Prabhat MBB-604 Assignments ppt.pptx drought

Assignment Presentation
On
Micro-propagation of commercially important plant species;
plant multiplication, hardening, and transplantation; genetic
fidelity; and bioreactors
Presented To
Dr. N. A. Khan
Professor & Head, Department of MB&B,
ANDUA&T, Kumarganj, Ayodhya-224229
Presented By
Mr. Prabhat Kumar Singh
Ph. D. Agril. Biotechnology
ID. No. A-10047/17/22
Course Title: Commercial Plant Tissue Culture
(MBB-604)
Dr. Hemant Kumar Yadav
Assistant Professor, Department of MB&B,
ANDUA&T, Kumarganj, Ayodhya-224229

Micropropagation: Stages, Types, Applications
 Micropropagation is the rapid vegetative propagation of plants under in vitro
conditions of high light intensity, controlled temperature, and a defined
nutrient medium.
 The technique has been applied to a substantial number of commercial
vegetatively propagated plant species.
 Plants can be propagated by sexual (through the generation of seeds) or
asexual (through multiplication of vegetative parts) means.
 Asexual reproduction through multiplication of vegetative parts is the only
method for the in vivo propagation of certain plants, as they do not produce
viable seeds e.g. banana, grape, fig, and chrysanthemum.

Stages of Micropropagation
The micropropagation process is conveniently divided into stages (based on
Murashige, and Debergh and Maene;
 Stage 0:
 This is the initial step in micropropagation and involves the selection and
growth of stock plants for about 3 months under controlled conditions.
 Stage I:
The explants are established in a suitable culture medium. This stage involves the
following steps:
Isolation of the explant
Surface Sterilization
Washing
The explant is established on an appropriate culture medium.

Conti...
 Stage II:
 It is in this stage, the major activity of micropropagation occurs in a defined culture
medium. Stage II mainly involves the multiplication of shoots or rapid embryo
formation from the explant. A growth chamber set at 20–24 °C is used, with a 2000- to
4000-lux light intensity, and a lighting period of 16 hours or so.
 Stage III:
 This stage involves the transfer of shoots to a medium for rapid development into
roots. Sometimes, the shoots are directly planted in soil to develop roots. In vitro
rooting of shoots is preferred while simultaneously handling a large number of species.
 Stage IV:
 This stage involves the establishment of plantlets in soil. This is done by transferring the
plantlets of stage III from the laboratory to the environment of the greenhouse. For
some plant species, stage III is skipped, and un-rooted stage II shoots are planted in
pots or in the suitable compost mix.

Applications and Advantages of Micropropagation
 Plants in large numbers can be produced in a short period.
 Large amounts of plants can be maintained in small spaces. This helps to save
endangered species and the storage of germplasm.
 The micropropagation method produces plants free of diseases.
 Increased yield of plants and increased vigor in floriculture species are
achieved.
 Fast international exchange of plant material without the risk of disease
introduction is provided. The time required for quarantine is lessened by this
method.
 The micropropagation technique is also useful for seed production in certain
crops as the requirement of genetic conservation to a high degree is
important for seed production.

Disadvantages of Micropropagation
The disadvantages of micropropagation are given below:
 The plants produced are not autotrophic.
 It cannot be implemented in all the crops.
 The plants find a problem acclimatizing to the natural environment.
Limitation of Micropropagation
 Micropropagation techniques require intensive labor and this often limits
their commercial application. Automation can reduce the labor required.

Genetic fidelity
 Genetic fidelity is the accurate transfer of genetic material from one genertaion
to next without any modifications. It is important in tissue culture techniques,
which can produce disease-free and true-to-type plants.
 It is important to test the genetic fidelity of micropropagated plants before
applying the micropropagation protocol on a commercial sacle.
 This is crucial for ensuring that the propagated plants are true-to-type and
consistent with the original plant variety.
 Ensuring high genetic fidelity is crucial for commercial plant tissue culture
because it guarantees that the plants produced are true-to-type, meaning they
retain the desired traits of the parent plants, such as disease resistance, yield,
and quality.

Key factors influencing genetic fidelity include
 Cellular and Molecular Stability:
 The culture conditions (like media composition and growth regulators) must be
optimized to prevent genetic mutations or somaclonal variation.
 Techniques for Verification:
 Techniques such as DNA fingerprinting, molecular markers, or karyotyping are
used to verify genetic stability and ensure that the tissue-cultured plants are
genetically identical to the parent plants.
 Management Practices:
 Proper management practices, including careful monitoring and control of
culture environments and regeneration protocols, help maintain genetic
fidelity.

Strategies to Ensure Genetic Fidelity
 Selection of Explants:
 Using explants from meristematic tissues (like shoot tips) reduces the risk of
somaclonal variation.
 Optimized Culture Conditions:
 Maintaining optimal media compositions, hormone levels, and culture
conditions to minimize stress, which can induce genetic variation.
 Molecular Markers:
 Techniques such as RAPD, SSR, and AFLP can be used to assess genetic fidelity
in vitro before plants are scaled up.
 Cryopreservation:
 Long-term preservation of germplasm through cryopreservation ensures that
the genetic material remains unchanged over time.

Scaling Up Tissue Culture
 Scaling up tissue culture for commercial production involves transitioning from
small-scale laboratory conditions to industrial-scale operations. Key
considerations include:
 Automation: Implementing automation in the culture process (e.g., automated
subculturing, bioreactors) can significantly increase throughput and
consistency while reducing labor costs.
 Bioreactors: Using liquid culture systems in bioreactors allows for large-scale
propagation of plantlets with reduced space requirements and enhanced
nutrient uptake, leading to faster growth.
 Standardization: Standardizing protocols for media preparation, explant
sterilization, and subculturing ensures uniformity and reduces variability in
large-scale operations.

Cost Reduction in Commercial Tissue Culture
 Bulk Media Preparation:
 Preparing media in bulk can reduce costs associated with reagents and
labor.
 Energy Efficiency:
 Utilizing energy-efficient lighting and climate control systems can reduce
the cost of maintaining growth chambers.
 Reusable Culture Vessels:
 Implementing reusable culture vessels instead of disposable ones can lead
to significant cost savings over time.
 Labor Optimization:
 Training workers in efficient techniques and utilizing semi-automated
systems can reduce labor costs.

Bioreactors using in micropropagation
 What Are Bioreactors?
 A bioreactor is a controlled environment designed to support the growth
and development of biological organisms, such as plant cells, tissues, or
organs, under sterile conditions. In micropropagation, bioreactors allow the
cultivation of large numbers of plantlets or plant tissues in a liquid or semi-
liquid medium.
 It provides optimum conditions like temperature, pH, substrate, oxygen, etc
required for the culturing of cells producing desired products. Simple
stirred-tank bioreactor and sparged stirred-tank bioreactor are the two
types of bioreactors used for this purpose.

Types of Bioreactors in Micropropagation
1. Temporary Immersion Bioreactors (TIB)
2. Continuous Immersion Bioreactors
3. Air-Lift Bioreactors
4. Bubble Column Bioreactors
5. Balloon-Type Bubble Bioreactors

Types of Bioreactors in Micropropagation
 Temporary Immersion Bioreactors (TIB):
These bioreactors allow the plant tissues to be periodically immersed in the liquid
nutrient medium. This approach reduces the risk of hyperhydricity (a condition
caused by excessive water uptake) and promotes better gas exchange, leading to
healthier and more vigorous plantlets.
 Continuous Immersion Bioreactors:
In these systems, plant tissues are constantly submerged in the liquid medium.
However, this can sometimes lead to issues like hyperhydricity, so it's less
commonly used than temporary immersion systems for plant tissues.

Conti...
 Air-Lift Bioreactors:
Air-lift bioreactors use air bubbles to circulate the medium and provide aeration.
The movement of air and medium helps prevent the settling of plant tissues and
ensures even nutrient distribution.
 Bubble Column Bioreactors:
Similar to air-lift bioreactors, bubble column bioreactors rely on the rise of air
bubbles to mix the medium and ensure proper oxygenation and nutrient
distribution.
 Balloon-Type Bubble Bioreactors:
These are modified bubble column bioreactors with a balloon-like structure at the
top, designed to increase the surface area for gas exchange, making them more
efficient in handling larger volumes.

Advantages of Using Bioreactors in Micropropagation
1. Scalability
2. Reduced Labor Costs
3. Improved Plant Quality
4. Efficient Use of Resources
5. Reduced Contamination Risk
6. Faster Growth Rates

Challenges and Considerations
1. Initial Setup Cost
2. Optimization of Conditions
3. Management of Hyperhydricity
4. Technical Expertise

Prabhat MBB-604 Assignments ppt.pptx drought

Prabhat MBB-604 Assignments ppt.pptx drought

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