Symborg NATURAL GROWTH Index

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Introduction of Symborg.

.': Reasons for the introduction of G. i. tenuihypharum in NZ.

A, Characteristics of G. i. tenuihypharum. ;% Mode of Action (MoA) G. i. tenuihypharum. _'; Benefits and adverse effects of introducing G. i. tenuihypharum in NZ.

.': Major concerns of introducing G. i. tenuihypharum in NZ.

What is Symborg?

J4., Symborg is a biotechnological company focused on the development of innovative agricultural technologies to help the farmers to maximise crop yield in a sustainable way. For Symborg, guarantee a sustainable future and improve the quality is a key point on the development of new products.

J",, It works everyday to promote the research and development of mycorrhizal technologies at international level, as member of the International Society (IMS). It is also part of the Biological Product Industry Alliance (BPIA) and International Biocontrol Manufactures Association (IBMA). It works actively to promote the concept of biological control in the sustainable agriculture systems by increasing the awareness about biopesticides as capable products and participating in the improvements in the regulation processes at international level.

J"% Symborg works with the exclusive and 100 % patented arbuscular mycorrhizal , Glomus iranicum var. tenuihypharum and possesses four different product formulations to help farmer in different agricultural systems.

Irrigation Seed coating Microgranules

Re% Resid. MG HC

MycoUp MycoUp Activ Resid HC Resid MG

Biological lnoculant Biological lnoculant Biological lnoculant Biological Inoculant Horticultural and woody Cereals and grains

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Symborg's distribution in the world

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7 subsidiaries: Spain, France, Turkey, China, USA, Mexico and Chile. Presence in more than 40 countries.

www.symborg.com Reasons for the introduction in NZ

.~% According to Stats NZ (www.stats.govt.nz):

`So' The agricultural land dedicated for crops, grain, nursery, vegetables and fruits has increased between 2012-2017, accounting with 617 000 ha. 181 Cereal crops are the crops with more area dedicated in New Zealand. The irrigable/irrigated land using spray and micro systems increased 25.5 %. 1$1 The irrigable/irrigated land using flood systems decreased 58.4 % in the same period.

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Glonins-based products are ideal for drip irrigation, seed coating and microgranules application systems

www.symborg.com Reasons for the introduction in NZ

.'% Symborg is growing and expanding to new markets, focusing on countries with strong intensive agricultural systems;

Our Glomus-products are designed to be applied exclusively in agriculture to: 4l' Maximize the crop yields and reduce the expenses in water and chemical fertilizers;

10' Allow the plants to resist better to abiotic stress (high pH and salinity, drought, etc.);

13' Improve soil's health and characteristics (increase microbial population, soil structure, etc.); `il' Contribute for a sustainable future and reduce the effects of chemical fertilizers in the environment;

www.symborg.com Glomus spp. distribution in the world

The Glomus spp. is a millennial microorganism (450 million years), naturally found in the soil; The Glomus spp. studies increased during the last decades; Most of the studies are located in Europe, USA and western Asia; J'4. Davidson et al., 2015 indicated that AMF endemism is low and can be found in all continents; .~!. Glomus iranicum was also found in Iran; .~% In Blaszkowski et al., 2010, the author mentioned that most of the undescribed AMF belong to Glomus spp., maybe due to: 181 a lack of or rare sampling of AMF in many terrestrial regions of Earth; `R, the few specialized and experienced

I I I I I I I I I I I I I mycologists that study of 18OW 15OW 120W 90W 60W 30W 0 30E 60E 90E 120E 150E 180E ; Source: Opik et al., 2013. `R, or to seasonal, rare or lack of sporulation by many AMF in the field.

www.symborg.com Characteristics Glomus iranicum var. tenuihypharum is an arbuscular mycorrhizal fungi (AMF);

.~% Creates symbiotic relationships (mycorrhizae) with plant ;

.~% It is an obligate symbiont, depending on their mycorrhizal association with plant roots to complete their life cycle;

Propagation is done by contact of the spores, the vesicles or the mycelium with the roots of a new host plant;

Cannot be cultured in the laboratory in the absence of a plant host;

The dissemination of arbuscular mycorrhizal fungus is low since they are lacking aerial spores;

.'~. Dissemination in the environment is therefore limited to the zone it is applied;

This strain is resistant to with high salinity (pH 9. 5);

.': It is not a genetically modified organism;

www.symborg.com MoA The G. i. var. tenuihypharum forms a symbiotic relationships that allows the plant to access through the extraradicular hyphae of the fungus to nutrients in exchange of C-based compounds from photosynthesis, and water. It secrete phosphatase to hydrolyse phosphate from organic P compounds and thus improve the crop productivity under P-deficient conditions. The mycorrhizal hyphae also increase the uptake of ammonium, immobile micronutrients (such as Cu and Zn), and other soil-derived mineral cations (K+, Ca'-+, Mg'-+, and Fe'+).

These are able to explore a great volume of soil, 1000 times higher than roots. The water absorption also directly improves due to a function of the mycorrhiza on the stomata's mechanisms.

The hyphae access the most intrinsic parts of the soil and the mycorrhizal formation increases considerably the ramification of the radicular system, making a better use of this resource. Allows the penetration in the soil of the CO, fixed by vegetables and the aggregation of fine particles by the release of (a glycoprotein in the rhizosphere), stabilising the soil's structure and promoting it's quality.

Also, provides resistance to abiotic stress by increasing the tolerance of the vegetables to heavy metals and salts.

www.symborg.com qF11l AN) i e MYCORRHIZAL SOIL SYMBIOTIC FUNGUS

• '.~Ilti Utv Lipids r • tru-i— lI Trehalose Sut 10 W • CARBON Nexose A Ammonium

IS Nitrate

NrttatB/Ammonium E Amino acids ♦ Urea

F Phosphate * Phosphates PHOSPHORUS Polyphosphate

1

4 .'% The fungus receives carbohydrates (sugars) and growth factors from the plant; The plants receives N- and P-based nutrients, improving growth and productivity; Benefits of symbiotic relationship .~% Extramatricial mycelium increases the roots systems; Tolerance to soils with inadequate plant growth conditions (high pH, heavy metals, etc....) increases; .': Reduces the drought stress, water and fertiliser needs;

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Benefits Adverse effects

.~% Environmental: .~: Environment: 1$1 Improve plant growth and performance; `k," Possible displacement of native AMF; 1$1 Resistance to abiotic stress; ti3' Possible association with invasive plant ; Environmental sustainability of farms; Erosion control; 18,Boost soil ecosystems through improved nutrient uptake by organisms; 11 Minor and could potentially be medium in effect.

.~% Economic: Economic: %$1 Long term benefits; No adverse effects were found. 181 Reduced farming input costs associated with fertilisers and water; `W Greater productivity through crop yield; `0, Shorter period of plant maturation for harvesting; `8, Estimated net benefit of approx. $114 M per year for maize silage and maize grain; 19,Minor at national scale but moderate on a local to regional scale.

www.symborg.com 11 Major Cause any significant displacement of any native species within its natural habitat concerns `S~' For a significant effect to occur to a native species in its natural habitat, G. i. var. tenuihypharum spores would need to disperse, germinate, and form a relationship with the roots of a plant before it could cause displacement of native AMF or other microorganisms living in the rhizosphere as it is an obligate symbiont incapable of living without a host. Further, multiple new relationships with plant hosts would need to be established in order to generate more than localized effects. Also, competence between species do not guarantee the full dominance of our AMF over the native species. Cause any significant deterioration of natural habitats Significant deterioration of natural habitats is considered unlikely as the intended application areas will be agricultural land and glasshouses. It must be considered that these areas are usually deteriorated, and microbial populations tends to be low. Furthermore, due to the type of application of the Glomus-based products, the affected area is minimal. There are also several abiotic and biotic factors that contribute to a species becoming invasive which further reduces the probability of significant deteriorations to natural habitats occurring from the use of this organism in agricultural soils. Cause significant adverse effects on human health and safety Glomus iranicum var. tenuihypharum has been used in research and applied to agricultural plants to boost their growth and development for the purposes of consumption. There are no known mechanisms of interaction between humans and G. i. var. tenuihypharum and there are no adverse effects to human health and safety. According to OCDE (1992) report on safety of microorganisms used as biofertilizers states that mycorrhizal fungi are not pathogenic and do not pose any evidential threat to animals or human beings.

www.symborg.com 12 .'; Cause significant effects on New Zealand's inherent genetic diversity Major It is difficult to quantify if the catalogued species of Glomus fungi in the New Zealand Fungal Database are concerns native or introduced. The scientific literature indicates that these Glomus species have been isolated in multiple continents spanning similar environments to New Zealand and over multiple timeframes which suggests a ubiquitous, global distribution. Furthermore, a literature research using the keywords ((TS = glomus) OR (TS = dominikia)) AND TS = Hybrid* was carried out, without any further restrictions, and did not retrieved any relevant information concerning that our Glomus spp. could hybridize with other arbuscular mycorrhizal fungi. .~% Cause disease, be parasitic, or become a vector for human, animal or plant disease The fungus is intended to improve agricultural production. There is no evidence to suggest or support Glomus iranicum var. tenuihypharum causing disease, being parasitic or for being a vector for human, animal or plant disease. .~% Establish undesirable self-sustaining populations The purpose of the application is to release a new organism that will establish and build self-sustaining populations. We expect that any population of G. i. var. tenuihypharum would not become undesirable on a national or regional scale as there is no evidence to support this organism being undesirable overseas. However, any indirect adverse effects on ecosystem processes and foodwebs would not have significant or permanent effects. Any effects are expected to take place within the vicinity of application. `S,' In any case, eradication could be feasible removing plants and applying hazardous substances such as fumigants to soils but that would not be a desirable course of action.

www.symborg.com 13

Efficiency of Glomus iranicum var. tenuihypharum

Continent Crop family Substrate Application System Product Maximum yield achieved Soil Irrigation MycoUp, MycoUp Activ 220/ Vegetables Hydropic Irrigation MycoUp, MycoUp Activ 33% Fruit trees Soil Irrigation MycoUp, MycoUp Activ 28% Soil Irrigation MycoUp, MycoUp Activ 7% Berries Hydropic Irrigation MycoUp, MycoUp Activ 40% Europe Cereals Soil Planting time Resid MG 27% Cereals Soil Seed treatment Resid HC 15% Corn Soil Planting time Resid MG 17% In seven years, the company has demonstrated constant and Cora Soil Seed treatment Resid HC 18% effective results, helping growers from all parts of the world Citrus Soil Irrigation MycoUp 19% Soil Irrigation MycoUp, MycoUp Activ 40% Vegetables to maximize their crop yields, increasing production in Hydropic Irrigation MycoUp, MycoUp Activ 27% almost all crop families. Fruit trees Soil Irrigation MycoUp, MycoUp Activ 23% Soil Irrigation MycoUp, MycoUp Activ 32% Berries America Hydropic Irrigation MycoUp, MycoUp Activ 21% The efficacy of the company's products has therefore been Cereals Soil Planting time Resid MG 16% shown in most major agricultural areas, climates, soils and Cereals Soil Seed treatment Resid HC 14% Com Soil Planting time Resid MG 19% agricultural systems, ranging from intensive to extensive. Com Soil Seed treatment Resid HC 16% Vegetables Soil Irrigation MycoUp, MycoUp Activ 22 % Soil Irrigation MycoUp, MycoUp Activ 9% Berries Hydropic Irrigation MycoUp, MycoUp Activ 33% Africa Cereals Soil Planting time Resid MG 13% Cereals Soil Seed treatment Resid HC 11% Com Soil Seed treatment Resid HC 18% Vegetables Soil Irrigation MycoUp 22% Vegetables Hydroponic Irrigation MycoUp 17% Citrus Soil Irrigation MycoUp 19% Eurasia Soil Irrigation MycoUp 19% Vegetables Hydroponic Irrigation MycoUp 34% Source: Femdndez et al., 2018.

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Effects in Date of Search: 17.10.2017 Human health Publication dates: 2007-2017 Keywords: TS="glomus iranicum" OR TS="glomus" AND TS="human*" AND (TS=adverse OR TS=allergic OR TS=sensitisation OR TS="infection*" OR TS="communicable disease*" OR TS="communicable" AND TS="disease*") NOT (TS="tumo*" OR TS="carotid")

The bibliographic research retrieved 65 publications and all of them were discharged after analysis. The articles mentioning the AMF were related to the beneficial effects caused on plants growth or with their ability to improve crop production in degraded/contaminated soils with heavy metals or other toxic elements. Also, great number of articles were not related with the AMF, being publications referring to the medical term "glomus tumor" (case studies and methodology of treatment and removal) even after the search being restricted for the medical terms "tumor" and "carotid". It was not found any article reporting toxicological effects on humans.

All the bibliographic research was done following the EFSA guide for Submission of scientific peer-reviewed open literature for the approval of pesticide active substances under Regulation (EC) No 1107/2009.

www.symborg.com 17 Fate and Date of Search: 19.10.2017 behaviour on Publication dates: 2007-2017 non-target Keywords: TS="glomus iranicum" AND (TS="fate*" OR (TS=behaviour)) NOT (TS=carotid OR TS=tumo*) OR organisms TS="glomus iranicum" AND TS=mobility OR TS="glomus iranicum" AND TS=soil AND (TS=motil* OR TS=concentration OR TS=population density) NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND TS=air OR TS="glomus iranicum" AND TS=water OR TS="glomus iranicum" AND (TS=lake OR TS=water)

From the bibliographic research, 6 publications were found but none of them referred to the persistence of Glomus ira.nicuni in the environment, being all the articles discharged after the analysis. The articles found were related to studies of the use of AMF in agriculture to improve crop production.

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Effects in the Date of Search: 18.10.2017 environment Publication dates: 2007-2017 Keywords: TS="glomus iranicum" AND (TS="bird*") NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND (TS="fish*" OR TS=daphid* OR TS=daphnia) NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND (TS="alga*" AND (TS=side effect* OR TS= adverse effect* OR TS= tox*)) OR TS="glomus iranicum" AND (TS="aquatic" AND TS=plant AND (TS=tox* OR TS=side effect*)) NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND (TS="bee") NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND (TS="arthropod*") AND ( TS=tox* OR TS=adverse effect*) NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND (TS="earthworm") NOT (TS=carotid OR TS=tumo*) OR TS="glomus iranicum" AND TS="soil' AND TS=microorg* AND (TS=tox* OR TS=side effect*) NOT (TS=carotid OR TS=tumo*)

From the 417 publications found using the keywords from above, no toxicological effects on any non-target organisms was found in literature. The articles mentioning the AMF were related to the beneficial effects on plants. According to literature, a combined use of earthworms and AMF, or the AMT alone, increases the availability of nutrients in the soil and indirectly creates an antagonistic environment for undesirable predators by increasing the beneficial microbial population of the soil.

www.symborg.com 19

Effects on Date of Search: 10.01.2018 native fungi Publication dates: No restrictions or invasive Keywords: ((TS = glomus AND TS= iranicum) OR (TS = dominikia AND TS = iranica)) AND (TS = parasit* OR TS = infect* OR TS = displac*) AND (TS = native OR TS = fung* OR TS = invas* OR TS = plant*) plants

From the 6 publications found using the keywords from above, any relevant information on the subject was retrieved concerning that our Glomus spp. improved the biological fitness of any invasive plant in ecosystems or that our Glomus spp. become invasive, parasitic or displaced any fungal species.

Hybridization Date of Search: 10.01.2018 with other Publication dates: No restrictions AMF Keywords: ((TS = glomus) OR (TS = dominikia)) AND TS = Hybrid*

From the 287 publications found using the keywords from above, any relevant infonnation on the subject was retrieved concerning that our Glomus spp. hybridize with other arbuscular mycorrhizal fungi.

www.symborg.com 20 Toxicity (Brief)

.'% No case of toxicity (oral, dermal, respiratory, intravenous, skin, and eye sensitivity) or pathogenicity on humans, livestock or mammals can be found in the literature;

:% No mention of a glomus toxin can be found in the literature;

:: No case of toxicity or pathogenicity on the non-target organisms can be found in the literature;

:% No case of persistence on the environment can be found in the literature; _'& No signs of toxicity have been seen in research or technical personnel working on the endomycorrhiza has been observed during personnel monitoring;

The Classification, Labelling and Packaging regulation (CLP) do not recognise the products based on microorganisms, however, our product based on the Glomus iranicum var tenuihypharum is classified as Non toxic;

www.symborg.com 21 Use of Web of Science database for bibliographic research:

- Web of Science Core Collection, FSTA — the Food Science Resource, KCI — Korean Journal Database, MEDLINE, Russian Science Citation Index, SciELO Citation Index and Zoological Record). Research according to:

- EFSA guidance — Submission of scientific peer-reviewed open literature for the approval of pesticide active substances under Regulation (EC) No 1107/2009 on the placing of Plant Protection Products on market.

- OECD Guidance for Industry Data Submissions for Microbial Pest Control Products and their Microbial Pest Control Agents (Dossier Guidance for Microbials), Series on Pesticides No. 23.

Published in: last ten years (2007-2017).

The active substances under assessment: - Glomus iranicum var. tenuihypharum

The information is divided in three categories:

- Effects on human health (toxicological and exposure data);

- Fate and behavior in the environment;

- Effects on non-target organisms (ecotoxicological studies, environmental impact).

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