The trap of the carnivorous plant Venus flytrap (Dionaea muscipula) catches prey by very rapid closure of its modified leaves. After the rapid closure secures the prey, repeated mechanical stimulation of trigger hairs by struggling prey and the generation of action potentials (APs) result in secretion of digestive fluid. Once the prey's movement stops, the secretion is maintained by chemical stimuli released from digested prey. We investigated the effect of mechanical and chemical stimulation (NH4Cl, KH2PO4, further N(Cl) and P(K) stimulation) on enzyme activities in digestive fluid. Activities of β-D-glucosidases and N-acetyl-β-D-glucosaminidases were not detected. Acid phosphatase activity was higher in N(Cl) stimulated traps while proteolytic activity was higher in both chemically induced traps in comparison to mechanical stimulation. This is in accordance with higher abundance of recently described enzyme cysteine endopeptidase dionain in digestive fluid of chemically induced traps. Mechanical stimulation induced high levels of cis-12-oxophytodienoic acid (cis-OPDA) but jasmonic acid (JA) and its isoleucine conjugate (JA-Ile) accumulated to higher level after chemical stimulation. The concentration of indole-3-acetic acid (IAA), salicylic acid (SA) and abscisic acid (ABA) did not change significantly. The external application of JA bypassed the mechanical and chemical stimulation and induced a high abundance of dionain and proteolytic activity in digestive fluid. These results document the role of jasmonates in regulation of proteolytic activity in response to different stimuli from captured prey. The double trigger mechanism in protein digestion is proposed.

Department of Plant Physiology Faculty of Natural Sciences Comenius University in Bratislava Bratislava Slovakia

Department of Plant Physiology Faculty of Natural Sciences Comenius University in Bratislava Bratislava Slovakia; Department of Biophysics Centre of the Region Haná for Biotechnological and Agricultural Research Palacký University Olomouc Czech Republic

Laboratory of Growth Regulators Centre of the Region Haná for Biotechnological and Agricultural Research Institute of Experimental Botany ASCR and Palacký University Olomouc Czech Republic

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Darwin C (1875) Insectivorous plants. John Murray, London.

Burdon-Sanderson JS (1873) Note on the electrical phenomena which accompany stimulation of the leaf of

Krol E, Dziubinska H, Stolarz M, Trebacz K (2006) Effects of ion channel inhibitors on cold- and ellectrically-induced action potentials in

Brown WH (1916) The mechanism of movement and the duration of the effect of stimulation in the leaves of

Hodick D, Sievers A (1988) The action potential of PubMed

Volkov AG, Adesina T, Jovanov E (2007) Closing of Venus flytrap by electrical stimulation of motor cells. Plant Signal Behav 2: 139–145. PubMed PMC

Volkov AG, Adesina T, Markin VS, Jovanov E (2008) Kinetics and mechanism of PubMed PMC

Volkov AG, Carrell H, Baldwin A, Markin VS (2009) Electrical memory in Venus flytrap. Bioelectrochemistry 75: 142–147. PubMed

Volkov AG, Carrell H, Markin VS (2009) Biologically closed electrical circuits in Venus Flytrap. Plant Physiol 149: 1661–1667. PubMed PMC

Hodick D, Sievers A (1989) On the mechanism of trap closure of Venus flytrap ( PubMed

Forterre Y, Skotheim JM, Dumais J, Mahadevan L (2005) How the Venus flytrap snaps. Nature 433: 421–425. PubMed

Markin VS, Volkov AG, Jovanov E (2008) Active movements in plants: mechanism of trap closure by PubMed PMC

Williams ME, Bennett AB (1982) Leaf closure in the Venus flytrap: an acid growth response. Science 218: 1120–1121. PubMed

Volkov AG, Harris SL, Vilfranc CL, Murphy VA, Wooten JD, et al. (2013) Venus flytrap biomechanics: Forces in the PubMed

Affolter JM, Olivo RF (1975) Action potentials in Venus's-flytraps: long term observations following the capture of prey. Am Midl Nat 93: 443–445.

Robins RJ (1976) The nature of the stimuli causing digestive juice secretion in PubMed

Lichtner FT, Williams SE (1977) Prey capture and factors controlling trap narrowing in

Ueda M, Tokunaga T, Okada M, Nakamura Y, Takada N, et al. (2010) Trap-closing chemical factors of the Venus flytrap ( PubMed

Escalante-Pérez M, Krol E, Stange A, Geiger D, Al-Rasheid KA, et al. (2011) A special pair of phytohormones controls excitability, slow closure, and external stomach formation in the Venus flytrap. PNAS USA 108: 15492–15497. PubMed PMC

Scala J, Iott K, Schwab DW, Semersky FE (1969) Digestive secretion of PubMed PMC

Robins RJ, Juniper BE (1980) The secretory cycle of

Takahashi K, Matsumoto K, Nishi W, Muramatsu M, Kubota K (2009) Comparative studies on the acid proteinase activities in the digestive fluids of

Athauda SBP, Matsumoto K, Rajapakshe S, Kuribayashi M, Kojima M, et al. (2004) Enzymic and structural characterization of nepenthesin, a unique member of a novel subfamily of aspartic proteinases. Biochem J 381: 295–306. PubMed PMC

Takahashi K, Suzuki T, Nishii W, Kubota K, Shibata C, et al. (2011) A cysteine endopeptidase (“Dionain”) is involved in the digestive fluid of PubMed

Schulze WX, Sanggaard KW, Kreuzer I, Knudsen AD, Bemm F, et al. (2012) The protein composition of the digestive fluid from the Venus flytrap sheds light on prey digestion mechanisms. Mol Cell Proteom 11: 1306–1319. PubMed PMC

Takahashi K, Nishii W, Shibata C (2012) The digestive fluid of

Ellison A (2006) Nutrient limitation and stoichiometry of carnivorous plants. Plant Biol 8: 740–747. PubMed

Pavlovič A, Krausko M, Libiaková M, Adamec L (2014) Feeding on prey increases photosynthetic efficiency in the carnivorous sundew PubMed PMC

Hlaváčková V, Krchňák P, Nauš J, Novák O, Špundová M, et al. (2006) Electrical and chemical signals involved in short-term systemic photosynthetic responses of tobacco plants to local burning. Planta 225: 235–244. PubMed

Glauser G, Grata E, Dubugnon L, Rudaz S, Farmer EE, et al. (2008) Spatial and temporal dynamics of jasmonate synthesis and accumulation in PubMed

Matušíková I, Salaj J, Moravčíková J, Mlynárová L, Nap JP, et al. (2005) Tentacles of in vitro-grown round-leaf sundew ( PubMed

Juniper BE, Robins RJ, Joel DM (1989) The carnivorous plants. London, UK: Academic Press.

Volkov AG, Pinnock MR, Lowe DC, Gay MS, Markin VS (2011) Complete hunting cycle of PubMed

Balotin NM, DiPalma JR (1962) Spontaneous electrical activity of PubMed

Scherzer S, Krol E, Kreuzer I, Kruse J, Karl F, et al. (2013) The PubMed

Dick CF, Dos-Santos ALA, Meyer-Fernandes JR (2011). Inorganic phosphate as an important regulator of Phosphatases. Enzyme Res 2011: ID 103980. PubMed PMC

Gallie DR, Chang SC (1997) Signal transduction in the carnivorous plant PubMed PMC

Eilenberg H, Pnini-Cohen S, Schuster S, Movtchan A, Zilberstein A (2006) Isolation and characterization of chitinase genes from pitchers of the carnivorous plant PubMed

Rottloff S, Stieber R, Maischak H, Turini FG, Heubl G, et al. (2011) Functional characterization of a class III acid endochitinase from the traps of the carnivorous pitcher plant genus, PubMed PMC

Mithöfer A (2011) Carnivorous pitcher plants: insight in an old topic. Phytochemistry 72: 1678–1682. PubMed

Hatano N, Hamada T (2012) Proteomic analysis of secreted protein induced by a component of prey in pitcher fluid of the carnivorous plant PubMed

Paszota P, Escalante-Perez M, Thomsen LR, Risør MW, Dembski A, et al. (2014) Secreted major Venus flytrap chitinase enables digestion of Arthropod prey. Biochim Biophys Acta 1844: 374–383. PubMed

Robins RJ, Juniper BE (1980) The secretory cycle of

Robins RJ, Juniper BE (1980) The secretory cycle of

Nishimura E, Kawahara M, Kodaira R, Kume M, Arai N, et al. (2013) S-like ribonuclease gene expression in carnivorous plants. Planta 238: 955–967. PubMed

Stephenson P, Hogan J (2006) Cloning and characterization of a ribonuclease, a cysteine proteinase, and an aspartic proteinase from pitchers of the carnivorous plant

Hatano N, Hamada T (2008) Proteome analysis of pitcher fluid of the carnivorous plant PubMed

Fisahn J, Herde O, Willmitzer L, Peňa-Cortés H (2004) Analysis of the transient increase in cytosolic Ca PubMed

Mousavi SAR, Chauvin A, Pascaud F, Kellenberger S, Farmer EE (2013) Glutamate receptor-like genes mediate leaf-to-leaf wound signals. Nature 500: 422–426. PubMed

Monshausen GB, Haswell ES (2013) A force of nature: molecular mechanisms of mechanoperception in plants. J Exp Bot 64: 4663–4680. PubMed PMC

Wasternack C, Hause B (2013) Jasmonates: biosynthesis, perception, signal transduction and action in plant stress response, growth and development. An update to the 2007 review in Annals of Botany. Ann Bot 111: 1024–1058. PubMed PMC

Nakamura Y, Reichelt M, Mayer VE, Mithöfer A (2013) Jasmonates trigger prey-induced formation of outer stomach in carnivorous sundew plants. Proc R Soc B 280: 20130228. PubMed PMC

Mithöfer A, Reichelt M, Nakamura Y (2014) Wound and insect-induced jasmonate accumulation in carnivorous PubMed

Staswick PE, Tiryaki I (2004) The oxylipin signal jasmonic acid is activated by an enzyme that conjugates it to isoleucine in PubMed PMC

Fonseca S, Chini A, Hamberg M, Adie B, Porzel A, et al. (2009) (+)-7-iso-Jasmonyl-L-isoleucine is the endogenous bioactive jasmonate. Nature Chem Biol 5: 344–350. PubMed

Sheard LB, Tan X, Mao H, Withers J, Ben-Nissan G, et al. (2010) Jasmonate perception by inositol-phosphate-potentiated COI-JAZ co-receptor. Nature 468: 400–407. PubMed PMC

Ballaré CL (2011) Jasmonate-induced defenses: a tale of intelligence, collaborators and rascals. Trends Plant Sci 16: 249–257. PubMed

Chung HS, Koo AJK, Gao X, Jayanty S, Thines B, et al. (2008) Regulation and function of PubMed PMC

Wang L, Allmann S, Wu J, Baldwin IT (2008) Comparisons of LIPOXYGENASE3- and JASMONATE-RESISTANT4/6-silenced plants reveal that jasmonic acid and jasmonic acid-aminoacid conjugates play different roles in herbivore resistance of PubMed PMC

Stintzi A, Weber H, Reymond P, Browse J, Farmer EE (2001) Plants defense in the absence of jasmonic acid: The role of cyclopentenones. PNAS USA 98: 12837–12842. PubMed PMC

Taki N, Sasaki-Sekimoto Y, Obayashii T, Kikuta A, Kobayashi K, et al. (2005) 12-oxo-phytodienoic acid triggers expression of a distinct set of genes and plays a role in wound-induced gene expression in PubMed PMC

Stelmach BA, Müller A, Weiler EW (1999) 12-oxo-phytodienoic acid and indole-3-acetic acid in jasmonic acid-treated tendrils of

Givnish TJ, Burkhardt EL, Happel RE, Weintraub JD (1984) Carnivory in the bromeliad

Pavlovič A, Demko V, Hudák J (2010) Trap closure and prey retention in Venus flytrap ( PubMed PMC

Pavlovič A, Slováková L', Pandolfi C, Mancuso S (2011) On the mechanism underlying photosynthetic limitation upon trigger hair irritation in the carnivorous plant Venus flytrap ( PubMed PMC

Jaffe MJ (1973) The role of ATP in mechanically stimulated rapid closure of the Venus's-flytrap. Plant Physiol 51: 17–18. PubMed PMC

Farnsworth EJ, Ellison AM (2008) Prey availability directly affects physiology, growth, nutrient allocation and scaling relationships among leaf traits in 10 carnivorous plant species. J Ecol 96: 213–221.

Pavlovič A, Singerová L, Demko V, Hudák J (2009) Feeding enhances photosynthetic efficiency in the carnivorous pitcher plant PubMed PMC

Pavlovič A, Slováková L', Šantrůček J (2011) Nutritional benefit from leaf litter utilization in the pitcher plant PubMed

Kruse J, Gao P, Honsel A, Kreuzwieser J, Burzlaff T, et al. (2014) Strategy of nitrogen acquisition and utilization by carnivorous PubMed

Gibson TC, Waller DM (2009) Evolving Darwin's ‘most wonderful’ plant: ecological steps to a snap-trap. New Phytol 183: 575–587. PubMed

Schulze W, Schulze ED, Schulze I, Oren R (2001) Quantification of insect nitrogen utilization by the Venus flytrap PubMed

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