Arun Sharma, In relation to the value of organic seed this is focused on seed from a certified organic system of management which is programmed and verified. The organic system must have a program of soil conservation and improvement and use biologically based pest management tools and mineral supplementation. The key to this is many of our issues are related to how monocultures based on synthetic inputs has degraded our soils. Mitchell group at the University of California show that under organic management the defensive flavonoid phytonutrients are enhanced because the synthetic inputs prevent the healthy responsive reaction found in the organic system. As such the nature of the product is enhances since these flavonoids are important in animal and human nutrition. Healthy nutrition is based on these components which are why fresh fruit and vegetables give optimized health for human and beast. The ability of humic substances to stimulate plant and animal performance is well documented and when the system of production is geared toward them the combination of withdrawing the negative effects of synthetic inputs and the augmenting of biological ones give a higher quality. Best the environment is optimized by organic matter and the natural stresses are stimulative of induced defenses an optimized nutritional outcome is a result. In relation to that work from Denmark shows both organic farmers and consumers are experiencing higher human sperm production which is a product of the elimination of the depletion of the soil and elimination of toxic inputs. 

This analysis make sense. Any how, what about doing biochemical testing for seeds. For example, protein content could be a good indicator for that. 

Ten-Year Comparison of the Influence of Organic and Conventional Crop Management Practices on the Content of Flavonoids in Tomatoes

Alyson E. Mitchell ,*† Yun-Jeong Hong ,† Eunmi Koh ,† Diane M. Barrett ,† D. E. Bryant ,§ R. Ford Denison ,#and Stephen Kaffka §

Department of Food Science and Technology and Department of Plant Sciences, One Shields Avenue, University of California Davis, Davis, California 95616, and Department of Ecology, Evolution and Behavior, University of Minnesota, St. Paul, Minnesota 55108

J. Agric. Food Chem., 2007, 55 (15), pp 6154–6159

DOI: 10.1021/jf070344+

Publication Date (Web): June 23, 2007

Copyright © 2007 American Chemical Society

Abstract

Understanding how environment, crop management, and other factors, particularly soil fertility, influence the composition and quality of food crops is necessary for the production of high-quality nutritious foods. The flavonoid aglycones quercetin and kaempferol were measured in dried tomato samples (Lycopersicon esculentum L. cv. Halley 3155) that had been archived over the period from 1994 to 2004 from the Long-Term Research on Agricultural Systems project (LTRAS) at the University of California Davis, which began in 1993. Conventional and organic processing tomato production systems are part of the set of systems compared at LTRAS. Comparisons of analyses of archived samples from conventional and organic production systems demonstrated statistically higher levels (P < 0.05) of quercetin and kaempferol aglycones in organic tomatoes. Ten-year mean levels of quercetin and kaempferol in organic tomatoes [115.5 and 63.3 mg g-1 of dry matter (DM)] were 79 and 97% higher than those in conventional tomatoes (64.6 and 32.06 mg g-1 of DM), respectively. The levels of flavonoids increased over time in samples from organic treatments, whereas the levels of flavonoids did not vary significantly in conventional treatments. This increase corresponds not only with increasing amounts of soil organic matter accumulating in organic plots but also with reduced manure application rates once soils in the organic systems had reached equilibrium levels of organic matter. Well-quantified changes in tomato nutrients over years in organic farming systems have not been reported previously.

Keywords: Tomato; organic agriculture; conventional agriculture; long-term research; flavonoid; flavonol; quercetin; kaempferol

Introduction

Fruits and vegetables are rich in dietary vitamins, minerals, and fiber and are the primary source of flavonoids in the diet. Epidemiological studies suggest that flavonoids protect against cardiovascular disease (1) and, to a lesser extent, against cancer (2) and other age-related diseases such as dementia (3). Flavonoids encompass a large group of phenolic secondary plant metabolites that demonstrate potent in vitro antioxidant activity (4, 5) and display free radical scavenging activity (6). Most attempts to assign health-promoting activity mechanistically to the antioxidant action of individual flavonoids in foods have been unsuccessful (7) and suggest that other mechanisms are likely involved in the health-promoting activities of these compounds.

Secondary plant metabolites such as the flavonoids function in plant defense mechanisms against herbivory, pathogen stress, and UV-B radiation (8). Many of these compounds are also the primary pigments responsible for attracting pollinators and seed dispersers. Flavonoids are produced through the phenylpropanoid pathway in plants. The key enzyme that catalyzes the biosynthesis is phenylalanine ammonia-lyase. Environmental stresses including nutrient deficiency, wounding, pathogens, and UV radiation are known to activate the biosynthesis of phenylpropanoid compounds (9, 10). Given the increasing data in support of a role for phenolic plant metabolites in the prevention of cardiovascular and other chronic diseases, efforts are underway to improve their levels in plant foods through improved cultivar selection and genetic manipulations. Alternatively, the phenolic content of plants may be influenced by manipulating the agronomic environment in which they are grown (11−13).

Fundamental differences between organic and conventional production systems, particularly in soil fertility management, may affect the nutritive composition of plants, including secondary plant metabolites. Organic systems emphasize the accumulation of soil organic matter and fertility over time through the use of cover crops, manures, and composts and rely on the activity of a diverse soil ecosystem to make nitrogen (N) and other nutrients available to plants. Conventional farms utilize fertilizers containing soluble inorganic nitrogen and other nutrients, which are more directly available to plants. The availability of inorganic nitrogen in particular has the potential to influence the synthesis of secondary plant metabolites, proteins, and soluble solids. Stamp proposed that increased crop growth and development rates and greater biomass accumulation in well-fertilized crops would also correlate with decreased allocation of resources toward the production of starch, cellulose, and non-nitrogen-containing secondary metabolites (14). Given that many secondary plant metabolites are produced for defense against herbivores and are inducible by pathogens or wounding, possible differences in pest pressure between conventional and organic systems might also influence levels in food crops.

Both conventional and organic agricultural practices include combinations of farming practices that vary greatly depending upon region, climate, soils, pests and diseases, and economic factors guiding the particular management practices used on the farm. Many of these influences change continuously, so a steady-state condition may never be achieved on most farms (15). The dynamic nature of agricultural systems also makes adequately controlled comparisons of produce quality, free from confounding influences, experimentally challenging. Comparisons of produce from different types of farms, even if they are near each other, are affected by the numerous factors mentioned. Reviews of studies comparing the nutritional quality of conventionally and organically produced vegetables demonstrate inconsistent differences with the exception of higher levels of ascorbic acid (vitamin C) and less nitrate in organic products (16, 17). However, these data are difficult to interpret because cultivar selection and agronomic conditions varied widely and different methods of sampling and analysis were used in the investigations cited. In contrast, cropping system comparisons using long-term research plots that have been managed consistently over time provide a means to overcome many of the confounding factors associated with farm-based sampling. Additionally, the effects of changes over time in cropping system behavior can be evaluated using archived soil and plant samples, and a reasonable estimate of the causes of those changes can be made.

The present study is an example of the use of long-term research to address complex processes operating in cropping systems. Its specific objective was to compare the content of the flavonoids quercetin, kaempferol, and naringenin in tomato samples (Lycopersicon esculentum L. cv. Halley 3155) produced in conventional and organic cropping systems that had been archived over the period from 1994 to 2004 from the Long-Term Research on Agricultural Systems project (LTRAS) at the University of California Davis, which began in 1993 (http://ltras.ucdavis.edu). LTRAS was designed to detect and estimate changes in crop productivity trends and other factors correlated with sustainability, which result from differing irrigation and fertilization practices (18). It includes an organic cropping system in which maize and tomatoes are grown in rotation and compared to the same crops produced conventionally. This archive of tomato samples is unique in California and perhaps the world.

This study focused on tomatoes because the per capita consumption of tomatoes (8.1 kg per capita in 2003) and tomato products (31.1 kg per capita in 2003) in the United States is very high, second only to the potato (19). Tomatoes provide an important and significant source of vitamin C (19 mg/100 g of fresh weight), vitamin A (623 IU/100 g of fresh weight), lycopene (3.0 mg/100 g of fresh weight), and flavonoids (20, 21). The main flavonoids found in tomatoes are quercetin, kaempferol, and naringenin (Figure 1). Quercetin predominates, with levels ranging from 0.03 to 2.77 mg/100 g in fresh tomatoes and 4.77 mg/100 g in processed tomato products (22). Levels of kaempferol range from 0.01 to 0.06 mg/100 g in fresh and processed tomatoes (22). Figure 1 Structures of the flavonoid aglycones of quercetin, keampferol, and naringenin.

Materials and Methods

Chemicals and Samples. Quercetin dihydrate (3,5,7,3‘,4‘-pentahydroxyflavone dihydrate, 98%), morin hydrate (3,5,7,2‘,4‘-pentahydroxyflavone hydrate, 95%), and tert-butylhydroquinone (TBHQ, 97%) were purchased from Aldrich Chemical Co., Inc. (Milwaukee, WI). Kaempferol (3,5,7,4‘-tetrahydroxyflavone, 95%) and naringenin (4‘,5,7-trihydroxyflavone, 95%) were obtained from Sigma Chemical Co. (St. Louis, MO), and Indofine Chemical Co., Inc. (Hillsborough, NJ), respectively. All solvents used were of HPLC grade. Ethanol was purchased from Spectrum Chemicals (Gardena, CA).

Tomato Cultivation in the LTRAS Cropping Systems. Following uniform cropping with Sudan grass (Sorghum vulgare L.) in 1992 and 1993, 10 different cropping systems were established in 1993 using 0.4 ha plots (18). Each cropping system was replicated three times, and both crops or phases of the two-year rotations were present each year. Plots were large enough to allow for the use of commercial-scale farm equipment. Irrigation amounts were measured using flow meters located at each irrigated plot. Systems differ in the amount of irrigation received (rain fed or irrigated), in the amounts of nutrients applied as fertilizers, and in organic matter applied to the soil as crop residues, winter legume cover crops, and/or composted manure. Conventional plots received herbicides and other pesticides as needed, whereas organic crops received only organically approved pesticides, such as sulfur and Bt compounds. Crop yields and total biomass were measured every year and analyzed for total N and C. Sample archiving included yearly plant and fruit samples from all cropping systems and time zero and subsequent soil samples collected every few years. Systems rather than single inputs were compared, so a valid comparison required that each system be managed carefully to achieve its potential yields. For example, both the conventional and organic maize/tomato systems had the same tomato cultivar (Halley 3155) and were irrigated similarly as needed, but the lower percent availability of organic N sources can require more total N input to meet crop N needs (24). The same cultivar was used throughout the 12-year period. The systems compared are model systems, chosen to include representative crops rather than more complex, changeable crop rotations. California farmers rarely follow fixed crop rotations as markets change, and organic farmers especially tend to have more complex rotations than the one studied at LTRAS.

Crop management practices follow best management practice guidelines in the region. Conventional tomatoes received 50 kg ha-1 of an N−P−K starter fertilizer and 118 kg ha-1 of ammonium nitrate as side dressing. A combination of tillage and herbicides was used for weed control. Aphids, mites, and stinkbugs occur periodically and were controlled as necessary, similar to practices in commercial fields. In both treatments, processing tomatoes were transplanted at the rate of 22500 plants ha-1 (10000 per acre). Transplanting in the organic treatment followed incorporation of a winter legume cover crop consisting of hairy vetch (Vicia villosa Roth) and field peas (Pisum sativum L.). Transplanting of all plots occurred within a period of 2−4 days in spring, commonly around the middle of April. Prior to incorporation of the cover crop, 9 Mg ha-1 of composted poultry manure currently is top-dressed and incorporated with the cover crop. The amount of N present in cover crops varies from year to year, but, typically, organic plots currently receive between 240 and 260 kg of N ha-1 year in addition to the N fixed by the legume cover crop (24). During the first three cropping cycles, to more rapidly increase soil organic matter levels and soil fertility, 19 Mg ha-1 of composted manure was added to tomato crops. This was reduced after organic matter levels had increased to a near constant level (24). Tomatoes were harvested each year when the field as a whole reached 90% ripe fruit. A commercial tomato harvester was used for main plot harvests. Similar methods of transplanting and plant populations were used for conventional plots as well, and transplanting of all plots occurred within a period of 2−4 days in spring, commonly around the middle of April.

Sampling and Preparation of Plant Material. Immediately prior to harvest of the main plot, samples for the archive were collected from four 3.1 m2 subsample areas (subplots) within the larger main plots. Total plant biomass and the yield of the red ripe fruit were determined for each subplot. A random sample of 20 ripe fruits from the four subplots was washed and oven-dried at 60 °C, ground, and stored in glass containers in the dark at 20 °C until use. Samples from conventional and organic plots from even numbered years (1994−2004) were chosen for flavonoid analysis. Three of the subplots (n = 3) for each treatment (three organic; three conventional) were analyzed each of the even years (1994−2004). These subplot samples were analyzed in triplicate.

Flavonoid Analysis. Flavonoid analysis followed the general method of Merken and Beecher (25) as modified by Chassy et al. for fresh processing tomatoes (26). Flavonoid glycosides were hydrolyzed to their corresponding agylcones, which were subsequently measured using HPLC. Briefly, 1 g of pestle-pulverized, air-dried tomato sample was combined with 100 mL of 1.6 g/L tert-butylhydroquinone, 1.2 N hydrochloric acid, and 50% methanol (MeOH). To this mixture was added 0.5 mL of a 1 mg/mL standard of luteolin as an internal standard. Internal standard recovery was 92−93% for luteolin. Samples were refluxed for 4 h at 100 °C in a 250 mL round-bottom flask. Four hours of reflux time, in 1.2 N hydrochloric acid, was determined as the optimal condition for maximal recoveries of quercetin, kaempferol, and naringenin in these samples. An aliquot was removed, diluted 50:50 v/v with MeOH (200 μL), and filtered through a 0.5 mL of 25 μm MC Ultrafree-MC filter (Millipore, Bedford, MA). Flavonoids were separated using a Hewlett-Packard 1090 HPLC equipped with a variable wavelength diode array detector and Chemstation (LC 3D rev. A.08.03) software (Agilent, Palo Alto, CA) monitoring 370 nm. Reversed phase HPLC was performed using a 250 × 4.6 mm i.d., 5 μm Zorbax XDB-C18 column and a 12.5 × 4.6 mm i.d. 5 μm Zorbax XDB-C18 precolumn (Agilent). The mobile phase consisted of 0.05% trifluoroacetic acid (TFA) in water (solvent A), 0.05% TFA in MeOH (solvent B), and 0.05% TFA in acetonitrile (solvent C). Separations were effected by a series of linear gradients using a flow rate of 1.0 mL/min as follows: 90−85% A, 6−9% B, 4−6% C, 0−5 min; 85−71% A, 9−17.4% B, 6−11.6% C, 5−30 min; 71−0% A, 17.4−85% B, 11.6−15% C, 30−60 min. The linear ranges of quantitation for quercetin and kaempferol were 0.5−10 and 0.1−10 μg/mL respectively.

Statistical Analysis. Data were analyzed using SAS software version 9.1 (SAS Institute, Cary, NC). Specifically, PROC Mixed and regression were used to analyze treatment differences and create single degree of freedom contrasts for differences between systems (27).

Results

The mean level of the flavonoids quercetin, naringenin, and kaempferol agylcones (mg g-1 of DM) were significantly higher (p < 0.001) in samples from the organic cropping system as compared to samples from the conventional cropping system (Table 1). Quercetin was the most abundant flavonoid in both organic (115.5 ± 8.0 mg g-1 of DM) and conventional (64.6 ± 2.4 mg g-1 of DM) tomatoes. In conventional tomatoes, kaempferol (32.06 ± 1.94 mg g-1 of DM) and naringenin (30.2 ± 1.57 mg g-1 of DM) levels were comparable, whereas levels of kaempferol (63.3 ± 5.21 mg g-1 of DM) were significantly higher than naringenin levels (39.6 ± 1.58 mg g-1 of DM) in organic tomatoes. Absolute differences in the flavonoids increased with time in both systems, with the largest differences observed in the last 4-year period (Figures 2 and 3). Figure 2 Quercetin (·), naringenin (○), and kaempferol (▾) levels in tomatoes derived from conventional cropping systems at LTRAS from 1994 to 2004 (mg g-1of DM). Values are given with standard deviations.

Figure 3 Quercetin (·), naringenin (○), and kaempferol (▾) levels in tomatoes derived from organic systems at LTRAS from 1994 to 2004 (mg g-1 of DM). Values are given with standard deviations.

Table 1. Mean Flavonoid Concentrations in Archived Samples from the LTRAS Experiment (1994−2004) and Single Degree of Freedom Contrasts between Conventional and Organic Farming Systems

mean (SD) (mg g-1 of DM)

flavonoid

conventional

organic

F

p

quercetin

64.6 (2.49)

115.5 (8.0)

108.16