Advantages of Artificial seeds
Artificial seeds have many advantages over the true seeds as follows:
1. Millions of artificial seeds can be produced at any time at low cost.
2. They provide an easy method to produce genetically engineered plants with desirable traits.
3. It is easy to test the genotype of plants.
4. They can potentially stored for long time under cryopreservation method.
5. Artificial seeds produce identical plants .
6. The period of dormancy of artificial seeds is greatly reduced, hence growth is faster with a shortened life cycle.

Micropropagation protocol for banana
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In vitro micropropagation of banana (Musa sp.)
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Suckers surface sterilized with 1% NaOCl for 30 minutes
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Apical meristems were isolated - cultured on MS basal medium supplemented with Benzylaminopurine (BAP) 10.0 mg/l and Indole Acetic Acid (IAA) 1.0 mg/l
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Induction of shoots within 168 days
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Kinetin 2.0 mg/l and Naphthalene Acetic Acid (NAA) 0.5 mg/l for root induction
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Acclimatization in polyhouse
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Followed by shade-house under 50% light for hardening process
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Tested for genetic Uniformity
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Transferred to field

Plant tissue culture techniques have several applications such as:
i. Improved hybrids production through somatic hybridization.
ii. Somatic embryoids can be encapsulated into synthetic seeds (synseeds). These encapsulated seeds or synthetic seeds help in conservation of plant biodiversity.
iii. Production of disease resistant plants through meristem and shoot tip culture.
iv. Production of stress resistant plants like herbicide tolerant, heat tolerant plants.
v. Micropropagation technique to obtain large numbers of plantlets of both crop and tree species useful in forestry within a short span of time and all through the year.
vi. Production of secondary metabolites from cell culture utilized in pharmaceutical, cosmetic and food industries.

Protoplasts are cells without a cell wall, but bounded by a cell membrane or plasma membrane. Using protoplasts, it is possible to regenerate whole plants from single cells and also develop somatic hybrids. The steps involved in protoplast culture are:
i) Isolation of protoplast: Small bits of plant tissue like leaf tissue are used for isolation of protoplast. The leaf tissue is immersed in 0.5% Macrozyme and 2% Onozuka cellulase enzymes dissolved in 13% sorbitol or mannitol at pH 5.4. It is then incubated overnight at 25°C. After a gentle teasing of cells, protoplasts are obtained, and these are then transferred to 20% sucrose solution to retain their viability. They are then centrifuged to get pure protoplasts as different from debris of cell walls.
ii) Fusion of protoplast: It is done through the use of a suitable fusogen. This is normally PEG (Polyethylene Glycol). The isolated protoplast are incubated in 25 to 30% concentration of PEG with Ca++ ions and the protoplast shows agglutination (the formation of clumps of cells) and fusion.
iii) Culture of protoplast: MS liquid medium is used with some modification in droplet, plating or micro-drop array techniques, Protoplast viability is tested with fluorescein diacetate before the culture. The cultures are incubated in continuous light 1000-2000 lux at 25°C. The cell wall formation occurs within 24-48 hours and the first division of new cells occurs between 2-7 days of culture.
iv) Selection of somatic hybrid cells: The fusion product of protoplasts without nucleus of different cells is called a cybrid. Following this nuclear fusion happen. This process is called somatic hybridization.

Protocol for virus free meristem tip culture
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Apical meristem tip with 1 or 2 leaf primordia are excised in sterile condition from the explant
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In vitro culture in 10 ml of solid MS medium supplemented with growth hormones
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Cultures are maintained at 24±1°C in dark for 3 days followed by normal illumination of 2400Lux
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Plantlets developed from meristem tip culture after organogenesis process, transferred to hardening process
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Transferred to field condition