BSPP News Summer 2001 - Online EditionThe Newsletter of the British Society for Plant Pathology
Number 39, Summer 2001
A week in the life of a molecular plant virologist
Sue Angell, Department of Disease and Stress Biology, John Innes Centre
I cycled to work to start the day off well with some fresh air and exercise, but arrived in a bad mood because a lunatic car driver tried to knock me off my bike; at least it wasn't raining.
My first task was to encourage my team, who are all studying plant-virus interactions, to go down to the glasshouse to check the state of our plants - expecting that all of our plants would be looking good, although I hoped that those inoculated with viruses would be looking horrible.
My next job was to chat with Ingela, my new post-doc from Sweden. Ingela is investigating changes in plant gene expression in response to virus infection. She knows exactly what she's doing and seems completely in control of her project and, to save questions, she's blonde and likes ABBA.
I then collected 240 Arabidopsis plants from the glasshouse to check under the microscope in the lab. These plants were inoculated last week with a virus tagged with the green fluorescent protein (GFP) reporter gene. Whenever the virus replicates, GFP is produced. So, by monitoring green fluorescence under UV light we can see where the virus has replicated and moved in the plant. This is one of my least favourite tasks as these plants have to be brought up to the microscope at least four times over a period of two weeks and each time the plants must be wrapped in plastic bags to provide biological containment and there are eight doors and a flight of stairs to negotiate. It's ridiculous that there is nowhere in the glasshouse to put our microscope and even crazier that I'm the one carting these plants around when I could make my research assistant do this for me - am I too nice to him?
At lunchtime it was our weekly lab meeting, held jointly with another research group. Each week someone gives an informal presentation of what they have been doing, and this week it was my turn. The scariest and most time-consuming part is the "cake baking" because "if you present a lab meeting, you make a cake for everyone else to eat". Most people worry about whether they have enough new data to present - I worried all weekend about what to make for Monday's presentation, and spent most of the weekend re-running video recordings of daytime cookery programmes. I hate it.
The afternoon was spent setting up cultures of Agrobacterium for virus inoculations and E. coli for various cloning experiments. Most of our viruses can be rub-inoculated onto leaves, but this won't work for some. We introduce these more awkward viruses into plants as infectious cDNA clones on T-DNA, via Agrobacterium. This procedure is termed "agro-inoculation"; I've learnt to hyphen this word since a friend thought we did "a groin oculation".
I left work at a reasonable hour as I had my carving class to get to. Most people at JIC seem to think I go calving on a Monday night, but, worryingly, don't seem surprised that I might spend my leisure time with a hand up a cow's bottom.
I cycled to work to start the day off well with some fresh air and exercise, but arrived in a bad mood because some idiotic car driver tried to knock me off my bike; at least it wasn't raining.
First, I had a chat with Dave, my PhD student. Dave is characterising genes controlling susceptibility to virus infection and is in his second year. His project is progressing well. However, at the end of our meeting I had to remind him of a job he was failing to keep up with, to whit, "when was he going to organise the next lab bowling trip?". Usually these evenings consist of ten-pin bowling, the odd beer, and a hot curry. Past outings have included a few extras: a student doing a paint stripping job on a taxi; a Terry Waite look-a-like doing a Tom Jones impersonation (complete with red knickers on head); and a very special invited guest ripping his clothes off outside Norwich railway station (in December!).
A distinguished virologist (picture blurred to protect his identity!)
enjoying an Angell lab evening on the town.
After lunch there was a Sainsbury Lab seminar on plant pathology and I had to do a virus preparation because I had noticed last week that our virion stocks were getting low. So, I rub-inoculated 60 plants with some infected sap and they now looked very sick. I ground up the symptomatic leaves to make a "green soup" and after a chloroform extraction and a couple of PEG precipitations I had a new virus prep that my team could use for inoculations. If only cooking was as straightforward.
I cycled to work to start the day off well with some fresh air and
exercise, but arrived in a bad mood because a homicidal car driver tried
to knock me off my bike; at least it wasn't raining. I soon cheered up
because a bowling trip had been organised for that evening! I congratulated
Dave on his achievements.
I made some E. coli competent cells to use in a DNA transformation. I wanted to clone the plant DNA flanking a T-DNA insertion because the Arabidopsis line carrying this particular insertion appeared to be resistant to virus infection. Last week I did thermal asymmetric interlaced polymerase chain reactions (TAIL-PCR) and now ligated the PCR products into a plasmid vector. Once in this plasmid I would be able to sequence the inserts and digest them back out again to go into other vector constructs. I put the transformation plates safely in the incubator overnight and offered a solemn prayer for colonies.
I wasn't feeling too good - I knew that bag of crisps last night was past its sell-by-date. I cycled to work to start the day off well with some fresh air and exercise, but arrived at work in a bad mood because my head was hurting and a car driver - doubtless party to the conspiracy - tried to knock me off my bike; at least it wasn't raining.
It was very gratifying to see colonies on my transformation plates and I set up cultures for DNA preps tomorrow. I was also delighted to see everyone else suffering from last night although the boys would never admit to frailty.
I collected those Arabidopsis plants again and after lunch I went back to the glasshouse to harvest leaves for RNA preparations.
Well, it was raining, so b****r the fresh air and exercise, I got in the warm, dry car and drove to work, avoiding suicidal cyclists, and arrived in a bad mood.
First, I had a chat with Anthony, my research assistant. I needed to catch up with what he was doing and discuss jobs for next week. Ant's been characterising plant proteins that interact with viral movement proteins. We have some interesting data from these experiments and I expect these will be published soon (another good excuse to go bowling?).
I then went to a weekly meeting for Project Leaders in my department where we discussed departmental issues and exchanged information from various committees, etc. I represent the department on the Controlled Environment, Biological Safety, and Capital Equipment Committees, but had nothing to report since there had been no committee meetings for me to attend that week.
After this, I went to the JIC weekly site seminar.
Lunch, and then I prepared plasmid DNA from cultures (set up yesterday), digested the DNAs, and cleaned up positive clones ready for sequencing. I set up the sequencing reactions and planned to pop in Saturday to clean up the reactions ready for Dave and Paddy (the JIC sequencing service) to put on Monday's sequencing run.
Finally, I set up cultures ready for Sunday, started to relax in front of my computer screen, and thought "not a bad week really". My screensaver came on and I watched Ryan Giggs score yet another amazing goal against Arsenal.
I've been a project leader for two years now and I guess it's a bit like being a football manager. You supervise a multi-national squad, you must score lots of goals (win grants, publish papers) to stay at the top, you can run around with your shirt off when you win and bawl your eyes out when you lose, and there's the occasional lousy refereeing. My ambition is for us to become the research equivalent of league champions, or at least get into the play-offs, because I'd like to be able to say "we done brilliant".