A secreted citrus protease cleaves an outer membrane protein of the Huanglongbing pathogen

A Alexander J. McClelland (The Sainsbury Laboratory, University of East Anglia) B Bin Hu Y Yuantao Xu X Xiaodong Fang (BGI Research, Hangzhou, China.) C Chunxia Wang (Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences) B Benjamin L. Koch (The Sainsbury Laboratory, University of East Anglia) A Amelia H. Lovelace (The Sainsbury Laboratory, University of East Anglia) E Eva Hawara (Department of Microbiology and Plant Pathology, University of California Riverside) Y Yuanchun Wang (Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences) Z Zhiqian Pang (Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences) A Agustina De Francesco (Department of Microbiology and Plant Pathology, University of California Riverside) S Suzanne P. van Wier (School of Chemistry, Pharmacy and Pharmacology, University of East Anglia) A Andrew M. Beekman (School of Chemistry, Pharmacy and Pharmacology, University of East Anglia) A Amit Levy (Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences) N Nian Wang (Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences) R Renier A. L. van der Hoorn (Plant Chemetics Laboratory, Department of Biology, University of Oxford) Q Qiang Xu (Key Laboratory of Material Simulation Methods & Software of Ministry of Education, College of Physics) W Wenbo Ma (The Sainsbury Laboratory, University of East Anglia)

Abstract

Plants secrete a variety of proteases as a defense response during infection by microbial pathogens. However, the relationship between their catalytic activities and antimicrobial functions remains largely unknown. Particularly, few biologically relevant substrates of these proteases have been identified. Huanglongbing (HLB) has been a major threat to the citrus industry worldwide. The HLB-associated bacterium, “ Candidatus Liberibacter asiaticus” (Las), was previously shown to deploy an inhibitor of papain-like cysteine proteases (PLCPs) to promote disease in citrus. In this study, we identified an outer membrane protein (OMP) of Las, LasOMP1, as a substrate of the citrus PLCP Cs RD21a. LasOMP1 is one of the most highly expressed genes in Las. Cs RD21a cleaves LasOMP1 and produces cleaved peptide products, which could be detected in vitro and in HLB-diseased citrus plants. We found that Cs RD21a targets the N-terminal portion of LasOMP1, potentially at an extracellular loop region. Importantly, transgenic sweet orange overexpressing Cs RD21a showed reduced Las populations and improved plant growth, highlighting that engineering this protease is a promising strategy to enhance HLB resistance in citrus. Together, our work reveals a pathogen-derived substrate of plant PLCPs and suggests bacterial OMPs may be direct targets of plant defense.

Article Details

Volume / Issue Vol. 123, Issue 15
Published April 14, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (18)

A

Alexander J. McClelland

The Sainsbury Laboratory, University of East Anglia

B

Bin Hu

Y

Yuantao Xu

X

Xiaodong Fang

BGI Research, Hangzhou, China.

C

Chunxia Wang

Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences

B

Benjamin L. Koch

The Sainsbury Laboratory, University of East Anglia

A

Amelia H. Lovelace

The Sainsbury Laboratory, University of East Anglia

E

Eva Hawara

Department of Microbiology and Plant Pathology, University of California Riverside

Y

Yuanchun Wang

Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences

Z

Zhiqian Pang

Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences

A

Agustina De Francesco

Department of Microbiology and Plant Pathology, University of California Riverside

S

Suzanne P. van Wier

School of Chemistry, Pharmacy and Pharmacology, University of East Anglia

A

Andrew M. Beekman

School of Chemistry, Pharmacy and Pharmacology, University of East Anglia

A

Amit Levy

Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences

N

Nian Wang

Citrus Research and Education Center, Department of Plant Pathology, Microbiology and Cell Science, University of Florida/Institute of Food and Agricultural Sciences

R

Renier A. L. van der Hoorn

Plant Chemetics Laboratory, Department of Biology, University of Oxford

Q

Qiang Xu

Key Laboratory of Material Simulation Methods & Software of Ministry of Education, College of Physics

W

Wenbo Ma

The Sainsbury Laboratory, University of East Anglia