citriphagetm fact sheet v4

2105 Dobler Ave. Baltimore, MD 21218

443.540.6594

CitriPhage™ Citrus Canker Disease Eradication

1. CitriPhage™ is a combination of bacteriophages (phages) that can eradicate citrus canker disease (Xanthomonas citri)—which is currently affecting the world’s larges citrus growing regions (U.S., Brazil, China, Mexico, Spain, etc.).

2. CitriPhage™ is a genetically engineered bacteriophage derived from naturally occurring organisms.

3. CitriPhage™ has been independently proven able to effectively suppress citrus canker growth in lab and field tests1.

4. CitriPhage™ supplants copper-based treatments, the current treatment protocol for Citrus Canker; thus reducing toxic (heavy metal) buildup in the environment.

5. CitriPhage™ has been shown to constrain the motility (bacterial spread in vitro) of Xanthomonas citri. (Takashi Yamada, Hiroshima University, Japan).

6. CitriPhage™ can be stored in regular -20–4°C (-4–39°F) refrigerator for at least 6 months.

7. CitriPhage™ is stable for up to 2 weeks at 20–40°C (68–104°F) after which it biodegrades. It can be modified to survive at higher temperatures.

8. CitriPhage™ is designed to infect citrus canker bacteria alone. 9. CitriPhage™ biodegrades if it fails to find its target (citrus canker), it will not

change microbial ecology, it is extremely safe in the environment. 10. CitriPhage™ cannot be reproduced by the customer or by a competitor without

the necessary bio-engineered feedstock / formulae. 11. CitriPhage™ is also effective at treating all strains of Xanthamonas, including:

Rice Blight, Black Rot, Bacterial Leaf Spot, Sugarcane Leaf Scald and Banana Wilt to name only a few.

Citrus Canker Disease

1. Citrus Canker bacterial infection causes unsightly lesions on the leaves, stems, and fruit of citrus trees, including lemon, lime, oranges, and grapefruit.

2. Citrus Canker significantly affects the viability of the citrus crops, causing leaves and fruit to drop prematurely. Infected fruit is unsuitable for sale.

3. Citrus Canker disease destroys citrus trees after just a few seasons, if left untreated.

4. Copper-based sprays (the current treatment protocol) are toxic to fish and aquatic invertebrates and will contaminate water through runoff2.

5. Copper-based pesticides have been shown to stimulate the growth of mite populations in the field3.

1 (Tsong-The Kuo, Academia Sinica, Taiwan; Takashi Yamada, Hiroshima University, Japan) 2 http://www3.epa.gov/pesticides/chem_search/ppls/091411-00002-20150507.pdf 3 http://www.bulletinofinsectology.org/pdfarticles/vol64-‐2011-‐069-‐072mao.pdf

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Figure 1. Morphology of a parental strain of CitriPhage™ under electron microscopy. (Kuo et al., 1987, Virology. 156:305-‐12).

Figure 2. A parental strain of CitriPhage™ inhibits citrus canker pathogen Xanthomonas citri division. (A) Unchanged cells division in non-‐infected bacteria, (B) CitriPhage™ -‐infected cells (arrow) are still undivided. (Kuo, et al, 1994; Arch Virol. 135:253-‐64.)

Figure 3. A parental strain of CitriPhage™ inhibits citrus canker pathogen Xanthomonas citri motility. Bacterial solution were inoculated in the swimming assay (A,B), swarming assay (C,D). A significant reduction in swimming and swarming motility was observed in XacF1-‐infected cells (Ahmad AA et al., 2014, Front Microbiol. 5:321).

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Figure 4. . A parental strain of CitriPhage™ inhibits canker symptom development. (A) Characteristic canker lesions occurred with uninfected cells, while no obvious cankers developed on CitriPhage™ -‐infected cells. (B) Canker lesions were smaller in CitriPhage™ -‐infected leaves. (C) Lesions on both lower and upper surfaces of leaves inoculated with the uninfected cells showed severe symptoms, expanding with time. No lesions formed on either surface of the leaves infected with CitriPhage™ -‐infected cells. (Ahmad AA et al., 2014, Front Microbiol. 5:321).

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Xanthamonas Sub-species that Can Be Treated with CitriPhage™

Species Disease Species Disease X. vasicola Banana wilt X. axonopodis Cassava – Bacterial

blight

X. axonopodis Bacterial pustule, leaf blight – Soybean (Glycine max)

X. sacchari Banana rot – Tanzania

X. arboricola Walnut blight / Strawberry blight

X. axonopodis Bacterial spot

X. translucens Black chaff – Italian ryegrass

X. campestris Bacterial leaf spot – Brassica oleracea

X. fragariae Angular leaf spot – Strawberry

X. oryzae Bacterial streak / leaf blight – Rice

X. axonopodis Bacterial blight – Cassava

X. hortorum Bacterial leaf blight – Carrot

X. fuscans Common bacterial blight – Bean

X. vesicatoria Bacterial spot – Tomato and Pepper

X. cassavae Bacterial necrosis – Cassava

X. perforans Bacterial spot – Tomato

X. citri Bacterial blight – Mexican lime / Cotton

X. gardneri Bacterial spot – Tomato and Pepper

X. citri Citrus canker – Mango X. campestris Black rot – Brassica X. citri Black spot – Mango X. campestris

(vasicola) Bacterial wilt (BXW) – Banana

X. axonopodis Bacterial blight – Pomegranate

X. campestris (vasicola)

Sugarcane gumming disease

X. axonopodis Cassava – Bacterial blight

X. campestris (vasicola)

Black Rot – Brassicaceae

X. axonopodis Citrus – Bacterial spot

X. fuscans Citrus canker

X. transclucens Black Chaff – Wheat X. albilineans Sugarcane leaf scald

X. campestris Black rot – Brassica oleracea var. capitata

X. oryzae Bacterial leaf blight – Rice

X. arboricola Leaf spot – Turkish hazel

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Biopesticides

Biopesticides represent an emerging approach to agricultural disease control, they are engineered to be used in small quantities and to specifically affect a target pest or organism; they are designed to decompose quickly, if they do not find their intended target(s). In contrast to broad-spectrum chemical pesticides, bio pesticides are inherently less toxic and thus, less likely to affect indigenous organisms, such as birds, insects, and mammals4. Thus, they are favored by the U.S. and E.U. environmental agencies.

4 http://www.epa.gov/oppbppd1/biopesticides/whatarebiopesticides.htm

citriphagetm fact sheet v4

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