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Parvati Pujer
UHS11PGM123
INTRODUCTION
 Many fruit crops are difficult to multiply by conventional
  propagation methods and improve through traditional breeding
  programmes

 Among the innovative techniques of micro propagation, the concept
  of somatic embryogenesis with synthetic seed production or artificial
  seed technology is very promising

 Synthetic seed is referred to as encapsulated somatic embryos, which
  functionally mimic seeds and can develop into seedling under
  suitable conditions
Contd……

 Besides, research has also been focused on the use of
  vegetative propagules like axillary buds, adventitious buds,
  shoot-tips, cormels, bulbs and protocorms for production of
  synthetic seeds

 In general, for seed propagated crops outstanding hybrids can
  be multiplied through tissue culture and propagated by using
  synthetic seeds rapidly
Advantages
 Direct delivery of tissue cultured plants to the field

 Propagation of desirable genotypes with genetic uniformity

 Reduction in cost of vegetative propagated elite plants

 Preservation of germplasm and convenience in germplasm exchange

 Reduction in dormancy period

 Large production identical embryos in short time

 It can be produced through out the year
Synthetic Seeds
 Synthetic seeds: It is living seed-like structure derived from
  somatic embryoids in vitro culture after encapsulation by a
  hydrogel.

 Such seed are encapsulated by protective gel like calcium
  alginate against microbes and desiccation
Fig 2: Flow diagram presenting the procedure of synthetic
       seed production
Fig 3: Different stage of embryo development
Table 1: Effect of endosperm and encapsulation on zygotic
         somatic embryo germination and conversion

    Embryos           Form          Germination       Conversion
                                        %                 %

                       Non              100                 100
     Zygotic       encapsulated

                   Encapsulated         100                 93.3


                       Non              93.3                93.3
    Somatic        encapsulated

                   Encapsulated         100                 100
Artificial seed coat


                                   Somatic embryo




Artificial endosperm

       Fig 4 : The concept of Artificial Seed
Table 2: Artificial seed production and plant conversion


In vitro propagules for                     Crop
encapsulation
                          Papaya, Brinjal, Mango, Carrot,
Somatic embryos           Sandalwood, Brassica, Alfalfa,
                          Lettuce, Celery,
Auxiliary buds/           Vitis, Eucalyptus, Citrus, Pineapple
Adventitious buds
Shoot tips                Banana, Cardamom, Apple, Kiwifruit
Table 4: Encapsulation for Synthetic Seed

    Gel (Concentration)            Complexing       Concentrat
          (% w/v)                    Agent           ion (µM)

Sodium alginate (0.5-5.0)      Calcium Salts          30-100

Sodium alginate       (2.0)    Calcium chloride       30-100
with gelatin          ( 5.0)

Carrageenan       (0.2-0.8)    Potassium chloride      500
                               Ammonium chloride
Locust Bean Gum (0.4-1.0)

Gelrite              (0.25)
Encapsulation methods for synthetic seed
A) Dropping procedure

1) The most useful encapsulation
  system.      Drip 2-3 % sodium
  alginate drops from at the tip of the
  funnel and the somatic embryos
  are inserted

2) Keep the encapsulated embryos
   complex in calcium salt for 20 min

3) Rinsed the capsules in water and
   then stored in a air tight container
B) Automate encapsulation process
This is the quick method of artificial
   seed production

A) Alginate solution with embryo is
   feed from supply tank

B) Alginate capsules were planted in
   speeding trays using a vacuum
   seeder.

C) The capsules are planted in the field
   using a stanhay planter

D) A hydrophobic coating is required
   for mechanical handling
BRIEF RESEARCH WORK ON SYNTHETIC SEEDS
Fig 5: Various developmental stages during plant regeneration
      via artificial seed in Carica papaya L.
    A- Somatic embryos in liquid culture after 30 days in maturation
         medium.
    B- Somatic embryo at the cotyledonary stage.
                                                        Castillo et al., 1998
C - Artificial seeds encapsulated in 2.5% sodium alginate.
D - Germinating artificial seed.


                                              Castillo et al., 1998
E - A plantlet germinated from an artificial seed on growth regulator-
   free basal MS medium.
F - A plantlet showing normal development after germination from an
   artificial seed. Culture tubes were 2.2 cm in diameter.
                                                    Castillo et al., 1998
Fig.8 : Plant regeneration from encapsulated somatic embryos of banana cv.
        Rasthali.
       A- Somatic embryos used for encapsulation.
       B- Encapsulated somatic embryos in 5% sodium alginate.

                                                    Ganapathi et al., 2001
C- Emergence of shoots after 2 week of culture from encapsulated somatic
   embryos.
D- Plantlet development from encapsulated somatic embryos on MS basal
   medium after 6 week of culture.
E-Two month old hardened plants derived from encapsulated somatic embryos
  growing in the polybags.                                Ganapathi et al., 2001
Synthetic seed production By- Parvati Pujar
Synthetic seed production By- Parvati Pujar
Synthetic seed production By- Parvati Pujar
Synthetic seed production By- Parvati Pujar

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Synthetic seed production By- Parvati Pujar

  • 1.
  • 3. INTRODUCTION  Many fruit crops are difficult to multiply by conventional propagation methods and improve through traditional breeding programmes  Among the innovative techniques of micro propagation, the concept of somatic embryogenesis with synthetic seed production or artificial seed technology is very promising  Synthetic seed is referred to as encapsulated somatic embryos, which functionally mimic seeds and can develop into seedling under suitable conditions
  • 4. Contd……  Besides, research has also been focused on the use of vegetative propagules like axillary buds, adventitious buds, shoot-tips, cormels, bulbs and protocorms for production of synthetic seeds  In general, for seed propagated crops outstanding hybrids can be multiplied through tissue culture and propagated by using synthetic seeds rapidly
  • 5. Advantages  Direct delivery of tissue cultured plants to the field  Propagation of desirable genotypes with genetic uniformity  Reduction in cost of vegetative propagated elite plants  Preservation of germplasm and convenience in germplasm exchange  Reduction in dormancy period  Large production identical embryos in short time  It can be produced through out the year
  • 6. Synthetic Seeds  Synthetic seeds: It is living seed-like structure derived from somatic embryoids in vitro culture after encapsulation by a hydrogel.  Such seed are encapsulated by protective gel like calcium alginate against microbes and desiccation
  • 7. Fig 2: Flow diagram presenting the procedure of synthetic seed production
  • 8.
  • 9. Fig 3: Different stage of embryo development
  • 10. Table 1: Effect of endosperm and encapsulation on zygotic somatic embryo germination and conversion Embryos Form Germination Conversion % % Non 100 100 Zygotic encapsulated Encapsulated 100 93.3 Non 93.3 93.3 Somatic encapsulated Encapsulated 100 100
  • 11. Artificial seed coat Somatic embryo Artificial endosperm Fig 4 : The concept of Artificial Seed
  • 12. Table 2: Artificial seed production and plant conversion In vitro propagules for Crop encapsulation Papaya, Brinjal, Mango, Carrot, Somatic embryos Sandalwood, Brassica, Alfalfa, Lettuce, Celery, Auxiliary buds/ Vitis, Eucalyptus, Citrus, Pineapple Adventitious buds Shoot tips Banana, Cardamom, Apple, Kiwifruit
  • 13. Table 4: Encapsulation for Synthetic Seed Gel (Concentration) Complexing Concentrat (% w/v) Agent ion (µM) Sodium alginate (0.5-5.0) Calcium Salts 30-100 Sodium alginate (2.0) Calcium chloride 30-100 with gelatin ( 5.0) Carrageenan (0.2-0.8) Potassium chloride 500 Ammonium chloride Locust Bean Gum (0.4-1.0) Gelrite (0.25)
  • 14. Encapsulation methods for synthetic seed A) Dropping procedure 1) The most useful encapsulation system. Drip 2-3 % sodium alginate drops from at the tip of the funnel and the somatic embryos are inserted 2) Keep the encapsulated embryos complex in calcium salt for 20 min 3) Rinsed the capsules in water and then stored in a air tight container
  • 15. B) Automate encapsulation process This is the quick method of artificial seed production A) Alginate solution with embryo is feed from supply tank B) Alginate capsules were planted in speeding trays using a vacuum seeder. C) The capsules are planted in the field using a stanhay planter D) A hydrophobic coating is required for mechanical handling
  • 16. BRIEF RESEARCH WORK ON SYNTHETIC SEEDS
  • 17.
  • 18. Fig 5: Various developmental stages during plant regeneration via artificial seed in Carica papaya L. A- Somatic embryos in liquid culture after 30 days in maturation medium. B- Somatic embryo at the cotyledonary stage. Castillo et al., 1998
  • 19. C - Artificial seeds encapsulated in 2.5% sodium alginate. D - Germinating artificial seed. Castillo et al., 1998
  • 20. E - A plantlet germinated from an artificial seed on growth regulator- free basal MS medium. F - A plantlet showing normal development after germination from an artificial seed. Culture tubes were 2.2 cm in diameter. Castillo et al., 1998
  • 21.
  • 22. Fig.8 : Plant regeneration from encapsulated somatic embryos of banana cv. Rasthali. A- Somatic embryos used for encapsulation. B- Encapsulated somatic embryos in 5% sodium alginate. Ganapathi et al., 2001
  • 23. C- Emergence of shoots after 2 week of culture from encapsulated somatic embryos. D- Plantlet development from encapsulated somatic embryos on MS basal medium after 6 week of culture. E-Two month old hardened plants derived from encapsulated somatic embryos growing in the polybags. Ganapathi et al., 2001