Bio fumigation can be an effective tool for soil sterilisation and augmentation of biocontrol control agents

1. Farmer's first choice since 1974
S.K. Ghosh, Nataraj. T. Usha Nandini and Asha
Multiplex Biotech Private Limited. A-420, 1st stage,
Peenya Industrial Estate. Peenya. Bangalore. Karnataka. India.
Email: trainingmbt@multiplexgroup.com ,
www.multiplexgroup.com

2. Farmer's first choice since 1974
Biofumigation process : A natural defense
mechanism of brassicas
Active ingredient
for Biofumigation

3. Farmer's first choice since 1974
• Brassica group represents a sustainable
technique for soil disinfection and fertility
management .
• Identified as alternative of methyl bromide
for soil sterilization.
• The technique consists of :
• Apart from improving soil fertility it has
allelopathic effect on pest and pathogens

4. Farmer's first choice since 1974
• Plant based approach found be impractical and a costly approach .
• Result is unpredictable and highly inconsistent .
• Concentration of active compound (Gls) and ability to suppress varies
• By season .
• Cover crops variety .
• Maturity at incorporation .
• Soil microbial diversity .
• Microbe population density .
• Commercial scope for the development of economically viable and
truly effective product based on votalite oil of mustard ( AITC) .

5. Farmer's first choice since 1974
• Allyl isothiocyanate - a predominant ITC in Indian mustard.
• Produced both naturally and synthetically.
• Highly pungent, strong lachrymator & skin irritant compound.
• Listed as “Generally Regarded as Safe” (GRAS) compound
• Approved by US Environmental Protection Agency (EPA),
Biopesticides and Pollution Prevention Division (BPPD) for
agricultural usage .
• Commonly used as a food ingredient and additive under USA
Food and Drug Administration (FDA) jurisdiction.
• Permitted in organic crop production by IOA (listing under Title
7 Code of Federal Regulations Part 205.601).

6. Farmer's first choice since 1974
Product
AITC
concentration
(mg/kg)
Brussels sprouts 0.10
Cabbage 3.00
Cauliflower 0.08
Horseradish 1,350
Mustard 400–15,000
Baked goods 25–100
Condiments 700–5,000
Fats, oils 50
Fish products 0.05–0.07
Gelatins, puddings 1.00–2.00
Meat products 35–60
Seasonings,
flavorings
6–30
Property Value /Description
Color Clear, colorless to light amber
Physical State Oily liquid
Molecular Formula CH2=CHCH2N=C=S (C4H5NS)
Molecular Weight (g/mol ) 99.15
Freezing Point, oC –80; –102.5
Boiling Point, oC 150–154
Density, g/mL 1.0126
Solubility in water at 20 oC, mg/L 2,000 (soluble)
Solubility in organic solvents Miscible in many organic solvents,
Soil Organic Carbon-Water Partition
Coefficient (Koc), mL/g
260
(Moderately mobile in soils)
Aerobic Soil Half-life (DT50) DT50 is 2 days
Hydrolysis Facile ( fully degraded within 80 minutes at pH 8)
Photodegradation Highly degradable (half-life = 2.4 hours at 25 o
C).
Vapor Pressure at 25 o
C, mm Hg 3.7
.

7. Farmer's first choice since 1974
Mustard seed meal powder (MSMP
• Extraction : Defatted MSM obtained by hexane Soxhlet
extraction process
• Grounded : Defatted -MSM was grounded by using ball mill at
450 rpm for an hour .
• Assess the particle size and AITC release by moistening MSMP
MSMP in pellet form
MSMP in Tablet form

8. Farmer's first choice since 1974
• AITC with 92.86% purity was used for developing formulation
• Commercially available as Volatile oil of mustard ( VOM)
Micro-encapsulation of AITC

9. Farmer's first choice since 1974
• Developed suspension concentrate
formulation of AITC ( 50% ) along
with MSMP .
• A free flow homogenous formulation
developed
• Readily soluble in water .

10. Farmer's first choice since 1974
CONTROL LOW MEDIUM HIGH
Fungicidal potency against Sclerotium rolfsii
Treatments After 24 hrs After 48 hrs After 72hrs
Control 10.5 cm 18.5cm 32.5cm
100 ppm Nil Nil Nil
150 ppm Nil Nil Nil
200 ppm Nil Nil Nil
50 % SC formulation of AITC could
suppress the growth of growing
colony S. rolfsii even at low dosage

11. Farmer's first choice since 1974
CONTROL LOW CONCENTRATION MEDIUM CONCENTRATION HIGH CONCENTRATION
Treatments After 24 hrs After 48 hrs After 72 hrs
Control 100% germination Normal growth Normal growth
Low : 100 ppm 100 % germinated Normal growth Normal Growth
Medium : 150 ppm 40 % non- germination Impaired growth Impaired growth
High : 200 ppm 100 % non-germinated Impaired growth Impaired growth
Against microsclerotia bodies of S. rolfsii
• AITC treated microsclerotia has
shown poor germination and
impaired growth
• Impact on microsclerotia is
concentration dependent .

12. Farmer's first choice since 1974
S. rolfsii infected sick soil
T-2: Sick Soil + Bio fumigated
T-3: Sterilized healthy soil
50
70
80
50
100
40.0
14.29
0.00
60
0.00
0
20
40
60
80
100
120
Low dosage Medium dosage High dosage Control Check
Biofumigation: Dosage Impact
Germination (%) Disease Incidences (%)
• T-1: Low dosage : Sick soil (5kg) + 10ml of 1% AITC dilution
• T-2: Medium Dosage : Sick soil (5kg) + 15ml of 1% AITC dilution
• T-3: High dosage : Sick soil (5kg) + 20 ml of 1% AITC dilution
• T-4: Control : Sick oil with S. rolfsii
• T-5: Check : Sterilized soil

13. Farmer's first choice since 1974
Low , medium and high : 10ml , 15ml and 20ml of 1% dilution of AITC / 100gm of soil
Method of application
ControlSick soil
Biofumigated
sick soil
Sick soil Biofumigated soil

14. Farmer's first choice since 1974

15. Farmer's first choice since 1974
Spot application method Drenching Method Drip irrigation method
• Dilution : 10 -fold dilution
• Application rate : 50ml / spot
• Distance : 10 ft apart
• Dilution : 1 % of AITC formulation
• Dosage : 500ml / acre
• Drenching of large area is a practical
constrain due to strong smell and proper
distribution
• Application rate : 500ml/ acre /
• Dilution rate : 1%
• AITC @ 25ml in 25L of water and
drip irrigated in each block

16. Farmer's first choice since 1974
• T-1: CONTOL Plot ( Non Biofumigated with Trichoderma + Pseudomonas )
• T-2: Biofumigated plot ( 4 bed )
• T-3: Bio fumigated plot with Trichoderma ( 4 bed)
• T-4: Biofumigated plot with Trichoderma + pseudomonas ( 4 bed)
• Transplantation done after 4 days of Biofumigation
• Number of seedling per bed : 75 and 300 per treatment
• Hybrid Variety: Indira.
• Bed size : 78 ft x 4ft = 312 sq ft
• Number of bed / treatment = 4
• Area of each treatment = 1248 sq ft

17. Farmer's first choice since 1974

18. Farmer's first choice since 1974
15
36
62
74.8
0
20
40
60
80
0 DAT 20 DAT 40 DAT 60 DAT
Plant height ( cm)
T-1 T-2 T-3 T-4
0 10 20 30 40 50 60 70 80
T-1
T-2
T-3
T-4
No. leaf/plant
60 DAT 40 DAT 20 DAT

19. Farmer's first choice since 1974
41.212.33
8.5
6.5
% Disease incidences
T-1
T-2
T-3
T-4
40.5
8.5 9.25
6.5
0
5
10
15
20
25
30
35
40
45
T-1 T-2 T-3 T-4
No of nematodes /gm of soil
0DAT 5 DAT 30DAT 60 DAT

20. Farmer's first choice since 1974
4 6
12
28
51
0
10
20
30
40
50
60
0 DAT 5 DAT 10DAT 30DAT 60 days
Assessment of Trichoderma population
( -x 10 4 /gm of soil)
T-1 T-2 T-3 T-4
3
9
17
22
36
0
5
10
15
20
25
30
35
40
0 DAT 5 DAT 10 DAT 30ADAT 60 days
Assessment of Pseudomans population
( -x 10 5 /gm of soil)
T-1 T-2 T-3 T-4
T-4 > T-1 > T-3 > T-2

21. Farmer's first choice since 1974
T-1 : CONTROL
(NON FUMIGATED WITH NISARGA + SPARSHA)
T-2 : BIOFUMIGATED
T-3 : BIOFUMIGATED + NISARGA T-4 : BIOFUMIGATED + NISHARGA+SPARSHA
28.8
37.33
43.2
46.33
0
10
20
30
40
50
60
T-1 T-2 T-3 T-4
a
a
b
c
P < 0 .001
Ø > 8 months crop duration
Ø Highest yield in T-4
( 60.86% ) > T-3 (50.00%) > T-2
( 29.61%)

22. Farmer's first choice since 1974
• Biofumigation an effective method for the sterilization of soil .
• Biofumigation of soil could enhance the proliferation of microbial BCA over non
fumigated soil with better growth and yield under protected cultivation of
capsicum.
• Biofumigation could be an effective complementary method to promote the usage
BCA in soil for their better performance under reduced competition of harmful
organisms

23. Farmer's first choice since 1974