I am working on Rhizopus oryzae. I am doing broth culture of Rhizopus oryzae, but when I inoculate it with pure culture, it forms clumps.
Hello Amit,
Have you tried titrating the amount of pure culture you start with? High amounts of starting culture per flask is more likely to cause clumping than using more flasks inoculated with lower amounts of the starting culture. The purpose for requiring isolated spores can make a difference in your experimental methods. Is the issue accuracy in counting spores or differences in metabolism or other considerations? The end-purpose is quite important for knowing what variables to adjust without compromising your primary experimental goal. Nutrients and calcium can influence clumping but may not be compatible with your experimental goals. Aeration can also be adjusted but may be impractical in a shared incubator. Trying different inoculum sizes, if this works for your purpose, has the advantage of avoiding more dramatic changes in other experimental variables such as medium composition.
Thank you very much Mam for adding answer to my question.Actually i am growing the Rhizopus oryzae pure culture on PDA culturing them for 7 days on and then transferring them to broth culture so that i can get the homogeneous culture of the same . When i have cultured them first time i got the homogeneous culture but when i have repeated the same it started following clump.If any method is there so that i can get the homogeneous culture of the same kindly tell me.
Please use a sterile cappilary tube and mount the spores on a slide and observe it under dissecting/light microscope and suck a single spore in the capillary tube. Further blow the single isolated spore in a sterile PDA broth. Try if it works. I have tried it successfully but it needs tremendous patience till a single spore is captured in a capillary tube.
We were also separating spores from Rhizopus oryzae. Through liquid culture, it was very difficult to separate the spores, traces of the media was contaminating. The only way is to culture the fungi in solid media and after sufficient growth, separation with spreader by adding a little bit water.
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