Embryo Culture In Animal Biotechnology Ppt

1. Animal Cell, Tissue And Organ Culture
   1. Embryo Transfer Lecture 7
2. Plant Tissue Culture Chapter 2 In Vitro Culture.
3. Animal Cell Culture Techniques

2 Embryo Culture Isolation & growth of an immature or mature embryo in vitro, with the goal of obtaining a viable plant. The first attempt to grow the embryos of angiosperms was made by Hannig (1904) from two Crucifers Cochleria & Raphanus.

4 Mature Embryo Culture It is the culture of mature embryos derived from ripe seeds. This type of culture is done when embryos do not survive invivo or become dormant for long periods of time. This culture is done to eliminate the inhibition of seed germination.

It is the culture of immature embryos to rescue the embryos of wide crosses. This type of culture is mainly used to avoid embryo abortion with the purpose of producing a viable plant.

Animal Cell, Tissue And Organ Culture

Mineral salts – K, Ca, N most important carbohydrate and osmotic pressure - 2% sucrose works well for mature embryos - 8-12% for immature embryos - transfer to progressively lower levels as embryo grows

Amino acids - reduced N is often helpful - up to 10 amino acids can be added to replace N salts, incl. glutamine, alanine, arginine, aspartic acid, etc. - requires filter-sterilizing a portion of the medium

Natural plant extracts - Coconut milk (liquid endosperm of coconut) - enhanced growth attributed to undefined hormonal factors and/or organic compounds - others – extracts of dates, bananas, milk, tomato juice

Historical Events In The Development Of Cell Culture

Plant Growth Regulators - globular embryos – require low conc. of auxin and cytokinin - heart-stage and later – moderate level of auxin & low level of cytokinin - GA and ABA regulate precocious germination

11 Embryo Rescue In immature embryo culture, wide hybrids often suffer from early spontaneous abortion due to several barriers which operate at pre- & post- fertilization levels. Pre- fertilization barriers include all factors that hinder effective fertilization . (due to inhibition of pollen tube growth)

12 Embryo Rescue Post fertilization barriers retard the development of the zygote after fertilization and normal development of the seed. (due to failure of endosperm) This is known as embryo-endosperm incompatibility where the endosperm produces toxins that kills the embryo.

Embryo Transfer Lecture 7

15 The Kernels of Maize has a pericarp of fruit fused with the seed coat and the entire Kernel is reffered as seed The Cob is close to a multiple fruit in structure, except that the individual fruits (Kernels) never fused to a single mass

Its Immature seeds (Kernels) were extracted out Surface sterilize the immature seeds by treating them with 0.1% mercuric chloride solution for 7 minutes Give three washings with autoclaved distilled water Dissect out the embryo from the seed under totally aseptic conditions Culture the embryos in the jars containing MS media Incubate the jars at temperature 28 ̊-30 ̊C

Production of haploids Prevention of embryo abortion in wide crosses Overcoming Seed dormancy Shortening of breeding cycle Prevention of embryo abortion with early ripening stone fruits

Plant Tissue Culture Chapter 2 In Vitro Culture.

Production of monoploids - useful for obtaining haploids of barley, wheat, other cereals. - the barley system uses Hordeum bulbosum as a pollen parent.

Hordeum vulgare is the seed parent. zygote develops into an embryo with elimination of HB chromosomes. eventually, only HV chromosomes are left embryo is rescued by culturing 10 PP to avoid abortion.

Embryo rescue from a cross between an early Japanese cv.- Sunago wase & a Chinese cv.- Yuhualu obtained “Zaoxialu” an extra early maturing peach cultivar.

Animal Cell Culture Lecture 1 Introduction

Resistance traits Potato leaf roll virus Triazine resistance Bacterial blight Crossing species 1. Solanum tuberosum x S. etuberosum 2. Brassica oleracea x B. napus 3.Oryza sativa x O. minuta

Embryo rescue technique is applied to break seed dormancy. Production of seedlings from seed of naturally vegetatively propagated plants such as Banana & Colocasia, whose seeds do not germinate in nature.

Under proper germination conditions, 3 years are required for seeds from mature berries to complete their embryonic development. The excised embryo goes through all the developmental sequences in vitro & plants can be obtained in 2-3 weeks time.

Animal Cell Culture Techniques

Some species produce sterile seeds that will not germinate under appropriate conditions e.g.- Peach, Nectarine, Plum. Embryo culture has been practised as a general method in horticultured crops including Peach, Nectarine & Plum.

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