IDENTIFICATION OF PHYTONEMATODES FROM VEGETABLE GROWING AREAS IN MALAKAND DIVISION OF

KHYBER PAKHTUNKHWA AND THEIR ORGANIC BIOCONTROL WITH PLEUROTUS SPECIES

By

Muhammad Rizwan Liaqat

Objectives

To identify plant parasitic nematodes from different vegetable growing areas of Malakand Agency.

To test in vitro bio-control efficacy of Pleurotus spp.

What are nematodes The Greek word nematode means "threadlike"

Nemat=thread ode=likeNematodes are unsegmented roundworms.Kingdom Animalia - Phylum Nematoda

WHY NEMATODES ARE IMPORTANT??

Nematodes are Economically Important to Agriculture!

"Annual crop loss estimates caused by plant parasitic nematodes are roughly $80 billion worldwide.

Most abundant animal life form... "If nothing but nematodes were present

there would be an outline of the Earth and many of its features

"Reproductive Potential (Egg production)

Anguina 25,000 eggs (2,000/wk)

Meloidogyne 500-1500 eggs

Nematode Groups

Marine, Freshwater, Soil Plant Parasites Microbivores ("free-living“) Fungivores Omnivores (feed on multiple sources) Predators (on nematodes, protozoa,etc.) Animal Parasites - Vertebrate (50 species attack humans) Nematode parasites of insects (entomopathogenic)

Root Knots

INTRODUCTION

During 1984 to 1988 the nematodes infestation increased to such a level that tomato and other solaneceious crops fail to grow in the whole Malakand and Swat Region (Ali khan and Haji Muhammad Agricultural Officers Dargai).

Chemical control of nematodes is not possible and not justifiable. As it were not useful against nematodes which are living inside the soil and also dangerous to living beings and environment.

Bio control is environment friendly and

durable method to control nematodes.

Different fungi were applied in past as bio control agent against nematodes (but harmful).

Pleurotus spp which are edible mushroom, have also used as good source of bio control agent against nematodes.

MATERIALS AND METHODS

Soil samples collection from fields. Processing of samples. Quantitative analysis Qualitative analysis Temporary Mounts Killing nematodes Permanent slide mounts

Photomicrograph of eelworm and cyst nematodes.

Collection of Pleurotus Species.

Inoculation of Pleurotus spp. in Malt Extract broth media.

Observation on growth of the Pleurotus spp.

Extraction of natural products. Application of the organic solvent to the nematodes in Petri

plates.

Data analysis.

MATERIALS AND METHODS

RESULTS

Table. 01. Qualitative and Quantitative analysis of plant parasitic nematodes

S. No. LocalityNo. of nematodes per

gramGenera isolated

1. Jabben Turbela Pull 0.86Pratylenchus Xiphenemalongidorous,

Tylenchorhynchus Helicotylenchus

2. Jabben 0.33Helicotylenchus

Pratylenchus Xiphinema

TylenchorhynchusTylenchus

longidorous,Hoplolaimus

3. Dobandy Varteer 0.16Ditylenchus

ParalongidorousTylenchorhynchusHelicotylenchus

Psilenchus

4. Dobandy 0.88Helicotylenchus

Pratylenchus XiphenemaTylechus

ParalongidorousDitylenchusAphelenchus

5. Dobandy 2 0.45Helicotylenchus

XiphenemaTylenchus

longidorous,HoplolaimusDitylenchusAphelenchus

6. Palona 0.34 HelicotylenchusXiphenema

7. Heroshah 0.43HelicotylenchusPsilenchusAphelenchusParalongidorousTylenchusXiphenema

8. Haryan Kohat 0.55HelicotylenchusXiphenemalongidorous,Paralongidorous

Table 02.Qualitative and percentage analysis of free-living and predacious nematodes in Dargi, Malakand.

S. NO.

Locality Free Living Nematodes

Percentage

1. Jabban PullDorylaimidsCephalobidsAcrobeloidesRhabditids

25.5%34.0%4.2%5.2%

2. JabbanDorylaimidsRhabditidsCephalobids

25%6.8%7.9%

3. Dobandy VertarDorylaimidsCephalobids

16.2%23.8%

4. DobandyMononchidsDorylaimids

8.9%15.8%

5. Dobandy 2DorylaimidsCephalobidsRhabditids

20%7.2%10.9%

6. PalonaDorylaimidsRhabditids

35.5%15.5%

7. HeroshahDorylaimidsRhabditids

16.2%8.1%

8. Haryan KohatDorylaimidsRhabditids

18.6%6.4%

Table 03. Frequency of occurrence of plant parasitic nematodes in each locality in Dargi, Malakand.

S. NO. Genera isolatedFrequency of

Occurrence (%)

1 Helicotylenchus 100%

2 Xiphinema 71%

3 Pratylenchus 57%

4 Ditylenchus 57%

5 Tylenchorhynchus 28%

6 Aphelenchus 28%

7 Hoplolaimus 27%

8 Tylenchus 25%

9 Psilenchus 16%

10 Longidorous, 13

11 Paralongidorous 11

12 Cyst nematodes 7%

Hoplolaimus seinhorsti Luc, 1958

Genus Helicotylenchus steiner, 1945

Genus Xiphinema Cobb, 1913

Genus Longidoroidea Thron, 1935

Genus Paralongidorus.major verma, 1973.

Genus Pratylenchus Filipjev, 1936

Genus Tylenchorhynchus

Genus Tylenchus Bastian, 1865

Genus Psilenchus de Man, 1921

Genus Aphelenchulus Cobb, 1920

Genus Ditylenchus Filipjev, 1936

Genus Mononchus Cobb. 1918.

Cyst (Heterodera spp.):

Effect of extracts from Pleurotus florida on nematodes mortality.

Concentration

Pleurotus florida Extracts

Fruiting bodyMushroom

wasteBroth

100 ppm  1 Hour (T1) 14 26 284 Hour (T2) 28 31 27

24 Hour (T3) 65 34 39Average 36 30 31200 ppm  

1 Hour (T1) 22 50 414 Hour (T2) 55 57 55

24 Hour (T3) 67 57 70Average 48 55 56300 ppm  

1 Hour (T1) 58 58 514 Hour (T2) 77 69 73

24 Hour (T3) 95 69 86Average 53 65 70400 ppm  

1 Hour (T1) 88 66 764 Hour (T2) 91 79 93

24 Hour (T3) 99 79 97Average 93 74 88

Effect of extracts from Pleurotus ostreatus on nematodes mortality.

Effect of extracts from Pleurotus citronopileatus on nematodes mortality.

Conclusion1) Our finding confirmed that Dargi region is heavily infested

by plant parasitic, cyst and root-knot nematodes.

2) They directly and indirectly infect crops (by acting

as a virus vectors).

3) Extracts from three species of Pleurotus (P. florida, P. ostreatus,

P.citronopileatus.) have been found effective against nematodes

in in vitro.

4) Extracts of P. citronopileatus was found even more effective in

nematode killing at lower level.

RECOMMENDATIONS AND FUTURE WORK: 

Pleurotus species can be used for the organic control of nematodes.

Extracts from these fungi should be further tested under field

conditions.

Molecular identification of the parasitic nematodes in the surveyed

area should be carried out. This will further confirm these results.

Chemical analysis would find the presence of the nematicidal

compounds in these fungi.

Theses extracts should be tested against different parasitic

nematodes under pot and field conditions.

Pleurotus species can be used for the organic control of nematodes.

Extracts from these fungi should be further tested under field

conditions.

Control Nematodes Save the Crops

ACKNOWLEDGMENTS

First of all I am thank full to Allah Almighty.

I am highly indebted to my Advisor Prof. Dr. Saifullah.

All Faculty members, staff and my student fellows, of Plant Pathology Department, The Agricultural University Peshawar, Khyber Pakhtunkhwa.

THANKYOU