Learning through mistakes

I contacted John Haggar regarding my problems with seed not germinated and he swiftly replied with some very interesting answers.

The reason for bleaching Dact seed is to seterilise it rather than breaking down the epidermis, so insufficient bleaching time is not likely to be an issue, which is a relief. If they do not germinate it indicates the seed is duff. Perhaps I did not sow it quickly enough and the seed had deteriorated in my less than perfect seed storage conditions...

The blackening and dying back of the protocorms is not liekly to be due to drying out (they are sealed and the containers show signs of condensation after all !) but could be due to a variety of reasons, John suggested:

• the oats medium could be too strong, casuing the B1 fungus to develop too strongly and behave as a pathogen: I used premixed HOS oats mix and I am failry certain that I mixed it properly, but I might be wrong.
• some sort of unrecognised infection - i don't know how I can identify this apart from leaving them to develop. 
• insufficient refrigeration time  - this is unlikely I have not removed them from the fridge since they were transplanted.
• John picked up on the fact that I had been chopping out a big chunk of medium with the protocorm and transplanting that as well. He suggested that this might be causing the fungus on the plate to develop in the new oats mix rather than the fungus IN the protocorms, which will effectively starve them. This was a bit of a eureka moment for me. I have been transplanting into pre prepared plates already innoculated with B1. I am fairly certain this is the cause of the problem - I should have been transplanting into . Although it is a shame to have lost all these plants, it is satisfying to work out why at least... I have replied to John and suggested this, so I'll see what he says.

So I need to get some forceps - I think cheap plastic ones are the ones to go for - and practice moving the protocorms about. I'll start by extracting any which look ok into new vessels. I will leave the rest which are in petri dishes just to see what will happen. And of course I can also leave the ungerminated seed as well to see whether they germinate. I will need to make some new flasks though.