Lab Summary 10/17/12

Neurospora crassa cross observations.  Perithicia were maturing but not yet ready (ascospores had not yet "shot" from the perithicia) to pick individual ascospores.

Neurospora crassa conidia

Crosses of Neurospora crassa 
resulting in perithiucia formation
on minimal Vogel's media

Squash mount of Neurospora crassa 
perithicium showing asci and ascospores

Field collected specimens were available for observation.  We were challenged to classify them as either Ascomycetes or Basidiomycetes.

Basidiomycete

Basidiomycete

Basidiomycete

Basidiomycete

Ascomycete

Ascomycete

Basidiomycete

Basidiomycete

Basidiomycete

Lab Practical 10/10/12

My comments on the lab practical.
I believe the practical was very fair.  There were 20 questions with 16 microscopes set up with different spore types to be identified to genus.  Perhaps there could have been bonus points for providing the species of the fungi observed.

Unknown fungal project :    "What is wrong with my plant?"

Unknown #2 (A SNEAK PEEK!!!) To be continued
Source: Hibiscus plant from nursery retailer submitted to TPDDL 

 

Hibiscus plant with flower blight submitted to TPDDL

Dissecting microscopic view of necrotic flower petals.

Sterile needle transfer of conidia to 1/2 strength PDA grown at 26C

 

Riddel mount set up

Microscopic view of the Riddel mount of Botrytis sp.

Lab Summary 10/3/12

1.  The following cultures were available for microscopic examination to aid in unknown identifications and the various different spore types, condiogenous cells, and hyphae.

Alternaria brassicicola, Aspergillus niger, Botrytis cinera, Colletotrichum coccodes, Curvularia sp., Epicoccum sp., Fusarium graminearum, Monilinia fructicola, Nigrospora sp., Pestilotia sp., Rhizoctonia solani, Thielaviopsis brasicola, and Trichoderma viride. 

Fusarium graminearum  macroconidia and microconidia

Rhizoctonia solani hypae with septation

 

 

 

 

 

Pigmented Pestilotia sp spores

Thielaviopsis brasicola in pansy roots (TPDDL sample) with aleuriospores 

 

 

 Alternaria brassicicola pigmented spores with cross and longitudinal septa

Neurospora crassa cross resulting
in the formation of Perithecia

2.  Neurospora crassa crosses (SmRP10 x SmRP 11, SmRP10 x CP-1 GFP, SmPR10 x NCACo21-1) were successful.  Perithicia were present however, the ostioles had not yet formed.  It was observed in some of the agar plates trichogynes (specialized hypae) of N. crassa were seeking the opposite mating type across the plate.

















 3.  The week before (9/26/12) 2 different corn varieties (Silver Queen and OPR2-1) were inoculated with the haploid strain of Utilago maydis Sg200.  A week later we observed tumor formation and proceeded to prepare the tissue for fungal observation.  Proceedure as follows:  Destain plant tissue with 2:1 acetic acid:ethanol for 24 hours, stain with trifan blue, and then briefly destain.

Detatining (24hr) of the corn leaf sections inoculated with Ustilago maydis
showing symptom expression (tumors)

4.  A week (inoculated on 9/26/12) after the dimorphic experiment was set up, all but one of the Mucor rouxii cups were contaminated with N. crassa and were slightly dried out.  I took a cross sections through the agar and did observe the two different stages via compound microscopy, the yeast stage and the filamentous stage (hyphae).

Lab Summary for 9/26/12

I was not present for this lab due to job commitment  but was updated as to what experiments were preformed.

1.   Neurospora crassa crosses were set up.
2.   Corn plants were needle inoculated with Utilago madys 
3.  A dimorphic gradient of Mucor sp. was set up.

Updates for the following experiments will be posted on the next lab summary 10/3/12

Lab Summary for 9/17/12

Discussed and viewed sexual and asexual life cycles videos for the protist - Phytophthora infestans, the Chytrid - Olpidium brassicae (causal organism of the club root pathogen), Saprolegnia, and  the Blastocladiales Allomyces (a really cool fungus).  

Baiting for Allomyces sp. from the soil surrounding the pecan pit in front of the Peterson building, utilizing non-viable heep seeds as the bait.  Observed zoospores (mitospores) from the initial microscopic examination.  Plated some of the water on YpSs agar plates.  Additionally, removed the heep seeds from the baited jar and placed the seeds into a small plastic petri dish with DS solution, parafilmed the petri dish and incubated for 48 hours at room temperature.  The initial YpSs agar plates were contaminated with bacteria and did not result in any fungal growth.  The heep seeds resulted in the amazing recovery of an Allomyces sp.  It was so incredible to see the zoospores (mitospores),  mitosporangia, meisporangia and rhizoids (photos below).  Transferred the fungal growth surrounding the heep seeds to YpSs agar plates in an attempt to recover a pure culture (update to come).  In addition, I took a video of the zoospores swimming in the petri dish (hopefully will be able to edit and post in the future).

Microscopic views of an Allomyces sp. 

A V8 agar plug of Phytophthora floating in soil water  was viewed microscopically to visualize the zoospores produced by the sporangia. I prepared a squash mount with a small piece of the agar plug stained with acid fucsion and observed via microscopic examination, coenocytic hypae, sporangia with zoospores present and  chlymadospores.

Briefly discussed Neurospora crasse crosses (mating) and will start those experiments next week (9/26/12).

Photo gallery update of the Riddell mounts from lab on 9/12/12

A. parasiticus

A. sojae

A. sojae

A. sojae

A. flavus

A. nidulans 

A. tamari

  

A. niger

A. oryzae

A. oryzae