Yes mmlr38 I only use 1/2 MS but will start to reduce this to about 1/3 with the next batch of media I mix up. 1/2 MS maybe a bit to strong for seedlings. I have had a bit of trouble with some seeds germinating but only growing for a week or two and then dieing off
I used leaf and tendril tissue from a developed pitcher Will try a younger leaf like you suggested thanks for the advice Fred.
I have experienced the dying off of seedlings as well when I used full strength MS for sarracenia seeds. They didn't like that much at all. They germinated and grew for maybe a week and then turned brown.
I just put a bunch of seeds into cultures with 1/2 MS. When (if) I notice that they start to germinate, I'll replate them quickly.
Hello I am no expert on TC however what is a good method to sterilze very small seeds like pingicula and utricularia forceps arent really an option because they are so small??? any help would be greatly appreciated!
I had the opportunity to go into a lab where they were working with Arabidopsis witch have pretty small seeds and they just made a little envelope out of paper and stapled it shut. When you submerse the packet I would also try to make sure I get all the air out. Good luck and let us know how it goes
is it beneficial to make a lot of callus then allow it to shoot? and are you using liquid media here ? looks like youve got it down packed! so perhaps 25% Ms would be the best option for nepenthes then ?
Yeah it does look a bit like liquid but I still use a gelling agent just didn’t add the right amount to the media this time still worked though. Yep I use 25% to 30% MS As for the callus I let it grow to a large size using BAP in the media then move it to another media with higher amounts of NAA it will start to grow shoots but the callus will continue to grow as well. after it has grow a lot of shoots I remove these shoots and place them into media containing a very high level of NAA and very low level of BAP shoots will start to form roots. As for the leftover callus I just cut it up and start again.
Wow thanks very kindly for the pictures and tips there, fascinating tc is, im so excited for my laminar flow hood to be built! and get tc ing here. its always inspiring to see pics like these! thanks again!!!!
Hi Maciey, I have PM an answer to you. But just for everyone, I change the amount of BAP and NAA I use dependent on nepenthes type some respond well to just BAP at 1ml Others I use 0.5ml BAP to 0.5ml NAA I would start with 0.5ml of each and see how things go, good old trial and error. Also ask around different people have success with different ratios. For shoot growth I add more BAP up to 2ml/L and reduce the NAA. For root growth increase NAA to 1ml/L and reduce the BAP This is a guide only like I said different nepenthes respond to different levels ask around to see what other people use
The extra water is only a problem when seeds are first started as it causes the seed to be washed around the container as well as water logging. I remove the water by draining the container just before I place the seed into i. I only do this in sterile conditions and only for seed if I am transferring plants to another container I leave the water in it but that’s just me.
Hi all hope everyone is growing plants and not mold like me. Had a bad run with contamination lately think i have been a bit careless. anyway still had some good results to here are a few photos of my latest batch of nepenthes hookeriana from callus to hardened off plants. The first is a small piece of plant with roots placed into media containing 2 ml of BAP per liter this plant material was taken from a larger callus
this photo shows the callus starting to form
once the callus has grow a bit and started to grow some large shoots i place it into plain media to allow the shoots to grow a bit more
this is one shoot placed into media with some added NAA about 0.5 ml per liter to allow plants to grow roots sometimes i dont even bother with the NAA the plants seem to grow roots without any NAA As you can see the one shoot placed into this container has formed several more shoots i cut these off and place them back into media to form roots
plants being hardened off in a mix of perlite and spag moss
This is the media formula i am currently using basicly the same as the old one just some of the volumes have been changed
: Murashige and Skoog basal medium with Vitamins (use at 1/3 strength) 1.47 grams : 1 lt of distilled water
after you make this use 50 ml of above mix and add 2ml of PPM this will give you a good sterilizing treatment. Soak seed for 2 to 4 hours in this don’t rinse just place seed strait on to media Ampullaria seed will need to be soaked for a lot longer up to 24 hours For the rest of the above media the 1 lt of MS you have made I add: : 1 ml of PPM : 30g sucrose (sugar) : About 5 grams of AGAR (test this out first with some plain water as some brands of agar require different ratios) I have been using grow gels for some time now they are better than agar : adjust PH to 5.5 - 5.68 range (use white vinegar to lower PH and bi-carb soda (baking soda) to raise PH) Add media to your containers or jars and sterilize as normal.