Page 30 - TriSociety Virtual Event

Abstracts

threat to sustainable crop production Fungal and their host plants are evolving together revealed an extensive catalogue of candidate
pathogens can persist in the environment by but as the climate changes fungal pathogens secreted efector proteins (CSEPs) employed
forming resistant spores A s such spore dor­ are adapting at an accelerated rate compared by C. lentis to mediate infection Among 38
mancy dispersal and germination are integral to their hosts We hypothesiZe that RNA­me­ CSEPs evaluated for gene expression during
to the spread of fungal diseases We use Ustila­ diated mechanisms enhance fungal adapta­ the necrotrophic phase only five showed
go maydis D C Corda as a model to study tion to stress We are using the model fungal expression at that stage and not in mycelium
molecular aspects of teliospore germination pathogen Ustilago maydis to investigate the Using a bait­and­capture approach we aim
Teliospores are thick­walled dormant diploid role that natural antisense transcripts (NATs) to identify L. ervoides host proteins targeted
cells that germinate and complete meiosis to play in modulating the stress response. NATs by these CSEPs Three CSEPs were success­
initiate new rounds of infection Early fungal are a subset of non­coding RNA s which have fully expressed as glutathione­S­transferase
spore studies indicated that all the necessar y regions of their sequence complementary to (GST)­tagged fusion proteins in Escherichia
components required for germination inclu­ an mRNA An RNA­seq comparison of three coli. Unfortunately heterologous expression
ding preformed messenger RNA s (mRNAs) smut fungi revealed that 349 of the 2617 NATs resulted in the aggregation of each fusion
are stored in the spore during dormancy We found in U. maydis are conser ved among all protein into insoluble inclusion bodies Mo­
hypothesiZe that some stored mRNA s are three smut species The conser vation of NATs difications of culture conditions (e g length
stabiliZed in the dormant teliospore through among these smut species suggests that and temperature of incubation period and
double­stranded RNA (dsRNA) formation and they have important functional roles Genes concentration of inducer) did not enhance
are unwound by RNA helicases during germi­ complementary to these conser ved NATs were solubility The successful solubiliZation of each
nation to be made available for translation investigated to identify those with previously natively folded fusion protein was media­
RNA helicases are highly conser ved enZymes documented roles in stress response Among ted by incubating the insoluble pellet with
that are capable of forming RNA clamps these genes the levels of complementary 10% sarcosyl following a published method
unwinding RNA­RNA duplexes and displacing NATs were assessed by reverse transcrip­ developed for GST­tagged fusion proteins
proteins We identified all RNA helicases in tion­PCR using RNA isolated from haploid Purification of the solubiliZed proteins requires
U. maydis and determined by RNA seq that U. maydis cells exposed to various stresses further optimiZation to improve yield Once
five RNA helicases had elevated transcript Seven NATs with altered expression levels established the proteome of L. ervoides will
levels in dormant teliospores and that these were identified and will be further characteri­ be screened for interacting host proteins The
levels decreased during germination Deletion Zed Future experiments will assess whether identification of protein isoforms targeted by
strains were created and their phenotypes expression of these NATs alters comple­ C. lentis CSEPs could be used to introgress
were assessed to gain insight into the function mentary mRNA stability stress responses or resistance from L. ervoides into L. culinaris
of two RNA helicases The information gained pathogenesis by the fungus Identifying and germplasm while removing undesirable traits
from this study enhances our understanding describing these RNA­mediated responses to from the wild species
of fungal spore germination which may lead environmental stress may provide new tools
to the development of novel disease control for combating the increasing prevalence and 11. Does trophic switch of Leptosphaeria
measures severity of fungal diseases maculans happen under both incompa­
tible and compatible interactions with
Session 3. Student oral competi­ 10. Good things come to those who bait: canola (Brassica napus)?
tion: Genetics, Biotechnology and using fungal effector proteins to identify K R E PADMATHILAKE Z ZOU AND W G
host targets
D FERNANDO Department of Plant Science,
Breeding M WENGLER J HALLIDAY A BEATTIE H University of Manitoba, Winnipeg, MB R3T2N2,
WANG AND S BANNIZA Department of Plant Canada.
9. Natural antisense transcripts and stress Science, University of Saskatchewan, 51 Cam­
response in Ustilago maydis pus Drive, S7N 5A8, Saskatoon, SK, Canada; Canola (Brassica napus) is a 29.6­billion­do ­
M K LARIVIERE AND B J SAVILLE Environ­ (H.W.) Department of Biology, University of llar crop in Canada. Leptosphaeria maculans
mental & Life Sciences Graduate Program, Trent Saskatchewan, 51 Campus Drive, Saskatoon, causes blackleg, is one of the most economi­
University, DNA Building, 2140 East Bank Drive, SK, S7N 5A8, Canada. cally significant diseases of canola. Since the
Peterborough, ON, K9J 7B8, Canada; (B.J.S) breakdown of resistance in B. napus genotype
Forensic Science Program, Trent University, DNA Of the foliar diseases that threaten lentil grown in Canada, it is pivotal to introduce new
Building, 2140 East Bank Drive, Peterborough, production anthracnose (caused by the he­ resistant genotypes. This study focused to
ON, K9J 7B8, Canada. mibiotrophic Colletotrichum lentis) poses the evaluate the potential of B. napus genotype,
greatest threat A wild relative of lentil Lens ‘01­23­2­1’ which carries Rlm7 to be the next
The climate is changing at an accelerated rate ervoides is a source of superior resistance to candidate gene to be introduced. 01­23­2­1
raising concern about global food security the virulent race currently dominating the pa­ line was inoculated with L. maculans isolate
While advances are being made to increase thogen population in Canada A combination (UMAvr7) carr ying an avirulence gene AvrLm7,
crop yields the threat of evolving pathogens of transcriptome and proteome analyses and and the CRISPR/Cas9 knockout AvrLm7
must also be considered Fungal pathogens the genome assembly of the pathogen has mutant (umavr7) of the same isolate to make

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