I'm taking a Tissue Culture course next semester, any advice?

[divaskid]

Hi,

I'm going to start a tissue culture course next semseter (lasts a year or longer) and I was wondering if anyone had any advice before I start. I'd like to be as successful as possible so the more information the better

I'll be doing TC on a nepenthes (unkown type so far) and also cephalotus. Both I've read aren't the easiest (especially the cephalotus), but the teacher gave me the go ahead on them. Does anyone have any advice or experience with these that would help me down the right path?

Thanks

 

[kulamauiman]

start them as seeds. Neps are a pain apparently to get explants cleaned....

assuming similar for the cephs....

 

[dionae]

Cephs are hard. I've only been successful once with several tries.

 

[BigBella]

Cephalotus are a breeze to culture, both from seed and explant. Use the PPM (Plant Preservative Media) process to clean the explant or seed and a dilute 1:3 MS or 2:3 Knudsen-C media with or without PGRs.

Check Matt's TC forum at www.flytrapcare.com; it is a valuable resource . . .

Cephalotus follicularis "Eden Black x Self" (seed)

 

[dionae]

What part of the ceph do you use? I'm 1 for 10 with leaves.

 

[BigBella]

What part of the ceph do you use? I'm 1 for 10 with leaves.

Leaves with the rhizomal -- white -- tissue remaining on the leaves and PPM for sterilization. The most common mistake involves killing the explant with alcohols and bleach . . .

 

[dionae]

Thats a beautiful culture. Thanks for the tips!

 

[pokie22]

What concentration of PPM and for how long? 8% for 3-4 hours?

 

[divaskid]

BigBella, great inspiration! Its getting me excited for school to start again. How long did it take to get your ceph that size?

Edit: is it true no one has done tissue culture of Nepenthes through explants, only by seeds?

 

[BigBella]

How long did it take to get your ceph that size?
Edit: is it true no one has done tissue culture of Nepenthes through explants, only by seeds?

In answer to your first question, that Cephalotus was established in Spring 2011; though the seed in culture had been previously cold-stratified for eight weeks . . .

In answer to the second: no, it can done; but the process often involves twenty-fours hours or more of soaking the explant in broad-spectrum, many now-illegal algal and fungicides -- and the partial dissection of the Nepenthes to allow the chemicals to reach the deeper-affected tissue (so as to kill the endosymbiotic algae). I have done it on a few occasions; but I was just as likely to kill the explant as to establish it; more so in fact.

In the short term, I would concern myself with seed; there is plenty to learn from the vagaries of in vitro germination . . .

 

[BigBella]

What concentration of PPM and for how long? 8% for 3-4 hours?

Eight percent for six hours at the very least, up to ten; and expect a good deal of phenolic bleeding afterwards. I generally replate twice before it ceases . . .

 

[Kinjie]

In answer to your first question, that Cephalotus was established in Spring 2011; though the seed in culture had been previously cold-stratified for eight weeks . . .

In answer to the second: no, it can done; but the process often involves twenty-fours hours or more of soaking the explant in broad-spectrum, many now-illegal algal and fungicides -- and the partial dissection of the Nepenthes to allow the chemicals to reach the deeper-affected tissue (so as to kill the endosymbiotic algae). I have done it on a few occasions; but I was just as likely to kill the explant as to establish it; more so in fact.

In the short term, I would concern myself with seed; there is plenty to learn from the vagaries of in vitro germination . . .

David, I was wondering what times your classes started in spring cause I would really like to sign up and learn this stuff from a master of TC. All your stuff always looks great!

 

[kulamauiman]

David, I was wondering what times your classes started in spring cause I would really like to sign up and learn this stuff from a master of TC. All your stuff always looks great!

new business, er front, for the real business

 

[BigBella]

David, I was wondering what times your classes started in spring cause I would really like to sign up and learn this stuff from a master of TC. All your stuff always looks great!

You're far too kind. I began in TC because I was the "new kid" at the time and low on the totem; the giant lab manuals were foisted upon me (the damn things looked like the Manhattan white pages); and I then had more patience and attention span than the guys regularly treating their "glaucoma" behind the lab . . .

 

[divaskid]

David, I was wondering what times your classes started in spring cause I would really like to sign up and learn this stuff from a master of TC. All your stuff always looks great!

I second that!