Nematodes are elongate, cylindrical, unsegmented worms. In elongate-cylindrical species the
anterior end is usually bluntly rounded with the oral aperture (mouth opening) terminal. The
posterior end may be bluntly rounded or pointed; sometimes it is tapered to a point or it may be long and filiform. Nematode identification is needed for the purposes of understanding nematode diversity, designing efficient management strategies, avoiding spreading of exotic nematodes in quarantine materials. So many methods of nematode identification has been generated over decades. Most of them are based on morphology which is also comes under classical identification techniques, but with the up-gradation of science several methods has been evolved. Among them different techniques has been mentioned here which are mainly based on the molecular tools for identification of the nematodes in a molecular level.
Role of micro nutrients and their deficiency symptoms in Mulberrymohd younus wani
Exponential growth in population has created unpredictable pressure on our agricultural land. On one had excessive cropping to cater to the food needs have assumed importance and on other hand no collective effort is being put to replenish our soils with the nutrients which we take out of it in the shape of agricultural produce. This state of affairs sequesters our soils and renders it deficient in various macro and micro nutrients. If this state of affairs continues, time is not for off when people will die of malnutrition and hunger. In order to restore the health of our soils and enrichment with nutrients is of vital importance irrespective of the crop which is being grown. Mulberry is one such economically important crop which is cultivated for the purpose of leaf which is fed to silkworms for cocoon production thereby revenue generation for the sizable number of population for their sustained livelihood. Micronutrients play a significant role in plant growth, photosynthesis, chlorophyll formation, cell wall development, water absorption, and xylem permeability, resistance to plant diseases and enzymatic reactions and important for activities of soil microorganisms. Increase in cocoon weight when Ni and Zn fortified leaves fed to silkworms. The modulatory effect of zinc chloride enriched mulberry leaves on various aspects of silkworm such as, Proteins in various tissues like silk gland, haemolymph and in muscles of the 5th instar silk worm larvae and also on the economic parameters of the cocoon. A model needs to be framed for maintaining continuous supply of micro nutrients to obtain desired quantity and quality of mulberry leaf for successful silkworm cocoon crop and increasing overall silk productivity.
Role of micro nutrients and their deficiency symptoms in Mulberrymohd younus wani
Exponential growth in population has created unpredictable pressure on our agricultural land. On one had excessive cropping to cater to the food needs have assumed importance and on other hand no collective effort is being put to replenish our soils with the nutrients which we take out of it in the shape of agricultural produce. This state of affairs sequesters our soils and renders it deficient in various macro and micro nutrients. If this state of affairs continues, time is not for off when people will die of malnutrition and hunger. In order to restore the health of our soils and enrichment with nutrients is of vital importance irrespective of the crop which is being grown. Mulberry is one such economically important crop which is cultivated for the purpose of leaf which is fed to silkworms for cocoon production thereby revenue generation for the sizable number of population for their sustained livelihood. Micronutrients play a significant role in plant growth, photosynthesis, chlorophyll formation, cell wall development, water absorption, and xylem permeability, resistance to plant diseases and enzymatic reactions and important for activities of soil microorganisms. Increase in cocoon weight when Ni and Zn fortified leaves fed to silkworms. The modulatory effect of zinc chloride enriched mulberry leaves on various aspects of silkworm such as, Proteins in various tissues like silk gland, haemolymph and in muscles of the 5th instar silk worm larvae and also on the economic parameters of the cocoon. A model needs to be framed for maintaining continuous supply of micro nutrients to obtain desired quantity and quality of mulberry leaf for successful silkworm cocoon crop and increasing overall silk productivity.
Characteristics of Soil Eroded due to water,Characteristics of Soil Eroded due to wind,Causes- Rain and rainwater runoff,Recreational Activities, Mining,farming ,Management:-
Crop rotation,Terraces,No-till planting
insect pest of medicinal plants and their control.pptxamandeepkaur363307
description of various insects damaging medicinal plants and their control. Infestation of pest can cause severe damage to plant and affects it quality and can also cause economic damages.
Molecular marker and its application in breed improvement and conservation.docxTrilokMandal2
Molecular markers have revolutionized the field of genetics and genomics by providing valuable tools for studying genetic diversity, identifying individuals, and characterizing traits of interest. This review paper aims to explore the applications of molecular markers in breed improvement and conservation. We discuss the various types of molecular markers commonly used, such as microsatellites, single nucleotide polymorphisms (SNPs), amplified fragment length polymorphisms (AFLPs), and many more. Additionally, we examine their applications in genetic diversity assessment, parentage analysis, marker-assisted selection (MAS), and conservation efforts. The paper highlights the importance of molecular markers in accelerating breed improvement programs and enhancing conservation strategies for maintaining genetic diversity within a population.Molecular markers have had a significant impact on breed development and conservation efforts, transforming genetics and offering vital insights into genetic diversity, lineage tracing, and genotype characterization. The importance of molecular markers in improving genetic gains, facilitating breeding programs, and preserving genetic diversity for the long-term sustainability of the animal population has been underlined in this review paper. Emerging advancements in molecular marker technology show enormous potential for improving and conserving breeds. Deeper insights into the genetic basis of complex traits will be provided through GWAS, CRISPR/Cas9, gene editing technologies, and sequencing technologies, resulting in faster genetic gains. Breeders and conservationists will be able to make more informed judgments thanks to these technologies. In conclusion, molecular markers have had a significant impact on breed conservation and enhancement. Their innovations have changed the industry and given both conservationists and breeders vital knowledge. We can pave the road for more effective and sustainable genetic improvement and the preservation of biodiversity for future generations by combining the power of molecular markers with conventional breeding and conservation techniques.
Characteristics of Soil Eroded due to water,Characteristics of Soil Eroded due to wind,Causes- Rain and rainwater runoff,Recreational Activities, Mining,farming ,Management:-
Crop rotation,Terraces,No-till planting
insect pest of medicinal plants and their control.pptxamandeepkaur363307
description of various insects damaging medicinal plants and their control. Infestation of pest can cause severe damage to plant and affects it quality and can also cause economic damages.
Molecular marker and its application in breed improvement and conservation.docxTrilokMandal2
Molecular markers have revolutionized the field of genetics and genomics by providing valuable tools for studying genetic diversity, identifying individuals, and characterizing traits of interest. This review paper aims to explore the applications of molecular markers in breed improvement and conservation. We discuss the various types of molecular markers commonly used, such as microsatellites, single nucleotide polymorphisms (SNPs), amplified fragment length polymorphisms (AFLPs), and many more. Additionally, we examine their applications in genetic diversity assessment, parentage analysis, marker-assisted selection (MAS), and conservation efforts. The paper highlights the importance of molecular markers in accelerating breed improvement programs and enhancing conservation strategies for maintaining genetic diversity within a population.Molecular markers have had a significant impact on breed development and conservation efforts, transforming genetics and offering vital insights into genetic diversity, lineage tracing, and genotype characterization. The importance of molecular markers in improving genetic gains, facilitating breeding programs, and preserving genetic diversity for the long-term sustainability of the animal population has been underlined in this review paper. Emerging advancements in molecular marker technology show enormous potential for improving and conserving breeds. Deeper insights into the genetic basis of complex traits will be provided through GWAS, CRISPR/Cas9, gene editing technologies, and sequencing technologies, resulting in faster genetic gains. Breeders and conservationists will be able to make more informed judgments thanks to these technologies. In conclusion, molecular markers have had a significant impact on breed conservation and enhancement. Their innovations have changed the industry and given both conservationists and breeders vital knowledge. We can pave the road for more effective and sustainable genetic improvement and the preservation of biodiversity for future generations by combining the power of molecular markers with conventional breeding and conservation techniques.
Advances in Molecular Cytogenetics: Potential for Crop Improvement.pptxKanshouwaModunshim
Title: Exploring Advances in Cytogenetics and Molecular Cytogenetics
Description:
Delve into the intricate world of cytogenetics and its cutting-edge counterpart, molecular cytogenetics, through this insightful presentation. Understand the profound relationship between chromosome structure, behavior, and gene function, with a particular focus on their relevance to crop improvement programs.
Key Points:
Introduction to Cytogenetics: Explore the fundamental principles of cytogenetics, its historical significance, and the recent influence of molecular tools, leading to the emergence of molecular cytogenetics.
Importance in Crop Improvement: Uncover the pivotal role of molecular cytogenetics in crop improvement programs, offering insights into the structural and functional organization of genomes within chromosomes.
Karyotyping: Gain a comprehensive understanding of karyotyping, its significance in identifying chromosomal abnormalities, and its applications in studying evolutionary relationships among different taxa.
Chromosome Identification and Sorting: Learn about the techniques involved in the identification and sorting of individual chromosomes, crucial steps in cytogenetics research for various crops.
Chromosome Banding Techniques: Explore different chromosome banding techniques, such as G-Banding and C-Banding, and understand their applications in detecting structural rearrangements.
CHIAS (Chromosome Image Analyzing System): Get insights into the CHIAS software and its role in mapping and identifying chromosomes automatically.
Flow Cytometry: Discover the applications of flow cytometry in detecting and measuring physical and chemical characteristics of cells, with a focus on its relevance in chromosome research.
In Situ Hybridization: Explore the technique of in situ hybridization, particularly the fluorescent variant, and its applications in precise localization of specific DNA segments.
Genomics and Whole Genome Sequencing: Delve into the realm of genomics and whole-genome sequencing, understanding the approaches like BAC to BAC and Whole Genome Shotgun.
Case Study: Uncover a case study involving the identification of a Wheat-Psathyrostachys huashanica ditelosomic addition line, showcasing the practical applications of the discussed techniques.
Conclusion: Summarize the key takeaways from the presentation, emphasizing the role of these techniques in advancing precision breeding and crop improvement.
Taxonomy is the branch of science concerned with the classification of organisms. A taxonomic designation is more than just a name. Ideally, it reflects evolutionary history and the relationship between organisms. Traditionally, taxonomic classification has relied upon morphological features and physiological characteristics. However, for bacterial taxonomy, phenotypic approaches have proven insufficient. Unrelated bacteria can exhibit identical traits, closely related bacteria can have divergent features, and methods for accurate identification may be too cumbersome for routine use. In contrast, molecular taxonomy approaches use data derived from hereditary material and provide a robust view of genetic relatedness. Advances in technology have been accompanied by improvements in the cost, speed, and availability of molecular methods. Here, we provide a brief history of approaches to prokaryotic classification and describe how molecular taxonomy is redefining our understanding of bacterial evolution and the tree of life.
Molecular Biology research evolves through the development of the technologies used for carrying them out. It is not possible to research on a large number of genes using traditional methods
Medcrave Group - Microfluidic technologiesMedCrave
Exosomes are cell-released small membrane vesicles derived from the endolysosomal pathway with a size range of 30-150 nm. Since the first discovery in 1981, exosomes have been found to be released from various cell types and present in many biological fluids, including blood, urine, erebrospinal fluid and ascites. Significant attention has been focused on exosome molecular components (e.g. roteins, mRNA and miRNA) which have been implicated in a variety of physiological functions and pathological disease states.
Cytogenetics is the diagnostic study of the structure and properties of chromosomes and cell division, which employs various methods, one of them being "karyotyping." It has made it possible to visualize undetected chromosomal anomalies such as small portions of chromosomes and translocations of tiny parts of chromosomes to one another. Because such procedures also enabled each pair of chromosomes to be distinguished individually, it has helped to further understanding the chromosomal basis of certain important genetic disorders.This seminar highlights the importance of "karyotyping" and its importance in the diagnosis of chromosomal disorders.
Introduction to HUMAN CHROMOSOME ANALYSIS: Conventional Karyotyping Method (G...SABARI KRISHNAN B. B.
This text is a report made on behalf of the training session organised by Dr M. Jeevan Kumar, PhD., Research Assistant Professor, Department of Genetic Engineering, SRM Institute of Science and Technology, Kattankulathur. The report covers the laboratory practices involved in karyotyping (GTG Banding) human chromosome from whole blood, the explanation to each step of karyotyping, the details about the functions of each reagent, reagent preparation protocols, etc. Karyotyping was done using GenASIs BandView software. The text involves invaluable information about the landmarks of each chromosome in a much simplified and organised way and several symbols approved by the ISCN used in karyotyping routine. Wishing all the very best to the readers and young scientists, for whom this text will find worthful.
A micro-array is a tool for analyzing gene expression that consists of a small membrane or glass slide containing samples of many genes arranged in a regular pattern.
This was made by me while I was in Masters. I have made few animations. I hope it makes understanding better.
The content is made by searching through internet and referencing books. I do not claim any content in whole presentation except the animations made on the subject.
Exosomes represent a novel mode of cell communication, involved in material transport, signal transmission, and gene expression. Studies have found that ncRNAs in exosomes such as miRNAs can mediate communication between diverse cell types, including immune cells. Cellular exchanges of RNA via exosomes contribute to adaptive and innate immune responses, influencing biological reproductive developmental processes and disease onset, including cancer, neurodegenerative diseases, and viral infections. Sequencing technology has advanced successfully to become one of the most used routine tools for life science research. Transcriptome profiling that uses sequencing technologies can supply valuable information. Here we bring you a standardized workflow and protocol for the isolation of exosomes and downstream analysis of exosomal RNA. https://www.cd-genomics.com/exosomal-rna-sequencing.html
If a microbiologist is studying bacteria that premeditate, or break down, toxic wastes and wants to know which specific genes are active when that bacterium is degrading, say, PCBs, he would likely use a tool called the DNA microarray.
Microarrays enable scientists to monitor the activities of hundreds or thousands of genes at once. All microarrays (also called DNA chips or gene chips) work on the basic principle that complementary nucleotide sequences in DNA (and RNA) match up like the two halves of a piece of Velcro coming together.
Pattern of gene activity on a microarray chip.
A microarray consists of an orderly arrangement of bits of genetic material in super-tiny spots laid down in a grid on a suitable surface, often a glass slide with a specially chemically treated surface.
Similar to Nematode identification techniques.pptx (20)
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
1. Recent Advancement in Nematode
Identification Techniques
Sabyasachi Ray
MSc. 1st year Second Semester
Dept. of Agricultural entomology
Bidhan Chandra Krishi Viswavidyalaya
Year- 2020
2. THE NEMATODES
• Aciliated
• Pseudocoelomic
• Unsegmented
• Bilaterally symmetrical
• Triploblastic
• Vermiform metazoa
• Generally dioecious
• Well developed digestive & reproductive and sensory systems
• A less developed excretory system
• Lacking in circulatory & respiratory systems
• Serpentine movement on the dorsoventral plane.
3. Purposes of nematode identification
To understand
nematode
diversity
To design efficient
management
strategies.
To avoid spreading
of exoticnematodes
in quarantine
materials
5. Classical Morphological Identification
• Nematode diagnosis and taxonomy have
traditionally relied on morphological and
anatomical characters using light microscopy
• Important morphological identification characters
in nematodes include shape of head, number of
annules, body length, length of stylet, shape of
stylet knob, structure of lateral fields,
presence/absence and shape of spermatheca,
shape of female tail terminus, shape and length of
spicule and gubernaculum.
7. Measurements for Identification
• n= Number of specimen
• L= Total bodylength in mm or µ
• a= Body length/greatest body width
• b= Body length/distance from anterior end to junction of
oesophagous and intestine
• b'= Body length / distance from anterior end to posterior
end of oesophageal glands
8. •c= Body length/tail length
•c'= Tail length / body width at anus or cloaca
•V= Distance of vulva from anterior end* 100/
body length
•T= Distance from cloaca to anterior part of
Testis* 100/body length
•Length of stoma or spear in µ
• Spicule length and gubernaculam length
9. The morphology of female perineal patterns has
been a character most frequently used in several
laboratories for the identification of Meloidogyne
species, a character located in the posterior body
region of adult females.
This area comprises the vulva-anus area
(perineum), tail terminus, phasmids, lateral lines,
and surrounding cuticular striae.
A more detailed account on root-knot nematode
perineal pattern development was given by Karssen
10. Comparison of
perineal patterns for 12
major Meloidogyne
spp. A, B: M. arenaria;
C, D: M. hapla; E, F:
M. incognita; G, H: M.
javanica; I: M.
acronea; J: M.
chitwoodi; K, L: M.
enterolobii; M: M.
ethiopica; N, O: M.
exigua; P: M. fallax;
Q, R: M. graminicola;
S, T: M. paranaensis
11. Advances in machine learning, also referred to as
deep learning or artificial intelligence (AI)
Machine learning for automated detection of
phenotypes takes place in multiple stages.
First, a large number of images (of nematodes, their
eggs, or cysts) is taken and independently annotated
by a group of experts to reduce subjectivity.
Then used to build an algorithm that learns (captures)
the salient features of the objects from the images in a
layer-wise hierarchy while masking (rejecting) the
noise in the background.
Machine learning
12. Cont.
The pattern of interest in the in-put images is then
reconstructed using a network model with a
supervised learning scheme.
Akintayo et al. designed a novel end-to-end
Convolutional Selective Autoencoder (CSAE) to
identify soybean cyst nematode (SCN) eggs in
dierent backgrounds to cover for variations in
background noise across samples from different
sources.
Another AI technique developed by Hakim et al.
using Caenorhabditis elegans called WorMachine
13. Qazi et al. demonstrated that eggs of different
helminths revealed characteristic fluorescence when
illuminated. at dierent wavelengths ranging from
white light to infrared
They also showed that differences in florescence
lifetime values (decay in florescence intensity) were
diagnostic of the species considered, Ascaris
lumbricoides and A. suum.
Qazi et al. concluded that spectroscopic features and
lifetime value measurements of autofluorescence in
nematodes are promising tools in the taxonomy of
these organisms.
Autofluorescence
14. As shown in Fig. a, the egg of Ascaris lumbricoides can
easily be visualized through light microscopy. Moreover,
they are fluorescent in the visible region when illuminated
with 390 nm (Fig. b), and 560 nm laser light (Fig. c).
15. A) Confocal map of Ascaris suum juvenile, showing red
fluorescence from outer membranes after 532 nm laser
excitation (B) Confocal image of Ascaris
lumbricoides juvenile, showing red fluorescence after
excitation with 561 nm.
16. LIMITATIONS
• Lack of clear variation among closely related taxa
• Differences of some of these morphological and morphometric
characters are subtle, subjective, and have overlapping characters
• Show intraspecific variation
• Requires well trained and experienced nematode
taxonomists
• Lack of interest of young scientists in classical taxonomy
17. Isozyme Analysis
First non-morphology-based method.
Involves the extraction of soluble proteins from whole nematodes
in buffer solutions, resolving the resulting extracts by starch or
polyacrylamide gel electrophoresis followed by staining for
specific enzymes.
Multi-locus Enzyme Electrophoresis (MEE), relies on the
migration patterns of isozymes, owing to differences in electrical
charge, molecular weight, and conformation stemming from slight
variations in amino acid compositions.
Several isozyme systems have been used, nonetheless,
carboxylesterase/esterase EST proved to be the most useful in
discriminating Meloidogyne species.
18. Two-Dimensional Gel Analyses
The technique allows resolution of complex protein mixtures by
charge using isoelectric focusing in one-dimension followed by
mass-based resolution in a dimension perpendicular to the first.
The resolution pattern is then compared among isolates to determine
similarities/dierences,
which can be scored as presence/absence for phenetic and/or
cladistic analyses of the resulting data matrix.
Navas et al. used 2-DGE to show proteomic variations among 18
root-knot nematodes representing four species.
They demonstrated that some of these variations were species-
specific,while other variations revealed evolutionary relationships
among the different species.
19. Mass Spectral Analysis
Matrix-assisted laser desorption/ionization (MALDI) is an ionization
technique, which uses laser energy-absorbing matrix to generate
gaseous ions from large molecules in solid state.
Time of flight mass spectrometer (ToF-MS) measures the time taken
by these ions to reach the detector as determined by the mass/charge
(m/z) values, with smaller and/or more charged ions travelling faster.
MALDI-ToF-MS is able to detect protein/peptide ions or protein
profiles that are diagnostic to the taxa being considered.
Perera et al. used intact second stage juveniles (J2s) and/or proteins
extracted from these using various organic solvents and discriminated
between Anguina tritici, A. funesta and M. javanica based on unique
peaks in their spectra and/or the spectral profiles.
20. Serological Analyses
The development of the hybridoma technique by Kohler and Milstein
raised the hope of the nematology community to develop mAbs for
diagnostic purposes.
The technique involved isolating mature B-cells from animals
immunized with nematode antigens, fusing these B-cells with mouse
lymphoid tumor cells to produce hybridomas that can be maintained
indefinitely in vitro for continuous production of the antibodies.
mAbs provide more specificity depending on the immunogen the
antibodies were raised against.
mAbs were raised against a variety of agriculturally important
nematodes including H. glycines, M. incognita, G. rostochiensis and
G. pallida using the hybridoma technique.
Schots et al. reported that some mAbs differentiated between G.
rostochiensis and G. pallida isolates.
21. DNA Based Analysis
Ribosomal DNA
A vast amount of examples of nematode diagnosis has
mostly been based on amplification of target DNA by
PCR using species-specific primers.
One of the approaches to design DNA-based markers that
can aided diagnosis of nematodes has been based usually
on conserved regions in the ribosomal DNA (rDNA)
cistron, i.e., the external transcribed spacer (ETS),
internal transcribed spacers 1 and 2 (ITS1 and ITS2), and
the intergenic spacer regions 1 and 2 (IGS1 and IGS2)
22. Cont.
Sequences that are divergent among nematode
species and conserved within several isolates
of a same species make ideal target for
designing species-specific primers.
Ribosomal DNA regions are multicopy genes
and provide sequences with enough variation
that can be used for diagnosis and
phylogenetic relationships among species.
23. Mitochondrial and satellite DNA
Small circular molecules ranging from 12 to 20 kilobases .
Divergences in mtDNA sequences due to insertions, deletions, and
accelerated ratio of mutations compared with nuclear DNA have
provided target markers suitable for discriminating nematode
species.
Satellite DNAs (satDNAs) are highly repeated tandem arrays of
short sequences ranging from 70 to 2000 bp.
It has different signature sequences, copy numbers, length, and
polymorphic regions that can be explored to find species-specific
markers.
Such PCR-based detection using satDNA markers in nematode
diagnosis represents a target option for designing diagnostic
primers.
24. RFLP, AFLP, RAPD, SCAR
(RFLP), a method that uses restriction enzymes to digest
whole genomic DNA or an amplified segment of it.
It generate DNA banding patterns according to divergences
in sequences among isolates.
This technique can also be coupled with DNA hybridization
with radioactive or nonradioactive labeled probes.
This method is less used nowadays due to technical
complexity and the need for a large amount of target DNA.
25. Cont.
DNA band obtained from random amplified
polymorphic DNA (RAPD) or amplified fragment
length polymorphism (AFLP) gels, with posterior
cloning and sequencing of bands differentiate across
related species and their conversion into species-
specific sequence characterized amplified region
(SCAR) markers.
SCAR-based markers and rDNA-based specific
primers have been used to diagnose nematodes with
either conventional or real-time PCR (q-PCR) .
26. Cont.
Numerous primers and approaches used for diagnosis of
nematodes using conventional and quantitative PCR
were designed based on several target regions in the
nematode genome(e.g., SCAR, rDNA, ITS, D2-D3
segment, IGS, among others).
In particular, successful SCAR markers have been
developed for diagnosing some of the major tropical
Meloidogyne spp. associated with important crops such
as coffee, guava, and grapevine, including M. arenaria ,
M. incognita , M. paranaensis, M. exigua , M.
enterolobii ,M. arabicida, M. izalcoensis , and M.
ethiopica etc.
27. Nematode species Target region Method
Meloidogyne spp. PCR
M. arabicida and M. izalcoensis SCAR PCR
M. arenaria SCAR PCR
M. chitwoodi IGS, SCAR PCR
M. exigua SCAR PCR
M. enterolobii mtDNA, SCAR PCR
M. ethiopica SCAR PCR
M. fallax IGS, SCAR PCR
M. graminis ITS PCR
M. hapla satDNA, SCAR, IGS PCR
M. incognita SCAR PCR
M. javanica SCAR PCR
M. marylandi 28S D2-D3 PCR
M. naasi ITS, 28S D2-D3 PCR
M. paranaensis SCAR PCR
28. Other Plant Parasitic Nematodes
Nematode species Target region Method
Bursaphelencus xylophilus satDNA PCR
B. xylophilus satDNA qPCR
B. xylophilus heat shock protein qPCR
Ditylenchus destructor D.
dipsaci
rDNA PCR/qPCR
H. glycines rDNA qPCR
H. schachtii ITS PCR
H. glycines SCAR qPCR
Pratylenchus penetrans rDNA qPCR
SCAR—sequence characterized amplified region; IGS—intergenic
spacer region; ITS—internal transcribed spacer; mtDNA—
mitochondrial DNA; satDNA—satellite DNA; PCR—polymerase
chain reaction; qPCR—quantitative real-time PCR.
29. qPCR and barcoding
• Quantitative PCR (q-PCR) is a technique that amplifies and quantifies
nucleic acids simultaneously
• Its advantage over conventional PCR is that it is fast, sensitive and does
not need post amplification processing of samples normally seen in
conventional PCR
• Application of q-PCR in nematode diagnosis using rDNA target or other
marker has been showed for major nematode species, including M.
incognita, M. chitwoodi, M. fallax, M. javanica, Bursaphelenchus
xylophilus, Globodera rostochiensis, and G. pallida.
• DNA barcode for nematode taxonomy related to a DNA sequence of a
particular region in the genome as a mean to give unique signature
(barcode) for the identification of nematode species .
• This approach has not been widely accepted since there has not been an
unique DNA locus that can define the limits of species boundary.
30. Concluding Remarks
The accurate identification of nematodes to species and
subspecies levels is essential for their control and is a
prerequisite to meaningful research.
Many nematode species are easily identified based on distinct
morphological characters and restricted host ranges.
Several species are difficult to identify due to their similarity to
other species or poor taxonomic descriptions.
The future prospects in nematode taxonomy and diagnostics are
dependent on molecular-based and biochemical based methods
and tools that will discriminate not only at the species level but
also at the level of host races, thereby opening up opportunities
for more focused management strategies.
31. References
• Carneiro, R. M. D. G., Lima, F. S. O. and Correia, V. R. Methods and
Tools Currently Used for the Identification of Plant Parasitic
Nematodes. Nematology - Concepts, Diagnosis and Control. Chapter 2;
19-35
• DiGennaro, P., Baniya, A. and Bogale, M. 2020. Nematode
Identification Techniques and Recent Advances. Plants 2020, 9, 1260;1-
15.
• Introductory Plant Nematology- P Parvatha Reddy
• Qazi, F., Khalid, A., Poddar, A., Tetienne, J. P., Nadarajah, A., Medina,
A. A., Shahsavari, I., Shukla, R., Prawer, S., Ball, A. S. and Hanic, S.
T.2020. Real-time detection and identification of nematode eggs genus
and species through optical imaging. Scientific Reports | (2020) 10:7219
• Textbook of Introductory Plant Nematology- Raman k. Walia & Harish
K. Bajaj