In vitro culture

techniques
Presented by: Dr. Alia Anwar
Invitro
• In vitro (meaning in glass, or in the glass) studies are
performed with microorganisms, cells, or biological
molecules outside their normal biological context.
• In vitro culture is a method applied for the growth and
development of plant cells, tissues, and organs that uses a
nutritive culture medium under controlled sterilized
conditions.
• This method is considered one of the most promising and
environmentally friendly biotechnological practices for the
sustainable supply of biofuels
• There are three main in vitro culture systems including
organogenesis (e.g., embryogenesis, direct and indirect shoot
regeneration), rhizogenesis, and callogenesis
organogensis
• Organogenesis is defined as the development of organs, like
roots, shoots, and flowers, either directly from an explant, or
from the callus culture. The embryo is not considered an organ
because of the absence of a vascular system and its
independent existence.
• There are three ways of organogenesis (by which adventitious
organs form):
• (1) from the callus culture,
• (2) from an explant, and
• (3) from the axillary bud.
organogenesis
• The process of organogenesis involves two steps:
dedifferentiation and re-differentiation. Dedifferentiation results
in the formation of callus from the explant tissue with
accelerated cell division. Whereas, re-differentiation causes the
development of primordia from a group of callus cells.
• The process of organogenesis is affected by three factors:
• (a) the inoculum,
• (b) the medium, and
• (c) the environmental conditions.
• So, the process can be regulated by the components of media,
the substances carried by the explant, and the endogenous
compounds produced in culture.
Rhizogenesis, and callogenesis
• Type of organogenesis by which only adventitious root
formation takes place in the callus tissues.
• Or ganoids: In some culture tissues, an error occurs in
development programming for organogenesis and an
anomalous structure is formed.
• Type of organogenesis by which only adventitious shoot bud
initiation take place in the callus tissue.
 Callus culture
• Plant callus : is a growing mass of unorganized
plant parenchyma cells. In living plants, callus cells are those cells
that cover a plant wound.
• In biological research and biotechnology callus formation is
induced from plant tissue samples (explants) after surface
sterilization and plating onto tissue culture medium in vitro (in a
closed culture vessel such as a Petri dish).
• The culture medium is supplemented
with plant growth regulators, such as 
auxin, cytokinin, and gibberellin,
to initiate callus formation or
 somatic embryogenesis.