RESUMEN
Oxidative burst, the rapid production of high levels of reactive oxygen species in response to external stimuli, is an early defense reaction against pathogens. The fungal elicitor chitosan causes an oxidative burst in the moss Physcomitrium patens (formerly Physcomitrella patens), mainly due to the peroxidase enzyme Prx34. To better understand the chitosan responses in P. patens, we conducted a screen of part of a P. patens mutant collection to isolate plants with less peroxidase activity than wild-type (WT) plants after chitosan treatment. We isolated a P. patens mutant that affected the gene encoding NAD(P)-binding Rossmann fold protein (hereafter, Rossmann fold protein). Three Rossmann fold protein-knockout (KO) plants (named Rossmann fold KO lines) were generated and used to assess extracellular peroxidase activity and expression of defense-responsive genes, including alternative oxidase, lipoxygenase (LOX), NADPH oxidase, and peroxidase (Prx34) in response to chitosan treatment. Extracellular (apoplastic) peroxidase activity was significantly lower in Rossmann fold KO lines than in WT plants after chitosan treatments. Expression of the LOX gene in Rossmann fold KO plants was significantly lower before and after chitosan treatment when compared with WT. Peroxidase activity assays together with gene expression analyses suggest that the Rossmann fold protein might be an important component of the signaling pathway leading to oxidative burst and basal expression of the LOX gene in P. patens. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.
Asunto(s)
Bryopsida , Quitosano , Lipooxigenasa/genética , Quitosano/farmacología , NAD , Bryopsida/genética , Peroxidasas/genética , Peroxidasa/genética , Peroxidasa/metabolismo , Plantas/metabolismoRESUMEN
Plant viruses are important pathogens able to overcome plant defense mechanisms using their viral suppressors of RNA silencing (VSR). Small RNA pathways of bryophytes and vascular plants have significant similarities, but little is known about how viruses interact with mosses. This study elucidated the responses of Physcomitrella patens to two different VSRs. We transformed P. patens plants to express VSR P19 from tomato bushy stunt virus and VSR 2b from cucumber mosaic virus, respectively. RNA sequencing and quantitative PCR were used to detect the effects of VSRs on gene expression. Small RNA (sRNA) sequencing was used to estimate the influences of VSRs on the sRNA pool of P. patens. Expression of either VSR-encoding gene caused developmental disorders in P. patens. The transcripts of four different transcription factors (AP2/erf, EREB-11 and two MYBs) accumulated in the P19 lines. sRNA sequencing revealed that VSR P19 significantly changed the microRNA pool in P. patens. Our results suggest that VSR P19 is functional in P. patens and affects the abundance of specific microRNAs interfering with gene expression. The results open new opportunities for using Physcomitrella as an alternative system to study plant-virus interactions.
Asunto(s)
Bryopsida/crecimiento & desarrollo , Bryopsida/genética , Bryopsida/virología , Interacciones Huésped-Patógeno/genética , Cucumovirus/genética , Cucumovirus/patogenicidad , Regulación de la Expresión Génica de las Plantas , Regulación Viral de la Expresión Génica , MicroARNs , Proteínas de Plantas/genética , Virus de Plantas/genética , Virus de Plantas/patogenicidad , Plantas Modificadas Genéticamente , Interferencia de ARN , Tombusvirus/genética , Tombusvirus/patogenicidad , Factores de Transcripción/genéticaRESUMEN
Drought and heat tolerance of the Sunagoke moss (Racomitrium japonicum) and the low thermal conductivity of the dry moss tissue offer novel greening and insulation possibilities of roofs and walls to mitigate the heat island phenomenon in urban environments. However, damage may appear in the moss panels under humid conditions in Japan. In this study we characterized fungi associated with the damaged areas of the Sunagoke moss panels. Fungi were identified by morphology and internal transcribed spacer (ITS) sequence analysis and tested for pathogenicity on R. japonicum (Grimmiaceae) and an unrelated moss species (Physcomitrella patens; Funariaceae) under controlled conditions. Alternaria alternata, Fusarium avenaceum and Fusarium oxysporum caused severe necrosis and death, whereas Cladosporium oxysporum and Epicoccum nigrum caused milder discoloration or chlorosis in both moss species. The fungi pathogenic on moss were closely related to fungal pathogens described from cultivated vascular plants. Ammonium increased severity of fungal diseases in moss. This study demonstrated that fungi can cause economically significant diseases in cultivated moss and hamper commercial use of the moss panels unless appropriate control methods are developed. Use of a single moss clone to cover large surfaces and the air pollutants such as ammonium may increase the risk for fungal disease problems.