annona squamosa as botanical pesticide
TRANSCRIPT
Term Paper PresentationAC 602
Advancement in botanical research with reference to Annona squamosa
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Prithusayak MondalRoll No. 10064
Introduction
• Importance of botanical pesticides
• Current botanicals (Neem, rotenone, pyrethrum, nicotine &essential oils etc.)
• Potential new botanical pesticide – Squamocin (Anonin)from the plant Annona squamosa
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Plant Profile
Kingdom Plantae – Plants
Subkingdom Tracheobionta – Vascular plants
Superdivision Spermatophyta – Seed plants
Division Magnoliophyta – Flowering plants
Class Magnoliopsida – Dicotyledons
Subclass Magnoliidae
Order Magnoliales
Family Annonaceae – Custard-apple family
Genus Annona
Species squamosa – Sugar apple
Scientific Name: Annona squamosa Linn.
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PropertiesIn
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Antifeedant, insecticidal, antidiabetic, anti-tumour,anticancer, antibacterial & antiviral activity (Pandey et al.,2011)
Plant parts Use
Bark Prevention of diarrhoea
Root Treatment of dysentry
Leaf Cold remedy, urine purification, treatment of hysteria
Fruit Haematinic, cooling, sedative, stimulant, expectorant & maturant tonic; treatment of anaemia & burning sensation;
making of ice cream & milk beverages
Seed Abortifacient & pesticidal activity
Chemical constituents
• The plant is reported to contain glycoside, alkaloids,saponins, flavonoids, tannins, carbohydrates, proteins,phenolic compounds, phytosterols, amino acids (Pandeyet al., 2011)
• The diterpenoid alkaloid atisine is the main component of the root
• Annonaceous acetogenins – Botanical pesticidesInd
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Preparation of Extract
Percentage practical yield of petroleum ether (40-60oC), chloroform,ethyl acetate, acetone and methanolic extracts were found to be3.85, 2.33, 2.39, 1.2, 7.07 % w/w respectively
• Shade drying of leaves & homogenization tocoarse powder
• Sohhlet extraction using following solvents:1) Petroleum ether (40-60°C), 2) Chloroform,3) Ethylacetate, 4) Acetone, 5) Methanol
• Concentration of extracts using rotary evaporatorat 40-50C & drying
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Agarwal et al., 2012
Preliminary phytochemical
screening
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HPTLC StudyIn
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Annonaceous Acetogenins
• Acetogenins have gained much attention recently,because of its wide range of bioactive spectra
• Series of C-35/C-37 natural products derived from C-32/C-34 fatty acids that are combined with a 2-propanol unit
• Characterized by a long aliphatic chain bearing a terminalmethyl-substituted α,β-unsaturated γ-lactone ring (orketolactone ring), with 1,2 or 3 THF rings located alongthe hydrocarbon chain and a number of oxygenatedmoities (hydroxyls, acetoxyls, ketones, epoxides) &/ordouble bonds
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AcOEt
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1) Petroleum ether2) Partition between petrolem ether & aquous methanol
Ground Annonasquamosa seeds
(2 kg)
Methanol layer
ResidueCrude acetogenin
fraction
Fr. 1 2 3 4 5 6 7 8 9 10
Petroleum ether layer
Silica gel chromatographyCHCl3/ AcOEt=2:1 to AcOEt/ MeOH= 20:1
Acetogenins Extraction
Araya et al., 2004
Structures of Acetogenins
R1=R2=R4=H, R3=OH: Squamocin
R1=R2=R3=H, R4=OH: Squamocin-C
R1=OH, R2=R3=R4=H: Squamocin-G
R1=R2=R3=R4=H: Squamocin-L
R1=R4=H,R2=R3=OH: Squamocin-O1,-O2
R 4
R 3 O H
O O
O
O H
R 2 R 1
O
2 4 1 5
3 7
R1=R2=H, R3=OH: Squamocin-D
R1=R3=H, R2=OH: Squamocin-F
R1=OH, R2=R3=H: Squamocin-H
R1=R2=R3=H,: Squamocin-M
R3
O O
O
OH
R2 R1
O
OH
24 15
37
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Squamocin-N
Squamosten-A
O O
O
OHO
24 15
37
OH
O
O
O H
O HO H
O
O H
2 3 1 5
3 7
Structures of Acetogenins …In
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R=OH: Squamocin-BR=H: Squamocin-J
R=OH: Squamocin-E
R=H: Squamocin-K
Squamocin-I
O O
2 2 1 3
O HO H
O
O
3 5
O O
2 2 1 3
O
O
3 5
O HRO H
O O
2 2 1 3
O
O
3 5
O HO H
R
Structures of Acetogenins …In
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R1=OH: Squamostatin-C
R1=H: Squamostatin-E
24
O O
O
OOH OH OH
R1
R2
19
12
37
R1=H, R2=OH: Squamostatin-A
R1=OH, R2=H: Squamostatin-B
R1=R2=H: Squamostatin-D
24
O O
O
OOH OH OH
R1
19
12
37
H
O
OO
15
18
Squamostanal-A
Structures of Acetogenins …In
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Structural Features
The fundamental structural features of acetogenins are:
(1) Having hydrocarbon chain, of C35 or C37 in length
(2) 1～3 tetrahydrofuran rings are present
(3) One γ-lactone is present at an end of hydrocarbonchain
(4) 2～8 hydroxyl groups (rarely carbonyl or acetoxygroup) are present
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Classification of Annonaceous
Acetogenins
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Structure Activity Relationship
(1) Generally, intensity of the bioactivity is (A) type (adjacent bis-THF) > (B) type (non adjacent bis-THF) > (C) type (mono-THF) >(D) type (non-THF)
(2) γ-Lactone is crucial for activity
(3) If all other structural features are identical, C35 acetogeninsare more potent than the C37 acetogenins
(4) Thirteen carbons space between the OH-flanked THF and γ-lactone is optimum for activity
(5) Three hydroxyl groups, two flanking the THF ring(s) andanother somewhere in the long hydrocarbon chain provide boththe optimal position and polarity needed for the most potentactivity, and for tetra-hydroxylated acetogenins the activity dropsdrastically
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Alali et al., 1999
Structure Activity Relationship …(6) Neither the 4-OH group nor the 10-OH group is essentialfor activity
(7) A ketone instead of a hydroxyl functional group decreasesthe activity
(8) Derivatives (acetates, chloride etc.) decrease the activity
(9) Ketolactone acetogenins are usually less active and moreselective than their parent compounds
(10) The THP ring compounds are as active as the THFcompounds and have the same mechanism of action
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Alali et al., 1999
Commercial Product
ANOSOM®It is a biological insecticide based on botanical extract of Annona squamosa(Custard Apple / Seethaphal). ANOSOM® contains squamocin (annonin) asactive ingredient and is formulated as 1% Emulsifiable Concentrate (10,000ppm). ANOSOM® is non phytotoxic and not toxic to higher animals.
Mode of Action
Annonaceous acetogenins present in ANOSOM® have pesticidal and/orinsect antifeedant properties.
Target Pests
Helicoverpa arimigera, Helicoverpa zea, Spodoptera litura, Spodopteraexigua, Earias spp. , Achaea janata, Nephotettix virescens, Bunch caterpillar,Green Leaf Hopper, Leaf Folder, Army worm, Cut Worm.In
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Commercial Product …Crops
ANOSOM® is suitable for application on cereals, millets, pulses, oilseeds,fibre crops, sugar crops, forage crops, plantation crops, vegetables, fruits,spices, flowers, medicinal crops, aromatic crops, orchards andornamentals.
Method of Application
Foliar application : Mix ANOSOM® @ 2ml/ L of water and spray on cropcanopy. The spray volume depends upon the crop canopy. It isrecommended that BioPesticides are sprayed in early hours of morning orlate afternoon for better results. UV radiation in peak sunny hours of theday may reduce the bio efficacy of BioPesticides.
Shelf Life
It is stable for a period of 24 months from the date of manufacturing.Ind
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Mode of Action
• They are potent inhibitors of NADH : ubiquinone oxidoreductase, whichis an essential enzyme in complex I of the electron transport system(ETS) which eventually leads to oxidative phosphorylation inmitochondria (Lewis et al., 1993).
• They act directly at the ubiquinone catalytic site(s) within complex 1and in microbial glucose dehydrogenase (Friedrich et al., 1994).
• They also inhibit the ubiquinone-linked NADH oxidase that is peculiarto the plasma membranes of cancerous cells and functions to permitcytosolic phosphorylation (substrate level phosphorylation) byrestoration of NAD levels.
Thus, the end result of both of these mechanisms is ATPdeprivation (Morre et al., 1995).
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