The catalytic activity of mitogen‐activated protein kinases (
Calmodulin (CaM) is an essential protein in cellular activity and plays important roles in many processes in insect development. RNA interference (RNAi) has been hypothesized to be a promising method for pest control. CaM is a good candidate for RNAi target. However, the sequence and function of CaM in
In the present study, two alternatively spliced variants of
- NSF-PAR ID:
- 10055203
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Pest Management Science
- Volume:
- 74
- Issue:
- 7
- ISSN:
- 1526-498X
- Page Range / eLocation ID:
- p. 1711-1719
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Summary MAPK s) is dynamically modified in plants. SinceMAPK s have been shown to play important roles in a wide range of signaling pathways, the ability to monitorMAPK activity in living plant cells would be valuable. Here, we report the development of a genetically encodedMAPK activity sensor for use inArabidopsis thaliana . The sensor is composed of yellow and blue fluorescent proteins, a phosphopeptide binding domain, aMAPK substrate domain and a flexible linker. Usingin vitro testing, we demonstrated that phosphorylation causes an increase in the Förster resonance energy transfer (FRET ) efficiency of the sensor. TheFRET efficiency can therefore serve as a readout of kinase activity. We also produced transgenic Arabidopsis lines expressing this sensor ofMAPK activity (SOMA ) and performed live‐cell imaging experiments using detached cotyledons. Treatment with NaCl, the synthetic flagellin peptide flg22 and chitin all led to rapid gains inFRET efficiency. Control lines expressing a version ofSOMA in which the phosphosite was mutated to an alanine did not show any substantial changes inFRET . We also expressed the sensor in a conditional loss‐of‐function double‐mutant line for the ArabidopsisMAPK genesMPK 3MPK 6MPK 3/6 are necessary for the NaCl‐inducedFRET gain of the sensor, while otherMAPK s are probably contributing to the chitin and flg22‐induced increases inFRET . Taken together, our results suggest thatSOMA is able to dynamically reportMAPK activity in living plant cells. -
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Premise Light is critical in the ability of plants to accumulate chlorophyll. When exposed to far‐red (
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SIG ) proteins contribute to promoter specificity of the plastid‐encodedRNA polymerase during chloroplast genome transcription. All six members of theSIG family, that is,SIG 1–SIG 6, are nuclear‐encoded proteins targeted to chloroplasts. Sigma factor 2 (SIG 2) is a phytochrome‐regulated protein important for stoichiometric control of the expression of plastid‐ and nuclear‐encoded genes that impact plastid development and plant growth and development. AmongSIG factors,SIG 2 is required not only for transcription of chloroplast genes (i.e., anterograde signaling), but also impacts nuclear‐encoded, photosynthesis‐related, and light signaling‐related genes (i.e., retrograde signaling) in response to plastid functional status. AlthoughSIG 2 is involved in photomorphogenesis in Arabidopsis, the molecular bases for its role in light signaling that impacts photomorphogenesis and aspects of photosynthesis have only recently begun to be investigated. Previously, we reported thatSIG 2 is necessary for phytochrome‐mediated photomorphogenesis specifically under red (R) and far‐red light, thereby suggesting a link between phytochromes and nuclear‐encodedSIG 2 in light signaling. To explore transcriptional roles ofSIG 2 in R‐dependent growth and development, we performedRNA sequencing analysis to compare gene expression insig2‐2 mutant and Col‐0 wild‐type seedlings at two developmental stages (1‐ and 7‐day). We identified a subset of misregulated genes involved in growth, hormonal cross talk, stress responses, and photosynthesis. To investigate the functional relevance of these gene expression analyses, we performed several comparative phenotyping tests. In these analyses, strongsig2 mutants showed insensitivity to bioactiveGA 3, high intracellular levels of hydrogen peroxide (H2O2) indicative of a stress response, and specific defects in photosynthesis, including elevated levels of cyclic electron flow (CEF ) and nonphotochemical quenching (NPQ ). We demonstrated thatSIG 2 regulates a broader range of physiological responses at the molecular level than previously reported, with specific roles in red‐light‐mediated photomorphogenesis.
