Joanne Adero is Research Assistant at The National Crops Resources Research Institute, Uganda and a CONNECTED Network member

Joanne successfully applied for a CONNECTED Training Voucher to visit The Institute of Molecular Plant Sciences, University of Edinburgh, UK, under the supervision of Prof. Gary Loake. The visit took place in Spring 2019.

Crossing Aradopsis under a microscope

During the training I was exposed to excellent research infrastructure. The laboratory has facilities that include plant growth rooms, transgenic greenhouses, and cutting-edge microscopy. Seeing these facilities therefore improved my knowledge of the current instrumentation required in plant molecular and tissue culture work.

I gained detailed knowledge in a number of areas including:

• Forward genetic screening working with Arabidopsis, screening for mutation in GSNOR deficient background of Arabidopsis. S-nitrosylation directly or indirectly regulates plant immunity proteins, with changes in protein activity, localization, interactions and stability
• Understanding how increased S-nitrosylation reduces salicylic acid accumulation and therefore increased susceptibility to pathogens
• I participated in genetic screening of GNSOR suppressor mutants, which supress the gsnor1 phenotype. This involved, using salicylic acid-induced luciferase. Expression of the PR1::Luc was screened with ultra-low light imaging after exogenous salicylic acid application. Candidate suppressor mutants that convey salicylic acid-induced PR1::Luc expression were selected for forward sequence analysis to identify possible second site mutations that can supress the gsnor1 phenotype.
• PR1::Luc was introduced into the GNOR background through back crossing of mutant parent lines with wild type genotypes, harvesting seed from mutant parent lines, washing and planting out seed in culture media, and phenotyping mutant Arabidopsis plants.
• Crossing Arabidopsis parental lines involved identification of potential flowers, opening of the recipient stigma, harvesting of anthers from identified male flowers and pollinating donor and recipient flowers. All of which is done under the microscope, thus highly improving my skills in microscopy.

I also carried out plant nucleic acid extractions for whole genome sequencing from using experimental protocols which differ from the protocols I am currently using. This new knowledge will improve the quality of nucleic acids I can obtain for my daily routine work for example in plant virus indexing.

Plating out seed in culture media, after harvesting seed from mutant parent and washing

There were also a lot of activities in tissue culture during my stay, which has improved my tissue culture techniques.

During my visit I had the opportunity to attend a symposium where scientists from Edinburgh University were presenting their current research. From this I got to learn about diverse research projects being carried out in different fields, and also managed to identify some research gaps. I was also able to attend Friday presentations by lab members on the progress of their ongoing research. These gave me an insight into research being done by the different research fellows in the laboratory, and presentation skills used.

Improving my future research

I lead the molecular and tissue culture activities and the training will improve my research in a number of ways, for example:

• My improved microscopy skills will be applied in plant meristem extraction work for virus elimination
• Improved techniques in tissue culture will be applied for routine tissue culture activities
• New techniques for extraction of nucleic acids will improve the quality of nucleic acids used in my daily routine work like virus indexing and genotyping
• I was also able to establish new connections with scientists from different parts of the world because the Institute has a wide range of national and international scientist and PhD students
• Most importantly, I was able to establish contacts with scientists doing transcriptome work. This is of direct interest to me because I am trying to learn this technique so that I can use it in studying gene expression in sweetpotato.

I am also involved in training and building capacity for technicians, undergraduate and post graduate students. Knowledge gained from the training will be passed onto to the different categories of personnel above during training and routine work.

I would like to thank CONNECTED and BBRSC for funding me and giving me this opportunity to gain such diverse knowledge during this training.

It was an exciting experience for me which I will recommend to as many early career researchers as I possibly can, and I look forward to participating in more activities and training.

Joanne Adero leads Plant Virology/Molecular Biology with the national sweetpotato breeding programme. Her duties and responsibilities include:

• Design and management of laboratory, screen house and field virology experiment
• Development and optimisation protocols for sweetpotato virus indexing, tissue culture, virus elimination and genomic DNA applications
• Conducting sweetpotato virus disease epidemiology experiments and surveys
• Data and results interpretation, analysis, report writing
• Training of undergraduate, postgraduate students and the community at large on sweetpotato pathology and diagnostics.

For information about, and to apply for, the second round of CONNECTED Training Vouchers, use this link