Biology subjects

Knight, N. L.

Publications and source records attributed to Knight, N. L..

4 recordsLinked to original sources

bioRxiv2025

Plasmid copy number variation impacts pathogenicity and quantification of Curtobacterium flaccumfaciens pv. flaccumfaciens infecting mung bean

In bacteria, plasmids can confer the ability to cause disease. Although they can potentially vary in copy number, little has been reported on the dynamics of plasmids in plant pathogenic bacteria. Pathogenicity of the bacterium Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which causes foliar disease on leguminous crops, including mung bean (Vigna radiata), has previously been linked to a plasmid. This study explored the variation in plasmid copy number among a genetically diverse collection of 25 Cff isolates using purposely designed quantitative PCR assays for chromosomal and plasmid DNA targets. Pathogenicity and virulence of six Cff isolates, including one plasmid-free isolate, were assessed on the susceptible mung bean cultivar Opal-AU using visual symptoms, trifoliate dry weights and Cff DNA quantities following stem inoculation. Plasmid copy numbers varied significantly (p <0.001) among plasmid-carrying isolates, ranging from 1.37 to 2.74. For subsequent quantification in planta, the single-copy chromosomal PCR target gene, gyrase b, was selected. Mung beans inoculated with plasmid-carrying isolates exhibited significant variation in visual symptoms (p <0.001), while trifoliate dry weights and Cff DNA quantities did not significantly differ. The treatment using a plasmid-free isolate was not significantly different from the negative control for each of the disease traits. Knowledge of plasmid dynamics in Cff populations lays a foundation for improved understanding of the inheritance and impact of plasmid-related traits. The quantification assays will be useful for monitoring Cff populations and the demonstrated variation in pathogenicity and virulence can assist efforts to breed host plant germplasm with reduced susceptibility to Cff.

pathology↗

bioRxiv2025

Monitoring fungicide resistance frequencies -- a case study of barley net blotch

Decreased sensitivity to fungicides impacts the effectiveness of fungicide applications for managing plant disease. Knowledge of the frequency of decreased sensitivity in field populations is critical for evaluating risks for disease control. This study has applied a droplet digital PCR detection approach to assess pathogen populations and quantify the frequencies of alleles associated with decreased sensitivity to either demethylation inhibitor or succinate dehydrogenase inhibitor fungicides in Pyrenophora teres causing net blotch on barley in Western Australia. Pyrenophora teres f. maculata was the most frequent form of the pathogen in the sampled region. Frequencies of decreased fungicide sensitivity alleles varied, being as great as 92% for the PtTi insertion in the CYP51A promoter, 88% for C-S75 and 28% for L489-2. Impacts of cultivar selection and weed hosts on the presence and survival of decreased fungicide sensitivity were observed. Determining the dynamics of alleles within different field populations of P. teres provides an important perspective on the impact of fungicides, the fitness associated with decreased fungicide sensitivity alleles and the susceptibility of barley cultivars.

pathology↗

bioRxiv2024

Clonal expansion from standing genetic variation underpins the evolution of an emerging plant pathogen in Australia

Pathogens can evolve rapidly, leading to the emergence of novel strains that can overcome commercially deployed host plant resistance. Understanding the genetic and phenotypic diversity and population dynamics of plant pathogens is crucial to inform breeding programs targeting resistance. Tan spot, otherwise known as bacterial wilt, caused by Curtobacterium flaccumfaciens pv. flaccumfaciens, is a destructive disease of mungbean in Australia. Since the 1990s, several mungbean cultivars with partial resistance to tan spot have been released; however, cultivars initially rated as moderately resistant were later rated as moderately susceptible to tan spot. This study investigated the genetic and phenotypic diversity and temporal evolutionary dynamics of C. flaccumfaciens pv. flaccumfaciens in Australian mungbean fields. Whole-genome sequencing of 119 isolates collected from mungbean and other legumes (1986-2019) enabled analyses of pathogen evolution in Australia and in a global context. Results revealed that clonal expansion from standing genetic variation, rather than introduction of novel genotypes, has driven the evolution of this pathogen in Australia since the 1980s. Glasshouse trials confirmed the role of a plasmid in pathogenicity of C. flaccumfaciens pv. flaccumfaciens but found no significant differences in aggressiveness between clonal lineages. This study underscores the importance of understanding pathogen diversity and aggressiveness to guide breeding programs for effective and durable disease resistance and highlights the need for further studies on the competitive fitness of isolates and characterisation of private alleles linked to dominant clonal lineages. SignificanceTan spot is an increasingly significant pathogen affecting beans and mungbean worldwide, with a broad host range and quarantine importance in many countries underscoring the need for effective breeding strategies to manage the disease. Our research provides insights into genetic and phenotypic diversity of this important plant pathogen and temporal changes in its population structure in Australia, which are crucial for developing informed breeding strategies and managing the spread and impact of the disease. This work also underscores the critical need for robust, validated screening protocols and selection of aggressive, locally representative isolates for resistance breeding to ensure accurate assessment of resistance in crops for effective and durable resistance.

evolutionary biology↗

bioRxiv2020

Disease responses of hexaploid spring wheat (Triticum aestivum) culms exhibiting premature senescence (dead heads) associated with Fusarium pseudograminearum crown rot.

Hexaploid spring wheat (Triticum aestivum) may exhibit significant crown rot disease responses to infection by Fusarium pseudograminearum, with a range of susceptibility levels available in commercial cultivars. Dry conditions during grain-fill may lead to the expression of prematurely senescing culms, which typically fail to set grain. Assessment of hexaploid spring wheat plants exhibiting both non-senescent and prematurely senescent culms was performed using visual discolouration, Fusarium pseudograminearum biomass, vascular colonisation and quantification of wheat DNA in culm sections sampled at three different heights above the crown and at the peduncle. A comparison of these parameters at four time points from milk development, when senescent culms are first observed, to maturity was conducted. Samples from six commercial cultivars were collected in 2014 from Narrabri and Tamworth, New South Wales and Wellcamp, Queensland. Prematurely senescent culms exhibited greater visual discolouration, Fusarium pseudograminearum biomass and vascular colonisation than non-senescent culms in each cultivar. Colonisation of xylem and phloem tissue was extensive in the basal portions of prematurely senescent culms (36 to 99%), and suggests significant impacts on water and nutrient movement during crown rot disease. Maturation coincided with significant changes in Fusarium pseudograminearum biomass and vascular colonisation. Wheat DNA content varied among cultivars, culm conditions, culm sections and sampling times. The variation in the severity of disease states between culms of the same plant suggests that the timing of initiation of infection in individual culms may vary.Competing Interest StatementThe authors have declared no competing interest.View Full Text

pathology↗