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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=like
Nematodes 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. Locality
No. of nematodes per
gram
Genera isolated
1. Jabben Turbela Pull 0.86
Pratylenchus
Xiphenema
longidorous,
Tylenchorhynchus
Helicotylenchus
2. Jabben 0.33
Helicotylenchus
Pratylenchus
Xiphinema
Tylenchorhynchus
Tylenchus
longidorous,
Hoplolaimus
3. Dobandy Varteer 0.16
Ditylenchus
Paralongidorous
Tylenchorhynchus
Helicotylenchus
Psilenchus
4. Dobandy 0.88
Helicotylenchus
Pratylenchus
Xiphenema
Tylechus
Paralongidorous
Ditylenchus
Aphelenchus
5. Dobandy 2 0.45
Helicotylenchus
Xiphenema
Tylenchus
longidorous,
Hoplolaimus
Ditylenchus
Aphelenchus
6. Palona 0.34 Helicotylenchus
Xiphenema
7. Heroshah 0.43
Helicotylenchus
Psilenchus
Aphelenchus
Paralongidorous
Tylenchus
Xiphenema
8. Haryan Kohat 0.55
Helicotylenchus
Xiphenema
longidorous,
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 Pull
Dorylaimids
Cephalobids
Acrobeloides
Rhabditids
25.5%
34.0%
4.2%
5.2%
2. Jabban
Dorylaimids
Rhabditids
Cephalobids
25%
6.8%
7.9%
3. Dobandy Vertar
Dorylaimids
Cephalobids
16.2%
23.8%
4. Dobandy
Mononchids
Dorylaimids
8.9%
15.8%
5. Dobandy 2
Dorylaimids
Cephalobids
Rhabditids
20%
7.2%
10.9%
6. Palona
Dorylaimids
Rhabditids
35.5%
15.5%
7. Heroshah
Dorylaimids
Rhabditids
16.2%
8.1%
8. Haryan Kohat
Dorylaimids
Rhabditids
18.6%
6.4%
Table 03. Frequency of occurrence of plant parasitic nematodes in each
locality in Dargi, Malakand.
S. NO. Genera isolated
Frequency 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 body
Mushroom
waste
Broth
100 ppm
1 Hour (T1) 14 26 28
4 Hour (T2) 28 31 27
24 Hour (T3) 65 34 39
Average 36 30 31
200 ppm
1 Hour (T1) 22 50 41
4 Hour (T2) 55 57 55
24 Hour (T3) 67 57 70
Average 48 55 56
300 ppm
1 Hour (T1) 58 58 51
4 Hour (T2) 77 69 73
24 Hour (T3) 95 69 86
Average 53 65 70
400 ppm
1 Hour (T1) 88 66 76
4 Hour (T2) 91 79 93
24 Hour (T3) 99 79 97
Average 93 74 88
Effect of extracts from Pleurotus ostreatus on nematodes mortality.
Effect of extracts from Pleurotus citronopileatus on nematodes mortality.
Conclusion
1) 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

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Identification of phytonematodes from vegetable growing areas in malakand division of khyber pakhtunkhwa and their organic biocontrol with pleurotus species

  • 1.
  • 2. IDENTIFICATION OF PHYTONEMATODES FROM VEGETABLE GROWING AREAS IN MALAKAND DIVISION OF KHYBER PAKHTUNKHWA AND THEIR ORGANIC BIOCONTROL WITH PLEUROTUS SPECIES By Muhammad Rizwan Liaqat
  • 3. Objectives  To identify plant parasitic nematodes from different vegetable growing areas of Malakand Agency.  To test in vitro bio-control efficacy of Pleurotus spp.
  • 4. What are nematodes The Greek word nematode means "threadlike" Nemat=thread ode=like Nematodes are unsegmented roundworms. Kingdom Animalia - Phylum Nematoda
  • 5. 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
  • 6. "Reproductive Potential (Egg production)  Anguina 25,000 eggs (2,000/wk)  Meloidogyne 500-1500 eggs
  • 7. 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)
  • 9. 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.
  • 10.  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.
  • 11. MATERIALS AND METHODS  Soil samples collection from fields.  Processing of samples.  Quantitative analysis  Qualitative analysis  Temporary Mounts  Killing nematodes  Permanent slide mounts
  • 12.  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
  • 13. RESULTS Table. 01. Qualitative and Quantitative analysis of plant parasitic nematodes S. No. Locality No. of nematodes per gram Genera isolated 1. Jabben Turbela Pull 0.86 Pratylenchus Xiphenema longidorous, Tylenchorhynchus Helicotylenchus 2. Jabben 0.33 Helicotylenchus Pratylenchus Xiphinema Tylenchorhynchus Tylenchus longidorous, Hoplolaimus
  • 14. 3. Dobandy Varteer 0.16 Ditylenchus Paralongidorous Tylenchorhynchus Helicotylenchus Psilenchus 4. Dobandy 0.88 Helicotylenchus Pratylenchus Xiphenema Tylechus Paralongidorous Ditylenchus Aphelenchus 5. Dobandy 2 0.45 Helicotylenchus Xiphenema Tylenchus longidorous, Hoplolaimus Ditylenchus Aphelenchus 6. Palona 0.34 Helicotylenchus Xiphenema
  • 15. 7. Heroshah 0.43 Helicotylenchus Psilenchus Aphelenchus Paralongidorous Tylenchus Xiphenema 8. Haryan Kohat 0.55 Helicotylenchus Xiphenema longidorous, Paralongidorous
  • 16. Table 02.Qualitative and percentage analysis of free-living and predacious nematodes in Dargi, Malakand. S. NO. Locality Free Living Nematodes Percentage 1. Jabban Pull Dorylaimids Cephalobids Acrobeloides Rhabditids 25.5% 34.0% 4.2% 5.2% 2. Jabban Dorylaimids Rhabditids Cephalobids 25% 6.8% 7.9% 3. Dobandy Vertar Dorylaimids Cephalobids 16.2% 23.8% 4. Dobandy Mononchids Dorylaimids 8.9% 15.8%
  • 17. 5. Dobandy 2 Dorylaimids Cephalobids Rhabditids 20% 7.2% 10.9% 6. Palona Dorylaimids Rhabditids 35.5% 15.5% 7. Heroshah Dorylaimids Rhabditids 16.2% 8.1% 8. Haryan Kohat Dorylaimids Rhabditids 18.6% 6.4%
  • 18. Table 03. Frequency of occurrence of plant parasitic nematodes in each locality in Dargi, Malakand. S. NO. Genera isolated Frequency 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%
  • 19. 9 Psilenchus 16% 10 Longidorous, 13 11 Paralongidorous 11 12 Cyst nematodes 7%
  • 28. Genus Psilenchus de Man, 1921
  • 33. Effect of extracts from Pleurotus florida on nematodes mortality. Concentration Pleurotus florida Extracts Fruiting body Mushroom waste Broth 100 ppm 1 Hour (T1) 14 26 28 4 Hour (T2) 28 31 27 24 Hour (T3) 65 34 39 Average 36 30 31 200 ppm 1 Hour (T1) 22 50 41 4 Hour (T2) 55 57 55 24 Hour (T3) 67 57 70 Average 48 55 56 300 ppm 1 Hour (T1) 58 58 51 4 Hour (T2) 77 69 73 24 Hour (T3) 95 69 86 Average 53 65 70 400 ppm 1 Hour (T1) 88 66 76 4 Hour (T2) 91 79 93 24 Hour (T3) 99 79 97 Average 93 74 88
  • 34. Effect of extracts from Pleurotus ostreatus on nematodes mortality.
  • 35. Effect of extracts from Pleurotus citronopileatus on nematodes mortality.
  • 36. Conclusion 1) 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).
  • 37. 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.
  • 38. 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.
  • 39.  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
  • 40. 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.