Anther culture

 WHAT IS ANTHER CULTURE........?

Anther :-   A  part stamen containing pollen .

Anther culture :- Culturing of anther obtained from unopened flower bud in the nutrient medium  under specific condition .

callus tissue or embryoids from anther ,that give rise to  haploids plantlets either  organogenesis or embryogenesis .

Principle of anther culture - 

1) To the production of haploid plant is to exploit totipotency of microspore.

2) In this process the normal development and function of the pollen cell become a male gamete is stopped and is diverted forcely to a new metabolic pathway for vegetative cell division .

Method of anther culture :- 

1) collection of unopened flower bud .

2) surface sterilized with 70 %  ethanol .

3) Anther excised from flower bud and kept separately .

4) Anther in first meiotic division is select by the acetocarmine test .

5) Inoculated in the medium containing glutamine , l-serine and inositol .

6) Incubated the culture at 25 C for 15 days ,here ,anther grow in to embryoids .

7) Embryoids transfer to rooting medium under 3000 lux illumination after 4-5 weeks the         embryoids  become plantlets .

8) For acclimatization , transfer to green house .

Factor affecting / influencing anther culture :-

1) Genotype of donor plats  - 

The genotype of the donor plant plays a significant role in determining the frequency of pollen production .

eg. Hordeum of each genotype differs with respect to androgenic response in anther culture.

2) Stage of the microspore - 

Anther are more productive when cultured at uninucleate microspore stage .

eg. Barley ,wheat ,rice ,etc.

3) Culture medium -

For anther culture medium requirments vary with genotype and the age of the anther as well as condition under with donor plant are grown .

Incorporation of activated charcoal into the medium has stimulated the induction of androgenesis .

potato extract ,coconuts milk and growth regulator like auxin and cytokinin are used for anther and pollen culture .

4) Temperature - 

Temperature enhance the induction frequency of microspore androgenesis .

The low temperature treatment to anther of flower bud enhance the haploid formation .

The low temperature effects the number of factor such as dissolution of microtubules ,lowering of abssicisic acid ,maintenance of higher ratio of viable pollen capable of embryogenesis .

5) Physiological status of donor plant -

Physiological status of donor plant such as water stress nitrogen requirement and age of donor plant highly affect the pollen embryogenesis .

plant staved of nitrogen may give more responsive anther compared to those that are well fed with nitrogen fertilizer .

 Advantages of anther culture -

1) simple 

2) less time consuming 

3) responsive 

Disadvantages -

1) Requires skill to removing anther without damage .

2) Not much successful in case of cereal crop .

3) Risk of chimera and callus formation from anther wall.

Application -

1) For mutation study 

2) For plant breeding and crop improvement .

3) To obtained the secondary metabolism .

    eg. Hyoscyamus niger  obtained by anther culture having higher alkaloid content .

4) Haploid are used in molecular biology and genetic engineering .