Matton, D. P. | Search bioRxiv

bioRxiv2024

Differential gene expression analysis identifies a group of defensin like peptides from Solanum chacoense ovules with in vitro pollen tube attraction activity

Solanum chacoense is a wild potato species with superior genetic resistance to diseases and pests that has been extensively used for introgression into cultivated potato. One determinant of crossing success between wild and cultivated potato species is the effective ploidy of the parents. However, little is known about whether other, prezygotic level, breeding barriers exist. We hypothesize ovular pollen tube guidance may serve as such a checkpoint. Tests for species-specific pollen tube guidance using semi-in vivo assays suggested a positive correlation between species-specificity and taxonomic distance. RNA-seq of ovules hand dissected from wild type plants at anthesis and two days before anthesis, as well as from a frk1 mutant lacking an embryo sac identified a list of 284 embryo sac-dependent genes highly expressed in mature ovules and poorly expressed in all other samples. Among these are 17 Solanum chacoense cysteine-rich proteins (ScCRPs), considered to be good candidates since CRPs are ovular pollen tube attractants in other species. A group of three cloned and purified ScCRP2 sequences belonging to the DEFL protein family showed moderate levels of in vitro pollen tube attraction activity in functional assays. We conclude that ScCRP2s are good candidates for ovular pollen tube guidance in S. chacoense. HighlightsThis work introduces key characteristics of pollen tube guidance in Solanum chacoense, identifies candidate attractants through transcriptome subtraction and reports in vitro pollen tube attraction activity for three defensin-like peptides.

plant biology↗

bioRxiv2022

A complete MAPK cascade, a calmodulin, and a protein phosphatase act downstream of CRK receptor kinases and regulate Arabidopsis innate immunity

Mitogen-activated protein kinase (MAPK) cascades are critical signal transduction modules in stress responses, but how their composition and mode of activation induces a stress response is poorly understood. We showed in Arabidopsis that CRK21, a cysteine-rich receptor-like protein kinase (CRK), phosphorylates MAPK kinase kinase 20 (MKKK20) and thus directly activates a novel MAPK cascade, consisting of MKKK20, the MAPK kinase MKK3, and the MAPK MPK6. Furthermore, the protein phosphatase PP2C76 and the calmodulin CaM7 were identified as negative and positive modulators of the cascade, respectively. Loss-of-function in components of the MAPK cascade or in CaM7 led to susceptibility to the bacterial pathogen Pseudomonas syringae and the fungal pathogen Botrytis cinerea. In contrast, loss-of-function of PP2C76 as well as transient overexpression of the genes in the MAPK cascade and CaM7 conferred resistance to the pathogens. Moreover, seven additional CRKs interacted with MKKK20 in vivo, and four of these were highly expressed after inoculation with P. syringae. In summary, our findings demonstrate that the novel CRK21-MKKK20-MKK3-MPK6 signaling pathway functions in immunity to fungal and bacterial pathogens and that CRKs may function in directly activating MKKKs.

plant biology↗