Uncovering the phosphoproteome of arbuscular mycorrhizal roots
EMSL Project ID
51375
Abstract
During the endosymbiotic association of plants and arbuscular mycorrhizal (AM) fungi, the fungus inhabits the root cortical cells where it delivers mineral nutrients to its plant host, and in return obtains carbon. Recent data indicate that kinases are involved in signaling that enables the root cell to accommodate the fungal symbiont. However, the substrates of these kinases, and the signaling pathways in which they are involved, are unknown. Deep transcriptomes of mycorrhizal roots are available, but there is only a single paper that reports phosphoproteome data of mycorrhizal roots and this was developed for citrus in association with Rhizophagus irregularis. There is no publicly available phosphoproteome data for mycorrhizal roots of Brachypodium distachyon, a flagship plant species on the DOE's phytozome portal, or for Medicago truncatula. The objective of this project is to identify and compare the phosphoproteomes of Brachypodium distachyon and Medicago truncatula mycorrhizal roots during association with the AM fungus, Diversispora epigaea, with their respective mock-inoculated controls. The experiments will include 3 genotypes of each host plant, the wild type and two kinase mutants. The kinase mutants are impaired in their ability to host intracellular fungal hyphae, indicating a role for the kinases in the endosymbiotic phase of the mycorrhizal association. Accessing the phosphoproteome of mycorrhizal roots is technically challenging because the colonized root cells comprise a small proportion of the mycorrhizal root, and the phosphoproteome comprises a small portion of the proteome. However, hand-dissected, mycorrhizal-enriched root samples combined with EMSL's protocols and advanced instrumentation, will enable these analyses. We anticipate phosphopeptide data will be obtained for both the plant and the fungal symbionts, although the former will dominate simply because fungal biomass comprises less than 10% of the mycorrhizal root. The data generated will provide a resource of immediate use for the research community where it will fill a major resource gap. The data from two host species, one monocot and one dicot, in association with the same fungal symbiont, provides valuable comparative opportunities. Data from wild type and kinase mutants will enable us to identify potential kinase targets for downstream investigation, and ultimately will advance an understanding of signaling during AM symbiosis.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2020-10-01
End Date
2023-08-25
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members