Abstract
Main conclusion
Genes of the PLAT protein family, including PLAT and ATS3 subfamilies of higher plants and homologs of liverwort, are involved in plant defense against insects.
Abstract
Laticifer cells in plants contain large amounts of anti-microbe or anti-insect proteins and are involved in plant defense against biotic stresses. We previously found that PLAT proteins accumulate in laticifers of fig tree (Ficus carica) at comparable levels to those of chitinases, and the transcript level of ATS3, another PLAT domain-containing protein, is highest in the transcriptome of laticifers of Euphorbia tirucalli. In this study, we investigated whether the PLAT domain-containing proteins are involved in defense against insects. Larvae of the lepidopteran Spodoptera litura showed retarded growth when fed with Nicotiana benthamiana leaves expressing F. carica PLAT or E. tirucalli ATS3 genes, introduced by agroinfiltration using expression vector pBYR2HS. Transcriptome analysis of these leaves indicated that ethylene and jasmonate signaling were activated, leading to increased expression of genes for PR-1, β-1,3-glucanase, PR5 and trypsin inhibitors, suggesting an indirect mechanism of PLAT- and ATS3-induced resistance in the host plant. Direct cytotoxicity of PLAT and ATS3 to insects was also possible because heterologous expression of the corresponding genes in Drosophila melanogaster caused apoptosis-mediated cell death in this insect. Larval growth retardation of S. litura occurred when they were fed radish sprouts, a good host for agroinfiltration, expressing any of nine homologous genes of dicotyledon Arabidopsis thaliana, monocotyledon Brachypodium distachyon, conifer Picea sitchensis and liverwort Marchantia polymorpha. Of these nine genes, the heterologous expression of A. thaliana AT5G62200 and AT5G62210 caused significant increases in larval death. These results indicated that the PLAT protein family has largely conserved anti-insect activity in the plant kingdom (249 words).
Abbreviations
- DEG:
-
Differentially expressed (uni)gene
- dph:
-
Days post-harvest
- EGFP:
-
Enhanced green fluorescent protein
- GO:
-
Gene ontology
- PR:
-
Pathogenesis-related
- rpkm:
-
Read counts per kilobase of unigene per million mapped reads
References
Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res 25(17):3389–3402. https://doi.org/10.1093/nar/25.17.3389
Armes NJ, Wightman JA, Jadhav DR, Ranga Rao GV (1997) Status of insecticide resistance in Spodoptera litura in Andhra Pradesh, India. Pesticide Sci 50(3):240–248. https://doi.org/10.1002/(SICI)1096-9063(199707)50:3%3c240::AID-PS579%3e3.0.CO;2-9
Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Series B (Statistical methodology) 57(1):289–300. https://doi.org/10.1111/j.2517-6161.1995.tb02031.x
Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118(2):401–415
Burghardt LT, Trujillo DI, Epstein B, Tiffin P, Young ND (2020) A select and resequence approach reveals strain-specific effects of Medicago nodule-specific PLAT-domain genes. Plant Physiol 182(1):463–471. https://doi.org/10.1104/pp.19.00831
Callus BA, Vaux DL (2007) Caspase inhibitors: viral, cellular and chemical. Cell Death Differ 14(1):73–78. https://doi.org/10.1038/sj.cdd.4402034
Castelblanque L, Balaguer B, Marti C, Orozco M, Vera P (2018) LOL 2 and LOL 5 loci control latex production by laticifer cells in Euphorbia lathyris. New Phytol 219(4):1467–1479. https://doi.org/10.1111/nph.15253
Castelblanque L, García-Andrade J, Martínez-Arias C, Rodríguez JJ et al (2020) Opposing roles of plant laticifer cells in the resistance to insect herbivores and fungal pathogens. Plant Commun. https://doi.org/10.1016/j.xplc.2020.100112
Chountala M, Vakaloglou KM, Zervas CG (2012) Parvin overexpression uncovers tissue-specific genetic pathways and disrupts F-actin to induce apoptosis in the developing epithelia in Drosophila. PLoS ONE 7(10):e47355. https://doi.org/10.1371/journal.pone.0047355
Coligan JE, Dunn BM, Ploegh HL, Speicher DW, Wingfield PT (1995) Current protocols in protein science. Wiley, New York
Darding M, Meier P (2012) IAPs: guardians of RIPK1. Cell Death Differ 19(1):58–66. https://doi.org/10.1038/cdd.2011.163
Fisher R (1922) On the interpretation of χ2 from contingency tables, and the calculation of P. J R Stat Soc 85(1):87–94. https://doi.org/10.2307/2340521
Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q, Chen Z (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29(7):644–652. https://doi.org/10.1038/nbt.1883
Hagel JM, Yeung EC, Facchini PJ (2008) Got milk? The secret life of laticifers. Trends Plant Sci 13(12):631–639. https://doi.org/10.1016/j.tplants.2008.09.005
Hay BA, Wassarman DA, Rubin GM (1995) Drosophila homologs of baculovirus inhibitor of apoptosis proteins function to block cell death. Cell 83(7):1253–1262. https://doi.org/10.1016/0092-8674(95)90150-7
Hilder VA, Gatehouse AM, Sheerman SE, Barker RF, Boulter D (1987) A novel mechanism of insect resistance engineered into tobacco. Nature 330(6144):160–163. https://doi.org/10.1038/330160a0
Holm S (1979) A simple sequential rejective multiple test procedure. Scand Stat Theory Appl 6:65–70
Van Hove J, De Jaeger G, De Winne N, Guisez Y, Van Damme EJ (2015) The Arabidopsis lectin EULS3 is involved in stomatal closure. Plant Sci 238:312–322. https://doi.org/10.1016/j.plantsci.2015.07.005
Huynh QK, Borgmeyer JR, Zobel JF (1992) Isolation and characterization of a 22 kDa protein with antifungal properties from maize seeds. Biochem Biophys Res Commun 182(1):1–5. https://doi.org/10.1016/S0006-291X(05)80103-2
Hyun TK, Albacete A, van der Graaff E, Eom SH, Großkinsky DK, Böhm H, Janschek U, Rim Y, Ali WW, Kim SY, Roitsch T (2015) The Arabidopsis PLAT domain protein1 promotes abiotic stress tolerance and growth in tobacco. Transgenic Res 24(4):651–663. https://doi.org/10.1007/s11248-015-9868-6
Hyun TK, van der Graaff E, Albacete A, Eom SH, Großkinsky DK, Böhm H, Janschek U, Rim Y, Ali WW, Kim SY, Roitsch T (2014) The Arabidopsis PLAT domain protein1 is critically involved in abiotic stress tolerance. PLoS ONE 9(11):e112946. https://doi.org/10.1371/journal.pone.0112946
Karnchanatat A, Tiengburanatam N, Boonmee A, Puthong S, Sangvanich P (2011) Zingipain, a cysteine protease from Zingiber ottensii Valeton rhizomes with antiproliferative activities against fungi and human malignant cell lines. Prep Biochem Biotechnol 41(2):138–153. https://doi.org/10.1080/10826068.2011.547347
Kitajima S, Aoki W, Shibata D, Nakajima D, Sakurai N, Yazaki K, Munakata R, Taira T, Kobayashi M, Aburaya S, Savadogo EH (2018) Comparative multi-omics analysis reveals diverse latex-based defense strategies against pests among latex-producing organs of the fig tree (Ficus carica). Planta 247(6):1423–1438. https://doi.org/10.1007/s00425-018-2880-3
Kitajima S, Kamei K, Taketani S, Yamaguchi M, Kawai F, Komatsu A, Inukai Y (2010) Two chitinase-like proteins abundantly accumulated in latex of mulberry show insecticidal activity. BMC Biochem 11(1):6. https://doi.org/10.1186/1471-2091-11-6
Kitajima S, Miura K, Aoki W, Yamato KT, Taira T, Murakami R, Aburaya S (2016) Transcriptome and proteome analyses provide insight into laticifer’s defense of Euphorbia tirucalli against pests. Plant Physiol Biochem 108:434–446. https://doi.org/10.1016/j.plaphy.2016.08.008
Kitajima S, Miura K, Yasuda J (2020) Radish sprouts as an efficient and rapidly available host for an agroinfiltration-based transient gene expression system. Plant Biotechnol 37:89–92. https://doi.org/10.5511/plantbiotechnology.19.1216a
Kitajima S, Taira T, Oda K, Yamato KT, Inukai Y, Hori Y (2012) Comparative study of gene expression and major proteins’ function of laticifers in lignified and unlignified organs of mulberry. Planta 235(3):589–601. https://doi.org/10.1007/s00425-011-1533-6
Kitajima S, Yamamoto Y, Hirooka K, Taki C, Hibino S (2013) Laticifers in mulberry exclusively accumulate defense proteins related to biotic stresses. Plant Biotechnol 30(4):399–402. https://doi.org/10.5511/plantbiotechnology.13.0326a
Konno K (2011) Plant latex and other exudates as plant defense systems: roles of various defense chemicals and proteins contained therein. Phytochemistry 72(13):1510–1530. https://doi.org/10.1016/j.phytochem.2011.02.016
Konno K, Hirayama C, Nakamura M, Tateishi K, Tamura Y, Hattori M, Kohno K (2004) Papain protects papaya trees from herbivorous insects: role of cysteine proteases in latex. Plant J 37(3):370–378. https://doi.org/10.1046/j.1365-313X.2003.01968.x
Langmead B, Salzberg SL (2012) Fast gapped-read alignment with bowtie 2. Nat Methods 9(4):357–359. https://doi.org/10.1038/nmeth.1923
López-García B, Hernández M, Segundo BS (2012) Bromelain, a cysteine protease from pineapple (Ananas comosus) stem, is an inhibitor of fungal plant pathogens. Lett Appl Microbiol 55(1):62–67. https://doi.org/10.1111/j.1472-765X.2012.03258.x
Mishra M, Knust E (2019) Analysis of the drosophila compound eye with light and electron microscopy. In: Weber BHF, Langmann T (eds) Retinal Degeneration. Methods in Molecular Biology, vol 1834. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8669-9_22
Munakata R, Kitajima S, Nuttens A, Tatsumi K, Takemura T, Ichino T, Galati G, Vautrin S, Bergès H, Grosjean J, Bourgaud F (2020) Convergent evolution of the UbiA prenyltransferase family underlies the independent acquisition of furanocoumarins in plants. New Phytol 225(5):2166–2182. https://doi.org/10.1111/nph.16277
Nakazaki A, Yamada K, Kunieda T, Sugiyama R, Hirai MY, Tamura K, Hara-Nishimura I, Shimada T (2019) Leaf endoplasmic reticulum bodies identified in Arabidopsis rosette leaves are involved in defense against herbivory. Plant Physiol 179(4):1515–1524. https://doi.org/10.1104/pp.18.00984
Nakazawa M, Matsubara H, Matsushita Y, Watanabe M, Vo N, Yoshida H et al (2016) The human Bcl-2 family member Bcl-rambo localizes to mitochondria and induces apoptosis and morphological aberrations in Drosophila. PLoS ONE 11(6):e0157823. https://doi.org/10.1371/journal.pone.0157823
Pislariu CI, Sinharoy S, Torres-Jerez I, Nakashima J, Blancaflor EB, Udvardi MK (2019) The nodule-specific PLAT domain protein NPD1 is required for nitrogen-fixing symbiosis. Plant Physiol 180(3):1480–1497. https://doi.org/10.1104/pp.18.01613
Ramos MV, Demarco D, da Costa Souza IC, de Freitas CDT (2019) Laticifers, latex, and their role in plant defense. Trends Plant Sci 24(6):553–567. https://doi.org/10.1016/j.tplants.2019.03.006
Ramos MV, Freitas CDT, Morais FS, Prado E, Medina MC, Demarco D (2020) Plant latex and latex-borne defense. Adv Bot Res 93:1–25. https://doi.org/10.1016/bs.abr.2019.09.002
Robinson MD, McCarthy DJ, Smyth GK (2010) edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26(1):139–140. https://doi.org/10.1093/bioinformatics/btp616
Sakai R, Suzuki M, Ueyama M, Takeuchi T, Minakawa EN, Hayakawa H et al (2019) E46K mutant α-synuclein is more degradation resistant and exhibits greater toxic effects than wild-type α-synuclein in Drosophila models of Parkinson’s disease. PLoS ONE 14(6):e0218261. https://doi.org/10.1371/journal.pone.0218261
Shin R, An JM, Park CJ, Kim YJ, Joo S, Kim WT, Paek KH (2004) Capsicum annuum tobacco mosaic virus-induced clone 1 expression perturbation alters the plant’s response to ethylene and interferes with the redox homeostasis. Plant Physiol 135(1):561–573. https://doi.org/10.1104/pp.103.035436
Szabo A, Tofaris GK (2019) Monitoring α-synuclein proteotoxicity in drosophila models. In: Bartels T (ed) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_15
Taira T, Toma N, Ishihara M (2005) Purification, characterization, and antifungal activity of chitinases from pineapple (Ananas comosus) leaf. Biosci Biotechnol Biochem 69(1):189–196. https://doi.org/10.1271/bbb.69.189
Takahashi Y, Hirose F, Matsukage A, Yamaguchi M (1999) Identification of three conserved regions in the DREF transcription factors from Drosophila melanogaster and Drosophila virilis. Nucl Acids Res 27(2):510–516. https://doi.org/10.1093/nar/27.2.510
Terras FR, Schoofs HM, Thevissen K, Osborn RW, Vanderleyden J, Cammue BP, Broekaert WF (1993) Synergistic enhancement of the antifungal activity of wheat and barley thionins by radish and oilseed rape 2S albumins and by barley trypsin inhibitors. Plant Physiol 103(4):1311–1319. https://doi.org/10.1104/pp.103.4.1311
van 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
Villard C, Larbat R, Munakata R, Hehn A (2019) Defence mechanisms of Ficus: pyramiding strategies to cope with pests and pathogens. Planta 249(3):617–633. https://doi.org/10.1007/s00425-019-03098-2
Yamada K, Goto-Yamada S, Nakazaki A, Kunieda T, Kuwata K, Nagano AJ, Nishimura M, Hara-Nishimura I (2020) Endoplasmic reticulum-derived bodies enable a single-cell chemical defense in Brassicaceae plants. Commun Biol 3:21. https://doi.org/10.1038/s42003-019-0739-1
Yamaguchi M, Hirose F, Inoue YH, Shiraki M, Hayashi Y, Nishi Y, Matsukage A (1999) Ectopic expression of human p53 inhibits entry into S phase and induces apoptosis in the Drosophila eye imaginal disc. Oncogene 18(48):6767–6775. https://doi.org/10.1038/sj.onc.1203113
Yamamoto T, Hoshikawa K, Ezura K, Okazawa R, Fujita S, Takaoka M, Mason HS, Ezura H, Miura K (2018) Improvement of the transient expression system for production of recombinant proteins in plants. Sci Rep 8:4755. https://doi.org/10.1038/s41598-018-23024-y
Acknowledgements
This study was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, Culture and Technology of Japan (to SK, No. 20K05768), the research grant for Mission Research on Sustainable Humanosphere from the Research Institute for Sustainable Humanosphere (RISH), Kyoto University (to SK), and Cooperative Research Grant of the Plant Transgenic Design Initiative (PTraD) by Gene Research Center, Tsukuba-Plant Innovation Research Center, University of Tsukuba (to SK, No 2002). We wish to thank Dr. Junji Shimabukuro, Kyoto Institute of Technology, for his generous aid in rearing S. litura.
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Savadogo, E.H., Shiomi, Y., Yasuda, J. et al. Gene expression of PLAT and ATS3 proteins increases plant resistance to insects. Planta 253, 37 (2021). https://doi.org/10.1007/s00425-020-03530-y
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DOI: https://doi.org/10.1007/s00425-020-03530-y