Artificial Seed

Study Material
M.Sc. Ag. IV semester

Course: Principles & Practices of Seed Production
Chapter: 06 (Seed Testing)
Course Teacher:
Dr. Shailendra Singh Gaurav

Professor & Head
Dept. of Genetics & Plant Breeding
Ch. Charan Singh University Campus, Meerut-250004 (UP)
E-mail: [email protected]; Mob.+91-0412782201
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Topic: SYNTHETIC SEEDS
Synthetic seeds are defined as artificially encapsulated somatic embryos, shoot buds, cell
aggregates, or any other tissue that can be used for sowing as a seed and that possess the
ability to convert into a plant under in vitro or ex vitro conditions and that retain this potential
also after storage. In simple words synthetic seed contains an embryo produced by somatic
embryogenesis enclosed within an artificial medium that supplies nutrients and is encased in
an artificial seed covering.
The technology designed to combine the advantages of clonal propagation with those of seed
propagation and storage. The first synthetic seeds were produced by Kitto and Janick in 1982
using carrot
Why Synthetic Seeds?

In some of the horticultural crops seeds propagation is not successful due to;
• Heterozygosity of seeds particularly in cross pollinated crops
• Minute seed size eg; orchids
• Presence of reduced endosperm
• Some seeds require mycorrhizal fungi association for germination eg: orchids
• No seeds are formed
Characteristics of Synthetic Seeds

1. Large scale propagation method
2. Maintains genetic uniformity of plants
3. Direct delivery of propagules to the field, thus eliminating transplants
4. Lower cost per plantlet
5. Rapid multiplication of plants
Advantages of Synthetic Seeds Over Somatic Embryos For Propagation
1. Ease of handling while in storage

2. Easy to transport
3. Has potential for long term storage without losing viability
4. Maintains the clonal nature of the resulting plants
5. Serves as a channel for new plant lines produced through biotechnological advances
to be delivered directly to the green house or field
6. Allows economical mass propagation of elite plant varieties
Types of synthetic seeds:
i) Desiccated: The desiccated synthetic seeds are produced from somatic embryos either
naked or encapsulated in polyoxyethylene glycol (Polyoxr) followed by their desiccation.
Desiccation can be achieved either slowly over a period of one or two weeks sequentially
using chambers of decreasing relative humidity, or rapidly by unsealing the petri-dishes and
leaving them on the bench overnight to dry. Such types of synseeds are produced only in
plant species whose somatic embryos are desiccation tolerant.
ii) Hydrated: hydrated synthetic seeds are produced in those plant species where the somatic
embryos are recalcitrant and sensitive to desiccation. Hydrated synthetic seeds are produced
by encapsulating the somatic embryos in hydrogel capsules.
Procedure for Synthetic Seed Production:

The somatic embryos for synthetic seeds are produced in the lab through culturing of somatic
cells and treating with different hormones to produce root and shoot. The following are the
different steps involved in artificial seeds production;
1. Establish somatic embryogenesis

2. Mature somatic embryos
3. Synchronize and singulate somatic embryos
4. Mass production of embryos
5. Encapsulation of matured somatic embryos
6. Desiccation
7. Field planting
Somatic Embryos
Somatic embryos are bipolar structure with both apical and basal meristematic regions which are
capable of forming shoot and root, respectively.
Somatic embryogenesis is the development of embryos form vegetative cells with in vitro systems.
Specific tissues have a capacity for somatic embryogenesis in cultural systems. This allows the clonal
propagation of normally seed-propagated crops analogous to the production of apomictic seedlings.
Somatic embryos develop through stages similar to zygotic embryos, however, the final size for the
cotyledons are usually reduced and there is no development of endosperm or seed coat.
After pollination, a zygotic embryo of a dicotyledonous species develops through a series of
morphological stages termed globular, heart and torpedo. Cotyledons develop and expand as the
storage reserves of protein, starch and/or oil are deposited. At some stages before the embryo achieves
its maximum weight, it acquires the ability to tolerate drying. Then, the seed's vascular connections to
the maternal plant are severed, it stops importing nutrients and it begins to lose water. Seeds of most
crop plants can survive drying and can be stored for several years. Once they are hydrated,
germination commences culminating in the emergence of a radicle and then the mobilization of the
storage reserves by the seedling.
Procedure of Somatic Embryogenesis:

1. Petiole explants plants are surface sterilized and cultured on SH medium (Schenk and
Hildebrandt, 1972) containing 2,4-D, kinetin and many other nutrients. 2,4-D
activates the cell cycle of many cells in the petiole - those in the vascular cambium
develop into a callus, whereas some sub-epidermal cells develop into a somatic
embryo.
2. The initial somatic embryos, which are only small dense cell clusters at this stage, are
embedded in a callus mass of non-differentiated cells.
3. To liberate these pro-embryonic structures, and to stimulate the formation of more
embryos, the callus is dispersed in a liquid medium to form a suspension culture
containing 2,4-D but not kinetin
4. After 7 days, the suspension is sieved and transferred to solid medium lacking 2,4-D
On this medium the embryos develop through morphological stages that appear to be
globular, heart and torpedo.
5. Once the majority of embryos reach the torpedo stage (7-10 days after sieving) they
are transferred to an enriched medium containing a high level of sucrose, nitrogen and
sulphur to prevent precocious germination and to enable deposition of storage
reserves. The embryos rapidly accumulate fresh and dry weight, reaching 1-2 mg dry
weight .
6. To induce the acquisition of desiccation tolerance, the somatic embryos are placed on
a modified medium containing abscisic acid (ABA) for 3 days. Then they are
removed from the medium, washed to remove sugar and other nutrients, and dried.
7. The standard method of drying is to place the somatic embryos in a sealed chamber
over a saturated salt solution designed to give specific relative humidifies. Daily for
one week, the embryos are transferred to a progressively lower relative humidity
chamber and finally are dried at ambient conditions. At this stage, the embryos have
reached approximately 15% moisture and can be stored for a year or more with good
viability.
Encapsulation of Matured Somatic Embryos

Somatic embryos produced naked embryos without storage materials and protective layer
(seed coat). This is very difficult for handling so this demand the encapsulation and coating.
The somatic embryos produced are encapsulated using gel agents like agar, alginate, polyco,
carboxy methyl cellulose, guar gum, sodium pectate etc. Among these alginate encapsulation
was found to be more suitable and practicable. Alginate hydrogel is frequently selected as a
matrix for synthetic seed because of its moderate viscosity and low spinnability of solution,
low toxicity for somatic embryos and quick gellation, low cost and bio-compatibility
characteristics. The use of agar as gel matrix was deliberately avoided as it is considered
inferior to alginate with respect to long term storage. Alginate was chosen because it
enhances capsule formation and also the rigidity of alginate beads provides better protection
to the encased somatic embryos against mechanical injury.
The somatic embryos are mixed with sodium alginate (2 %) and the suspension is dropped
into the calcium salts solution (200mM). The principle involved is when sodium alginate
dropped into the calcium salt solutions it from round firm beads due to the ion exchange
between Na+ in sodium alginate and Ca2+ in calcium slat solutions and sodium alginate form
calcium alginate in 20-30 minutes. Since somatic embryos lack seed coat and endosperm the
matrix of encapsulation can be added with nutrients and growth regulator, which will serves
as an artificial endosperm. This will increase the efficiency of germination and viability of
seeds. Addition to these nutrients other useful materials fungicides, pesticides, antibiotics and
micro organism can also be incorporated.
Application of Synthetic Seeds
By combining the benefits of a vegetative propagation system with the capability of long-
term storage and with the clonal multiplication, synthetic seeds have many diverse
applications
1. Multiplication of non-seed producing plants, ornamental hybrids or polyploids plant

2. Propagation of male or female sterile plants for hybrid seed production
3. Germplasm conservation of recalcitrant species
4. Multiplication of transgenic
Limitations
1. Limited production of viable micro-propagules that are useful in synthetic seed

producer
2. Asynchrous development of somatic embryos
3. Improper maturation of somatic embryos that makes them inefficient for germination
and conversion in to normal plants
4. Lack of dormancy and stress tolerance in somatic embryos that limit the storage of
synthetic seeds
5. Somo clonal variations which may alter the genetic constituent of the embryos
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Shailendra Singh Gaurav

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Study Material

M.Sc. Ag. IV semester

Dr. Shailendra Singh Gaurav

Topic: SYNTHETIC SEEDS

Why Synthetic Seeds?

Characteristics of Synthetic Seeds

1. Ease of handling while in storage

Types of synthetic seeds:

Procedure for Synthetic Seed Production:

1. Establish somatic embryogenesis

Procedure of Somatic Embryogenesis:

Encapsulation of Matured Somatic Embryos

1. Multiplication of non-seed producing plants, ornamental hybrids or polyploids plant

1. Limited production of viable micro-propagules that are useful in synthetic seed

Shailendra Singh Gaurav

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