Type of in vitro culture. Culture of intact plants (Seed orchid culture) Embryo culture (embryo rescue) Organ culture 1. shoot tip culture 2. Root culture 3. Leaf culture 4. anther culture Callus culture Cell suspension and single cell culture Protoplast culture. Seed culture.
1. shoot tip culture
2. Root culture
3. Leaf culture
4. anther culture
( separated ) and cultured.
Haploid plants are derived from microspores (pollen) cultured individually or in anthers
First report of Haploid plant from anther and pollen culture is by eminent Indian embryologists Guha & Maheshwarifrom the plant Daturastramonium.
Wheat, Rice, Maize, Rye, Tobacco, Potato, Brassicas has been used in pollen & anther culture to release the cultivars having superior chts. , high yield & disease resistance.
Bajaj, Y.P.S. 1983. In D.A. Evans, W.R. Sharp, P.V. Ammirato, and Y. Yamada (eds.), Handbook of Plant Cell Culture. Volume 1. Techniques for Propagation and Breeding. MacMillan, New York. p. 228-287.
Embryogenesis Ammirato, and Y. Yamada (eds.), Handbook of Plant Cell Culture. Volume 1. Techniques for Propagation and Breeding. MacMillan, New York. p. 228-287.
Applications of plant cell culture
Cell without cell wall is called protoplast.
For tissue culture application protoplast can be isolated from almost any part of the plant but leaves are preferentially used for this purpose as these are easy to handle.
Technique involves following steps;
A. Sterilization of the explant by 2% sodium hypochloride for about 30 minutes.
B. Peeling of the epidermal cells.
C. Digestion of cell wall by enzymes, macerozymes, cellulase, hemicellulase and pectinase.
D. Washing of the released protoplasts.
The common methods employed for the fusion of the released protoplasts is as:
A. Spontaneous fusion of protoplast.
B. Mechanical induction fusion.
C.NaNO3 induced fusion.
D. High p H and Ca++ induced fusion.
E. Poly Ethylene Glycol induced fusion (PEG).
F. Electro fusion.
Haploid - fusion.Gametic number of chromosomes, n which may not be equivalent to x (x is one genomic complement)
Monoploid - haploid derived from a diploid, x(one genomic complement)
Polyhaploid - haploid from a polyploid (nx), prefix indicates genome complement number, e.g. tobacco is a dihaploid
Agricultural applications for haploids - fusion.Rapid generation of homozygous genotypes after chromosome doubling