Written by Amina Sultana. This is the report from a 2020 BSPP Lockdown Bursary. Click here to read more/apply for one yourself.
During the summer of 2020 I was fortunate enough to be awarded a BSPP undergraduate lockdown bursary. My bursary project was to investigate oilseed rape yield losses caused by two closely related phoma stem canker pathogens Leptosphaeria maculans and L. biglobosa. The L. maculans is generally associated with stem base canker while L. biglobosa is associated with upper stem lesions. Therefore, L. maculans is considered more damaging than L. biglobosa. However, recent research showed that L. biglobosa can cause both basal stem cankers and upper stem lesions. Therefore, the contribution of Lb to oilseed rape yield losses has been underestimated. In the UK both L. maculans and L. biglobosa are present in natural pathogen populations, it is difficult to distinguish the yield loss caused by L. maculans from L. biglobosa in field experiments. Therefore, the experiment was done in a glasshouse. Two oilseed rape cultivars were inoculated with L. maculans only, L. biglobosa only and both L. maculans and L. biglobosa. Plants inoculated with water were used as control. The inoculated plants were assessed for stem canker severity. After canker assessment, each individual plants were harvested for yield. The yield components, such as number of branches, number of pods on the main stem and total seed yield of each plant were assessed.
After the glasshouse work (e.g. disease and yield components assessment) and remote work (e.g. data entering and analysis), I really want to learn some molecular techniques related to plant pathology. Due to the COVID-19 climate there were certain issues for lab-based work. Thanks to the Dean of School of Life and Medical Sciences, Dr Richard Southern, I had permission to access laboratories at the University of Hertfordshire to facilitate with my learning. The laboratories were set up with enough space for us to work with social distancing measures in place. I had my personal PPE and my own supply of chemicals and equipment required for the practical work.
Over the course of the project I was able to learn many new techniques. I was introduced to disease severity on stems and assessing yield losses as a result of the disease, this involved some remote learning as well. I was also introduced to the methods used for processing spore tapes from a spore sampler, being able to identify and count these spores as well as DNA extraction from the spore tapes by following a protocol. I was also taught how to carry out RNA extraction from leaves and mycelia by also following a protocol. I was shown how to conduct qPCR to detect and quantify L. maculans from L. biglobosa from spore samples. I learned how to measure DNA or RNA quality and concentration using the Qubit machine. Finally, I was shown agarose gel electrophoresis to check PCR products or overall quality of the RNA samples.
Overall, this project was a fantastic opportunity for me to learn all kinds of useful new techniques which will benefit me in my future years in research. I would like to thank the BSPP and my supervisor Dr Yongju Huang for granting me with this enjoyable experience. I would like to thank Dr Chinthani Dewage, PhD student James Fortune for their help and guidance and bursary student Evren Bingol for support and friendship.
Amina Sultana
University of Hertfordshire