Hey all,
After over a year of delay, I finally managed to get some sterile cultures going last March and in early May. It sure is convenient to have access to a lab with flow hoods and ethanol/supplies! The jars and gel/nutrients are all purchased by me though.
I was in san diego all summer so i made these cultures a week before i left, placed them under my lights with the rest of the plants in the basement (~80-85F in summer), and then I came back after 3 months to see what happened .
I decided to use a ton of media in each of the jars so that they could hopefully live longer after germinating.
I should've used more gelrite since the gel was almost liquid after it solidified...so after 3 months, the plants used up all the nutrients and the gel became entirely liquid for many, even though I stirred the gelrite/nutrient melted mix really well before pouring most of the jars. Tips would be appreciated.
but here's what i did if you're interested (I used 1.5-mL microcentrifuge tubes and vortexed them on the vortex genie):
70% etoh rinse for all seeds, 2mins
Larger seed size group- do 6 min vigorous vortexing in 10% bleach (ie D. capensis, D. indica)
Med seed size group- do 6 min vigorous vortex in 5% bleach
Small seed size group- do 4 min vigorous vortex in 1% bleach (ie for D. burmannii or sessifolia)
My mix:
1/3 MS with vitamins (full conc. Is 4.4g/L)
30g/L sucrose
2.0g/L Gelrite
After spreading the seeds out with a pipetteman, I sealed the jars with micropore surgical tape, commonly used at ISU in tc labs.
About 1/4 didn't germinate and 1/6 had contamination. hopefully i will get better with time. I will also be taking a graduate level horticulture class to learn about sterile protoplast culture, pollen culture (i think) and other tc techniques, which should be amazing!!!
I am extremely appreciative of everyone's help- especially Dudo, Matt, and anyone who helped answer my annoying questions
So enough talk...here are the pics:
D. anglica
D. anglica var. 2
D. ascendens
not all the plants made it...this jar didn't have enough media in it to keep them alive
D. brevifolia
D. collinsiae
D. indica from my 1st attempt:
I should've stored it in the fridge, cuz it ran out of nutrients
D. nidiformis
D. sessifolia gone wild!
D. tokaeinsis jar 1 (just a little bit overcrowded )
D. tokaeinsis jar 2
D. tomentosa
VFT (the only seed that germinated)
typical D. capensis, grown in my 1st attempt and stored in the fridge for 3 months
All sundews will be used for/by the ISU CP Club that my friend (forum name peat) and I started.
Thanks for looking!
Aaron
After over a year of delay, I finally managed to get some sterile cultures going last March and in early May. It sure is convenient to have access to a lab with flow hoods and ethanol/supplies! The jars and gel/nutrients are all purchased by me though.
I was in san diego all summer so i made these cultures a week before i left, placed them under my lights with the rest of the plants in the basement (~80-85F in summer), and then I came back after 3 months to see what happened .
I decided to use a ton of media in each of the jars so that they could hopefully live longer after germinating.
I should've used more gelrite since the gel was almost liquid after it solidified...so after 3 months, the plants used up all the nutrients and the gel became entirely liquid for many, even though I stirred the gelrite/nutrient melted mix really well before pouring most of the jars. Tips would be appreciated.
but here's what i did if you're interested (I used 1.5-mL microcentrifuge tubes and vortexed them on the vortex genie):
70% etoh rinse for all seeds, 2mins
Larger seed size group- do 6 min vigorous vortexing in 10% bleach (ie D. capensis, D. indica)
Med seed size group- do 6 min vigorous vortex in 5% bleach
Small seed size group- do 4 min vigorous vortex in 1% bleach (ie for D. burmannii or sessifolia)
My mix:
1/3 MS with vitamins (full conc. Is 4.4g/L)
30g/L sucrose
2.0g/L Gelrite
After spreading the seeds out with a pipetteman, I sealed the jars with micropore surgical tape, commonly used at ISU in tc labs.
About 1/4 didn't germinate and 1/6 had contamination. hopefully i will get better with time. I will also be taking a graduate level horticulture class to learn about sterile protoplast culture, pollen culture (i think) and other tc techniques, which should be amazing!!!
I am extremely appreciative of everyone's help- especially Dudo, Matt, and anyone who helped answer my annoying questions
So enough talk...here are the pics:
D. anglica
D. anglica var. 2
D. ascendens
not all the plants made it...this jar didn't have enough media in it to keep them alive
D. brevifolia
D. collinsiae
D. indica from my 1st attempt:
I should've stored it in the fridge, cuz it ran out of nutrients
D. nidiformis
D. sessifolia gone wild!
D. tokaeinsis jar 1 (just a little bit overcrowded )
D. tokaeinsis jar 2
D. tomentosa
VFT (the only seed that germinated)
typical D. capensis, grown in my 1st attempt and stored in the fridge for 3 months
All sundews will be used for/by the ISU CP Club that my friend (forum name peat) and I started.
Thanks for looking!
Aaron