Welcome, Guest. Please login or register.
Did you miss your activation email?

Username: Password:
Pages: [1] 2 3 ... 10
 1 
 on: Today at 06:34:50 am 
Started by MRTree - Last post by bosqueberg
b. muricata flowered this year, in a 15 gallon pot. Plant is around 5+ years old. No special treatment, kind of neglected. Rarely gets fertilized.

 2 
 on: Today at 06:31:12 am 
Started by Roze - Last post by bosqueberg
That is amazing. Interspecies plant communicator! Widening my perspective on this issue

 3 
 on: Today at 01:30:13 am 
Started by MRTree - Last post by Dirt
Side ask; what is meant by “old caapi”?

 4 
 on: February 23, 2019, 09:20:07 pm 
Started by Roze - Last post by ONandONandON

 5 
 on: February 23, 2019, 09:08:58 pm 
Started by Ian Morris - Last post by ONandONandON
Quote
Experiment 5: Anther Culture
 
Background
 
 The purpose of anther and pollen culture is the production of haploid plants by the induction of embryogenesis from repeated divisions of microspores or immature pollen grains (Dodds and Roberts, 1985).  The chromosome complement of these haploids can be doubled by colchicine treatment or other techniques to yield fertile homozygous diploids.  The resultant diploids can be used in plant breeding to improve crop plants (Sunderland and Cocking, 1978).
168    Plant Tissue Culture
 
 The haploid nature of embryoids should be determined by standard chromosome staining procedures (acetocarmine or Feulgen reaction) prior to colchicine treatment.  Similarly, the effect of colchicine on chromosome doubling needs to be monitored by chromosome staining.
 
Materials
 
 tobacco buds  sharp pointed forceps, surgical scissors  95% ethanol  petri dishes of culture medium (1/2 strength MS, 2% sucrose 0.8% agar, glutamine [800 mg/liter], serine [100 mg/liter])
 
 
Methods 
 
1. Obtain two buds at the appropriate stage.  This occurs in tobacco when the sepals and the petals in the bud are the same length.
 
2. Holding the bud by the pedicel between the thumb and first finger, dip the entire bud in 95% ethanol for 15 seconds
 
3. Remove bud and allow excess alcohol to drip off.
 
4. With a pair of sterile forceps, remove the outer layer of tissue, the sepals.
 
5. Next, remove the inner layer of tissue, the petals, exposing the anthers.
 
6. Open the petri dish containing the medium for the induction of haploids.  Remove each anther from the bud and drop it onto the medium.  Do not damage the anther or include any filament tissue.
 
7. Repeat for another bud.
 
8. When finished, seal the plates and place in incubator (25°C).
 
9. In 2–3 weeks examine for somatic embryo initiation.  Embryoid-forming cells are characterized by dense cytoplasmic contents, large starch grains and a relatively large nucleus.  Embryoids appear opaque among translucent cells.  Embryoids also exhibit high dehydrogenase activity and can be detected by tetrazolium staining (Dodds and Roberts, 1985).
 
Acknowledgements
 
 Paul Bottino (Botany Department, University of Maryland, College Park) provided the sterilization procedure for Experiment 2 and Methods for Experiments 4 and 5.
 
 Plant Tissue Culture        169
 
 
Literature Cited
 
Bottino, P. J.  1981.  Methods in plant tissue culture.  Kemtec Educational Corp., Kensington, Maryland, 72 pages. Butcher, D. N., and D. S. Ingram.  1976.  Plant tissue culture.  Arnold, London, 67 pages. Dodds, J. H., and L. W. Roberts.  1985.  Experiments in plant tissue culture.  Second edition.  Cambridge University Press, New York, 232 pages. Street, H. E.  1973.  Plant tissue and cell culture.  Blackwell Scientific Publications, Oxford, 320 pages. Sunderland, N., and E. C. Cocking.  1978.  Plant tissue culture in China—major change ahead?  Nature, 274:643-44. Sunderland, N., and M. Roberts.  1977.  New approach to pollen culture.  Nature, 270:236-238. Ting, I. P.  1982.  Plant physiology.  Addison-Wesley, Reading, Massachusetts, 642 pages. Wetherell, D. F.  1982.  Introduction to "in vitro" propagation.  Avery Publishing Group Inc., Wayne

 6 
 on: February 23, 2019, 08:48:58 pm 
Started by Mangrove - Last post by koacaia
Definitely remove the jelly, 3% should be fine just rinse them again maybe.

 7 
 on: February 23, 2019, 08:33:47 pm 
Started by dEEcor - Last post by ONandONandON
https://en.wikipedia.org/wiki/Galantamine

 8 
 on: February 21, 2019, 05:28:01 pm 
Started by Mangrove - Last post by Mangrove
If you wash the seeds and rinse them with hydrogen peroxide you can put them in a bag of moss and send them fresh.

Should I use a 3% solution of peroxide to wash them or a diluted solution of it? Should I remove/scrape the jelly like outer layer before they're washed with peroxide?

 9 
 on: February 20, 2019, 07:59:10 am 
Started by Mangrove - Last post by koacaia
If you wash the seeds and rinse them with hydrogen peroxide you can put them in a bag of moss and send them fresh.

 10 
 on: February 18, 2019, 07:07:00 pm 
Started by Mangrove - Last post by XDX
I saved a bunch of seeds from a fruit last summer
They have a jelly-like skin layer that should be removed before planting
I cleaned mine for the most part
I was a lot of seeds, so I didn’t individually inspect each or anything

I let the seeds soak in water for months,
For got about them a few times,
Water would smell funky anerobic,
Would rinse them and change the water and forget about them again
Eventually I got around to planting,
Nearly all sprouted.

So deff good for months if kept moist
Idk if they are the same once dried

Pages: [1] 2 3 ... 10