Agrobacterium tumefaciens-mediated transformation and expression of GFP in Ascochyta lentis to characterize ascochyta blight disease progression in lentil

Autor: Johannes W. Debler, Robert C. Lee, Christina R. Grime, Bernadette M. Henares, Lina M Farfan-Caceres
Jazyk: angličtina
Rok vydání: 2019
Předmět:
0106 biological sciences
0301 basic medicine
Leaves
Fungal Structure
Confocal Microscopy
Germ tube
Plant Science
Pathology and Laboratory Medicine
01 natural sciences
Fluorescence Microscopy
Gene Expression Regulation
Plant
Genes
Reporter
Mesophyll
Medicine and Health Sciences
Pathogen
Fungal Pathogens
Microscopy
Multidisciplinary
biology
Plant Anatomy
Plant Fungal Pathogens
Light Microscopy
Agrobacterium tumefaciens
Ascochyta
Medical Microbiology
Host-Pathogen Interactions
Medicine
Lens Plant
Pathogens
Research Article
Agrobacterium
Plant Cell Biology
Science
Green Fluorescent Proteins
Plant Pathogens
Mycology
Research and Analysis Methods
Microbiology
03 medical and health sciences
Transformation
Genetic
Ascomycota
Blight
Microbial Pathogens
Plant Diseases
Appressorium
Inoculation
fungi
Biology and Life Sciences
Cell Biology
Plant Pathology
biology.organism_classification
030104 developmental biology
Confocal Laser Microscopy
010606 plant biology & botany
Zdroj: PLoS ONE, Vol 14, Iss 10, p e0223419 (2019)
PLoS ONE
ISSN: 1932-6203
Popis: The plant immune system is made up of a complex response network that involves several lines of defense to fight invading pathogens. Fungal plant pathogens on the other hand, have evolved a range of ways to infect their host. The interaction between Ascochyta lentis and two lentil genotypes was explored to investigate the progression of ascochyta blight (AB) in lentils. In this study, we developed an Agrobacterium tumefaciens-mediated transformation system for A. lentis by constructing a new binary vector, pATMT-GpdGFP, for the constitutive expression of green fluorescent protein (EGFP). Green fluorescence was used as a highly efficient vital marker to study the developmental changes in A. lentis during AB disease progression on the susceptible and resistant lentil accessions, ILL6002 and ILL7537, respectively. The initial infection stages were similar in both the resistant and susceptible accessions where A. lentis uses infection structures such as germ tubes and appressoria to gain entry into the host while the host uses defense mechanisms to prevent pathogen entry. Penetration was observed at the junctions between neighbouring epidermal cells and occasionally, through the stomata. The pathogen attempted to penetrate and colonize ILL7537, but further fungal advancement appeared to be halted, and A. lentis did not enter the mesophyll. Successful entry and colonization of ILL6002 coincided with structural changes in A. lentis and the onset of necrotic lesions 5–7 days post inoculation. Once inside the leaf, A. lentis continued to grow, colonizing all parts of the leaf followed by plant cell collapse. Pycnidia-bearing spores appeared 14 days post inoculation, which marks the completion of the infection cycle. The use of fluorescent proteins in plant pathogenic fungi together with confocal laser scanning microscopy, provide a valuable tool to study the intracellular dynamics, colonization strategy and infection mechanisms during plant-pathogen interaction.
Databáze: OpenAIRE
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