TITLE

EFFECT OF LIGHT AND TEMPERATURE ON IN VITRO

CULTURE OF PEPPERS.

Hamna Sehar
2016-ag-1246
M.phil. Biotechnology
Supervisor:
Dr.Shah Nawaz-ur-Rehman
Introduction
Tissue Culture
 Growth of tissues or cells separate
from the organism.
 Tissue culture is an important tool for
the study of the biology of cells from
multicellular organisms.
 Micropropagation technique is used in
tissue culture.
Plant Tissue Culture (PTC)
 PTC is a collection of technique used to maintain
or grow plant cells, tissues or organs, Under sterile
condition on a nutrient culture medium of known
composition.
A member of nightshade family. Solanaceae
Capsicum annuum, bell, sweet, or chili
pepper
Cultivated varieties are perennial
herbaceous plants in the Solanaceae
Plant description:
multi-branched plant
growing up to 75 cm in
cultivated varieties
shrubby in appearance
Plant Characteristics
 Herbaceous perennial
 Species: annuum longum group
 Leaves: ovate, smooth, medium green in color,
alternate arrangement
 White colour self pollinated flower
 TYPES OF PTC
Seed culture Meristem culture
Embryo culture

Cell culture Plant tissue culture Protoplast

culture

Callus culture Organ culture

Pollen culture
Overall process of PTC
 Selection of explants.
 Sterilization of explant.
 Plant transferred to solid culture
media.
 Addition of hormone.
 Placed in another media.
 Transplantation.
Selection of explant
 The selected explant is either haploid or diploid
 The plant growth can be achieved in two ways:
 1. shoot directly by appropriate media
 2. By somatic embryogenesis
Sterilization
 SterilizationMethods Used in Tissue
 Culture Laboratory – All the
materials, e.g., vessels, instruments,
medium, plant material, etc., used in
culture work must be free from
microbes
STERILIZATION TECHNIQUES
 Sterilization is achieved by one of the
following approaches:
 Dry heat treatment
 Flame sterilization
 Autoclaving
 Filter sterilization
 Wiping with 70% ethanol
 Surface sterilization.
Culture media
Explants are then usually placed on the
surface of a solid culture medium, but are
sometimes placed directly into a liquid
medium, when cell suspension cultures are
desired
Culture media are generally composed of
inorganic salts plus a few organic nutrients,
vitamins and plant hormones.
Culture media cont..
 As cultures grow, pieces are typically
sliced off and transferred to new
media (subcultured) to allow for
growth or to alter the morphology of
the culture.
Steps in plant tissue culture
technique
Selection of plant

Isolation of explant

Sterilization of explant

Inoculation of explant
 Incubation

Initiation of callus

Sub culturing

Regeneration

Hardening

Transfer of plantlets to
Green house or open
field
The impact of temperature
ranges on Pepper plant:
 Temperature plays an important role for
plant organ formation at maximum for five
months with consecutive light provisions
 Soil temperature for germination is 28o C
for Growth and Development
 Temperature ranging up to 35oC for anther
dependent calli and development phases in
tissue regenerations
Influence of temperature
and light:
Red and blue light leads to the growth
parameters of leaf surface. Red lights
proved to be effective for enhanced leaf
lengths compared to blue
Applications of PTC
Can conserve endangered
species.
Can obtain viral free plant.
Hybrids can be regenerated by
protoplasm fusion.
Plant can be grown in less time.
Advantage of PTC
 Exact copy of the plant is obtained.
 Production of multiple plant without
pollinator.
 Whole plant can be genetically
modified.
 Totipotency can be used i.e whole
plant can grown by single cell.