Agave Victoriae-reginaes

artificial seeds
Gabriela Galicia
What is an artificial seed?
Artificial seed technology envolves the production
of tissue culture derived somatic embryos
encased in a protective coating.
Advantages of artificial seeds
Large scale propagation and mixed genotype
plantations

Easy handling during storage
Transportation and planting and inexpensive
transport reason being their small size
Storage life comparable to natural seeds
Why Agave victoriae-reginae?
Mexico is one of the 17 Megadiverse Countries
Endemic species of Chihuahuas Desert
Coahuila, Durango, Nuevo Len (North of
Mexico)
High Ornamental Value
Endangered Species

Endangered species
Key species
Nutrient providers for pollinators.
Illegal obtention ($10USD to $2000 USD)
NOM-059-ECOL-2010

Basic Requirements for production of
Artificial Seeds
High quality, vigorous somatic embryos
Easy to obtain them
Well identified
Establish somatic
embryogenesis
Production of somatic embryos
Mature somatic embryos
Encapsulation
Mass production of syntetic
seeds
Syntetic seeds storage
Green house and field planting
I.
PRODUCTION OF SOMATIC EMBRYOS
PLANT TISSUE CULTURE
What is Plant Tissue Culture?
Biotechnological tool
Tissue culture is the in vitro aseptic culture of
cells, tissues, organs or whole plant under
controlled nutritional and environmental
conditions
Dates back to Cell Theory in 1838.
What for?
Large scale plant multiplication, mainly
industrial importance
Plant improvement, disease elimination
Secondary metabolites production
Research tool
Conservation of endangered species
The basic concepts that explain
Vegetal Tissue culture are:

- Vegetal totipotency
Initiate cell division from almost any tissue on the
plant.
- Cell dedifferentiation redifferentiation
- Medium culture
- Growth regulators.

Medium
Macronutrients, Micronutrients
Vitamins
Plant Growth regulators
Carbon source -- Sugar
Gelling agent --- Agar
MS 50%
Micropropagation Stages
Stage 0. Obtention of mother plant
Stage 1. Sterilization of explants and culture in
laminar flux cabinet.
Stage 2. Plant tissue propagation
Stage 3. Formation of roots
Stage 4. Aclimatization
Plant tissue culture
Plant regeneration
Somatic Embryogenesis Organogenesis
Directly or indirectly (callus)
Embryo initiation: High
concentration of 2,4-D
Embryo production: No or
very low 2,4-D levels

Plant organs roots, shoots,
leaves
Directly, indirectly

Agave victorias-reginaes plantlets
Agave victoriae-reginaes somatic embryos were
obtained through MS 50% with added 2-4D and
BAP.
Formation of somatic embryo was observed after
6 months.

II.
Encapsulation process
Encapsulation process
b)
c)
a)
d)
a) Sodium alginatum solution
b) Calcium chloride solution
c) Seeds obtained
d) Pasteur pipette

Alginate Matrix
Alginate
CaCl2 H2O
Ion exchange between Na
+
and Ca
2+
Form round and firm beads
100mL MS 50%
1) Added Alginate 3%
2) 100mM CaCl2 H2O (1.47g)
Production of syntetic seeds
Seeds storage
Results
475 encapsulated somatic embryos
439, Aprox. 80% manteined good conditions
10 capsules were contaminated
36 seeds germinated prematurely
Syntetic Seeds
Optimal
conditions
Contamination
Early
germination
Conclusions
A. victoriae-reginae being an endangered
endemic species, it is very important to develop
means of conservation.

Encapsulation is a viable way to generate a
germplasm bank, as both the contamination and
early germination percentages are very low.