Management of Plant Parasitic Nematode Through Bio-agents

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Fungal Bio-agents and Plant Extracts for the Management of Plant

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Biological control is defined as the action of parasites, predators or pathogen in maintaining another organism’s population at a lower average than occur in their absence. Biological control aims at increasing the parasites, predators and pathogens of nematodes in the soil, in order to increase the mortality of PPN. BIOLOGICAL CONTROL & BIOPESTICIDES Fungal antagonist of nematodes have been studies since the first observation of the nematophagus habit of the fungus Harposporium anguillulae by Lohde in 1874.Nematophagus fungi play an important role in regulating nematode population dynamics

DESIRABLE CHARACTERISTICS OF BIOLOGICAL CONTROL AGENTS:

DESIRABLE CHARACTERISTICS OF BIOLOGICAL CONTROL AGENTS Host specific Parasitism is always lethal Easily manipulated in laboratory Can be mass produced Easily disseminated with standard equipment Potential for establishment & recycling Provides control for extended period Not harmful to environment Shelf-life of at least one year No hyperparasites Compatibility with others

TYPE OF FUNGI 1.PREDACIOUS FUNGI 2.OPPORTUNISTIC FUNGI Nematode Trapping Fungi Adhesive Networks Adhesive Knobs Non-constricting Rings Constricting Rings Adhesive Conidia Zoosporic Fungi Egg Parasite :

TYPE OF FUNGI 1.PREDACIOUS FUNGI 2.OPPORTUNISTIC FUNGI Nematode Trapping Fungi Adhesive Networks Adhesive Knobs Non-constricting Rings Constricting Rings Adhesive Conidia Zoosporic Fungi Egg Parasite

TYPE OF FUNGI 1.PREDACIOUS FUNGI :

TYPE OF FUNGI 1.PREDACIOUS FUNGI 1)Nematode-trapping fungi (predators, predaceous fungi): predacious fungi are those which produce some type of capturing organs/ devices to consume their prey i.e., the nematode. These fungi are classified as : a)Those which produce sticky hypha / knobs or branches e.g., Stylopage , Dactylella cionopaga and D. gephyropaga b)those which produce constricting rings. Caught nematode in the ring by bulging the inner walls of lumen e.g., Dactylella bembicodes, Arthrobotrys Dactyloides

Arthrobotrys oligospora trapping a nematode :

Arthrobotrys oligospora trapping a nematode

Arthrobotrys spp. developing constricting ring traps. :

Arthrobotrys spp. developing constricting ring traps.

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Adhesive network of Arthrobotrys oligospora with captured nematode Adhesive columnar branches One- to three-celled hyphal branches covered with adhesive substance. Examples: Monacrosporium cionopagum Monacrosporium gephyropagum Dactylella lobata

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The nematophagus fungus Arthrobotrys candida Formation of a net-like trap by Arthrobotrys oligospora

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Nematode captured by the constricting rings of the predatory fungus Arthrobotrys anchonia .  Note that the ring cells "cushion" around the body of the victim but have not yet constricted the body.  This is a very early stage after capture. Nematode was captured by non-constricting rings of Dactylella

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Rings constrict by fungus

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Adhesive three-dimensional networks covered with adhesive substance. Loop diameter 20-59 µm. High trapping quota in vitro because of their three-dimensional structure. Arthrobotrys superba Arthrobotrys oligospora Arthrobotrys dactyloides Dactylaria brochopaga Monacrosporium doedycoides Constricting rings Stalked, three-celled rings. Cells expand towards the inner side when touched there and ensnare the prey. Trapping Mechanism

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Non-constricting rings Stalked, three-celled rings. µm. Prey is entangled when trying to pass. Dactylaria candida Dactylella leptospora Hourglass-shaped adhesive knobs Multinuclear syncytia covered with adhesive substance. Short, non-cellular stalks. Very adhesive Nematoctonus geogenius Nematoctonus leiosporus

ENDOPARASITIC / ENDOZOIC FUNGI:

ENDOPARASITIC / ENDOZOIC FUNGI These fungi produce either simple or flagellate adhesive spores which upon contacting a nematode quickly and firmly attached to its cuticle. It then germinates and enter the nematode body through germ tube. Simple spore are ingested by nematode, reach the oesophagous / buccal cavity where they germinate e.g.. Harposporium anguillulae while sticky spores stick to the nematode body & germinate and penetrate directly through the nematode cuticle, produce infective hyphae e.g. Meria coniospora & Nematoctonus bisporus . Example of flagillated spore is Cateneria auxilliaris.( infecting Heterodera avenae & Globodera rostochiensis) The infection of endozoic fungi has been observed on Heterodera schachtii (Khun, 1877); H. avenae and G. rostochiensis ( Kerry, 1975; Kerry et al.., 1976)

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Zoosporangia of Catenaria in a nematode body

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A female of cereal cyst nematode, Heterodera avenae , and the cyst-parasitic fungus, Nematophthora gynophila

Parasitic Fungi: Zoosporic:

Parasitic Fungi: Zoosporic Infection process: Consumption of conidia of Harposporium sp. Sticky conidia: Cephalosporium spp. and Verticillium spp. Zoospores: Chytridiales: Catenaria auxiliaris Oomycota: Nematophthora gynophila

EGG PARASITIC FUNGUS:

EGG PARASITIC FUNGUS These fungi may be of particular significance with those nematodes that aggregate their eggs and fungal food source in masses. Particularly root-knot & cyst nematodes. Paecilomyces lilacinus: It shown some effect in controlling Meloidogyne incognita eggs on potato roots and also found invading mature females of cyst of Globodera pallida. This fungus also found to increase the growth of brinjal plants and reduced root galling. This fungus @ 4 & 8 g per kg soil significantly reduced M. incognita population. Dickson and Mitchell (1985) however , have reported that Paecilomyces lilacinus is largely effective for the management of M. javanica on tobacco in micro plots. Verticillium chlamydosporium : This fungus found to parasitize M. arenaria females and also found in the cysts of Heterodera glycines . It has been observed that young white females are not attacked but those in which eggs have been susceptible to invasion. It has potential to prevent the egg hatching of M. arenaria and to colonize eggs by hyphal penetration. This fungus play a crucial role as a part of INM when used along with oil cakes and other fungi.

Effect of V. chlamydosporium:

Effect of V. chlamydosporium

Fungal Parasites of Sedentary Stages :

Fungal Parasites of Sedentary Stages Cylindrocarpon destructans Dactylella oviparasitica Paecilomyces lilacinus Verticillium chlamydosporium Verticillium lamellicola Verticillium lecanii Verticillium leptobactrum Verticillium suchlasporium Rapid growing Culturable May produce disruptive enzymes and toxic metabolites

Paecilomyces lilacinus :

Paecilomyces lilacinus PAECIL™, BIOACT®, NEMACHEK Technological Innovation Corporation, Australia BIOSTAT, Laverlam S.A., Colombia

Trichoderma spp.:

Trichoderma spp. Trichoderma spp. The filamentous fungi have been widely studied for their effectiveness in controlling nematodes. The mechanisms involved in this is direct through antibiosis and parasitism. Trichoderma grows around the target and releases toxic compounds and different lytic enzymes mainly chitinases, glucanases and proteases. These proteins facilitate Trichoderma penetration into the nematode and the utilization of the nematode components for nutrition. The role of these lytic enzymes has been proved in nematode control.

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Table. Effect of different plant extracts on root-knot nematode, M. incognita infesting tomato , brinjal and Okra S.No Treatments Okra Brinjal Tomato Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant 1 Allium cepa 5% 39.67 23.67 35.33 26.00 56.33 39.00 2 Allium cepa 10% 32.00 15.00 28.00 23.00 59.67 51.00 3 Allium cepa 20% 31.67 21.67 30.67 19.67 53.67 31.33 4 Zingiber officinalis 5% >100 >100 >100 >100 >100 >100 5 Zingiber officinalis 10% >100 >100 >100 >100 >100 >100 6 Zingiber officinalis 20% >100 >100 >100 >100 >100 >100 7 Allium sativum 5% 30.33 21.33 35.67 21.33 49.33 38.33 8 Allium sativum 10% 23.33 16.00 25.33 11.00 40.33 27.00 9 Allium sativum 20% 16.33 13.00 27.33 13.00 39.33 27.00 10 Baugainvillia glabra 5% >100 >100 >100 >100 >100 >100 11 Baugainvillia glabra 10% >100 >100 >100 >100 >100 >100 12 Baugainvillia glabra 20% >100 >100 >100 >100 >100 >100 13 Curcuma longa 5% >100 >100 >100 >100 >100 >100 14 Curcuma longa 10% >100 >100 >100 >100 >100 >100 15 Curcuma longa 20% >100 >100 >100 >100 >100 >100 16 Control (water) >100 >100 >100 >100 >100 >100

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Table. Effect of different plant extracts on root-knot nematode, M. incognita infesting tomato and brinjal S. No Treatments Brijal Tomato Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant 1 Sadabahar (Vinca major ) 5% > 50 > 45 >50 >50 2 Sadabahar (Vinca major) 10% 35.33 26.33 37.00 35.00 3 Sadabahar (Vinca major) 15% 34.33 27.00 41.00 35.00 4 Amaltash ( Cassia foitida) 5% >100 >100 >100 >100 5 Amaltash ( Cassia foitida) 10% >100 >100 >100 >100 6 Amaltash ( Cassia foitida) 15% >100 >100 >100 >100 7 Jasmin ( Jasminum sp) 5% >100 >100 >100 >100 8 Jasmin ( Jasminum sp) 10% >100 >100 >100 >100 9 Jasmin ( Jasminum sp) 15% >100 >100 >100 >100 10 Tulsi ( Ocimum sanctum) 5% 35.33 29.00 38.00 30.00 11 Tulsi ( Ocimum sanctum) 10% 29.33 24.00 31.00 27.33 12 Tulsi ( Ocimum sanctum) 15% 26.00 23.00 29.00 27.00 13 Control >100 >100 >100 >100

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S.No. Treatments Okra Brijal Tomato Average Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant Galls / Plan t Egg masses / Plant Galls / Plant Egg masses / Plant 1 Neem 20% 23.00 17.00 26.67 21.33 52.67 42.33 34.12 26.89 2 Dhak 10% 48.67 37.67 51.33 43.67 67.33 59.00 55.78 46.78 3 Dhak 20% 49.33 40.33 48.00 41.33 59.67 42.67 52.34 41.45 4 Jatropha 10% 31.67 27.00 36.33 31.67 49.33 42.33 39.11 33.67 5 Jatropha 20% 21.33 18.33 32.67 28.00 42.00 40.00 32.00 28.78 6 Mahua 10% 23.33 21.33 27.33 21.00 39.00 30.67 29.89 24.34 7 Mahua 20% 19.67 13.67 25.67 20.33 30.33 28.33 25.33 20.78 8 Control 91.33 76.00 101.00 81.67 121.00 87.67 104.45 81.78 9 Average 38.55 31.42 43.63 36.13 57.67 46.63 _- - Table. Effect of different plant extracts on root-knot nematode, M. incognita infesting tomato , okra and brinjal

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S.No. Treatment Okra Brijal Tomato Average Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant Galls / Plant Egg masses / Plant 1 Neem 10% 38.67 22.67 37.33 21.00 66.33 49.00 47.45 30.89 2 Neem 20% 27.00 15.00 28.67 20.00 69.67 58.67 41.78 31.23 3 Dhatura 10% 31.67 21.67 30.67 19.67 53.67 31.33 38.67 24.23 4 Dhatura 20% 26.00 17.67 19.00 10.33 49.67 27.00 31.56 18.34 5 Calotropis10% 31.33 19.33 26.67 11.67 85.00 40.33 47.67 23.78 6 Calotropis20% 23.33 18.33 17.67 8.33 47.00 24.33 29.34 17.00 7 Guarpatha 10 30.33 21.33 36.67 21.33 49.33 38.33 38.78 27.00 8 Guarpatha 20 16.33 13.00 27.33 13.00 39.33 27.00 27.67 17.67 9 control 73.33 60.67 92.33 60.33 116.67 87.00 94.11 69.34 10. Average 33.11 23.30 35.15 20.63 64.08 42.56 Table. Effect of different plant extracts on root-knot nematode, M. incognita infesting tomato , okra and brinjal

Vesicular arbuscular mycorrhiza (VAM):

Vesicular arbuscular mycorrhiza (VAM) The association of VAM fungi with plant nematodes and the beneficial effect of mycorhizal symbiosis on plant growth has led to investigations into the potential of VAM fungi to limit yield losses due to nematodes. e.g. Glomus fasiculatum & others

HOW THEY WORKS:

HOW THEY WORKS The hypothesis proposed to explain AMF effects on soil borne pathogen generally considered to have either a physical or physiological basis. They work efficiently under stress conditions. They considered to be a microbial communities changer, resulting influence the growth and health of plants. The major mechanism is nutritional, because plants with high phosphorus status are less sensitive to pathogen damage.

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Physical mechanism: Morphological changes in roots and root tissue Physiological mechanism: Nutritional changes Competition for host photosynthesis and infection Biochemical changes or changes in chemical constituents of plant tissue Biological mechanisms Microbial changes in the mycorrhizosphere

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MODE OF ACTION 1)Attraction 2)Attachment 3)Penetration 4)Pathogenicity of fungi to nematodes

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1.Biological agents may effects directly over the pathogen include inhibition by microbial compounds (antibiosis), competition for colonization sites and nutrients, degradation of pathogenicity factors and parasitism. 2.Indirec mechanisms include improvement of plant nutrition and damage compensation , change in root system anatomy, microbial changes in the rhizosphere and activation of plant defense mechanisms leading to enhance plant resistance . For an effective bio-control agent combination of both way is more beneficial

CONCLUSION ::

CONCLUSION : Most of the research indicated under green house conditions that bio-control agent have the potential to nullify the damage due to phytonematodes, but intensive investigation reed to be carried out under field condition to asses their practical utilization. Bioagents with highest potential for the management of PPN are likely to be those that are endemic in agriculture soils can be induced to proliferate by methods compatible with conventional farming practices The opportunistic fungus which infect eggs and females belonging to the genera Paecilomyces , Trichoderma and Verticillium seen to be most promising. These fungi spread rapidly are present abundantly in the rhizosphere. They are well adopted to the environment, being successful competitors and are able to survive host free period. Besides they also have the added advantages of being easily grown in axenic culture and introduced into soil.

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