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Inhibition of BpEIN3 causes plaques in leaves of Betula platyphylla × B. pendula

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In this study, constructing BpEIN3 overexpression, inhibition expression vector, we transferred to birch to obtain BpEIN3 transgenic lines and study the function of this gene.

Abstract

EIN3 is an important transcription factor in the ethylene signal transduction pathway. It initiates the expression of a series of genes that regulate ethylene, positively regulating ethylene reaction, which affects the lesion mimic of plants through interacting ethylene signal and other signals in the signal transduction pathways. In order to reveal the function of BpEIN3 in Betula platyphylla, this study used birch as a genetic transformation receptor. Constructing BpEIN3 overexpression, inhibition expression vector, we transferred to birch to obtain BpEIN3 transgenic lines and study the function of this gene, which provided a reference for further revealing the molecular mechanism of BpEIN3 regulation of senescence and programmed cell death of birch leaves. In this study, the plant height, diameter and photosynthetic parameters of BpEIN3 transgenic lines were measured. The number of leaf lesions, histochemical staining, antioxidant enzymes, endogenous hormones and the expression of disease-related genes were also recorded. Results: We analyzed the expression of BpEIN3 by qRT-PCR, which was down-regulated in BpEIN3-inhibited expression lines and up-regulated in overexpression lines. The 2-year-old BpEIN3-RNAi lines exhibited brown spots on the leaves, leading to premature senescence and defoliation. The number of leaf spots is significantly different between different transgenic lines based on solid microscope. Photosynthetic parameters of BpEIN3-RNAi lines such as photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) were significantly lower than wild type. The accumulation of H2O2 in the leaves of transgenic lines was more than WT. The content of endogenous salicylic acid (SA), abscisic acid (ABA), superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in BpEIN3-RNAi lines was higher than that in WT, except jasmonic acid (JA) content. Meanwhile, BpEIN3-RNAi lines showed strong resistance to Alternaria alternata. We analyzed the expression of genes which were involved in ABA, JA and SA biosynthesis, metabolism and senescence. The results showed that BpPYR9, BpPYL4, BpNPR1 and BpJAZ10 were up-regulated and BpSAG12, BpNAC2, BpPR1a and BpPR1b were abundantly expressed in the 6th and 7th leaves of the transgenic lines. We concluded that the suppression of BpEIN3 could lead to lesion and promote premature senescence.

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References

  • Berrocal-Lobo M, Molina A (2004) Ethylene response factor 1 mediates arabidopsis resistance to the soilborne fungus Fusarium oxysporum. Mol Plant Microb Interact 17:763–770

    Article  CAS  Google Scholar 

  • Bruggeman Q, Raynaud C, Benhamed M, Delarue M (2015) To die or not to die? Lessons from lesion mimic mutants. Front Plant Sci 6:24

    Article  Google Scholar 

  • Chen H, Xue L, Chintamanani S et al (2009) Ethylene insensitive 3 and ethylene insensitive 3-like 1 repress salicylic acid induction deficient 2 expression to negatively regulate plant innate immunity in Arabidopsis. Plant Cell 21:2527–2540

    Article  CAS  Google Scholar 

  • Chini A, Fonseca S, Fernández G et al (2007) The JAZ family of repressors is the missing link in jasmonate signalling. Nature 448:666–671

    Article  CAS  Google Scholar 

  • Coll NS, Epple P, Dangl JL (2011) Programmed cell death in the plant immune system. Cell Death Differ 18:1247–1256

    Article  CAS  Google Scholar 

  • Dangl JL, Dietrich RA, Richberg MH (1996) Death don’t have no mercy: cell death programs in plant-microbe interactions. Plant Cell 8:1793–1807

    Article  CAS  Google Scholar 

  • Gan SS, Hörtensteiner S (2013) Frontiers in plant senescence research: from bench to bank. Plant Mol Biol 82:503–504

    Article  CAS  Google Scholar 

  • Harkenrider M, Sharma R, De Vleesschauwer D, Tsao L, Zhang X, Chern M, Canlas P, Zuo S, Ronald PC, Wang Z (2016) Overexpression of rice wall-associated kinase 25 (OsWAK25) alters resistance to bacterial and fungal pathogens. PLOS ONE 11(1):e0147310

    Article  Google Scholar 

  • Hibi T, Ohashi Y (2007) Involvement of EIN3 homologues in basic PR gene expression and flower development in tobacco plants. J Exp Bot 58:3671–3678

    Article  CAS  Google Scholar 

  • Huang QN, Yang Y, Shi YF et al (2010) Spotted-leaf mutants of rice (Oryza sativa). Rice Sci 17:247–256

    Article  Google Scholar 

  • Huang H, Wang S, Jiang J et al (2014) Overexpression of BpAP1 induces early flowering and produces dwarfism in Betula platyphylla × B. pendula. Physiol Plant 151:495–506

    Article  CAS  Google Scholar 

  • Hyo JK, Sung HH, You WK (2014) Gene regulatory cascade of senescence-associated NAC transcription factors activated by ethylene-insensitive2-mediated leaf senescence signalling in Arabidopsis. J Exp Bot 65:4023–4036

    Article  Google Scholar 

  • Lawton KA (1994) Acquired resistance signal transduction in Arabidopsis is ethylene independent. Plant Cell Online 6:581–588

    Article  CAS  Google Scholar 

  • Li H, Jianzhen L, Qin P et al (2003) Research progress on NPR1 gene. J Guangxi Normal Univ 21

  • Li R, Chen S, Liu G et al (2017) Characterization and identification of a woody lesion mimic mutant lmd, showing defence response and resistance to Alternaria alternate in birch. Sci Rep 7:11308

    Article  Google Scholar 

  • Lin ZF, Li SS, Lin GZ, Sun GC, Guo JY (1984) Superoxide dismutase activity and lipid peroxidation in relation to senescence of rice leaves. Acta Bot Sin 26:605–615

    CAS  Google Scholar 

  • Liu JG, Wang YQ, Zhang HS et al (2013) Research progress of ERF transcription factors in plant biotic and abiotic stress responses. Acta Agric Boreali Sin

  • Liu J, Shen J, Xu Y et al (2016) Ghd2, a CONSTANS-like gene, confers drought sensitivity through regulation of senescence in rice. J Exp Bot 67:5785–5798

    Article  CAS  Google Scholar 

  • Mcgrann GRD, Steed A, Burt C et al (2015) Differential effects of lesion mimic mutants in barley on disease development by facultative pathogens. J Exp Bot 66:3417–3428

    Article  CAS  Google Scholar 

  • Miura K, Tada Y (2014) Regulation of water, salinity, and cold stress responses by salicylic acid. Front Plant Sci 5:4

    Article  Google Scholar 

  • Morris K, Mackerness SAH, Page T et al (2000) Salicylic acid has a role in regulating gene expression during leaf senescence. Plant J 23:677–685

    Article  CAS  Google Scholar 

  • Noushina I, Khan NA, Antonio F et al (2017) Ethylene role in plant growth, interaction with other phytohormones. Frontiers in Plant Science, Development and Senescence

    Google Scholar 

  • Qi T, Wang J, Huang H, Liu B, Gao H, Liu Y, Song S, Xie D (2015) Regulation of jasmonate-induced leaf senescence by Antagonism between bHLH subgroup IIIe and IIId factors in Arabidopsis. Plant Cell 27(6):1634–1649

    Article  CAS  Google Scholar 

  • Santino A, Taurino M, Domenico SD et al (2013) Jasmonate signaling in plant development and defense response to multiple (a)biotic stresses. Plant Cell Rep 32:1085–1098

    Article  CAS  Google Scholar 

  • Séverine L, Vailleau F, Claudine B et al (2003) Lesion mimic mutants: keys for deciphering cell death and defense pathways in plants? Trends Plant Sci 8:0–271

    Google Scholar 

  • Shoumin X (1997) Changes in photosynthetic capacity and related enzyme activities during soybean (Glycine max) leaf senescence. J Northeast Normal Univ (Nat Sci Ed) 4:70–73

    Google Scholar 

  • Solano R, Stepanova A, Chao Q et al (1998) Nuclear events in ethylene signaling: a transcriptional cascade mediated by ETHYLENE-INSENSITIVE3 and ETHYLENE-RESPONSE-FACTOR1. Genes Dev 12:3703–3711

    Article  CAS  Google Scholar 

  • Thines B, Katsir L, Melotto M et al (2007) JAZ repressor proteins are targets of the SCFCOI1 complex during jasmonate signalling. Nature 448:661–665

    Article  CAS  Google Scholar 

  • Thomma BP, Penninckx IA, Cammue BP et al (2001) The complexity of disease signaling in Arabidopsis. Curr Opin Immunol 13:63–68

    Article  CAS  Google Scholar 

  • Vos MD, Zaanen WV, Koornneef A et al (2006) Herbivore-induced resistance against microbial pathogens in Arabidopsis. Plant Physiol 142:352–363

    Article  Google Scholar 

  • Wang Pengjie W, Dianxing SX (2018) Research progress on rice senescence-related mutants. Nucl Agron J 32:0497–0505

    Google Scholar 

  • Williams B, Dickman M (2008) Plant programmed cell death: can’t live with it; can’t live without it. Mol Plant Pathol 9:531–544

    Article  CAS  Google Scholar 

  • Yan Y, Stolz S, Chetelat A et al (2007) A downstream mediator in the growth repression limb of the jasmonate pathway. Plant Cell Online 19:2470–2483

    Article  CAS  Google Scholar 

  • Yang G, Chen S, Wang S et al (2015) BpGH3.5, an early auxin-response gene, regulates root elongation in Betula platyphylla × Betula pendula. Plant Cell Tissue Org Cult 120(1):239–250

    Article  CAS  Google Scholar 

  • Zhang CL, Guo HW (2012) Study on ethylene signal transduction pathway. Nature 34(4):219–227 (In Chinese)

    Google Scholar 

  • Zhang YJ, Zhao JZ, Zhang KW (2014) Research progress on mechanisms of phytohormones regulating leaf senescence. Plant Physiol J

  • Zhang W, Wei R, Chen S et al (2014) Functional characterization of Cinnamoyl-CoA Reductase (CCR) in birch (Betula platyphylla × Betula pendula) through overexpression and suppression analysis. Physiol Plant 154

  • Zhu ZQ, An FY, Feng Y (2011) Derepression of ethylene-stabilized transcription factors (EIN3/EIL1) mediates jasmonate and ethylene signaling synergy in Arabidopsis. PNAS 108:12539–12544

    Article  CAS  Google Scholar 

Download references

Acknowledgements

I thank Professor Jiang Jing and Professor Liu Guifeng for their guidance and help in the experiment. At the same time, I would like to thank Wang Chu and Li Ranhong for their help in experimental skills. I also acknowledge the help of Associate Professor Chen Su and Professor Yu Qibin in writing.

Funding

This study was supported by the following foundations: 1. National Natural Science Foundation of China (NSFC) (Grant No. 31670673); 2. The 111 Project (B16010); 3. National Natural Science Foundation of China (NSFC) (Grant No. 31800558).

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Authors and Affiliations

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin, China

    Xiaoyuan Li, Chu Wang, Su Chen, Jing Jiang & Guifeng Liu

  2. Department of Life Science and Technology, Mudanjiang Normal University, Mudanjiang, China

    Ranhong Li

  3. University of Florida, Gainesville, FL, USA

    Qibin Yu

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  1. Xiaoyuan Li
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  2. Ranhong Li
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  3. Chu Wang
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Correspondence to Jing Jiang.

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Li, X., Li, R., Wang, C. et al. Inhibition of BpEIN3 causes plaques in leaves of Betula platyphylla × B. pendula. Trees 34, 483–495 (2020). https://doi.org/10.1007/s00468-019-01930-6

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  • DOI: https://doi.org/10.1007/s00468-019-01930-6

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