This document discusses techniques for preserving plant specimens infected with nematodes. It begins with an introduction to nematodes, describing their characteristics. It then discusses types of plant-parasitic nematodes and their feeding behaviors. The document outlines objectives of preserving specimens and describes various techniques for preserving aerial and root parts of plants. It discusses methods such as artificial drying, air drying, microwave drying, freeze drying, glycerin, formalin, and alcohol for long-term specimen storage. The document emphasizes properly storing specimens to prevent damage and moisture to allow access as reference collections.

1. Presented To: Dr. M. Zeeshan Mansha
Presented By: Muhammad Zeeshan (05)
Course Title Plant Disease Diagnosis (403)
Department of Plant Pathology
BZU, Bahadur Campus Layyah
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Preservation of plant specimen caused by
Nematode

2. Outlines
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 Introduction
 Definition of Nematode
 Types of PPN
 Preservation Techniques
 Objectives of preservation
 Storage

3. Introduction
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 Nematodes are a diverse group of worm-like animals. They are found
in virtually every environment, both as parasites and as free-living
organisms
 They are generally minute, but some species can reach several meters
in length
 They feed above or below ground, most nematodes spend at least
part of their life cycle in the soil. While they can’t move very far
under their own power, they can swim freely in water and they move
more quickly in moist and sandy soil

4. Nematodes
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 Nematodes are
 Thin and elongated (hair-like)
 Tubular in appearance
 Majority are microscopic while a few can be viewed
with the naked eye
 Head and tail are not visually distinct
 They have a cuticle
 They have a nervous system that controls the muscle
cells

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6. Types of PPN
 Plant parasitic nematodes can be separated into
i. Aerial parasites – those feeding on above-ground parts of plants
ii. Root and tuber parasites – those feeding on below-ground parts
On the basis of their feeding behavior and motility three main groups:
• Migratory endoparasites e.g Scutellonema
• Sedentary endoparasites e.g. Heterodera spp.
• Ectoparasites e.g. Helicotylenchus spp.
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cyst
Nematode
Tylenchulus spp.
Scutellonema
Heterodera spp.

8. What is plant specimens preservation
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Specimen preservation mean long term preservation of
organisms either plant or animal in the best possible
condition so that it can be accessed in future as reference
collection for scientific purposes
Plant Preservation:
It is a process to keep plant safe from decomposition or
destruction and
Reserve them for personal or special use

9. Objectives of preservation
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 Preserved plant specimens provide us with important information
about plant diversity and distribution
 Serves as evidence of a plant's existence in time and space
 If these specimens are properly preserved and maintained, they can
last for many years
 For morphological study of particular organism

10. Preservation Techniques
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Two types of preservation
 Aerial part preservation
Root part preservation

11. Aerial part preservation
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 Aerial part preservation is mostly used for
preserving the upper or above ground parts of the
plant e.g. leaves, stem etc.
Following are the types of aerial part
preservation
 Artificial Drying
 Air Drying
 Microwave Drying
 Freeze drying

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Deformed heads of wheat and barley
due to seed gall nematode Anguina tritici
White tip disease of rice caused
by Aphelenchoides besseyi
Patchy distribution and reduced
tillering in wheat attacked by the root
lesion nematode Pratylenchus neglectus

13. Aerial part preservation
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1. Artificial Drying
 The principle of drying leaves and roots is the same to
remove moisture slowly
 while at the same time maintaining as much of the original
shape and texture as possible
2. Air Drying
 This is by far the simplest and least expensive method used
to dry leaves
 It takes little time and skill, and nearly always produces
satisfactory results

14. Aerial part preservation
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3. Microwave Drying
Using a microwave oven for drying
specimen is another method to preserve
specimen and other plant materials
 Microwave drying, which takes only a
few minutes (2-3min) in the oven,
provides material that looks fresher and
more colorful than that obtained by other
methods
 Use only glass, microwave-safe paper, or
other microwave-approved containers to
hold the specimen

15. Aerial part preservation
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 Always place a small cup of water in the
oven before cooking to prevent excessive
drying
 Pressing
 The colors and forms of many leaves and
some flowers can be preserved by placing
them between layers of newspaper and
weighting the top with a heavy flat object

16. Aerial part preservation
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4. Freeze drying
 Freeze drying plants Specimens typically
results in the most natural-looking preserved
materials
 However, this approach requires specialized
and expensive equipment and is best
accomplished by professionals

17. Root specimen preservation
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Following technique are use to preserve root
 Glycerin
 Formalin
 Alcohol

18. Glycerin
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 Glycerin, available through drug stores, can be
used to preserve root specimen
 Although roots turn brown in this process, they
will remain flexible indefinitely
 Cut root when they are dry and place them 2-4
inches deep in a mixture of 1% glycerin to 2%
water
 When the entire root is brown, remove it from the
glycerin
 Average time for this treatment is 2 to 3 weeks

19. Formalin
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 A strength of 10% formalin is best for most purposes
 If the original strength is 40%, it should be mixed at a ratio of
nine parts water to one part formalin
 The advantages of formalin over other preservatives are:
 it is inexpensive
 It is available at drugstores and chemical supply houses

20. Alcohol
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 For storage of plant specimen it should be used in the
proportion of 3 parts 95% alcohol to 1 part water
 Alcohol which has been stored in open
containers loses its strength rapidly due to
evaporation
 Strength may be tested with an alcoholmeter
Specimens which have been fixed in alcohol
should be carefully watched for signs of rotting
 it is necessary that liquid preservative be
introduced into the roots , leaves and stem
within as short a time as possible after the
nematode have been killed Alcoholmeter

21. Storage
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 When plant parts have been preserved, take
utmost care to prevent them from being
damaged
 Pack specimens in closed boxes or in sealed
plastic bags containing mothballs
 Put packets of silica gel in the boxes to absorb
any moisture in the air
 Dried plant materials are highly flammable, so
take precautions to prevent fire hazards

22. References
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 Collecting and preserving nematodes, A safrinet manual for
nematology, Edited by K.P.N. Kleynhans, Compiled by the
Biosystematics Division, ARC–PPRI, South Africa
 https://edis.ifas.ufl.edu/ep004
 Practical plant nematology, A field and laboratory guide, D.L. Coyne,
J.M. Nicol and B. Claudius-Cole

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