Agrobacterium-Mediated Transformation to Create an Insertion Library in Magnaporthe grisea
-
Last Update: 2020-11-16
-
Source: Internet
-
Author: User
Search more information of high quality chemicals, good prices and reliable suppliers, visit
www.echemi.com
Magnaporthe grisea
is the causal agent of rice blast disease and represents a model organism for the study of fungal plant-pathogen interactions. Pathogenicity is a complex phenotype, which is carefully orchestrated by the fungus and begins with recognition and infection of the host plant, followed by growth within the plant cells, and finally dissemination to the next host and continuation of the fungal life cycle. Certain genes must condition the ability of a pathogenic fungus to infect and cause disease symptoms. To learn more about the infection process and the genes that are involved in the complex interplay between
M. grisea
and rice, we used an insertional mutagenesis approach to attempt to randomly disrupt all genes in the fungal genome. Two transformation approaches were used to build a library of insertion strains in
M. grisea
. Polyethylene glycol/CaCl
2
-mediated protoplast transformation was the initial method we used and resulted in the generation of just more than 17,000 insertion strain lines. Later
Agrobacterium tumefaciens
-mediated transformation was adopted and the final number of insertional mutant strains of
M. grisea
strain 70-15 generated was more than 57,000. Here, we describe the methods used for
A. tumefaciens
-mediated transformation of
M. grisea
and the optimized protocols we have developed to enable high-throughput fungal transformation. Further details of this optimization can be found elsewhere.
This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only.
This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of
the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed
description of the concern or complaint, to
service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content
will be removed immediately.
