Can Replating be Skipped?...

[mohi]

Hi... I'm going to start sowing my seeds as soon as my pods ripen... I guess Within 3-4 months for a phal...
The first barrier on my path is replating the protocorms... Can this step be skipped by putting few seeds on large flasks and adding sterile water to spread them evenly?... And perhaps adding water/agar-less medium if the flasks get dry?... Please help...
Thanks...

[margo]

Sorry I can't help with any info about that. I just made my very first steps for flasking and so far I have one contaminated flask (...I don't really have it enymore...it went through the drain) and two others that are sitting and doing nothing. It's been over two weeks now and I still don't see anything there. One step at a time... If I get to replating then I'll think about it... Honestly- I think I'll have to start over...LOL

[mohi]

I'm starting with phal hybrids... Those showy ones we see as cut flowers... I think they produce protocorms much faster... IMHO skipping the replate step reduces the risk of contamination...
I've done lots of mycology stuff... Grabbing ONE spore from agar medium and growing it on PDA to make a clean colony so working with sterile stuff is no problem... But think of a thousand protocorms waiting to be replated... Sounds a bit scary to me!...

[Brutal_Dreamer]

It sounds like it would work, but you having worked in mycology should know better than me for sure! I do hope if you try this that you will post the results and process.

Cheers,
BD

[Gin]

That is where I ran into a problem replating . I made up the new flasks and let them sit with out the seedlings , they were contaminated . I think it was how they were vented .
From what I have read the replate medium is different then what is used in the mother flasks, more nutrients for better growth . If a person could get away with leaving them in the mother flask would be a lot easier , my seedlings are a species Phal. cross equestris X stobartiana . I took them out early and lost a lot of them . Please keep us posted on what happens ... Gin

[mohi]

Ok... Thanks for the information... Looks that my crosses were successful... The flowers are wilting and the basal part is swelling up... I did the same with my Catt and I think I did it correctly...
em... What happens if the medium of the mother flask has the same (high level) nutritions?... I'm going to use a mixture of MS (with extra hormones and vitamins) and NA (nutrient agar)... And perhaps a little charcoal and banana...

[wetfeet101b]

I think one of the main reasons for replating is to encourage the different progressions in the protocorm growth.

The mother flask medium is made up of nutrients to encourage seed germination and protocorm development.
The first replate introduces nutrients that encourage root development. And then another replate for leaf growth.

For mass producers, it is more economical for them on a large scale to use only nutrients that are necessary for that particular growth stage.

I would think that if your mother flask medium has all the nutrients needed for all stages of growth, and if the nutrients would not expire before they are needed, then you might be able to just use a single flask without replating.
The only other problem is that the protocorms would be so crowded in the mother flask and you would need to replate anyway just to keep them from choking each other.

[mohi]

Thanks John... I think I'll do the sowing in petri dishes... And then transfer them into jars...

[John D.]

I'd leave out the hormones if they are the ones used commonly for root growth etc. They are usually used in a mulitplication medium for growing many plants of the same type from a small piece of tissue (meristem culture). Starting with seed you don't need them and they may cause the protocorms to proliferate (grow into masses of tissue) instead of differentiate (into a small plant with roots and leaves). The banana and potato that people add to replate medium is to encourage the differentiation, not needed in mother flasks.