Tuesday, October 12, 2010

in vitro part 2: seeds seeds seeds!

I was most fortunate to receive seeds of the following Phalaenopsis species (click on the names for links to images): P. cornu-cervi, P. aphrodite, P. gigantea, P. taenialis, P. lindenii, P. celebensis, P. violacea forma mentawai and P. heiroglyphica. Many of these species are certainly difficult if not impossible to get in South Africa so I am most pleased to have the opportunity to hopefully raise some by seed.

I purchased some Phytamax P6668 and P1056 from Sigma in anticipation of the seeds and for later re-plating respectively. A colleague of mine uses P6668 at half strength with added 20g sucrose so I prepared two sets of flasks with both full strength and half strength P6668 with added sucrose (10g and 20g respectively). I finished my media preparation and sterilisation on the evening of 11 October and  I did not have the time to work though all of the above species but did flask P. gigantea, P. cornu-cervi, and P. violacea forma mentawai. I also threw in some Microcoelia exilis for good measure but have no idea if the tiny pods were carrying any viable seed... we will see. The rest of the species were done on 12 October along with some Disa crassicornis and D. atricapilla.

I must admit sitting down in front of the sterile transfer chamber on the first night with all these species was quite a rush! I had one chance to get it right and hopefully I did not get any contamination. I followed the advice of my colleague who said I should sterilise the seeds with 0.5% sodium hypochlorite (active %, not v/v concentration) for only 2 minutes before rinsing and sowing. This to me seemed a bit short and in the back of my mind I am hoping it was long enough to inactivate any potential bacteria and fungal spores. He has done this before with good results though so lets have faith. I should know within 3 days or so and will either be sadly disappointed or happy with the results. I certainly hope all goes well.

I thought I would also add a brief update on my Phalaenopsis hybrid seed pods that are progressing very nicely. The selfing of the large pink from Woolworth and the coss with P. golden wonder are half way now. They will be harvested in mid December. Other pods are at varying stages of developemnt and I have had good success with most crosses with only a few rejections.

Phalaenopsis seed pod - 2 months on

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