Mike Blatt

Dr. Jonas Alvim

Postdoctoral Fellow, researches the mechanisms of ion channel interactions and regulation with membrane vesicle traffic

Martina Klejchova

Daphne Jackson Research Fellow

Mr. William Carroll

PhD Student, is pursuing the structure of K⁺ channels associated with channel clustering and gating

Ms. Naomi Donald

Research Technician, supports activities across the Laboratory of Plant Physiology and Biophysics

Mr. Adrian Hills

Research Computing Officer, develops software, including the HENRY suite for electrophysiology data acquisition and analysis and the ONGUARD platform for system modelling and analysis of stomatal function

Dr. Cecile Lefoulon

Postdoctoral Fellow, has broad interests in K⁺ channel structure and regulation, and in channel regulation in CAM plants

Dr. Binh-Ahn Thu Nguyen

Postdoctoral Fellow, is researching the mechanics of K⁺ channel gating and its regulation by external K⁺

Dr. Thanh-Hao Nguyen

Postdoctoral Fellow, studies the role of subsidiary cell ion transport in stomatal function

Ms. Amparo Ruiz-Prado

Glasshouse Technician, supports activiteis across the Laboratory of Plant Physiology and Biophysics

Dr. Fernanda Silva-Alvim

Postdoctoral Fellow, is pursuing the structure and function of novel ion channels in stomata of C4 plants

Research

Ion channel structure and function

Ion channel structure and function is at the centre of much of what we do. Current research focuses on how outward-rectifying K⁺ channels of plants respond to K⁺ outside, even though K⁺ carrying channel current comes from inside the cell. Our recent discovery of a key domain determining the clustering of the GORK channel (Eisenach et al 2014 Plant J 78,203; Horaruang et al 2022 Nature Plants 8,1262; Blatt & Alvim 2022 Nature Plants 8,1216) has yielded important clues to how clustering and gating are interconnected. We are collaborating with structural biologists in current research to uncover the detail behind these mechanisms.

Key structural features of plant K⁺ channel subunits

Guard cell membrane transport

Guard cell membrane transport is a major driver of stomatal movements and determines the responsiveness of stomata to environmental change. We continue to develop and make use of the OnGuard platform to understand how ion transport is coordinated in order to maximise stomatal responsiveness in C3, C4 and CAM plants. Recent work with OnGuard uncovered a ‘carbon memory’ in responsiveness (Jezek et al 2021 Nature Plants 7,1301) and has helped guide successful efforts to enhance stomatal responsiveness by accelerating K⁺ flux through engineering K⁺ channel gating (Horaruang et al 2022 Nature Plants 8,1262). We are combining structural information about guard cell ion channels to further enhance stomatal responsiveness and are looking to translate our findings to crops

Engineering GORK channel gating enhances plant growth
and water use efficiency in a variable environment (from Horaruang et al 2022)

Vesicle traffic

Vesicle traffic underpins plant membrane transport by regulating the populations of ion channels and pumps that are available at the membrane. We have found that several trafficking proteins also bind directly with ion channels for mutual co-regulation (Honsbein et al 2009 Plant Cell 21,2859; Grefen et al 2015 Nature Plants 1,15108; Waghmare et al 2019 Plant Physiol 181,1096). These findings are only a first glimpse of a far more extensive network that we now pursue, to understand how traffic and transport are coordinated, whether for cell growth or for stomatal movements.

Cover image from Grefen et al (2015)

Funders

Resources

We offer material resources of broad interest to the scientific community, including vectors and markers for plant and yeast transformation, as well as specialist seed lines. A small fee will be charged for markers and seeds to cover handling and postage. Contact us or see here for more information.

Sequence information is provided in genebank (*.gb) files; you can either open them directly with DNA software (VectorNTI, clc workbench, editseq) or simply use a text editor tool.

UBQ10 Vectors

Promoter-driven Gateway vectors for fluorescent tagging of proteins.

For the use of UBQ10 Promoter Driven Vectors please cite:

Grefen,C., Donald,N., Hashimoto,K., Kudla,J., Schumacher,K., and Blatt,M.R. (2010) “A ubiquitin-10 promoter-based vector set for fluorescent protein tagging facilitates temporal stability and native protein distribution in transient and stable expression studies.” Plant J. 64: 355-65

Yeast Vectors

Vectors and strains for split-ubiquitin screening and assays.

For the use of Gateway SUS Vectors please cite:

Grefen C, Obrdlik P and Harter K (2009) “The determination of protein-protein interactions by the mating-based split-ubiquitin system (mbSUS).” Methods Mol. Biol. 479:217-33.

For the use of SUS Yeast Strains please cite:

Obrdlik, et al. (2004) “K⁺ channel interactions detected by a genetic system optimized for systematic studies of membrane protein interactions.” PNAS 101:12242-7.

Grefen C, Lalonde S and Obrdlik P (2007) “Split-ubiquitin system for identifying protein-protein interactions in membrane and full-length proteins.” Curr. Protoc. Neurosci. Chapter 5: Unit 5.27.

For the use of the cytoSUS vector designed for soluble baits please cite:

Karnik, R., Zhang, B., Waghmare, S., Aderhold, C., Grefen, C., and Blatt, M.R. (2015). Binding of SEC11 indicates its role in SNARE recycling after vesicle fusion and identifies two pathways for vesicular traffic to the plasma membrane. Plant Cell 27, 675-694.

For use of the GPS vector system for soluble baits please cite:

Ben Zhang, Rucha Karnik, Naomi Alice Donald, Mike R. Blatt. Detecting protein interactions at the membrane surface. Plant Physiol. Jul 2018, pp.00577.2018; DOI: https://doi.org/10.1104/pp.18.00577

Bicistronic and Multicistronic Vectors

Bicistronic vectors for fluorescence co-expression tagging of proteins in Arabidopsis.

Multicistronic vectors for Secretory Traffic Assay

For the use of the Bicistronic Vector please cite:

Chen,Z.H., Grefen,C., Donald,N., Hills,A., and Blatt,M.R. (2011) A bicistronic, Ubiquitin-10 promoter-based vector cassette for transient transformation and functional analysis of membrane transport. Plant Cell and Environment, 34:554-564.

For the use of the Multicistronic Vectors please cite:

Karnik, R., Grefen, C., Bayne, R., Honsbein, A., Kohler, T., Kioumourtzoglou, D., Williams, M., Bryant, N.J., and Blatt, M.R. (2013). Arabidopsis Sec1/Munc18 Protein SEC11 Is a Competitive and Dynamic Modulator of SNARE Binding and SYP121-Dependent Vesicle Traffic. Plant Cell 25, 1368-1382.

Secretory Traffic Vectors

Gateway compatible vector for stable fluorescence co-expression and tagging of proteins in Arabidopsis.

For the use of Dex inducible secretory traffic vector please cite:

Grefen, C., Karnik, R., Larson, E., Lefoulon, C., Wang, Y., Waghmare, S., Zhang, B., Hills, A., and Blatt, M.R. (2015). A vesicle-trafficking protein commandeers Kv channel voltage sensors for voltage-dependent secretion. Nature Plants 1, 15108.

FRET Vectors

Binary 2-in-1 FRET vector set for dynamic protein interaction studies.

Binary 2-in-1 Vector for detecting voltage dependent conformation changes by FRET.

For the use of 2-in-1 FRET vectors please cite:

Hecker, A., Wallmeroth, N., Peter, S., Blatt, M.R., Harter, K., and Grefen, C. (2015). Binary 2in1 Vectors Improve in Planta (Co)localization and Dynamic Protein Interaction Studies. Plant Physiol 168, 776-787.

For the use of the pVd FRET vector please cite:

Vector Conversion

Conversion Vectors for 2-in-1 cloning.

For the use of 2-in-1 Conversion Vectors please cite:

Grefen,C., and Blatt,M.R. (2012) “A 2in1 cloning system enables ratiometric bimolecular fluorescence complementation (rBiFC).” Biotechniques 53,311-14.

Download*

rBiFC Vectors

Binary 2-in-1 vector set enabling ratiometric Bimolecular Fluorescence Complementation (rBiFC).

For the use of rBiFC Vectors please cite:

Grefen,C., and Blatt,M.R. (2012) “A 2in1 cloning system enables ratiometric bimolecular fluorescence complementation (rBiFC).” Biotechniques 53,311-14.

Download*

Software

We continue to address specialist needs with the development of industrially robust software packages for electrophysiology, imaging, biometrics and systems biology. All our software is available to the academic community free of charge, and can be downloaded by following the relevant links on this page.

Electrophysiology

Henry’s EP Suite is a complete, self-contained package for electrophysiologists. This software can be used in multiple environments.

Plant Biometrics

In a collaborative project we have developed the EZ-Rhizo application, which allows for fast and accurate semi-automatic measurement of plants’ root system architecture.

Modelling

OnGuard is a groundbreaking platform for interactive, mechanistic modelling of stomatal dynamics and guard cell membrane transport with readouts in stomatal conductance, whole-plant water relations and carbon assimilation.

SDM-Assist

SDM-Assist is a stand-alone application which allows SDM primer design in just 3 clicks without the need for an internet connection and will work on almost any machine.