Answer: Yes! Experiment Growing tips of fungi emerging from leaf tissue is excised and transplanted to a new culture plate. If culture is pure (by visual inspection), thrice-autoclaved barley seeds are added to the fungi. The fungi overgrow and colonize the barley seeds which are then stored at -70oC. To recover the fungi, a colonized
Project 3) Can we store and recover fungi for long-term storage? Read More »
Endophyte 1.0 (Fall semester- early Spring 2014) ANSWER: YES! We see some morphotypes consistently emerging from Magnolia leaf tissue. Experiment: Our first experiments were exploratory in nature. We adapted published techniques to conditions in our lab (Bascom-Slack et al., 2012 and supplementary material; Appendix). We explored various ways of consistently producing equivalent size pieces of tissue
Project 2) Can we culture any endophytes from Magnolia leaf tissue? Read More »
ANSWER: All kinds of stuff, but very different from those we find in Magnolia. Experiment: MEA (malt extract agar) or PDA (potato dextrose agar) plates were left open for 16-24 hours in the lab, sealed with parafilm and incubated at room temperature for 4-5 days. Results: These fungi are a different population than those
Project 1) What’s in the Air? Read More »
We had a very successful year. It is worth noting that this project did not exist a year ago. The hard work of the Tague lab crew is evident. We have successfully cultivated endophytic fungi from Magnolia grandiflora. We have isolated purified cultures for some of these fungi and established long-term stocks. We have obtained consistent
First Year Conclusions and Directions Read More »
Endophyte 3.1 (BWT) ANSWER: Smaller is better, but the 2.5 cork borer may be the best to use for consistent results. The modified protocol is fine, but it is more difficult to keep the tissue sterile during the cork boring process. It is much easier to cut the disks first and then surface-sterilize them. Experiment:
Project 6) Does size matter with the modified protocol? Read More »
Endophyte 3.0 (SH, AQ, more?) Answer: Maybe. Experiment We compared our Standard protocol to a modified protocol. From the same leaf, we 1) cut disks → surface sterilize as usual → plated AND 2) cut a large piece of leaf → surface sterilized → THEN cut disks with a (sterile) cork borer. Results The data are scattered, particularly in terms
Project 5) Is the sterilization protocol too harsh? Read More »
Endophyte 2.0 Answer: Yes! But in a different way than Gamboa et al (2002). The population density of culturable fungi in our (temperate) Magnolia seems to be much lower than those found in tropical species. Consistently, a substantial number of leaf pieces have no fungi emerging from them. This might allow us to have a simple
Project 4) Does size (of the piece of leaf tissue) matter? Read More »
