Patent Application
20080072494
The micronutrient of the present invention is an all-natural plant amendment derived from chitin/chitosan and is 100% water soluble, whereas chitin/chitosan is not water soluble. Chitin/chitosan occurs naturally in a range from 100% chitin to 100% chitosan as a mixed polymer. By contrast, an NMR analysis of the micronutrient of the present invention revealed characteristics of approximately 20% chitin and approximately 80% chitosan. Below is data showing that the micronutrient of the present invention outperforms chitin/chitosan as an elicitor of self-protecting enzymes.
One of the classical responses to elicitation of plants is induction of certain enzyme activities. These may
Enzyme activity measurements relate to the level of a given enzyme protein in the plant tissue. As an example of enhanced enzyme activity, β-1,3-glucanase was measured. The enzyme, β-1,3-glucanase, was assayed using laminarin (a soluble β-1,3-glucan) as substrate. Crude homogenates of the seedlings from treated seeds yielded the data in FIG. 1. Increased β-1,3-glucanase activity compared to controls (without seed treatment) was obtained in the micronutrient of the present invention treatments ten days following germination. Elicitation of mung beans seeds that were treated with the micronutrient of the present invention were compared to those treated with two types of elicitors. First, various concentrations of purified colloidal chitin/chitosan were used. The dose response to chitin/chitosan concentrations of 9, 0.9 and 0.09 mg/seed followed no regular pattern. A nearly equivalent concentration of the micronutrient of the present invention (1 mg/seed) elicited five times as much β-1,3-glucanase enzyme activity. Secondly, lower concentrations of the chitin oligosaccharide containing six glycan moieties, N-acetylchitohexaose were studied. The importance of the chitin oligosaccharide is that short chains of chitin have been found optimal in elicitation of many types of plants. The dose response relationship to the oligosaccharide concentrations of 0.5, 0.05 and 0.005 mg/seed is negative; i.e. higher doses resulted in lower specific enzyme activities. Comparisons similar to those with chitin/chitosan could be made between the performance of 1 mg/seed micronutrient of the present invention and lower concentrations of the more optimal oligosaccharide.
A dose response for the micronutrient of the present invention in induction of elevated β-1,3-glucanase activity in adzuki beans is demonstrated by data in FIG. 2. Induction of this enzymatic activity increases with quantity of micronutrient of the present invention applied to the seeds. Comparison of elicitation between treatments with 0, 0.5, 1.0 and 2.0 mg/seed and controls in specific enzyme activity was evaluated in both hypocotyl and epicotyl tissues.
The specific enzyme activities in both tissues increased with dosage 21 days after germination. The differences become significant in root tissue using 2.0 mg/seed with twice the level of activity, compared to controls.
Further analysis of the micronutrient of the present invention revealed that the if using the micronutrient of the present invention for irrigation treatment, application on the order of 1-10 mg micronutrient per gallon of water is a suitable concentration and use of about one pint of this mixture per acre is sufficient to protect most crops. The same concentration of about 1-10 mg micronutrient per gallon of water is a suitable concentration for foliar treatment as well as a seed dip. Use of the micronutrient of the present invention as an irrigation or foliar treatment provides contact of the micronutrient with receptors on the plant cell surface, which initiates signal transduction pathways and enhances vigor of seedlings. These processes lead to earlier and more robust root systems, earlier and more robust foliage, which provide more development in a growing period and produce greater crop yields.
The signal transduction brought about by contact of the micronutrient of the present invention with cell surface receptors on a plant further enhance growth and crop yield by inducing the plant to generate protective enzymes and phytoalexins for resistance to bacteria, fungi, entomologic attack, other pathogens and suppression of parasitic nematodes.
The signal transduction brought about by contact of the micronutrient of the present invention with cell surface receptors on a plant further enhance growth and crop yield by allowing the plant to stimulate production of chemical engines in the golgi bodies and mitochondria which enhance the plant's ability to withstand and overcome environmental stress such as mineral imbalances, hail, drought, wind and pathogenic and entomologic stresses.
The signal transduction brought about by contact of the micronutrient of the present invention with cell surface receptors on a plant further enhance growth and crop yield by increasing the effective growing period by delaying senescence by suppressing the action of ethylene, thereby allowing more complete crop development before harvest.
Use of the micronutrient of the present invention as a seed treatment enhances seed germination by increasing the rate of germination as well as the proportion of seeds germinating by increasing enzyme activity, such as alpha-amylase, for example, which degrades polymers in the seed coat. The site of this enzyme activity resides in the aleurone cells, which reside beneath the seed coat.
Additionally, the present invention does not demonstrate a negative physiological impact on field crops. Crops are not hurt by the elicitation or suffer physiological damage or impairment of growth. Only positive results have been observed. Thus the switch in this manner behaves in a positive manner.
Furthermore there is a positive effect on diseases also. Parasitic nematodes did not increase number but decreased in number (Becker, 2005). Thus the switch in this manner is negative as regards disease pressure.
The following discussion describes how the invention works to elicit various chemical engines within field crops. Pathogen associated molecular patterns for nematodes are defined as either good( i.e., beneficial nematode) or bad (i.e., parasitic nematodes).
The signal transduction inner-polarity as elicited by the invention, is a polarity switch or sets of switches comprised to activate the chemical engine(s) within the plant. Some of these may be of the same pole to activate specific enzyme pathways for resisting disease and pathogens but able to not harm good nematodes. The chemical switch in the present invention is of opposite poles; one positive pole activation for resisting infection (Linden, 1998) and a negative pole for not resisting beneficial nematodes (Becker, 2005).
The inner-pole is not limited to opposite elicitation features. The signal transduction inner-pole activations include combinations of polarities. Thus the switch's poles may both be “positive”, eliciting beneficial chemical engines resulting in physiological improvement (increased biomass and yields, higher quality, increased shelf life, etc). We demonstrated under controlled greenhouse conditions the soybean seed counts increased 78% and seed weight increased of 81% over control with seed and irrigation treatments (Linden, 2006).
The switch poles may both be “negative” for elicitation of chemical engines, when the physiological aspects that ward off insects from leaves and stems as well as resisting rhizoctonia disease infection (CSU Potato Field Trials 1995).
Signal transduction inner-polarity can be elicited where there is a combination of multiplexed switches of various poles, each acting independently of one another. In cell biology these concepts are referred to as upregulation, downregulation and signalling crosstalk. Chemical engines result in a wide range of physiological enhancements as well as defending, resisting and overcoming disease and pathogen pressures.
Repeated application of the invention can cause cascading signal transduction inner-polarity activations for greater power of the chemical engines resulting in significant yield increases between multiple applications (Linden, 2006).
The micronutrient of the present invention is applied at different rates depending upon seed size (grams per seed) as shown below. The smaller the seed the less is required for the signal transduction response.
Results for Potato May 5, 2006
Results for Soybean May 30, 2006
It is important to note that amounts of the micronutrient of the present invention required to elicit the signal transduction response range from 0.003 to 0.01 ml per gram of seed (depending upon seed size, i.e., seed potatoes weigh more than soybean seeds).
Below is supporting data on a wide range of field crops.
Soybean
Central Illinois Agricultural Research Farms, Inc., 1229 W. Edwards, Springfield, Ill. 62704-1634, 800-497-1525 conducted the following experiment. This experiment was conducted at the Henry White Experimental Farm, Field 4, Soybeans, treated and control, Sep. 1, 2005, Lab. No. 25109 and 25106, composite samples from four replications.
Comments: The most limiting nutrient is Iron (Fe). 8 ratios out of 40 are good. The average deviation is 129 for the treated soybeans and 125 for the control. The deviation is high and indicates that several nutrients are out-of-balance and/or this is a disease scenario. The Becker Nematode Index (BNI) is 83 and 103 respectively. The higher BNI in the control suggests that there are more nematode problems in those strips. Nematode assays were conducted after harvest.
Oct. 11, 2005
Average soybean counts per foot of row in the treated strips=920
Average soybean counts per foot of row in the control strips=776
Nov. 16, 2005
Results of Nematode Assays from the Henry White Experimental Farm Field 4
Nematode Counts, Total and Parasitic of 100 ml of soil. The treatment is 1 pint of the micronutrient of the present invention per acre and there were 4 replications in a paired comparison design.
The strips treated with the micronutrient of the present invention averaged 11.0% parasitic nematodes while the control strips averaged 16.7% parasitic nematodes. The two most common parasitic nematodes were lance and lesion. Yield losses can be expected when parasitic levels are higher than 10%.
Soil profile examinations showed compaction problems between 3 and 12 inches deep. Root development was restricted and yields were affected. Control strips averaged 52.2 bushels per acre and the treated strips averaged 53.4 bushels per acre.
A review of the above data shows that the plant signal transduction defense response induced by the micronutrient of the present invention destroys harmful parasitic nematodes. The micronutrient of the present invention has no negative impact on beneficial nematodes and other beneficial micro-organisms.
Soybean
Set forth below is greenhouse data on soybean yields conducted at Colorado State University. This data shows a combination of the micronutrient elicitor of the present invention seed and a foliar treatment had 41% increase in yield. Also the yield seeds weighed more than the control seeds by as much as 49%. Treatment #1 is control. Treatment #2 is untreated seed with two irrigated applications. Treatment #3 is treated seed. Treatment #4 is treated seed with two irrigated applications.
Field data results for other crops. note—peanuts had 56% increase in yield).
Tomatoes
A comparison of poor soil conditions for tomatoes found that treatment with the micronutrient of the present invention yielded a 37% increase over control in poorer fields where soil and environmental conditions reduce output. In higher quality fields, where soil and environmental conditions produce higher output, treatment with the micronutrient of the present invention yielded a 24% increase over control. (Project 030410A)
Potato
Potato yields from fungus infected soils from greenhouse and fields in Mexico: In normal soil plants treated with the micronutrient of the present invention had a 27.84% increase in daughter tuber yields over the control group. Treated plants grown in infected soil had a 35.37% increase in daughter tuber yield over the control group.
Control group: applied chemicals/pesticide per manufacturer's recommendations.
Treated group was treated with 1 liter micronutrient of the present invention/1000 liters of water/hectare.
Set forth below are the results of an experiment on the fields of Sr. Ernesto Ortegon Cervera. The crop planted was potato, date of burning of the field was Nov. 27, 2001, date of sowing was Nov. 27, 2001, and the date of harvest was Apr. 4, 2002. The fields were irrigated by rolling irrigators and the fertilizer used was “Propia.” Ortegon is comprised of 0.5 parts Agrimicin, 1.0 part Confidor, 8.0 parts Pentaclor, 5.0 parts Temir and 0.6 parts Tecto 60. The cost of application of Ortegon was $345.68 per hectare while the cost of application of the micronutrient of the present invention was $175.03 per hectare. Units are in tons per hectare.
Set forth below are the results of an experiment on the fields of Sr. Salvador Zazueta (Chava). The crop planted was 135 day Snowden (peas), date of burning of the fields was Apr. 8, 2001. date of sowing was Nov. 22, 2001, and the date of harvest was Apr. 18, 2002. The fields were irrigated by aspersion and the fertilizer used was “Propia. ”Sr. Zazueta applied material to his crops which comrpised 1.5 parts Fuvadan 350, 10.0 parts Captan, 5.0 parts Vitamin, 10.0 parts Carbovit, 0.15 parts giberelic acid and 0.8 parts Tecto 60. The cost of application of this mixture was on the order of $265 per hectare while the cost of application of the micronutrient of the present invention was $175.03 per hectare. Units are in tons per hectare.
Set forth below are the results of an experiment on the fields of Sr. Enrique Free Pacheco. The crop planted was potato, date of burning of the fields was Mar. 7, 2002, date of sowing was Nov. 22, 2001, and the date of harvest was Apr. 4, 2002. The fields were irrigated by aspersion and the fertilizer used was “Propia.” Sr. Pacheco applied material to his crops which comrpised 2.5 parts Manzate 200, 3.8 parts Cercobin M, 0.75 parts Coprimicin, 19.0 parts Pcnb 80 and 1.75 parts Nuvacron. The cost of application of this mixture was $315.05 per hectare while the cost of application of the micronutrient of the present invention was $175.03 per hectare. Units are in tons per hectare.
It is also seen in citrus where the presence of the micro-nutrient of the present invention decrease ethylene production and increased sugar content. The micronutrient of the present invention can also increase shelf life of citrus. Application of 16 oz per acre of the micronutrient of the present invention to the crops, citrus resulted in 10% reduction in citrus decay in packing house resulting in 32% increase in juice grade yields after 5 days of storage.
With respect to the above description, it is to be realized that the optimum relationships for the components of the invention, to include variations in composition, proportion and manner of use, are deemed readily apparent and obvious to one skilled in the art, and all equivalent relationships to those described in the specification are intended to be encompassed by the present invention.
Therefore, the foregoing is considered as illustrative only of the principles of the invention. Further, since numerous modifications and changes will readily occur to those skilled in the art, it is not desired to limit the invention to the exact composition and operation shown and described, and accordingly, all suitable modifications and equivalents may be resorted to, falling within the scope of the invention.
