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Plant antiherbivore defenses in Fabaceae species of the Chaco

Defesas das plantas anti-herbivoria em espécies de Fabaceae do Chaco

Abstract

The establishment and maintenance of plant species in the Chaco, one of the widest continuous areas of forests in the South American with sharp climatic variations, are possibly related to biological features favoring plants with particular defenses. This study assesses the physical and chemical defenses mechanisms against herbivores of vegetative and reproductive organs. Its analyses of 12 species of Fabaceae (Leguminosae) collected in remnants of Brazilian Chaco shows that 75% present structural defense characters and 50% have chemical defense – defense proteins in their seeds, like protease inhibitors and lectins. Physical defenses occur mainly on branches (78% of the species), leaves (67%), and reproductive organs (56%). The most common physical characters are trichomes and thorns, whose color represents a cryptic character since it does not contrast with the other plant structures. Defense proteins occur in different concentrations and molecular weight classes in the seeds of most species. Protease inhibitors are reported for the first time in seeds of: Albizia niopoides, Anadenanthera colubrina, Mimosa glutinosa, Prosopis rubriflora, and Poincianella pluviosa. The occurrence of physical and chemical defenses in members of Fabaceae indicate no associations between defense characters in these plant species of the Chaco.

Keywords:
structural characters; protease inhibitors; lectins; defense mechanisms

Resumo

O estabelecimento e a manutenção de espécies no Chaco, uma planície semi-árida da América do Sul com variações climáticas importantes, possivelmente estão relacionados a características biológicas que favorecem as plantas detentoras de defesas particulares. Este estudo teve como objetivos avaliar os mecanismos de defesa física e química anti-herbivoria em órgãos vegetativos e reprodutivos. Analisamos 12 espécies da família Fabaceae (Leguminosae) obtidas em remanescentes de Chaco brasileiro. Observamos que 75% das espécies estudadas apresentam atributo de defesa física e 50% possuem defesa química – proteínas de defesa nas sementes, como inibidores de protease e lectinas. As defesas físicas ocorrem principalmente nos ramos (78% das espécies), nos órgãos reprodutivos (56% das espécies) e nas folhas (67%). Os atributos físicos mais frequentes são tricomas e espinhos, cuja coloração não contrastante com as demais estruturas das plantas representa um caráter críptico. Proteínas de defesa ocorrem nas sementes da maioria das espécies, com diferentes concentrações e classes de pesos moleculares. Inibidores de protease nas sementes estão sendo relatados pela primeira vez em: Albizia niopoides, Anadenanthera colubrina, Mimosa glutinosa, Prosopis rubriflora e Poincianella pluviosa. A ocorrência de defesas física e química entre os membros de Fabaceae indica que não há associações entre as características de defesa das espécies de plantas avaliadas no Chaco.

Palavras-chave:
características estruturais; inibidores de protease; lectinas; mecanismos de defesa

1 Introduction

When present in plants, physical and chemical defenses affect the development and survival of their attackers – herbivores (Hanley et al., 2007Hanley, M.E., Lamont, B.B., Fairbanks, M.H. and Rafferty, C.M., 2007. Plant structural traits and their role in anti-herbivore defence. Perspectives in Plant Ecology, Evolution and Systematics, vol. 8, no. 4, pp. 157-178. http://dx.doi.org/10.1016/j.ppees.2007.01.001.
http://dx.doi.org/10.1016/j.ppees.2007.0...
). Physical defenses act as barriers to herbivory through rigid protuberances and structures as thorns and/or spines, trichomes, leaf rigidity, formation of minerals - raphides and druses - in vegetal tissues, and seeds protected by hard testae (Dickison, 2000Dickison, W.C., 2000. Integrative plant anatomy. Burlington: Academic Press. 533 p.; Valverde et al., 2001Valverde, P.L., Fornoni, J. and Núñez-Farfán, J., 2001. Defensive role of leaf trichomes in resistance to herbivorous insects in Datura stramonium.Journal of Evolutionary Biology, vol. 14, no. 3, pp. 424-432. http://dx.doi.org/10.1046/j.1420-9101.2001.00295.x.
http://dx.doi.org/10.1046/j.1420-9101.20...
). Spinescence, raphides, and druses are mainly associated with protection from mammals, while pubescence and sclerophylly essentially thwart the access of insects (Wagner, 1991Wagner, G.J., 1991. Secreting glandular trichomes: more than just hairs. Plant Physiology, vol. 96, no. 3, pp. 675-679. http://dx.doi.org/10.1104/pp.96.3.675. PMid:16668241.
http://dx.doi.org/10.1104/pp.96.3.675...
; Hanley et al., 2007Hanley, M.E., Lamont, B.B., Fairbanks, M.H. and Rafferty, C.M., 2007. Plant structural traits and their role in anti-herbivore defence. Perspectives in Plant Ecology, Evolution and Systematics, vol. 8, no. 4, pp. 157-178. http://dx.doi.org/10.1016/j.ppees.2007.01.001.
http://dx.doi.org/10.1016/j.ppees.2007.0...
).

Chemical defenses against herbivores are characterized by the synthesis of primary metabolites, as defense proteins, and secondary metabolites, as terpenoids and nitrogen-containing and phenolic compounds, resulting from the production of compounds that are essential to plants (Chen, 2008Chen, M.-S., 2008. Inducible direct plant defense against insect herbivores: a review. Insect Science, vol. 15, no. 2, pp. 101-114. http://dx.doi.org/10.1111/j.1744-7917.2008.00190.x.
http://dx.doi.org/10.1111/j.1744-7917.20...
). Direct defenses thwart herbivores by producing secondary metabolites or defense enzymes that act directly against attackers, while indirect defenses involve producing volatile compounds that attract natural enemies of herbivores (Pieterse et al., 2012Pieterse, C.M.J., Van Der Does, D., Zamioudis, C., Leon-Reyes, A. and Van Wees, S.C.M., 2012. Hormonal modulation of plant immunity. Annual Review of Cell and Developmental Biology, vol. 28, no. 1, pp. 489-521. http://dx.doi.org/10.1146/annurev-cellbio-092910-154055. PMid:22559264.
http://dx.doi.org/10.1146/annurev-cellbi...
). Among plant defense proteins are protease inhibitors (PIs) and lectins, which are found in vegetal tissues, mainly in reserve organs, and usually act against insects, bacteria and fungi (Peumans and Van Damme, 1995Peumans, W.J. and Van Damme, E.J.M., 1995. Lectins as plant defense proteins. Plant Physiology, vol. 109, no. 2, pp. 347-352. http://dx.doi.org/10.1104/pp.109.2.347. PMid:7480335.
http://dx.doi.org/10.1104/pp.109.2.347...
; Dunaevsky et al., 2005Dunaevsky, Y.A.E., Elpidina, E.N., Vinokurov, K.S. and Belozersky, M.A., 2005. Protease inhibitors in improvement of plant resistance to pathogens and insects. Molecular Biology, vol. 39, no. 4, pp. 608-613. http://dx.doi.org/10.1007/s11008-005-0076-y.
http://dx.doi.org/10.1007/s11008-005-007...
). Lectins and PIs affect the digestive process of insects by reducing the breakdown of the ingested proteins into amino acids (Murdock and Shade, 2002Murdock, L.L. and Shade, R.E., 2002. Lectins and protease inhibitors as plant defenses against insects. Journal of Agricultural and Food Chemistry, vol. 50, no. 22, pp. 6605-6611. http://dx.doi.org/10.1021/jf020192c. PMid:12381159.
http://dx.doi.org/10.1021/jf020192c...
; Macedo et al., 2004Macedo, M.L.R., Castro, M.N. and Freire, M.G.M., 2004. Mechanisms of the insecticidal action of TEL (Talisia esculenta Lectin) against Callosobruchus maculatus (Coleoptera: Bruchidae). Archives of Insect Biochemistry and Physiology, vol. 56, no. 2, pp. 84-96. http://dx.doi.org/10.1002/arch.10145. PMid:15146543.
http://dx.doi.org/10.1002/arch.10145...
; Vandenborre et al., 2011Vandenborre, G., Smagghe, G. and Van Damme, E.J.M., 2011. Plant lectins as defense proteins against phytophagous insects. Phytochemistry, vol. 72, no. 13, pp. 1538-1350. http://dx.doi.org/10.1016/j.phytochem.2011.02.024. PMid:21429537.
http://dx.doi.org/10.1016/j.phytochem.20...
).

The preponderance of legumes (Fabaceae) in different plant formations worldwide suggests that some biological features favor their establishment. In these species, the main physical characters related to herbivory are thorns, spines, tector trichomes, secretory trichomes, and seeds with hard testae. Fabaceae species also produce chemical compounds of different classes that have distinct functions against herbivores (Levin, 1976Levin, D.A., 1976. The chemical defenses of plants to pathogens and herbivores. Annual Review of Ecology and Systematics, vol. 7, no. 1, pp. 121-159. http://dx.doi.org/10.1146/annurev.es.07.110176.001005.
http://dx.doi.org/10.1146/annurev.es.07....
; Kortt and Jermyn, 1981Kortt, A.A. and Jermyn, M.A., 1981. Acacia proteinase inhibitors: purification and properties of the trypsin inhibitors from . Acacia elata seedEuropean Journal of Biochemistry, vol. 115, no. 3, pp. 551-557. http://dx.doi.org/10.1111/j.1432-1033.1981.tb06238.x. PMid:7238519.
http://dx.doi.org/10.1111/j.1432-1033.19...
). In this family, defense proteins have been studied in species as Adenanthera pavonina L., Bauhinia bauhinioides (Mart.) J.F.Macbr., Dimorphandra mollis Benth., Inga laurina (Sw.) Willd., Plathymenia foliolosa Benth., among others (Macedo et al., 2002Macedo, M.L.R., Mello, G.C., Freire, M.G.M., Novello, J.C., Marangoni, S. and Matos, D.G.G., 2002. Effect of a trypsin inhibitor from seeds on the development of Dimorphandra mollisCallosobruchus maculatus.Plant Physiology and Biochemistry, vol. 40, no. 10, pp. 891-898. http://dx.doi.org/10.1016/S0981-9428(02)01441-9.
http://dx.doi.org/10.1016/S0981-9428(02)...
, 2004Macedo, M.L.R., Castro, M.N. and Freire, M.G.M., 2004. Mechanisms of the insecticidal action of TEL (Talisia esculenta Lectin) against Callosobruchus maculatus (Coleoptera: Bruchidae). Archives of Insect Biochemistry and Physiology, vol. 56, no. 2, pp. 84-96. http://dx.doi.org/10.1002/arch.10145. PMid:15146543.
http://dx.doi.org/10.1002/arch.10145...
, 2011Macedo, M.L.R., Freire, M.G.M., Franco, O.L., Migliolo, L. and Oliveira, C.F.R., 2011. Practical and theoretical characterization of Kunitz inhibitor on the control of Inga laurinaHomalinotus coriaceus.Comparative Biochemistry and Physiology, vol. 158, no. 2, pp. 164-172. http://dx.doi.org/10.1016/j.cbpb.2010.11.005. PMid:21094272.
http://dx.doi.org/10.1016/j.cbpb.2010.11...
; Ramos et al., 2008Ramos, V.S., Silva, G.S., Freire, M.G.M., Machado, O.L.T., Parra, J.R.P. and Macedo, M.L.R., 2008. Purification and characterization of a trypsin inhibitor from . Plathymenia foliolosa seedsJournal of Agricultural and Food Chemistry, vol. 56, no. 23, pp. 11348-11355. http://dx.doi.org/10.1021/jf802778b. PMid:18991455.
http://dx.doi.org/10.1021/jf802778b...
; Sumikawa et al., 2010Sumikawa, J.T., Brito, M.V., Macedo, M.L.R., Uchoa, A.F., Miranda, A., Araujo, A.P.U., Silva-Lucca, R.A., Sampaio, U.M. and Oliva, M.L.V., 2010. The defensive functions of plant inhibitors are not restricted to insect enzyme inhibition. Phytochemistry, vol. 71, no. 2-3, pp. 214-220. http://dx.doi.org/10.1016/j.phytochem.2009.10.009. PMid:19939420.
http://dx.doi.org/10.1016/j.phytochem.20...
).

Only found in South America, the Chaco is a semi-arid grassland with extreme climatic conditions that encompasses parts of Argentina, Bolivia, Paraguay, and of the southwestern region of the state of Mato Grosso do Sul, Brazil. In the Brazilian Chaco, Fabaceae stands out by the richness and diversity of its species, many of which present physical defenses as thorns and trichomes (Alves and Sartori, 2009ALVES, F.M. and SARTORI, A.L.B., 2009. Caesalpinioideae (Leguminosae) de um remanescente de Chaco em Porto Murtinho, Mato Grosso do Sul, Brasil. Rodriguésia, vol. 60, pp. 531-550.; Noguchi et al., 2009NOGUCHI, D.K., NUNES, G.P. and SARTORI, A.L.B., 2009. Florística e síndromes de dispersão de espécies arbóreas em remanescentes de Chaco de Porto Murtinho, Mato Grosso do Sul, Brasil. Rodriguésia, vol. 60, pp. 353-365.). Yet no investigations have ever considered the chemical characters of legumes growing in the Brazilian Chaco.

The establishment and maintenance of species in this region must require biological features favoring plants with particular characters. The occurrence of legumes with thorns, spines, and seeds with hard testae and the report of defense proteins in seeds of some species of this family suggest a possible relation between these defense mechanisms, an aspect not yet studied. This work thus assesses the physical defenses of legumes and the presence of defense proteins in their seeds.

2 Methods

Botanical material was collected in remnants of wooded steppic savanna (Wooded Chaco), forested steppic savanna (Forested Chaco), and transition areas between Chaco and Cerrado, Porto Murtinho municipality, western part of the state of Mato Grosso do Sul, Brazil (21°40’S, 57°52’W), from April to September 2011. Seeds of tree and shrub legume were collected simultaneously from at least three individuals sampled for each species, according to their availability in the environment. Were assessed 12 species, six Mimosoideae and six Caesalpinioideae (Table 1).

Table 1
Fabaceae species from the Brazilian Chaco classified in subfamilies showing data on spinescence, trichome types, thorn color, protease inhibitors, and lectins.

Physical defenses were assessed on vegetative (leaves and stem) and reproductive (flowers and fruits) organs. They include the following characters: spinescence, trichome types, and the color of thorns/spines compared to that of branches (Ronel and Lev-Yadun, 2012Ronel, M. and Lev-Yadun, S., 2012. The spiny, thorny and prickly plants in the flora of Israel. Botanical Journal of the Linnean Society, vol. 168, no. 3, pp. 344-352. http://dx.doi.org/10.1111/j.1095-8339.2011.01211.x.
http://dx.doi.org/10.1111/j.1095-8339.20...
). Physical characters like spinescence and color of thorns were analyzed on field specimens, while trichome types on herbarium and field specimens. Botanical material was deposited in the Herbarium of Universidade Federal de Mato Grosso do Sul (CGMS Herbarium). Family nomenclature follows Lewis et al. (2005)Lewis, G.P., Schrire, B., Mackinder, B. and Lock, M., 2005. Legumes of the world. Kew: Royal Botanic Gardens. 577 p..

As for chemical defenses in seeds, the presence or not of protease inhibitors (PI) and lectin was assessed. The collected seeds of each species were macerated into fine granulated flours that were subjected to delipidation and protein extraction. Protein were extracted with (0.1M, pH 7.6) potassium phosphate buffer and (0.15 N) NaCl buffer in PI and lectin assays, respectively. Protein concentration was then estimated according to the method of Bradford (1976)Bradford, M.M., 1976. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of dye binding. Analytical Biochemistry, vol. 72, no. 1-2, pp. 248-254. http://dx.doi.org/10.1016/0003-2697(76)90527-3. PMid:942051.
http://dx.doi.org/10.1016/0003-2697(76)9...
, with absorbance measured at 595 nm.

The presence of PIs was observed through the method of Erlanger (Erlanger et al., 1961Erlanger, B.F., Kokowsky, N. and Cohen, N., 1961. Preparation and properties of two new chromogenic substrates of trypsin. Archives of Biochemistry and Biophysics, vol. 95, no. 2, pp. 271-278. http://dx.doi.org/10.1016/0003-9861(61)90145-X. PMid:13890599.
http://dx.doi.org/10.1016/0003-9861(61)9...
). For each species, 5 µg of proteins were used per µL of crude extract and assays were carried out with Tris(hydroxymethyl) aminomethane buffer, N-benzoyl-DL-arginine-p-nitroanilide (BApNA) substrate, and bovine trypsin, and absorbance was read at 410 nm. Inhibitory activity was defined by the following formula: IU = (T - A) / (0.250 × Vassay), where: IU = inhibition unit; T = trypsin reading; A = sample reading; and Vassay = volume of sample used in the assay. The occurrence of PIs was considered as a chemical defense mechanism when concentration exceeded 100 IU g–1.

The presence of lectin was observed through hemagglutinating activity (HA) using microtitration plates. For each species, a 100 μL sample was assayed in triplicate serial dilutions and homogenized. Then, 100 μL of a 2%, suspension of red blood cells prepared with human blood (type A Rh positive) were added and red blood cell agglutination was observed 60 minutes later. Results were expressed in hemagglutination units (HU), defined as the reciprocal of the highest dilution in which hemagglutination was observed.

To estimate the apparent molecular weight, the proteins extracted from each species were analyzed by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), according to Laemmli (1970)Laemmli, V.K., 1970. Cleavage of structural proteins during the assembly of the bacteriophage T4. Nature, vol. 227, no. 5259, pp. 680-685. http://dx.doi.org/10.1038/227680a0. PMid:5432063.
http://dx.doi.org/10.1038/227680a0...
. A molecular mass marker with six proteins: lysozyme (14 kDa), β-lactoglobulin (18 kDa), trypsinogen (24 kDa), pepsin (34 kDa), and albumin (66 kDa) (SIGMA) was used. All chemical analyses were performed at the Laboratório de Purificação de Proteínas e suas Funções Biológicas of Universidade Federal de Mato Grosso do Sul.

3 Results

Among the 12 species assessed, nine (75%) presented physical defenses and six (50%), chemical defenses (Table 1). In most species (67%) different types of physical defenses occurred on more than one plant structure, predominantly on vegetative organs. The most frequent were tector trichomes (67%) and spinescence (33%). Stems and branches presented most defenses (78%), followed by leaves (67%) and reproductive organs (56%).

Leaves only presented tector trichomes, while reproductive organs bore tector and glandular trichomes, and branches had spinescence and both tector and glandular trichomes (Table 1). Spinescence was observed in four species, three of which had spinescent stipules. Thorns were mainly brown and only presented vinaceous ends in Mimosa glutinosa Malme.

All the species of Mimosoideae assessed presented some kind of physical or chemical defense. Mimosa glutinosa and Prosopis rubriflora Hassl. had both, Mimosa hexandra Micheli and Microlobius foetidus (Jacq.) M.Sousa & G.Andrade only showed physical defenses, and Albizia niopoides (Spruce ex Benth.) Burkart and Anadenanthera colubrina (Vell.) Brenan only presented chemical defenses.

Of the six species of Caesalpinioideae studied, only Libidibia paraguariensis (D. Parodi) G.P.Lewis had none of the defenses assessed. Poincianella pluviosa (DC.) L.P.Queiroz and Peltophorum dubium (Spreng.) Taub. presented both types. Parkinsonia praecox (Ruiz and Pav. ex Hook.) J. A. Hawkins, Pterogyne nitens Tul., and Senna occidentalis (L.) Link, only presented physical defenses.

Lectin, although in low concentration (4 HU), was only found in the seeds of Albizia niopoides. Six species presented high concentrations of PIs (above 100 IU g–1) and three very low concentrations (1-10 IU g–1), as shown in Table 1. The molecular weights of the soluble proteins found in the seeds varied from 10 to 66 kDa. In Albizia niopoides and Anadenanthera colubrina, protease inhibitors ranged 18-24 kDa; Prosopis rubriflora and Poincianella pluviosa, 10-24 kDa; Mimosa glutinosa, 10-18 kDa; while in Peltophorum dubium 66 kDa.

4 Discussion

Trichomes on leaves, branches, and reproductive organs protect plants against herbivores, pathogens, excess of heat, and water loss (Wagner, 1991Wagner, G.J., 1991. Secreting glandular trichomes: more than just hairs. Plant Physiology, vol. 96, no. 3, pp. 675-679. http://dx.doi.org/10.1104/pp.96.3.675. PMid:16668241.
http://dx.doi.org/10.1104/pp.96.3.675...
; Agrawal and Fishbein, 2006Agrawal, A.A. and Fishbein, M., 2006. Plant defense syndromes. Ecology, vol. 87, no. 7, (suppl.), pp. S132-S149. http://dx.doi.org/10.1890/0012-9658(2006)87[132:PDS]2.0.CO;2. PMid:16922309.
http://dx.doi.org/10.1890/0012-9658(2006...
). Physical defenses, as spinescence, play a more important role to protect stems and branches than reproductive organs (Ronel and Lev-Yadun, 2012Ronel, M. and Lev-Yadun, S., 2012. The spiny, thorny and prickly plants in the flora of Israel. Botanical Journal of the Linnean Society, vol. 168, no. 3, pp. 344-352. http://dx.doi.org/10.1111/j.1095-8339.2011.01211.x.
http://dx.doi.org/10.1111/j.1095-8339.20...
). Among the features analyzed, it is worth highlighting that in spinescent species thorns color tends to be a cryptic character, i.e. it is the same as that of the structure that bears it.

In addition to constituting a physical barrier to herbivory, in some species, thorns and spines may have warning coloration to mammals herbivores and may be associated with pathogenic microorganisms (Lev-Yadun, 2001Lev-Yadun, S., 2001. Aposematic (warning) coloration associated with thorns in higher plants. Journal of Theoretical Biology, vol. 210, no. 3, pp. 385-388. http://dx.doi.org/10.1006/jtbi.2001.2315. PMid:11397139.
http://dx.doi.org/10.1006/jtbi.2001.2315...
). Chemical and physical defense mechanisms can act together to potentiate defenses against insects-plagues in plants of economic interest, like PIs expression in leaf trichomes of transgenic plants, which increases density and ramification of trichomes, resulting in extra resistance mechanism (Liu et al., 2006Liu, J., Xia, K.F., Zhu, J.C., Deng, Y.G., Huang, X.L., Hu, B.L., Xu, X. and Xu, Z.F., 2006. The nightshade proteinase inhibitor IIb gene is constitutively expressed in glandular trichomes. Plant & Cell Physiology, vol. 47, no. 9, pp. 1274-1284. http://dx.doi.org/10.1093/pcp/pcj097. PMid:16926166.
http://dx.doi.org/10.1093/pcp/pcj097...
; Luo et al., 2009Luo, M., Wang, Z., Li, H., Xia, K.F., Cai, Y. and Xu, Z.F., 2009. Overexpression of a weed (Solanum americanum) proteinase inhibitor in transgenic tobacco results in increased glandular trichome density and enhanced resistance to Helicoverpa armigera and Spodoptera litura.International Journal of Molecular Sciences, vol. 10, no. 4, pp. 1896-1910. http://dx.doi.org/10.3390/ijms10041896. PMid:19468345.
http://dx.doi.org/10.3390/ijms10041896...
).

Plants respond to herbivory through several strategies, which lead to different interpretations for the evolution of plant defense (Agrawal, 2006Agrawal, A.A., 2006. Macroevolution of plant defense strategies. Trends in Ecology & Evolution, vol. 22, no. 2, pp. 103-109. http://dx.doi.org/10.1016/j.tree.2006.10.012. PMid:17097760.
http://dx.doi.org/10.1016/j.tree.2006.10...
; Moreira et al., 2016Moreira, X., Abdala-Roberts, L., Rasmann, S., Castagneyrol, B. and Mooney, K.A., 2016. Plant diversity effects on insect herbivores and their natural enemies: current thinking, recent findings, and future directions. Current Opinion in Insect Science, vol. 14, pp. 1-7. http://dx.doi.org/10.1016/j.cois.2015.10.003. PMid:27436639.
http://dx.doi.org/10.1016/j.cois.2015.10...
). There are species that invest in given defenses according to resource availability (Almeida-Cortez et al., 2004Almeida-Cortez, J.S., Shipley, B. and Arnason, J.T., 2004. Growth and chemical defense in relation to resource availability: tradeoffs or common responses to environmental stress? Brazilian Journal of Biology = Revista Brasileira de Biologia, vol. 64, no. 2, pp. 187-194. http://dx.doi.org/10.1590/S1519-69842004000200002. PMid:15462290.
http://dx.doi.org/10.1590/S1519-69842004...
; Hanley et al., 2007Hanley, M.E., Lamont, B.B., Fairbanks, M.H. and Rafferty, C.M., 2007. Plant structural traits and their role in anti-herbivore defence. Perspectives in Plant Ecology, Evolution and Systematics, vol. 8, no. 4, pp. 157-178. http://dx.doi.org/10.1016/j.ppees.2007.01.001.
http://dx.doi.org/10.1016/j.ppees.2007.0...
). Moreover defense mechanisms antiherbivore may complement each other, favoring the presence of mixed defense (Carmona and Fornoni, 2013Carmona, D. and Fornoni, J., 2013. Herbivores can select for mixed defensive strategies in plants. The New Phytologist, vol. 197, no. 2, pp. 576-585. http://dx.doi.org/10.1111/nph.12023. PMid:23171270.
http://dx.doi.org/10.1111/nph.12023...
).

Plant species growing in similar environments converge on suites of co-varying defense characters, according to theory of plant defense syndromes (Agrawal and Fishbein, 2006Agrawal, A.A. and Fishbein, M., 2006. Plant defense syndromes. Ecology, vol. 87, no. 7, (suppl.), pp. S132-S149. http://dx.doi.org/10.1890/0012-9658(2006)87[132:PDS]2.0.CO;2. PMid:16922309.
http://dx.doi.org/10.1890/0012-9658(2006...
). We verified that in the Brazilian Chaco there is no pattern in the occurrence of physical and chemical defense characters between members of Fabaceae. This suggests that in Chaco, plants must maximize their resources since they are subject to weather extremes as severe variations in temperature and water availability. Therefore, the preponderance of a physical or chemical defense mechanism is possibly not viable.

About defense proteins, molecular weight prevailing suggests inhibitors of the Kunitz type, except Mimosa glutinosa, that requires further investigations. Among legumes, the most studied families of proteinase inhibitors are of the Kunitz (20 kDa) and Bowman-Birk (8-10 kDa) types, which are frequently found in their seeds (Oliva et al., 2010Oliva, M.L.V., Silva, M.C.C., Sallai, R.C., Brito, M.V. and Sampaio, M.U., 2010. A novel subclassification for Kunitz proteinase inhibitors from leguminous seeds. Biochimie, vol. 92, no. 11, pp. 1667-1673. http://dx.doi.org/10.1016/j.biochi.2010.03.021. PMid:20363284.
http://dx.doi.org/10.1016/j.biochi.2010....
; Macedo and Freire, 2011Macedo, M.L.R. and Freire, M.G.M., 2011. Insect digestive enzymes as a target for pest control. Invertebrate Survival Journal: ISJ, vol. 8, pp. 190-198.; Macedo et al., 2011Macedo, M.L.R., Freire, M.G.M., Franco, O.L., Migliolo, L. and Oliveira, C.F.R., 2011. Practical and theoretical characterization of Kunitz inhibitor on the control of Inga laurinaHomalinotus coriaceus.Comparative Biochemistry and Physiology, vol. 158, no. 2, pp. 164-172. http://dx.doi.org/10.1016/j.cbpb.2010.11.005. PMid:21094272.
http://dx.doi.org/10.1016/j.cbpb.2010.11...
). High concentrations of protease inhibitors in the seeds in five legumes: Albizia niopoides, Anadenanthera colubrina, Mimosa glutinosa, Prosopis rubriflora and Poincianella pluviosa are data published for the first time.

Acknowledgements

We thank CNPq for funding the Casadinho/Procad research project; the Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul (FUNDECT - Foundation for the development of Teaching, Science and Technology of the state of Mato Grosso do Sul) for the fellowship granted to the first author; and the Programa de Pós-graduação em Biologia Vegetal da Universidade Federal de Mato Grosso do Sul (Post-graduation Program in Vegetal Biology of the Federal University of Mato Grosso do Sul) for its logistical support.

References

  • Agrawal, A.A. and Fishbein, M., 2006. Plant defense syndromes. Ecology, vol. 87, no. 7, (suppl.), pp. S132-S149. http://dx.doi.org/10.1890/0012-9658(2006)87[132:PDS]2.0.CO;2 PMid:16922309.
    » http://dx.doi.org/10.1890/0012-9658(2006)87[132:PDS]2.0.CO;2
  • Agrawal, A.A., 2006. Macroevolution of plant defense strategies. Trends in Ecology & Evolution, vol. 22, no. 2, pp. 103-109. http://dx.doi.org/10.1016/j.tree.2006.10.012 PMid:17097760.
    » http://dx.doi.org/10.1016/j.tree.2006.10.012
  • Almeida-Cortez, J.S., Shipley, B. and Arnason, J.T., 2004. Growth and chemical defense in relation to resource availability: tradeoffs or common responses to environmental stress? Brazilian Journal of Biology = Revista Brasileira de Biologia, vol. 64, no. 2, pp. 187-194. http://dx.doi.org/10.1590/S1519-69842004000200002 PMid:15462290.
    » http://dx.doi.org/10.1590/S1519-69842004000200002
  • ALVES, F.M. and SARTORI, A.L.B., 2009. Caesalpinioideae (Leguminosae) de um remanescente de Chaco em Porto Murtinho, Mato Grosso do Sul, Brasil. Rodriguésia, vol. 60, pp. 531-550.
  • Bradford, M.M., 1976. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of dye binding. Analytical Biochemistry, vol. 72, no. 1-2, pp. 248-254. http://dx.doi.org/10.1016/0003-2697(76)90527-3 PMid:942051.
    » http://dx.doi.org/10.1016/0003-2697(76)90527-3
  • Carmona, D. and Fornoni, J., 2013. Herbivores can select for mixed defensive strategies in plants. The New Phytologist, vol. 197, no. 2, pp. 576-585. http://dx.doi.org/10.1111/nph.12023 PMid:23171270.
    » http://dx.doi.org/10.1111/nph.12023
  • Chen, M.-S., 2008. Inducible direct plant defense against insect herbivores: a review. Insect Science, vol. 15, no. 2, pp. 101-114. http://dx.doi.org/10.1111/j.1744-7917.2008.00190.x
    » http://dx.doi.org/10.1111/j.1744-7917.2008.00190.x
  • Dickison, W.C., 2000. Integrative plant anatomy. Burlington: Academic Press. 533 p.
  • Dunaevsky, Y.A.E., Elpidina, E.N., Vinokurov, K.S. and Belozersky, M.A., 2005. Protease inhibitors in improvement of plant resistance to pathogens and insects. Molecular Biology, vol. 39, no. 4, pp. 608-613. http://dx.doi.org/10.1007/s11008-005-0076-y
    » http://dx.doi.org/10.1007/s11008-005-0076-y
  • Erlanger, B.F., Kokowsky, N. and Cohen, N., 1961. Preparation and properties of two new chromogenic substrates of trypsin. Archives of Biochemistry and Biophysics, vol. 95, no. 2, pp. 271-278. http://dx.doi.org/10.1016/0003-9861(61)90145-X PMid:13890599.
    » http://dx.doi.org/10.1016/0003-9861(61)90145-X
  • Hanley, M.E., Lamont, B.B., Fairbanks, M.H. and Rafferty, C.M., 2007. Plant structural traits and their role in anti-herbivore defence. Perspectives in Plant Ecology, Evolution and Systematics, vol. 8, no. 4, pp. 157-178. http://dx.doi.org/10.1016/j.ppees.2007.01.001
    » http://dx.doi.org/10.1016/j.ppees.2007.01.001
  • Kortt, A.A. and Jermyn, M.A., 1981. Acacia proteinase inhibitors: purification and properties of the trypsin inhibitors from . Acacia elata seedEuropean Journal of Biochemistry, vol. 115, no. 3, pp. 551-557. http://dx.doi.org/10.1111/j.1432-1033.1981.tb06238.x PMid:7238519.
    » http://dx.doi.org/10.1111/j.1432-1033.1981.tb06238.x
  • Laemmli, V.K., 1970. Cleavage of structural proteins during the assembly of the bacteriophage T4. Nature, vol. 227, no. 5259, pp. 680-685. http://dx.doi.org/10.1038/227680a0 PMid:5432063.
    » http://dx.doi.org/10.1038/227680a0
  • Levin, D.A., 1976. The chemical defenses of plants to pathogens and herbivores. Annual Review of Ecology and Systematics, vol. 7, no. 1, pp. 121-159. http://dx.doi.org/10.1146/annurev.es.07.110176.001005
    » http://dx.doi.org/10.1146/annurev.es.07.110176.001005
  • Lev-Yadun, S., 2001. Aposematic (warning) coloration associated with thorns in higher plants. Journal of Theoretical Biology, vol. 210, no. 3, pp. 385-388. http://dx.doi.org/10.1006/jtbi.2001.2315 PMid:11397139.
    » http://dx.doi.org/10.1006/jtbi.2001.2315
  • Lewis, G.P., Schrire, B., Mackinder, B. and Lock, M., 2005. Legumes of the world. Kew: Royal Botanic Gardens. 577 p.
  • Liu, J., Xia, K.F., Zhu, J.C., Deng, Y.G., Huang, X.L., Hu, B.L., Xu, X. and Xu, Z.F., 2006. The nightshade proteinase inhibitor IIb gene is constitutively expressed in glandular trichomes. Plant & Cell Physiology, vol. 47, no. 9, pp. 1274-1284. http://dx.doi.org/10.1093/pcp/pcj097 PMid:16926166.
    » http://dx.doi.org/10.1093/pcp/pcj097
  • Luo, M., Wang, Z., Li, H., Xia, K.F., Cai, Y. and Xu, Z.F., 2009. Overexpression of a weed (Solanum americanum) proteinase inhibitor in transgenic tobacco results in increased glandular trichome density and enhanced resistance to Helicoverpa armigera and Spodoptera litura.International Journal of Molecular Sciences, vol. 10, no. 4, pp. 1896-1910. http://dx.doi.org/10.3390/ijms10041896 PMid:19468345.
    » http://dx.doi.org/10.3390/ijms10041896
  • Macedo, M.L.R. and Freire, M.G.M., 2011. Insect digestive enzymes as a target for pest control. Invertebrate Survival Journal: ISJ, vol. 8, pp. 190-198.
  • Macedo, M.L.R., Castro, M.N. and Freire, M.G.M., 2004. Mechanisms of the insecticidal action of TEL (Talisia esculenta Lectin) against Callosobruchus maculatus (Coleoptera: Bruchidae). Archives of Insect Biochemistry and Physiology, vol. 56, no. 2, pp. 84-96. http://dx.doi.org/10.1002/arch.10145 PMid:15146543.
    » http://dx.doi.org/10.1002/arch.10145
  • Macedo, M.L.R., Freire, M.G.M., Franco, O.L., Migliolo, L. and Oliveira, C.F.R., 2011. Practical and theoretical characterization of Kunitz inhibitor on the control of Inga laurinaHomalinotus coriaceus.Comparative Biochemistry and Physiology, vol. 158, no. 2, pp. 164-172. http://dx.doi.org/10.1016/j.cbpb.2010.11.005 PMid:21094272.
    » http://dx.doi.org/10.1016/j.cbpb.2010.11.005
  • Macedo, M.L.R., Mello, G.C., Freire, M.G.M., Novello, J.C., Marangoni, S. and Matos, D.G.G., 2002. Effect of a trypsin inhibitor from seeds on the development of Dimorphandra mollisCallosobruchus maculatus.Plant Physiology and Biochemistry, vol. 40, no. 10, pp. 891-898. http://dx.doi.org/10.1016/S0981-9428(02)01441-9
    » http://dx.doi.org/10.1016/S0981-9428(02)01441-9
  • Moreira, X., Abdala-Roberts, L., Rasmann, S., Castagneyrol, B. and Mooney, K.A., 2016. Plant diversity effects on insect herbivores and their natural enemies: current thinking, recent findings, and future directions. Current Opinion in Insect Science, vol. 14, pp. 1-7. http://dx.doi.org/10.1016/j.cois.2015.10.003 PMid:27436639.
    » http://dx.doi.org/10.1016/j.cois.2015.10.003
  • Murdock, L.L. and Shade, R.E., 2002. Lectins and protease inhibitors as plant defenses against insects. Journal of Agricultural and Food Chemistry, vol. 50, no. 22, pp. 6605-6611. http://dx.doi.org/10.1021/jf020192c PMid:12381159.
    » http://dx.doi.org/10.1021/jf020192c
  • NOGUCHI, D.K., NUNES, G.P. and SARTORI, A.L.B., 2009. Florística e síndromes de dispersão de espécies arbóreas em remanescentes de Chaco de Porto Murtinho, Mato Grosso do Sul, Brasil. Rodriguésia, vol. 60, pp. 353-365.
  • Oliva, M.L.V., Silva, M.C.C., Sallai, R.C., Brito, M.V. and Sampaio, M.U., 2010. A novel subclassification for Kunitz proteinase inhibitors from leguminous seeds. Biochimie, vol. 92, no. 11, pp. 1667-1673. http://dx.doi.org/10.1016/j.biochi.2010.03.021 PMid:20363284.
    » http://dx.doi.org/10.1016/j.biochi.2010.03.021
  • Peumans, W.J. and Van Damme, E.J.M., 1995. Lectins as plant defense proteins. Plant Physiology, vol. 109, no. 2, pp. 347-352. http://dx.doi.org/10.1104/pp.109.2.347 PMid:7480335.
    » http://dx.doi.org/10.1104/pp.109.2.347
  • Pieterse, C.M.J., Van Der Does, D., Zamioudis, C., Leon-Reyes, A. and Van Wees, S.C.M., 2012. Hormonal modulation of plant immunity. Annual Review of Cell and Developmental Biology, vol. 28, no. 1, pp. 489-521. http://dx.doi.org/10.1146/annurev-cellbio-092910-154055 PMid:22559264.
    » http://dx.doi.org/10.1146/annurev-cellbio-092910-154055
  • Ramos, V.S., Silva, G.S., Freire, M.G.M., Machado, O.L.T., Parra, J.R.P. and Macedo, M.L.R., 2008. Purification and characterization of a trypsin inhibitor from . Plathymenia foliolosa seedsJournal of Agricultural and Food Chemistry, vol. 56, no. 23, pp. 11348-11355. http://dx.doi.org/10.1021/jf802778b PMid:18991455.
    » http://dx.doi.org/10.1021/jf802778b
  • Ronel, M. and Lev-Yadun, S., 2012. The spiny, thorny and prickly plants in the flora of Israel. Botanical Journal of the Linnean Society, vol. 168, no. 3, pp. 344-352. http://dx.doi.org/10.1111/j.1095-8339.2011.01211.x
    » http://dx.doi.org/10.1111/j.1095-8339.2011.01211.x
  • Sumikawa, J.T., Brito, M.V., Macedo, M.L.R., Uchoa, A.F., Miranda, A., Araujo, A.P.U., Silva-Lucca, R.A., Sampaio, U.M. and Oliva, M.L.V., 2010. The defensive functions of plant inhibitors are not restricted to insect enzyme inhibition. Phytochemistry, vol. 71, no. 2-3, pp. 214-220. http://dx.doi.org/10.1016/j.phytochem.2009.10.009 PMid:19939420.
    » http://dx.doi.org/10.1016/j.phytochem.2009.10.009
  • Valverde, P.L., Fornoni, J. and Núñez-Farfán, J., 2001. Defensive role of leaf trichomes in resistance to herbivorous insects in Datura stramonium.Journal of Evolutionary Biology, vol. 14, no. 3, pp. 424-432. http://dx.doi.org/10.1046/j.1420-9101.2001.00295.x
    » http://dx.doi.org/10.1046/j.1420-9101.2001.00295.x
  • Vandenborre, G., Smagghe, G. and Van Damme, E.J.M., 2011. Plant lectins as defense proteins against phytophagous insects. Phytochemistry, vol. 72, no. 13, pp. 1538-1350. http://dx.doi.org/10.1016/j.phytochem.2011.02.024 PMid:21429537.
    » http://dx.doi.org/10.1016/j.phytochem.2011.02.024
  • Wagner, G.J., 1991. Secreting glandular trichomes: more than just hairs. Plant Physiology, vol. 96, no. 3, pp. 675-679. http://dx.doi.org/10.1104/pp.96.3.675 PMid:16668241.
    » http://dx.doi.org/10.1104/pp.96.3.675

Publication Dates

  • Publication in this collection
    05 Sept 2016
  • Date of issue
    Apr-Jun 2017

History

  • Received
    18 Aug 2015
  • Accepted
    08 Jan 2016
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