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WO2000033860A1 - Enrichissement au selenium de plantes alliaceae et brassicaceae - Google Patents

Enrichissement au selenium de plantes alliaceae et brassicaceae Download PDF

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Publication number
WO2000033860A1
WO2000033860A1 PCT/US1999/029454 US9929454W WO0033860A1 WO 2000033860 A1 WO2000033860 A1 WO 2000033860A1 US 9929454 W US9929454 W US 9929454W WO 0033860 A1 WO0033860 A1 WO 0033860A1
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selenium
plant
plants
plant tissue
seo
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William M. Ii Randle
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University of Georgia Research Foundation Inc UGARF
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/30—Oligoelements
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
    • A23L27/14—Dried spices
    • A23L27/16—Onions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/16—Inorganic salts, minerals or trace elements

Definitions

  • the field of this invention is the area of plant cultivation, in particular as applied to the selenium fertilization of plants, especially those in the families Alliiaceae and Brassicaceae.
  • the present invention relates to increasing the selenium content of plants, especially for use as food and/or nutritional supplements.
  • Se selenium
  • Se Se.
  • S Se and sulfur
  • Selenium and S ions are present in the environment in 2 " , 0, 4 + , and 6 + oxidation states [Mikkelsen et al. ( 1989) Amer. Soc. Agron.-Soil Sci. Soc. Amer. p. 65-94].
  • Selenium and S compete for the same binding sites in plants during active absorption [Bryant and Laishly (1988) Can. J. Microbiol. 34:700-703].
  • Sulfate (SO 4 ⁇ 2 ) and selenate (SeO 4 2 ) may be in direct competition for a high-affinity S permease [Shrift and Ulrich (1969) Plant Physiol. 44:893-896]. Most plants are able to utilize and substitute Se for S due to their chemical similarity [Brown and Shrift (1982) Biol. Rev. 57:59- 84].
  • Selenium can be absorbed by plants as SeO 4 "2 , selenite (SeO 3 "2 ), or organic Se complexes
  • Selenite is incorporated into various selenoether amino acids, such as Se-methylselenocysteine, selenocystathionine, and Se-methylselenomethionine [Anderson and Scarf (1983) Metals and Micronutrients: Uptake and Utilization by Plants, p. 241-275].
  • Se-amino acids replace corresponding S-amino acids and are incorporated into proteins.
  • the toxic effect of Se to plants results mainly from interference of Se with S metabolism [Mikkelsen et al. (1989) Amer. Soc. Agron.-Soil Sci. Soc. Amer. p. 65-94].
  • the "primary indicators” are plant species which accumulate from 100 to 10,000 mgSe-kg “1 dry mass and are the suspects in acute selenosis of range animals. Some “primary indicators”, such as species of Astragalus and Neptunia, show a potential Se requirement for optimal growth [Ernst (1982) convinced ⁇ kologie und mineralstoff Play p. 511-519]. High S-containing plants, the brassicas and other species of
  • Selenium is an essential trace element in mammalian nutrition, but it has not been classified as an essential plant micronutrient [Mayland et al. (1989) Amer. Soc. Agron.-Soil Sci.
  • Se as a nutrient.
  • the health benefits of supplemental Se include carcinoma suppression, immune system enhancement, and reduced cardiovascular disease [Levander (1982) Clinical, Biochemical, and Nutritional Aspects of Trace Elements p. 345-368].
  • Selenium status in a population is determined by the geochemical environment of the primary food producing area. Thus, Se status corresponds highly with the Se content of locally produced crops [Combs (1989) Nutrition and Cancer Prevention: Investigating the Role ofMicronutrients p.389-419]. In areas of the world with low Se, Se fertilization is practiced to avoid Se deficiencies in animals and humans [Gissel-Nielsen et al. (1984) Adv. Agron. 37:397-461].
  • Se can compete with sulfur (S) during nutrient uptake. Se can also substitute for S in different amino acids and proteins, and Se can replace S in the non-protein amino acids associated with onion (Allium cepa L.) Flavor. Because onions are primarily consumed for their unique flavors, and because onions have been suggested as a good Se delivery source for the human diet, the present inventors studied the effects of Se uptake on the S-compounds in the onion flavor pathway. Four onion cultivars were grown to maturity in modified nutrient solutions with or without 2.0 mg Na 2 SeO 4 added per liter. Selenium did not affect total flavor precursor content (ACSO) in three cultivars. However, Se affected several individual ACSOs and precursor intermediates.
  • ACSO total flavor precursor content
  • Flavor changes are expected when onions are grown in a high Se environment; however, specific changes may be cultivar dependent.
  • Onion (Allium cepa L.) flavors are dominated by a unique group of S-compounds derived from the precursors collectively known as S-alk(en)yl-L-cysteine sulfoxides (ACSO).
  • the flavor precursors are non-protein S amino acids.
  • the starting point of the proposed onion biosynthetic flavor pathway is the cysteine containing tripeptide glutathione [Lancaster and Boland (1990) Onions and Allied Crops. Vol. 3, p. 33-72]. Glutathione, may become a temporary storage compound for reduced S [Rennenberg (1984) Annu. Rev. Plant Physiol. 35:121-153].
  • MCSO (+)-S-methyl-L-cysteine sulfoxide
  • PCSO (+)-S-propyl- L-cysteine sulfoxide
  • PrenCSO trans-(+)-S-(l-propenyl)-L-cysteine sulfoxide
  • the vegetable Alliums were determined to be an excellent delivery system for Se that suppressed mammary cancer in mice [Ip et al. (1992) Nutr. Cancer 17(3):279-286; Ip and Lisk (1994) Carcinogenesis 15:1881-1885]. A synergistic effect of the Allium S-compounds with the Se in cancer suppression was also reported. Selenium-flavor analogues exist in onion and garlic [Cai et al. (1994) J. Agr. Food Chem. 42:2081-2084]. Moreover, the Se-enriched Alliums had minimal Se accumulation in body tissues which were normally found with the standard Se supplements, such as selenomethionine and selenite. The Se-enriched Alliums also did not alter the production and function of mammalian selenoenzyme [Ip and Lisk (1994) Carcinogenesis
  • An object of the present invention is to provide methods for the cultivation of plants of the Alliaceae and/or Brassicaceae families, especially edible plants within those families.
  • Plants of the family Alliaceae which can be cultivated according to these methods include Allium sativum (garlic), Allium cepa (onions) and chives.
  • Vegetable brassicas useful in the present invention include cabbage, cauliflower, brussel sprouts, kale, mustard, collards, turnip, rutabaga, kohlrabi, radish, and broccoli, among others.
  • Those plants are fertilized with solutions comprising selenium in the form of selenate or selenite salts and sulfur, desirably in the form of sulfate salts, where the molar ratio of selenium to sulfur is equal to or less than about 1 :2 and where the sulfate concentration in the fertilization solution or the hydroponic growth medium is from about 0J mM to about 10 mM, about 0.5 mM to about 5 mM, or desirably about 1 mM.
  • the selenium concentration is adjusted to enhance plant accumulation in a variety of forms while avoiding significant inhibition of cell and dry matter accumulation.
  • a desirable mode of cultivating plants according to the present methods is hydroponic plant cultivation because hydroponics allows close control over the selenium and sulfur contents of the plant growth medium.
  • Plants cultivated according to the methods of the present invention are enriched in selenium content, desirably containing from about 100 mg selenium compounds per kg dry weight of plant tissue to about 5000 mg selenium compounds per kg dry weight of plant material, depending on growth conditions and plant tissue type.
  • Roots and leaves are desirably ingested as raw or cooked vegetable matter or forage, or formulated as nutritional supplements in dry form.
  • the plants or dried plant material cultivated according to the methods of the present invention can be ingested by humans or animals to supplement selenium intake in the diet.
  • the present invention further provides a method for enriching tissues of species in the Alliiaceae and Brassicaceae plant families with endogenous seleno-compounds or seleno-analogs of naturally occurring sulfur compounds.
  • This method provides selenium in both organic and inorganic forms at high plant tissue concentrations. At least 50% of the available selenium is protein bound and 25%> is available in water-soluble seleno-compounds.
  • the plants are grown in either liquid or solid media and fertilized with periodic or continuous application of selenite or selenate in combination with other nutrients.
  • the ratio of SeO 4 "2 or SeO 3 "2 to SO 4 "2 is less than or equal to 1 selenium to 2 sulfur ions in the nutrient mix.
  • Harvested tissues can be used fresh, cooked or dried and made into a tablet or encapsulated.
  • the process provides a unique multi-form selenium dietary supplement.
  • the supplement is formulated from plants grown by the selenium fertilization methods of the present invention.
  • the supplement contains inorganic selenium, selenoproteins, and selenium analogs of naturally occurring sulfur compounds. It is believed that currently available selenium supplements are single form sources composed of either selenite, selenate, colloidal selenium, or yeast derived selenomethionine. Single-form supplements increase the risk of toxic exposure. While selenium is an essential mammalian micronutrient, it can be toxic if ingested in high quantities for sustained periods.
  • a multiple- form selenium supplement will lessen the risk of particular metabolites accumulating in any single form.
  • naturally occurring therapeutic compounds in the Brassicaceae and Alliiaceae can act in a synergistic manner with the endogenous seleno-compounds to augment supplement's health value.
  • the method used to grow the plants allows selenium to accumulate in much higher concentrations than found in conventionally grown plants.
  • An advantage is that the present selenium enrichment cultivation method results in a variety of selenium compounds in the plant tissues. Concentrating selenium in plant tissues is necessary if plant matter is to be used as a dietary supplement.
  • Another aspect of the present invention are nutritional supplements formulated from the plants cultivated according to the methods of the present invention.
  • Plants of the edible Alliiaceae and/or Brassicaceae comprising from about 100 to about 5000 mg/kg dry weight can be formulated for nutritional supplements.
  • These nutritional supplements can be administered to humans or animals to derive health benefits from the selenium compounds therein and/or whose diets are deficient in selenium.
  • Animals benefitting from such supplements include, without limitation, cattle, sheep, poultry and fish, for example, catfish, and other farmed fish and/or shellfish.
  • Figures 1A-1B show typical chromatograms for S-alk(en)yl-L-cysteine sulfoxides (ACSO) of 'Granex 33' onions grown with (Fig. IB) and without (Fig. 1 A) 2.0 mg Na 2 SeO 4 per liter.
  • ACSO S-alk(en)yl-L-cysteine sulfoxides
  • peaks are: 1) (+)-S-methyl-L-cysteine sulfoxide (MCSO), 2) trans-(+)-S-(l- propenyl)-L-cysteine sulfoxide (PrenCSO), 3) (+)-S-propyl-L-cysteine sulfoxide (PCSO), 4) butyl-L-cysteine sulfoxide (BCSO(S)), 5) butyl-L-cysteine sulfoxide (BCSO(R)).
  • Se uptake was positively correlated with leaf, stem, and root Se (Table 4).
  • Selenium accumulation in leaf, stem, and roots of the B. oleracea RCBP were highly correlated with leaf S and stem S.
  • Total fresh mass was negatively correlated with leaf, stem, and root Se, and leaf and stem S.
  • Total dry mass was also negatively correlated with leaf, stem, and root Se, and leaf and stem S.
  • the vegetable alliums e.g. onions and garlic, have been identified as good delivery systems for organic Se in human diets [Ip and Lisk (1994) Carcinogenesis 15:1881-1885; Ip et al. (1992) Nutr. Cancer 17(3):279-286].
  • the health benefits of Se include carcinoma suppression, immune system enhancement, and cardiovascular disease reduction [Levander (1982) Clinical, Biochemical, and Nutritional Aspects of Trace Elements p. 345-368].
  • Organic Se delivered through enriched vegetables retains the health benefits, but also reduces the risk of excessive Se accumulation in muscle and organ tissues [Ip and Lisk (1994) Carcinogenesis
  • glucosinolates have fungistatic, bacteriostatic, and anti-tumor properties and selenoglucosinolates may have nutritional and medicinal applications [Nyberg (1991) Plant Foods Human Nutr. 41 :69-88].
  • the chemical form and dose of Se consumed are the most important factors determining its biological activity [Ip et al. (1991) Cancer Res. 51 :595-600] .
  • enriched Brassica and Allium vegetables could deliver Se in a multitude of forms [Block et al. (1996) Pure Appl. Chem. 68(4):937-944], thereby reducing the risks associated with any one form.
  • Brassica species remove Se from soils, they make ideal candidates for phyroremediation of contaminated areas, and plant wastes could be used as animal feed to supplement Se in mammalian diets [Banuelos and Meek (1990) J. Environ. Qual. 19:772-777]. Animal diets need to contain 0J to 0J mg Se-kg "1 body mass [Mayland (1994) Selenium in the
  • the reproductive characteristics of the RCBPs may be used as a model for the identification of the genetic parameters of Se accumulation, and the eventual transfer of desirable traits to vegetable varieties.
  • the metabolism of Se by the brassicas holds promise for the delivery of beneficial and varied forms of Se in mammalian diets.
  • Short-day onion cultivars were chosen for testing with the selenium fertilization regimes of the present invention.
  • onions grown in the Se supplemented nutrient solutions accumulated significant levels of Se.
  • Bulb Se was not significantly different among cultivars, and averages ranged from 143.2 to 156J ⁇ g Se per g dry mass with
  • 'Granex 33' and 'Sweet Success' MCSO content significantly increased with 2.0 mg Na 2 SeO 4 per liter (7 3.05; Table 6).
  • MCSOs were 44%, 48%, 47% and 40% of the total ACSO content for 'Granex 33', 'Pegasus', 'Primavera' and 'Sweet Success', respectively.
  • MCSO content increased to 62%, 49%, 55%, and 65%o of total ACSO for 'Granex 33', 'Pegasus', 'Primavera' and 'Sweet Success', respectively. Even though S was maintained at a level which should have been sufficient for proper growth and flavor development in both nutrient solutions, the elevated MCSO content with 2.0 mg
  • Significant decreases in PrenCSO content with 2.0 mg Na 2 SeO 4 per liter were found for all of the cultivars tested (Table 6).
  • PrenCSO was 46%, 41%, 43%, and 50% of the total ACSO content for 'Granex 33', 'Pegasus', 'Primavera' and 'Sweet Success', respectively.
  • PrenCSO decreased to 30%, 27%, 36%, and 25% of the total ACSO content for 'Granex 33', 'Pegasus',
  • ⁇ GPECSO is the penultimate compound leading to PrenCSO synthesis.
  • Selenium significantly decreased ⁇ GPECSO content for 'Granex 33', 'Primavera', and 'Sweet Success' (Table 6). Decreases in ⁇ GPECSO content should cause a decrease in PrenCSO content.
  • Values for EPY and SSC were within the range previously reported for onions grown under similar Na 2 SeO 4 treatments [Kopsell and Randle (1997b) Euphytica 96:385-390]. 'Sweet Success', however, was the only cultivar in which EPY and SSC differed significantly between Na 2 SeO 4 treatments (Table 5).
  • Onions have the ability to accumulate large amounts of Se which, in turn, affects the S- based flavor precursor and precursor intermediates.
  • the effect of Se on the flavor pathway was cultivar dependent, but Se generally decreased PrenCSO and increased MCSO content.
  • Short-day onion cultivars were chosen as representative members of the Alliiaceae for testing.
  • Other members desirably of the genus Allium, including onions (A. cepa), garlic (A. sativum) and chives, can also be used in the practice of the current invention.
  • Supplementation can be in the form of vegetable consumption, or as nutritional supplements (dried powders, capsules, tablets, enteric-coated tablets or capsules, among others, extracts and/or the like).
  • nutritional supplements dried powders, capsules, tablets, enteric-coated tablets or capsules, among others, extracts and/or the like.
  • the data presented herein are for the selenium fertilized hydroponic growth of onions and broccoli, other vegetable members of the Brassicaceae, including Brassica spp.
  • crucifers such as cabbage, cauliflower, brussel sprouts, turnip, radish, rutabaga, kohlrabi, kale, collards and mustard, among others, can serve as sources of selenium supplementation for humans and animals.
  • Supplementation can be in the form of vegetable consumption, or as nutritional supplements (dried powders, tablets, extracts and/or the like).
  • “Functional foods” are prepared from the edible portions of the vegetable alliums and/or brassicas cultured according to the methods of the present invention. This selenium-enriched plant material can be consumed raw or cooked.
  • Nutritional supplements are generally formulated from dried plant material, grown using the selenium fertilization methods described herein. Where alliums such as garlic, onions or chives are grown, the leafy material can be mowed and harvested, leaving the bulbs to grow new leaves with further cultivation. Dried material can be milled and formulated into pills, capsules or enteric-coated tablets or capsules (to prevent undesirable odors or flavors apparent to the consumer). Such formulation methods are well known to the art, as are binders, fillers (if needed) and enteric coatings.
  • the plant tissues were placed in paper bags and dried in a forced-air oven at 70°C (model G01350A; Lindberg/Blue M, Asheville, NC) for no less than 36 h.
  • the dried plant material was then ground through a 0.5-mm screen (Cyclotec Sample Mill; model 1093; Tector, H ⁇ ganas, Sweden).
  • a wet acid digest was used for Se analysis. Ground tissues were placed into 125-mL flask with 10 mL of concentrated nitric acid (10% HNO 3 ) and placed on a hot plate (model 2200;
  • Tanks (2.74 x 1.22 x 0.18 m) with prefabricated black liners were each filled with 500 liters of a half-strength Hoagland's nutrient solution [Hoagland and Arnon (1950) Calif. Agr. Expt. Sta. Circ. 347].
  • the S concentration was 246.5 mg MgSO 4 JH 2 O per liter of nutrient solution (1 mM).
  • the solutions were circulated and aerated using a Teel pedestal pump (Model 3P61 IE, Dayton Electric Mfg. Co., Chicago, IL) attached to PVC piping. Solution levels were maintained in the tanks with a mechanical float attached to a deionized water source.
  • Styrofoam insulation boards were placed in the tanks to support the plants. The boards were suspended over the solutions with a PVC pipe framework. Wire fencing was placed 20J cm over the boards to support the foliage.
  • the combined wedge groups were placed into paper bags and allowed to dry at 60 °C in a forced- air drying oven (Model 630, National Appliance Co., Portland, OR) for no less than 48 h.
  • the dried tissue was then ground to pass through a 0.5 mm screen in a Cyclotec mill grinder (Model 1093, Tector, H ⁇ ganas, Sweden).
  • a wet acid digest was used.
  • One g of onion powder was placed into a 125 ml flask with 10 ml of concentrated nitric acid (70%> HNO 3 ) and set on a hot plate (Type 2200, Thermolyne, Dubuque, IA) for 4 h at 165 °C.
  • a funnel test tube was inserted into each flask, and when filled with water, acted as a condenser to reduce sample loss during digestion. The flasks were then allowed to cool to room temperature and brought up to a final volume of 50 ml with deionized water. The solutions were filtered through Whatman #1 filter paper (Maidstone, England). Total Se was measured by graphite furnace atomic absorption spectrophotometry (GFAA; Model 4100ZL, Perkin Elmer Corp., Norwalk,
  • Total S in the plant tissue samples was determined using a Leco Sulfur Determinator (Model SC-232, St. Joseph, MI).
  • Leco Sulfur Determinator Model SC-232, St. Joseph, MI
  • vanadium pentoxide accelerator Leco Corp., St. Joseph, MI
  • Total S was measured as SO 2 with an infrared cell detector and quantified against S standards.
  • Total SO 4 "2 was measured by a modification of the turbidimetric method for total S determination [Gaines and Mitchell (1979) Univ. of Georgia coastal Plain Expt. Sta. Agron. Hdbk. 1 :1-105].
  • Magnesium nitrate was not added to the onion powder prior to ashing so only tissue SO 4 "2 was detected.
  • Precursor intermediates and ACSOs were extracted using a 12 methanol : 3 water solution, and an 80% ethanol solution according to Lancaster and Kelly [( 1983) J. Sci. FoodAgr.
  • Percent recovery throughout ion exchange fractionation, derivitization, and analysis was quantified by addition S-methyl glutathione (MeGTH; at 0.5 mg-g ' fresh mass), ⁇ - L-glutamyl-L-glutamic acid ( ⁇ gG; at 0.2 mg-g "1 fresh mass), and ( ⁇ )-S-l -butyl-L-cysteine sulfoxide (BCSO; at 1.0 mg-g "1 fresh mass, as internal standards.
  • An extracted sample representing one g fresh mass of bulb tissue (15 ml of extract) was placed into a test tube, along with each internal standard, and blown to dryness using ambient air, and redissolved in one ml of deionized/distilled water.
  • TEA triethymine
  • PITC phenyhsothiocyanate
  • a Waters (Milford, MA) 2690 HPLC separator module with a 996 photodiode array detector was used for analysis.
  • a Sphri-5 RP-18 5 micron 250 x 4.6-mm column (Applied Biosystems) fitted with a 15 x 3J-mm 7 micron guard column (RP-18 Newgard; Applied Biosystems) was used for separation.
  • the column temperature was maintained at 30 ° C.
  • Eluted compounds were detected at 254 nm, and data were collected and recorded using a personal computer with Millenium chromatography manager software (version 2J 5.01 ; Waters, Milford, MA).
  • Solvents were A) aqueous acetonitrile (60%) and B) 0J4 M sodium acetate with 0.05% TEA buffered to pH 6.5 using glacial acetic acid. Solvents were filtered through a 0.45- ⁇ m Magna nylon filter (MSI, Westboro, MA). Forty ⁇ l sample volumes were injected onto the column. A flow rate of 1.0 ml-min "1 was used. The solvent gradient used was 15% A for 1 JO min, 15% to 45% A over 21 J min, 45% to 100% A over 1 min, and a hold at 100% A for 14 min. The flow was returned to the initial solvent gradient over 1 min, and the column was re- equilibrated for 12.0 min before the next sample was injected.
  • Onion pungency for each treatment combination was determined using the pyruvic acid method of Randle and Bussard [(1993) HortSci. 28(1):60].
  • Enzymatically formed pyruvic acid (EPY) is routinely used to measure an onion's gross flavor intensity [Lancaster and Boland (1990) Onions and Allied Crops. Vol. 3, p. 33-72], and it has been used to measure onion pungency in response to increasing Se fertility [Kopsell and Randle (1997a) J. Amer. Soc. Hort. Sci. 122(5):721-726]. Wedges from each eight bulb group were juiced in apneumatic press and subjected to the analysis.
  • Flavor precursors and precursor intermediates 1 (mg per gram fresh mass) of four
  • z MCSO (+)-S-methyl-L-cysteine sulfoxide
  • PrenCSO trans(+)-S-(l-propenyl)-L-
  • ⁇ GPECSO ⁇ -L-glutamyl-S-(l-propenyl)-L-cysteine sulfoxide, 2-carb

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Abstract

La présente invention concerne des procédés de culture de plantes, en particulier de plantes des familles Alliaceae et Brassicaceae, par fertilisation au sélénium, le rapport molaire entre sélénate ou sélénite et soufre (de préférence sous forme de sulfate) étant inférieur ou égal à 1/2 et le contenu en soufre représentant environ 0,1 mM à environ 5mM. Des plantes cultivées selon le système de fertilisation au sélénium décrit ici s'en trouvent améliorées et ce, par rapport à des plantes cultivées selon les techniques classiques car il se produit une accumulation importante de sélénium dans le tissu de la plante, des concentrations représentant, avantageusement, environ 100 mg/kg du poids sec du tissu de la plante à environ 5000 mg/kg du poids sec du tissu de la plante.
PCT/US1999/029454 1998-12-09 1999-12-08 Enrichissement au selenium de plantes alliaceae et brassicaceae Ceased WO2000033860A1 (fr)

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CN105523786A (zh) * 2014-10-01 2016-04-27 饶明高 一种黄豆富硒植物生长素的制备方法

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