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Horticulture
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UPCATET PG / M. Sc. Agriculture

Molecular markers & genomics

Molecular Marker: A DNA sequence associated with a specific gene or trait, used to identify individuals with that trait.

Types of Markers:

  • Morphological markers – Color, shape
  • Biochemical markers – Isozymes
  • Molecular markers – RFLP, RAPD, SSR, SNP

Marker

Full form

Key Point

RFLP

Restriction Fragment Length Polymorphism

First DNA marker used

RAPD

Random Amplified Polymorphic DNA

Fast, less reliable

SSR

Simple Sequence Repeat (Microsatellite)

Highly reproducible

SNP

Single Nucleotide Polymorphism

Single base change marker

Applications:

  • Genetic diversity analysis
  • Marker-Assisted Selection (MAS)
  • Gene mapping
  • Identification of hybrids

Fact:

  • Human Genome Project completed in 2003.
  • Rice genome sequencing completed in 2005 (IRGSP).

 

Biofertilizers & biopesticides

Biofertilizers: Microorganisms that enhance nutrient availability to plants.

Type

Microorganism

Function

Nitrogen fixers

Rhizobium, Azospirillum

N fixation

Phosphate solubilizers

Pseudomonas, Bacillus

P solubilization

Potassium solubilizers

Bacillus mucilaginosus

K availability

Mycorrhizae

Glomus spp.

P absorption

Cyanobacteria

Anabaena, Nostoc

N fixation in rice fields

 

Biopesticides: Biological agents used to control pests/diseases.

Type

Example

Target

Bacterial

Bacillus thuringiensis

Lepidopteran insects

Viral

NPV, GV

Caterpillars

Fungal

Beauveria bassiana, Metarhizium

Soil insects

Botanical

Neem (Azadirachtin)

Broad-spectrum repellent

 

Molecular diagnostics

Used for:

  • Pathogen detection
  • Genetic purity testing
  • Disease resistance screening

Techniques:

  • PCR (Polymerase Chain Reaction) – Amplifies DNA.
  • ELISA (Enzyme Linked Immunosorbent Assay) – Detects proteins/antigens.
  • Southern/Northern/Western Blotting – Detect DNA/RNA/proteins respectively.

Fact: PCR discovered by Kary Mullis (1983).

 

Genome editing technologies

CRISPR-Cas9 Technology: A genome editing tool for precise modification of DNA sequences.

  • Discovered from bacterial immune system.
  • Components:
    • Cas9 enzyme – acts as molecular scissors.
    • Guide RNA (gRNA) – directs Cas9 to target DNA.
  • Advantages: High accuracy, cheap, easy to use.

Other Genome Editing Tools:

  • ZFN (Zinc Finger Nuclease)
  • TALEN (Transcription Activator-like Effector Nuclease)

 

Biosafety and regulatory bodies (india)

Agency

Full Name

Function

DBT

Department of Biotechnology

Policy & funding

GEAC

Genetic Engineering Appraisal Committee

Approval of GM crops

RCGM

Review Committee on Genetic Manipulation

Monitors lab-level trials

IBSC

Institutional Biosafety Committee

Safety at institutional level

NBA

National Biodiversity Authority

Regulates genetic resources

Fact: Biosafety Guidelines released by DBT in 1990, 1998, 2017.

 

Important scientists & discoveries

Discovery

Scientist

Term “Biotechnology”

Karl Ereky (1919)

Restriction enzyme

Hamilton Smith

Recombinant DNA

Stanley Cohen & Herbert Boyer

Agrobacterium-mediated transformation

Jeff Schell & Marc Van Montagu

Ti plasmid

Agrobacterium tumefaciens

PCR

Kary Mullis (1983)

Golden Rice

Ingo Potrykus & Peter Beyer

Bt toxin gene

Bacillus thuringiensis

CRISPR

Jennifer Doudna & Emmanuelle Charpentier

 

Key facts for exam

  • First transgenic plant: Tobacco (1983).
  • First GM crop approved in India: Bt Cotton (2002).
  • Ti plasmid is a natural vector used in plant genetic engineering.
  • Selectable markers: nptII, hpt, bar genes.
  • Gene gun (biolistics) used for direct gene transfer into plants.
  • Agrobacterium-mediated transformation – indirect gene transfer method.
  • Cry genes in Bt – provide insecticidal properties.
  • Golden Rice – Biofortified with β-carotene (Pro-vitamin A).
  • Bt Brinjal – First GM food crop approved in India (not commercially released).
  • Somaclonal variation – Genetic variation among tissue culture plants.
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