Bacillus subtilis, Strain QST 713 Use in Integrated Pest … · 2015-09-17 · Bacillus subtilis strain QST 713, the active ingredient of biofungicide product Serenade® was voted

D. W. EdgecombD. Manker

AgraQuest, Inc. Davis, CA

Bacillus subtilis, Strain QST 713Use in Integrated Pest Management

Bacillus subtilis, Strain QST 713Use in Integrated Pest Management

Isolated from Soil – CA Organic Peach OrchardRod-shaped, Gram Positive, Aerobic, Motile Bacterium Naturally Occurring, No Genetic ModificationsUnique – Patented Bacillus StrainDistinguished by Previously Unknown MetabolitesUS - EPA Registration – July 2000First “Effective” Broad Spectrum Bio-fungicidePresidential Green Chemistry Award – 2003World Technology Award for Environment - 2004Now Registered Globally – Trade Name Serenade®

Isolated from Soil – CA Organic Peach OrchardRod-shaped, Gram Positive, Aerobic, Motile Bacterium Naturally Occurring, No Genetic ModificationsUnique – Patented Bacillus StrainDistinguished by Previously Unknown MetabolitesUS - EPA Registration – July 2000First “Effective” Broad Spectrum Bio-fungicidePresidential Green Chemistry Award – 2003World Technology Award for Environment - 2004Now Registered Globally – Trade Name Serenade®

Bacillus subtilis, Strain QST 713

0

5

10

15

20

25

30

Cum

mul

ativ

e T

otal

1999 2000 2001 2002 2003 2004 2005 20060

5

10

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30

Cum

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1999 2000 2001 2002 2003 2004 2005 2006GermanySpainGreece AustraliaCanadaPeruThailandChinaUK

FranceItalyKoreaEcuadorColumbia

Guat.Hond.Switz.TurkeyArgen.

JapanIsraelPhilip.

C.RicaMexicoN.Z.P.Rico

U.S.Chile

Global Registration StatusGlobal Registration StatusGlobal Registration Status

Bold = Currently registered

Bacillus subtilis (Strain QST 713)Bacillus subtilis (Strain QST 713)

EU Annex 1 InclusionBacillus subtilis strain QST 713, the active ingredient of biofungicide product Serenade® was voted for inclusion into Annex 1 of Directive 91/414/EEC at the EU Standing Committee on the Food Chain and Animal Health meeting on July 14, 2006.

EU Annex 1 InclusionBacillus subtilis strain QST 713, the active ingredient of biofungicide product Serenade® was voted for inclusion into Annex 1 of Directive 91/414/EEC at the EU Standing Committee on the Food Chain and Animal Health meeting on July 14, 2006.


Registration Milestone

Grapes Gray Mold (Botrytis cinerea)Powdery mildew (Uncinula necator)

Bananas Black Sigatoka (Mycosphaerella fijiensis)Yellow Sigatoka (Mycosphaerella musicola)

Tomato / Pepper Bacterial Leaf Spot (Xanthomonas spp.)Early Blight (Alternaria solani)P.Mildew (Leveillula taurica)

Cucurbits P.Mildew (Erysiphe / Sphaerotheca spp.)Lettuce Leaf Drop (Sclerotinia spp.)Pome Fruit Fire Blight (Erwinia amylovora)

Scab (Venturia inaequalis)Mango Anthracnose (Colletotrichum gloeosporioides)Beans White Mold (Sclerotinia sclerotiorum)

Major Global Crops / DiseasesBacillus subtilis, Strain QST 713

Formulations: (AS) Aqueous Suspension(WP) Wettable PowderActive Ingredients Spec. = Minimum 1 X 109 cfu / gramOrganic Formulations (OMRI, IMO, BCS, JAS)> Two Year Storage Stability Excellent SuspensibilityPhysical / Biological Tank Mix Compatibility with standard fungicides

Formulations: (AS) Aqueous Suspension(WP) Wettable PowderActive Ingredients Spec. = Minimum 1 X 109 cfu / gramOrganic Formulations (OMRI, IMO, BCS, JAS)> Two Year Storage Stability Excellent SuspensibilityPhysical / Biological Tank Mix Compatibility with standard fungicides


B.B. subtilissubtilis spores cover leaf spores cover leaf surface & prevent fungal surface & prevent fungal spores from penetrating spores from penetrating and infecting the plant.and infecting the plant.

LipopeptideLipopeptide metabolitesmetabolitesbreak down pathogen cell break down pathogen cell membranes, causing membranes, causing pathogen to collapse & die.pathogen to collapse & die.

Lipopeptides prevent Lipopeptides prevent pathogens from multiplying.pathogens from multiplying.

Applied Just Like Applied Just Like Other ProductsOther Products Novel Modes of ActionNovel Modes of ActionNovel Modes of Action

SAR - Role in MOA Academic or Significant

Contribution?

SARSAR -- Role in MOA Role in MOA Academic or Significant Academic or Significant

Contribution?Contribution?

No Special Equipment / Handling or Storage

No Special Equipment / No Special Equipment / Handling or StorageHandling or Storage


RT: 0.01 - 30.01

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30Time (min)

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10

20

30

40

50

60

70

80

90

100

Rel

ativ

e Ab

unda

nce

24.58

7.478.88

23.8010.91 23.52

25.7111.229.30

20.1019.0217.79 20.196.27 11.92 26.905.50 16.7515.414.93 12.62 28.933.731.66

NL:1.07E8Base Peak F: + c Full ms

IturinsIturins

Agrastatins/PlipastatinsAgrastatins/Plipastatins

SurfactinsSurfactins

LC-MS Profile of QST 713LC-MS Profile of QST 713

CH2

CH2

COOH

CH2

CH2

CH2

NH2

CH2

O

CH

CO

CH2

CH3

CH2

CH2

COOH

CH3

N

CH2CH2

OH

CH2

CO-NHCHCO-NHCHCO-NHCHCO-NHCHCO-NHCHCO-NHCHCO

OCCHNH-OCCHNH

HO

CONH2

CH2

CHCH3

CH3

OCCHNH

CHOH(CH2)12

CH3

Orn

Tyr

Thr Glu Ala

GlnTyrVal

Pro

Agrastatin 1

Agrastatin 2,Ala replaced by Val

CH2

CONH2

CH2

N CO

CONH2

HC

OH

CH2

CH3

CO-NHCHCO-NHCHCO-NHCHCO

Asn Tyr

CH2

CONH2

Asn

Gln

NH-OCCHNH-OCCHNH-OC

CH

CH2

OH

HN

CH CH2

CH2

Ser ProAsn

CH(CH3)2CHCH2CH3

CONH2

CH2

R(CH2)8~12

Iturin A R= CH3

Profile of B. subtilis, QST 713 Metabolites

Botrytis 0% inhibitionIturins 10ppm orAgrastatins 50ppm

Botrytis 90% inhibitionIturins 10ppm + Agrastatins 2.5ppmSynergistic Action of Metabolites

B.s. spores – Colonize and Out-compete PathogenFungicidal Metabolites – Lipopeptides Present in Formulation / Produced by Colonizing B.s.Destroy Pathogen Spore & Mycelium MembranesProvide Barrier to Infection by PathogenOptimal Use = Preventative Control ProgramNew / Unique Resistance Management Tool

B.s. spores – Colonize and Out-compete PathogenFungicidal Metabolites – Lipopeptides Present in Formulation / Produced by Colonizing B.s.Destroy Pathogen Spore & Mycelium MembranesProvide Barrier to Infection by PathogenOptimal Use = Preventative Control ProgramNew / Unique Resistance Management Tool

Novel / Multiple Modes of ActionNovel / Multiple Modes of Action


Use in Integrated Programs with Conventional

Fungicides

Use in Integrated Programs with Conventional

Fungicides


98.4Std FungicideQST 713Std FungicideQST 713Chemical + Biological

98.6QST 713Std FungicideQST 713Std FungicideChemical + Biological

86.9QST 713QST 713QST 713QST 713BiologicalRepeating

98.8Std FungicideStd FungicideStd FungicideStd FungicideChemicalRepeating

---WaterWaterWaterWaterUntreated

% Control20 Day13 Day7 Day0 Day

IPM Program for Cucurbit Powdery Mildew (Sphaerotheca fuliginea)

50% Reduction in Chemical Fungicide

Chemical standard fungicide = Quinomethionate

Bacillus subtilis, Strain QST 713Use in Integrated Disease Control Program - Japan

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74

87

89

6/20

94

66

10

4

6/25

38

10

5

4

6/20

Untreated control

6/256/14

---17

305QST 713 at 4 lbs

891QST 713 at 4 lbs (5 applns); followed by Azoxystrobin at 0.15 lb ai (3 applns)

962Chlorthalonil (2.25 lbs ai)alt with Azoxystrobin (0.25 lb ai)

% Control% Powdery MildewTreatment (Amt/100 gal/acre)*

*Applied weekly on 5/3, 5/8, 5/15, 5/22, 5/30, 6/6, 6/13, and 6/19

Bacillus subtilis, Strain QST 713Cucurbit Powdery Mildew (Sphaerotheca fuliginea) - USA

Bacillus subtilis Strain QST 713 - Bananas

Black Sigatoka (Mycosphaerella fijiensis)

Bacillus subtilis Strain QST 713 - Bananas

Black Sigatoka (MycosphaerellaMycosphaerella fijiensisfijiensis)

Requirement = 6 Functional Leaves at HarvestRequirement = 6 Functional Leaves at Harvest

4.9 b3.4 bUntreated

8.5 a4.1 aStandard ProgramMancozeb – 1.8 liters

7.4 a3.9 aB.subtilis + mancozeb2 + 0.9 liters

7.8 a3.9 aB.subtilis 2 liters

Functional LeavesAt Harvest

Visible StreaksOn Leaves

Treatment(rate / hectare)

Bacillus subtilis, Strain QST 713 – “Sustainable” Black Sigatoka (Mycosphaerella fijiensis) Control Program in Bananas

Mindanao – The Philippines (March to August 2003)Small Plot / RCB Trial MindanaoMindanao –– The Philippines (March to August 2003)The Philippines (March to August 2003)Small Plot / RCB Trial Small Plot / RCB Trial

Product Positioning – FeaturesEffective “Non-Chemical” Option for Disease Control ProgramsBroad Spectrum Disease Control – Fungal and Bacterial PathogensTank mix and Rotation with Protectants / Systemics in ProgramsMRL Management / Sustainable / Organic Production Tool Resistance Management

Single Site Compounds – e.g., Strobilurin and Triazole FungicidesExempt From Tolerance / 0-Day PHI / No CODEX- Late Season, Pre-Harvest Option / No ResiduesIntegrated Pest ManagementNot toxic to Non-target Organisms -Honeybees, Lady Beetles, Lacewings, Parasitic Wasps, and EarthwormsSafe to Workers / Environment

Product Positioning – FeaturesEffective “Non-Chemical” Option for Disease Control ProgramsBroad Spectrum Disease Control – Fungal and Bacterial PathogensTank mix and Rotation with Protectants / Systemics in ProgramsMRL Management / Sustainable / Organic Production Tool Resistance Management

Single Site Compounds – e.g., Strobilurin and Triazole FungicidesExempt From Tolerance / 0-Day PHI / No CODEX- Late Season, Pre-Harvest Option / No ResiduesIntegrated Pest ManagementNot toxic to Non-target Organisms -Honeybees, Lady Beetles, Lacewings, Parasitic Wasps, and EarthwormsSafe to Workers / Environment


Documents

Bacillus subtilis, Strain QST 713 Use in Integrated Pest … · 2015-09-17 · Bacillus subtilis strain QST 713, the active ingredient of biofungicide product Serenade® was voted