BIOTIC N 2 Cultivating New Opportunities

1. BIOTIC N2 Cultivating New Opportunities

2. INTRODUCTION • Background Information of Biotic N2 • Who uses Biotic N2 • Crops with Biotic N2 • Mechanisms • Procedures and applications • Biotic N2 Microbes: behavior after application • -1st Mechanism of Action. • -2nd Mechanism of Action • Trials and Results (See attachments). • Benefits of Biotic N2 • Physical and Chemical properties • Stability and Reactivity • Toxicological Information • Ecological Information • Disposal Considerations • First-aid measures • Conclusions • Contacts • Note: See attachments for MSDS information and experiments with results from the product.

3. BIOTIC N2 • This is a unique product that reduces the amount of N fertilizer and increases the plant productivity. • It is a freeze dry microbial product . • Microbes know as Diazotrophs (bacteria that fix atmospheric • nitrogen). • Produced in modern and sterile fermentation facilities. • It colonizes the rhizosphere(soil zone near roots). • Microbes can be applied to the foliage or the root system. • After the application they will multiply to exit in the plant • foliage, stem and roots. • >1011 cfu/ha High concentration of microbes (Azorhizobium Caulinodans, Azoarcus Indigensand Azospirillum Brasilense) • Reliable shelf life is 12 months at cool temperature of (4 oC) storage • Containers: comes in 1, 5, 10, and100 ha packs

4. This Product is Appropriate for: • Organic farmers wanting to improved N nutrition • and better crop growth. • Biological farmers wanting a reliable microbial N • fixer and soil improver • Conventional farmers wanting to reduce N use • and increase profits. • Crops and Biotic N2 • Broad-acre /field crops: cereals, corn, cotton, wheat, barley, oats, sorgum, soybean, rice , • sugarcane, lupins, mug beans and others. • Intensive temperate and tropical pastures/forage/lucerne • Temperate fruits apples, stone fruits, blueberries, • Tropical tree fruits, avocados, bananas, citrus, coffee, tea • Macadamian, pecan, almonds • Wine and table grapes • Vegetables: tomatoes, celery, spinach, potatoes, onions, cauliflower and many others • Note: For more information in regards to trials and results please see the attached materials to this presentation.

5. MECHANISMS • Biotic N2 will improve the crop performance by using three main mechanism. • Biotic N2converts N2 from the air into a steady supply of plant available N through the entire crop season. • Biotic N2produces a large, more efficient root system due to production of Plant Growth Factors (PGFs) that gives improved capture of applied N fertilizers (N use efficiency). • Biotic N2improves soil health and structure. • Longer term use of Biotic N2 lowers soil disease pressure and builds soil carbon. • Biotic N2 microbes assist in mobilizing bound P. Nitrogenase N2 +8 H + 8e 2NH3 + H2

6. CaHPO4 +H (+) H2PO4 + Ca (2+) 2NH3 + H2 Ca5 (PO4) 3(OH) +4H (+) 3HPO4 + 5Ca (2+) + H2 O Elemental Composition of Bacterial

7. BIOTIC N2 PROCEDURE • Do not vary this rate: • Use a 1 ha pack for 1 ha, a 5 ha pack for 5ha, or a 10 ha pack for 10 ha. • Step 1: REHYDRATING THE MICROBES • Fill the plastic rehydration container with 50 or 100 ml of non-chlorinated water (shake to mix in the microbe food). • Remove the sealed lid of the vial of microbes. • Pour in some water from the rehydration container and place a thumb over the top. • Shake to dissolve the microbes and pour them back into the rehydration container (repeat this until all microbes are in the rehydration container- for 10 ha packs repeat this with the second vial). • Screw the lid on firmly and shake thoroughly for 30 seconds to 1 minute. • Wait approximately 5 minutes then shake thoroughly again. • The microbes are now ready to mix into the application tank to apply to the crop. • The biotic N2 solution can be kept in the fridge for 1 week taking good care not to let them freeze.

8. Step 2:APPLICATION TO THE CROP Application methods needs to deliver the Biotic N2 solution into the moist root zone. Contact with foliage is not a problem, but the bulk of the application should contact the root zone to maximize efficacy. Do not tank mix with fertilizers or agrochemicals. Application: Via overhead irrigation in enough water to take the microbes into the root zone. Via any fertigation system (micro-sprinkles, drip irrigation, trickle tape). Via liquid injection from a tine that cuts into the root zone (water supply 100l/ha). Via water run in furrow irrigation (flood irrigation). Via boom spray onto moist soil. Use very coarse or streaming nozzles and as much water as is practical. Note: Calculate the amount of water to be delivered to the crop area and mix in the correct amount of biotic N2 microbes solution after shaking it to suspend the microbes. Apply the tank missed microbes within 24 hours and stir to re-suspend the microbes in the tank if necessary. Chlorine water can be de-chlorinated by adding 2g of sodium thiosulfate (STS) per 100L. Pre –dissolve in a small volume of water and mix into the tank water thoroughly wait approximately 5 minutes before adding biotic N2 .

9. Biotic N2 Run in-furrow irrigation Any Fertigation system (Micro-sprinkle, drip irrigation, trickle tape etc.) Boom spray onto moist soil Liquid Injection from a tine that cuts into the root zone. Overhead irrigation

10. Leaves & branches ENDOPHYTES Biotic N2 Microbes: Behavior after the Application. • When the Biotic N2 is applied via soil application they • colonize the rhizosphere. • They also move up into the plant tissue and end up throughout • the plant. • Azospirillum sp on rice • Root. • Biotic N2 microbes get carbohydrates and nutrients as root • exudates and from plant tissue. In return they provide N2, increase • root efficiency and root / soil health- a true symbiosis. X Y L EM Twin N microbes colonize roots Root exudates

11. 1st Mechanism of Action • It fixes the atmospheric nitrogen into nitrogenous compounds that are immediately available to the plants. • NH3 is supplied steadily right through the season. • This helps keep total leaf N at optimum levels without high nitrates which reduces fruit • quality and shelf life. N2 From the atmosphere N Fixing Bacteria Nitrogenase enzyme N2+ 8H++ 8e- 2NH3+H2 Nitrogenfor Crop

12. 2ndMechanism of Action • Biotic N2produces large root systems due to • auxin synthesis (IAA ). • Greatly increased root hair density gives better • nutrients capture of all nutrients. • Improved capture of mineralized and applied N • increasing nitrogen use efficiently. • More vigorous root growth help crops fight back • from root damage from pest and diseases. NOTE : Picture is from a DPI- measured comparison and root mass rating increased >13% in biotic N2 plots . This effect is mainly from increased root hairs increasing the root ball. Improved N nutrients is due to a combination of N fixation plus better N use efficiency via the effect of biotic N2 on roots.

13. Trials and Results a) No Biotic N2 b) 2 months after the application of Biotic N2 Information: Organic Avocado Location : Australia Results: As you can see on the picture to the right there is a greatly increase in root density. No Biotic N2 b) 120 days after applying Biotic N2 Information: Organic Cherry Location: Chile Results: the application in here was via fertigation, as we can see roots showed much greater secondary root development.

14. No Biotic N2 and N deficiency is evident b)21 days after applying Biotic N2 Information: Organic Blueberries Location: Chile Results: 21 days after applying Biotic N2 we can see that we can obtain a healthier leaf color.

15. Benefits of Biotic N2 • Biotic N2 can reduce populations of some pathogenic microbes. • Biotic N2encourages growth of beneficial microbes that help keep soil pathogens in check. • Biotic N2increases root nodulation in legumes. • Biotic N2enables reduces N application rates. • Lowers impact of urea on soil organic carbon. • Avoids problems with soil pH. • Lowers impact of synthetic N on soil structure. • Biotic N2increases nutrient availability. • Biotic N2 microbes release organic acids improving in this way the availability of P (P solubilization). • Increases the availability of some micronutrients in soils. • Improves profitability • Increase the yields (when the crops values are high). • It will decrease input cost from N fertilizers (specially when the N prices are high and the crops prices are low).

16. Benefits of Biotic N2 • Increase sustainability of production. • Biotic N2improves the soil and productivity and crop vigor. • Decrease synthetic nitrogen effects on the environment • Decrease carbon foot print of production (e.g bio-ethanol). • Enable compliance with various legislation to limit Nitrogen Fertilizers applications- without the normal yield penalties from N application.

17. Physical and Chemical Properties: • VAPOR DENSITY: No data • EVAPORATION RATE (n-BuAc=1): No data • SPECIFIC GRAVITY @20o C: No Data • SOLUBILITY IN WATER: Completely soluble • VAPOR PRESSURE, mm Hg @20o C (68oF): No Data • pH:6-7 • APPEARANCE, ODOR and COLOR: This product is a light brown/grey powder with a very slight odor. Stability and Reactivity • STABILITY: Stable • DECOMPOSITION PRODUCTS: thermal degradation produces carbon dioxide, carbon monoxide and ammonia gases. • MATERIAL WITH WHICH SUBSTANCE IS INCOMPATIBLE: None known • POSSIBILITY OF HAZARDOUS REACTIONS: Will not occur • CONDITIONS TO AVOID: None

18. Toxicological Information • TOXICITY DATA: • CAS: No Data • SUSPECTED CANCER AGENT: The following components of this products are listed by agencies tracking the carcinogenic potential of chemical compounds as follows: None known • IRITANCY OF PRODUCT: Contact of this product can cause slight skin and eye irritation. • SENSITIZATION TO THE PRODUCT: These products are not known to cause human skin or respiratory sensitization. • REPRODUCTIVE TOXICITY INFORMATION: Listed below is information concerning the effects of this product and its components on the human reproductive system. • Mutagenicity: The components of this product are not reported to produce mutagenic effects in humans. • Embryotoxicity: The components are not reported to produce embryotoxic effects in humans. • Teratogenicity: The components of this product are not reported to produce teratogenicity effects in humans. • Reproductive Toxicity: The components of this product are not reported to produce reproductive effects in humans.

19. Ecological Information • ALL WORK PRACTICES MUST BE AIMED AT ELIMINATING ENVIRONMENTALS CONTAMINATION. • ENVIRONMENTAL FATE: This is not expected to be hazardous to the environment. • ENVIRONMENTAL TOXICITY: This material is not expected to be toxic to aquatic life. Disposal Considerations Beneficios de Biotic N2 • PREPARING WASTE FOR DISPOSAL: Waste disposal must be in accordance with appropriate U.S. federal, State and local regulations, those of Canada, Australia, EU member States and Japan. • Note: Please review the Material Safety Data Sheet (MSDS)for further information.

20. First-Aid Measurements • INHALATION FIRST AID: Not expected as a result of this product, however if breathing problems occur remove victim to fresh air. If persistent breathing occurs, get medical attention. • SKIN CONTACT FIRST AID: Wash skin with plenty of soap and water after handling or contact. Wash clothing before reuse. Get medical attention if irritation persist. • EYE CONTACT FIRST AID: If contact with eyes, flush eyes with plenty of water for at least 15 minutes lifting upper and lower eyelids occasionally .Get medical attention if irritation persist. • INGESTION: Routine uses of this product is not expected to cause any situation which could lead to indigestion. If this product is swallowed, CALL PHYSICIAN OF POISON CONTROL CENTER FOR MOST CURRENT INFORMATION. • If professional advice is not available, do not induce vomiting. Never induce vomiting or give diluents (milk or water) to someone who is unconscious, having convulsions, or unable to swallow. • MEDICAL CONDITIONS AGGRAVATED BY EXPOSURE: None known • RECOMMENDATIONS TO PHYSICIANS: treat symptoms and eliminate overexposure.

21. Conclusions

22. Yields • Basically biotic fertility mimics natural fertility; however , it provides an interrupted • nutrient supplied to crops. • Since the Biotic N2improves the nutrients absorbance to the roots the crops grow up • faster and healthier than any other fertility. • Even though UREA has also an excellent yield the cost and extra care that has to be • provided to the crops affect in the cost of the production. • Quality • Biotic fertility specially Biotic N2has shown in the past decade a higher over –all • quality than Crops grown with other fertilizers like UREA and many others. • If too much N2is provided to crops this will affect the quality of the product and dry • or even kill the plants. Biotic N2 provides the right amount of nutrients needed by the plants. • Cost • UREA fertilizer is costly because you have to buy additional pesticides, fungicides and • herbicides that causes a large atmospheric impact leading to the pollution and • erosion of the soil. • In the future more money will have to be invested in order to restore the soil for future crops.

23. Sustainable fertility • Biotic fertility constantly amends and re-mineralizes depleted soils. • Grower no longer worries about the exhaustion of soils (Now and the future). • As a result, the soil become healthier , therefore, the products become richer in nutrients. • *Eventually growers are able to back off the amount applied after their soils are • restored to a level of fertility (Remember : in conventional fertility they deplete • the organic matter in the soil. • Surface erosion • Biotic fertility does not upset topsoils and the C:N balance in the soil. • When this in-balance occur the loss of vital soil acids gels which act to hold the • surface of the soil in place resulting in erosion.

24. Environmental Impact • Biotic fertility will greatly reduce the amount of green house gasses volatilized into the atmosphere than any other method. • Many companies around the world are attracted to this fairly new trend of becoming more eco-friendly, now it is the turn in the agricultural field. • Our soils are very poor in nutrients and everyday they are dramatically depleting. We must take actions in this matter and Biotic N2 is a great source.

25. Contacts Raúl Muñoz Authorized Distributor Phone :9933-7597 (Chile) (201) 942-0400 (EU) Address : Avenida Pocuro 2521 Providencia Santiago, Chile E-mail: RHMunoz@purelyOrganicSA.com Gladys Michelle Reyes Biochemist Phone: (201) 963-1028 Ext:104 (EU) E-mail :gmichellereyes@gmail.com