Nero' the following research objectives will be met. Examine the continued effect of organic-based nitrogen rates on plant nutritional status (leaf samples), fruit yield, brix, juice pH and anthocyanin/polyphenol content and character. Determine differences, if any, between use of conventional nitrogen and organic nitrogen on nutritional status (leaf samples), fruit yield, brix, juice pH and anthocyanin/polyphenol content and character. Determine the effect of plant spacing on nutritional status (leaf samples), fruit yield, brix, juice pH and anthocyanin/polyphenol content and character. These information obtained from these objectives will provide a basis for publications, extension programs and initial data for submission of larger grant proposals.
Objectives – Using the presently established orchard with the two cultivars Aronia mitschurinii 'Viking' and Aronia mitschurinii 'Nero' the following research objectives will be met. Examine the continued effect of organic-based nitrogen rates on plant nutritional status (leaf samples), fruit yield, brix, juice pH and anthocyanin/polyphenol content and character. Determine differences, if any, between use of conventional nitrogen and organic nitrogen on nutritional status (leaf samples), fruit yield, brix, juice pH and anthocyanin/polyphenol content and character. Determine the effect of plant spacing on nutritional status (leaf samples), fruit yield, brix, juice pH and anthocyanin/polyphenol content and character. These information obtained from these objectives will provide a basis for publications, extension programs and initial data for submission of larger grant proposals.
Project Methods
Research Plan – The Orchard at Wye Research and Education Center An active Aronia research orchard, established in May of 2006 will give this research immediate results towards proposed goals. The orchard contains three sections, 1, 2 and 3. Each section contains four rows (A, B, C and D) approximately 23 meters in length with approximately 3 meters of turf between rows. Only Organic Materials Review Institute fertilizers and pesticides have been utilized in sections 1 and 2. Section 1 consists of 100 Aronia plants in four rows of 25 plants at approximately one meter spacing between plants, and containing the cultivars Aronia 'Viking' and Aronia 'Nero'. Twenty-four Aronia 'Viking' plants were planted as two year-old bare root plants in groups of six, distributed randomly within the four rows. Seventy-four Aronia 'Nero' plants, less than 12 months old, were planted with the Aronia 'Viking' plants so that the two cultivars are distributed in randomized groups within the rows. Each plant is considered an experimental unit. Treatment groups of 4 to 6 plants were randomly assigned within the rows and plants on the outside of the treatment group have been considered guard plants from which no data are utilized. Initial fertility consisted of 6 grams of nitrogen (N) per plant the first year of planting. Presently an ongoing organic N rate treatment is being studied, but to date, only yield weight and soluble sugar content has been measured at harvest. Since 2009, the plants have been consistently treated with 4 N levels of 0, 3, 7 and 14 grams per plant per year (for Nero) and 2 N levels of 0 and 7 grams per plant per year (for Viking), randomly applied within the treatment groups of each row. This experimental design allows for a Randomized Complete Block Design with the potential to nest treatments within the rows, which serve as blocks. Section 1 serves as our original orchard and as the main source of information on yield, soluble sugar content and pest monitoring for the Extension program. Section 2 contains 80 Aronia 'Viking” plants, planted in June of 2010 as 12-month old liner plants at a spacing of 1.2 meters between plants. Initial fertility at planting was 7 grams of N per plant. In 2011 and 2012, N fertility was split into 14 grams per plant per year on rows A and C and 3 grams N per plant per year in rows B and D. No viable data has been taken from Section 2 as of yet. However, in this forth growing season, plant will yield their second harvest for data collection. Section 3 was planted in 2011 with 44 Aronia 'Viking' plants at spacing of 2.1 meters between plants with the intent of quantifying differences in fruit quality as related to canopy sunlight interception and in contrast to Sections 1 and 2. No initial fertilization was given to Section 3 plants at planting nor since then. However, this year, Section 3 plants will be fertilized by either the conventional chemical fertilizer (Scotts® with IBDU and sulfur coated urea with 50% WIN activity) or with OMRI certified Boost® Natural and rock phosphate. Potassium will also be added to the conventional N source to balance K amongst treatments. Rows will be divided into fertility rates similar to Section 2 with either 3 or 14 g of N per plant. Also 0.3 g or 1.4 g of P and 1.6 g or 7.4 g K per plant will be applied to maintain the NPK ratio. Each row will be split in half. Plants in one randomly chosen half-row will receive conventional chemical fertilizer and the other half will receive the OMRI certified organic fertilizers. N rates will remain the same within the row. Plants in Sections 1 and 2 have been fertilized in March of each year and plants in Section 3 will be fertilized in March of this year and thereafter. Leaf Samples For Objectives 1, 2 and 3, all leaves will be harvested in the same manner for all Sections. Leaf samples will be taken 1 month before harvest to determine nutritional status of plants based on fertility and spacing treatments. A total of approximately 15 grams of leaves (first fully expanded leaves from different locations on the plant) will be harvested from each experimental unit and dried at 60 oC for two days. Samples will be milled and then analyzed for nutrient status at the University of Delaware Soils Laboratory, Newark, DE. Harvest For Objectives 1, 2 and 3, all fruit will be harvested in the same manner for all Sections. Fruit will be harvested in mid to late August. Fruit fresh weight will be recorded from each experimental unit (each plant). Guard-plant fruit will not be used for analyses. Approximately 100 grams of fruit will be randomly taken as a grab from each experimental unit after weighing. Brix will be measured on ten fruit from this grab and values averaged. The rest of the grab will be frozen in individual packages and sent to the laboratory of Dr.Victoria Volkis at University of Maryland Eastern Shore for chemical analyses. Detailed methods for chemical analyses are outlined below. Sample Preparation Two types of samples from Aronia will be prepared from each experimental unit: Freeze Dried Fruit Samples and Juice Preparation Anthocyanin Determination Sample Preparations and Analyses Calculations of Anthocyanin Pigment Calculation of anthocyanin pigment concentration (APC), expressed as cyanidin-3-glucoside equivalents, will be done as follows: APC (cyanidin-3-glucoside equivalents, mg/L) = A* MW * 103/ ? x 1 where A = (A520nm – A700nm)pH 1.0 – (A520nm – A700nm)pH 4.5; MW (molecular weight) = 449.2 g/mol for cyanidin-3-glucoside (cyd-3-glu); DF = dilution factor established above (100X); l = pathlength in cm; ? = 26 900 molar extinction coefficient, in L & mol-1 & cm-1, for cyd-3-glu; and 10×3 = factor for conversion from g to mg. Results will be reported as monomeric anthocyanin, expressed as cyanidin-3-glucoside equivalents in mg/L. Total Polyphenols Analyses Total Polyphenols Sample Preparation Preparation of Gallic Acid stock solution: 0.55291g of gallic acid monohydrate was dissolved in in 10mL of ethanol and diluted to volume of 100mL with water. Preparation of Sodium Carbonate solution: 233.962g of sodium carbonate monohydrate was dissolved in 800mL of water, brought to a boil, and left to settle for 24hrs. Then, the solution was filtered and water was added to bring the volume to 1L. Prior to sample measurements, a calibration curve will be created. To prepare a calibration curve, 0, 1, 2, 3, 5, and 10 mL of the gallic acid stock solution was added into 100 mL volumetric flasks and diluted to volume with water. These solutions then had the phenol concentrations of 0, 50, 100, 150, 250, and 500 mg/L gallic acid. Parameter data from fruit assays will be analyzed according to the experimental procedures of which are RCBD's. Inference from all results will rely on statistical analyses performed as RCBD with ANOVA by SAS Proc Mixed (SAS Institute Inc., Cary, N.C.). Block effect will be determined and used if significant in explaining variation. If treatment interaction is not significant, main effects will be reported and discussed. However, if treatment interactions are significant, simple effects (the effect of a variable at a specific level of another variable) will be reported and discussed. Data showing unequal variation will be transformed and reanalyzed. Significant results will tell us how cultural management parameters as treatments were effective in increasing phytochemcial production or yield or optimally, both. Inferences from this information will be made and will be disseminated by the methods perviously described in Products.
Research Plan – The Orchard at Wye Research and Education Center An active Aronia research orchard, established in May of 2006 will give this research immediate results towards proposed goals. The orchard contains three sections, 1, 2 and 3. Each section contains four rows (A, B, C and D) approximately 23 meters in length with approximately 3 meters of turf between rows. Only Organic Materials Review Institute fertilizers and pesticides have been utilized in sections 1 and 2. Section 1 consists of 100 Aronia plants in four rows of 25 plants at approximately one meter spacing between plants, and containing the cultivars Aronia 'Viking' and Aronia 'Nero'. Twenty-four Aronia 'Viking' plants were planted as two year-old bare root plants in groups of six, distributed randomly within the four rows. Seventy-four Aronia 'Nero' plants, less than 12 months old, were planted with the Aronia 'Viking' plants so that the two cultivars are distributed in randomized groups within the rows. Each plant is considered an experimental unit. Treatment groups of 4 to 6 plants were randomly assigned within the rows and plants on the outside of the treatment group have been considered guard plants from which no data are utilized. Initial fertility consisted of 6 grams of nitrogen (N) per plant the first year of planting. Presently an ongoing organic N rate treatment is being studied, but to date, only yield weight and soluble sugar content has been measured at harvest. Since 2009, the plants have been consistently treated with 4 N levels of 0, 3, 7 and 14 grams per plant per year (for Nero) and 2 N levels of 0 and 7 grams per plant per year (for Viking), randomly applied within the treatment groups of each row. This experimental design allows for a Randomized Complete Block Design with the potential to nest treatments within the rows, which serve as blocks. Section 1 serves as our original orchard and as the main source of information on yield, soluble sugar content and pest monitoring for the Extension program. Section 2 contains 80 Aronia 'Viking” plants, planted in June of 2010 as 12-month old liner plants at a spacing of 1.2 meters between plants. Initial fertility at planting was 7 grams of N per plant. In 2011 and 2012, N fertility was split into 14 grams per plant per year on rows A and C and 3 grams N per plant per year in rows B and D. No viable data has been taken from Section 2 as of yet. However, in this forth growing season, plant will yield their second harvest for data collection. Section 3 was planted in 2011 with 44 Aronia 'Viking' plants at spacing of 2.1 meters between plants with the intent of quantifying differences in fruit quality as related to canopy sunlight interception and in contrast to Sections 1 and 2. No initial fertilization was given to Section 3 plants at planting nor since then. However, this year, Section 3 plants will be fertilized by either the conventional chemical fertilizer (Scotts® with IBDU and sulfur coated urea with 50% WIN activity) or with OMRI certified Boost® Natural and rock phosphate. Potassium will also be added to the conventional N source to balance K amongst treatments. Rows will be divided into fertility rates similar to Section 2 with either 3 or 14 g of N per plant. Also 0.3 g or 1.4 g of P and 1.6 g or 7.4 g K per plant will be applied to maintain the NPK ratio. Each row will be split in half. Plants in one randomly chosen half-row will receive conventional chemical fertilizer and the other half will receive the OMRI certified organic fertilizers. N rates will remain the same within the row. Plants in Sections 1 and 2 have been fertilized in March of each year and plants in Section 3 will be fertilized in March of this year and thereafter. Leaf Samples For Objectives 1, 2 and 3, all leaves will be harvested in the same manner for all Sections. Leaf samples will be taken 1 month before harvest to determine nutritional status of plants based on fertility and spacing treatments. A total of approximately 15 grams of leaves (first fully expanded leaves from different locations on the plant) will be harvested from each experimental unit and dried at 60 oC for two days. Samples will be milled and then analyzed for nutrient status at the University of Delaware Soils Laboratory, Newark, DE. Harvest For Objectives 1, 2 and 3, all fruit will be harvested in the same manner for all Sections. Fruit will be harvested in mid to late August. Fruit fresh weight will be recorded from each experimental unit (each plant). Guard-plant fruit will not be used for analyses. Approximately 100 grams of fruit will be randomly taken as a grab from each experimental unit after weighing. Brix will be measured on ten fruit from this grab and values averaged. The rest of the grab will be frozen in individual packages and sent to the laboratory of Dr.Victoria Volkis at University of Maryland Eastern Shore for chemical analyses. Detailed methods for chemical analyses are outlined below. Sample Preparation Two types of samples from Aronia will be prepared from each experimental unit: Freeze Dried Fruit Samples and Juice Preparation Anthocyanin Determination Sample Preparations and Analyses Calculations of Anthocyanin Pigment Calculation of anthocyanin pigment concentration (APC), expressed as cyanidin-3-glucoside equivalents, will be done as follows: APC (cyanidin-3-glucoside equivalents, mg/L) = A* MW * 103/ ? x 1 where A = (A520nm – A700nm)pH 1.0 – (A520nm – A700nm)pH 4.5; MW (molecular weight) = 449.2 g/mol for cyanidin-3-glucoside (cyd-3-glu); DF = dilution factor established above (100X); l = pathlength in cm; ? = 26 900 molar extinction coefficient, in L & mol-1 & cm-1, for cyd-3-glu; and 10×3 = factor for conversion from g to mg. Results will be reported as monomeric anthocyanin, expressed as cyanidin-3-glucoside equivalents in mg/L. Total Polyphenols Analyses Total Polyphenols Sample Preparation Preparation of Gallic Acid stock solution: 0.55291g of gallic acid monohydrate was dissolved in in 10mL of ethanol and diluted to volume of 100mL with water. Preparation of Sodium Carbonate solution: 233.962g of sodium carbonate monohydrate was dissolved in 800mL of water, brought to a boil, and left to settle for 24hrs. Then, the solution was filtered and water was added to bring the volume to 1L. Prior to sample measurements, a calibration curve will be created. To prepare a calibration curve, 0, 1, 2, 3, 5, and 10 mL of the gallic acid stock solution was added into 100 mL volumetric flasks and diluted to volume with water. These solutions then had the phenol concentrations of 0, 50, 100, 150, 250, and 500 mg/L gallic acid. Parameter data from fruit assays will be analyzed according to the experimental procedures of which are RCBD's. Inference from all results will rely on statistical analyses performed as RCBD with ANOVA by SAS Proc Mixed (SAS Institute Inc., Cary, N.C.). Block effect will be determined and used if significant in explaining variation. If treatment interaction is not significant, main effects will be reported and discussed. However, if treatment interactions are significant, simple effects (the effect of a variable at a specific level of another variable) will be reported and discussed. Data showing unequal variation will be transformed and reanalyzed. Significant results will tell us how cultural management parameters as treatments were effective in increasing phytochemcial production or yield or optimally, both. Inferences from this information will be made and will be disseminated by the methods perviously described in Products.
MID ATLANTIC ARONIA GROWERS ASSOCIATION 