Proteome analysis and phenotypic characterization of the lesion mimic mutant bspl in barley
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Characterization and molecular mechanism dissection for lesion-mimic mutation can help to gain an in-depth insight into regulatory mechanisms of programmed cell death in plants. In present study, the barley lesion mimic mutant bspl (barley spotted 1eaf) was used to investigate the formation of the bspl lesion mimic phenotype in barley using proteomics and quantitative real-time (qRT)-PCR techniques. The experiment showed that the formation of the bspl phenotype was directly related to light. Cell necrosis spontaneously formed in multiple parts of the leaf, and H2O2 accumulated continuously along with the formation of lesions. A total of 31 differentially expressed protein spots were identified using two-dimensional electrophoresis and mass spectrometry analysis of the expressed proteins between the bspl and the wild-type. These differentially expressed proteins are involved in different biological processes, mainly related to defense, photosynthesis, nitrogen acid metabolism, and carbohydrate metabolism. The expression of 15 genes was analyzed by qRT-PCR, and the results showed that the transcription level was basically consistent with the change in the protein level. The decreased expression of ATPase β subunit, cytochrome B6-F complex, photosystem I reaction center subunit VI, and other proteins may lead to the blockage of electron transport during photosynthesis, leading to cell death. Moreover, the overexpression of chitinase, thaumatin-like protein, PR-1a and other proteins, and the increased expression of NADPH-thioredoxin reductase and dehydroascorbate reductase may increase the disease resistance and stress resistance of bspl. This study will provide a new insight into the molecular mechanism underlying bspl-induced cell death in plants.
KeywordsBarley Lesion mimic mutant Phenotype Proteome qRT-PCR
This work is funded by National Key Technology Research and Development Program (2015BAD01B02) to DX and the National Natural Science Foundation of China (31401316) to XZ. We would like to thank LetPub (http://www.letpub.com) for providing linguistic assistance during the preparation of this manuscript. The authors would also like to thank Professor Longbiao Guo for improving English language.
DX and XZ conceived the research work. XZ, BT, YF, TT and JZ conducted experiments, and prepared the figures and tables. DX, XZ and BT wrote the paper.
Compliance with ethical standards
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
- Cornelissen BJ, Hooft van Huijsduijnen RA, Van Loon LC, Bol JF (1986) Molecular characterization of messenger rnas for ‘pathogenesis related’ proteins la, lb and lc, induced by tmv infection of tobacco. EMBO J 5:37–40. https://doi.org/10.1002/j.1460-2075.1986.tb04174.x CrossRefGoogle Scholar
- Han XY, Yang Y, Chu-Lang YU, Zhang WH, Sheng-Hai YE, Chen B (2014) A proteomic study on a disease-resistance-enhanced rice lesion mimic mutant. Chin J of Rice Sci 28:559–569. https://doi.org/10.3969/j.issn.1001-7216.2014.06.001 Google Scholar
- Hinkle PC, Mccarty RE (1978) How cells make ATP. Sci Am 238:104. https://doi.org/10.1038/scientificamerican0378-104 CrossRefGoogle Scholar
- Hooft van Huijsduijnen RA, Cornelissen BJ, van Loon LC, van Boom JH, Tromp M, Bol JF (1985) Virus-induced synthesis of messenger rnas for precursors of pathogenesis-related proteins in tobacco. Embo J 4:2167. https://doi.org/10.1002/j.1460-2075.1985.tb03911.x CrossRefGoogle Scholar
- Kosslak RM, Dieter JR, Ruff RL, Chamberlin MA, Bowen BA, Palmer RG (1996) Partial resistance to root-borne infection by phytophthora sojae in three allelic necrotic root mutants in soybean. J Hered 87:415–422. https://doi.org/10.1093/oxfordjournals.jhered.a023030 CrossRefGoogle Scholar
- Lim PO, Kim HJ, Nam HG (2007) Leaf senescence. Annu Rev Plant Biol 58:115–136. https://doi.org/10.1146/annurev.arplant.57.032905.105316 CrossRefGoogle Scholar
- Liu S, Cheng Y, Zhang X, Guan Q, Nishiuchi S, Hase K (2007) Expression of an nadp-malic enzyme gene in rice (Oryza sativa L.) is induced by environmental stresses; over-expression of the gene in arabidopsis confers salt and osmotic stress tolerance. Plant Mol Biol 64:49–58. https://doi.org/10.1007/s11103-007-9133-3 CrossRefGoogle Scholar
- Loon LC, Rep M, Pieterse CM (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162. https://doi.org/10.1146/annurev.phyto.44.070505.143425 CrossRefGoogle Scholar
- Lu L (1996) Chinese barley science. China Agriculture Press, BeijingGoogle Scholar
- Shirano Y, Kachroo P, Shah J, Klessig DF (2002) A gain-of-function mutation in an arabidopsis toll interleukin1 receptor-nucleotide binding site-leucine-rich repeat type R gene triggers defense responses and results in enhanced disease resistance. Plant Cell 14:3149–3162. https://doi.org/10.1105/tpc.005348 CrossRefGoogle Scholar
- Sun Y, Wenyi LU, Zhang Y, Wang G, Sun J, Xue D (2014) Genetic and physiological analyses of barley lesion mimic mutant bspl. J Hangzhou Norm Univ 13:602–605. https://doi.org/10.3969/j.issn.1674-232X.2014.06.008 Google Scholar
- Tang S (2014) Study on the light adaptation of leaves in alternate breathing pathways. Dissertation/master’s thesis. Northwest Normal University, Lan ZhouGoogle Scholar
- Van LL, Rep M, Pieterse CM (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162. https://doi.org/10.1146/annurev.phyto.44.070505.143425 CrossRefGoogle Scholar
- Zhang XT, Xiong YM (2005) The progress in studies of thioredoxin reductase. Foreign Med Sci-Sect Medgeogr 26:148–151Google Scholar
- Zhang XQ, Xue DW, Zhou WH, Fei-Bo WU, Zhang GP (2011) Screening and identification of the mutants from two-row barley cultivar zju3 induced by ethyl methane sulfonate (EMS). J Zhejiang Univ 37:169–174. https://doi.org/10.3785/j.issn.1008-9209.2011.02.008 Google Scholar