Toxicity toChrysomela tremulae (Coleoptera: Chrysomelidae) of transgenic poplars expressing a cysteine proteinase inhibitor

Abstract

The aim of this study was to test the potential of proteinase inhibitors to control Chrysomela tremulae, a beetle that causes severe damage in young plantations and in short-rotation intensive culture (SRIC) of poplar. As a first step, cysteine proteinases were determined to be the major digestive proteinases of C. tremulae and oryzacystatin OCI, a cysteine proteinase inhibitor, was shown to inhibit this activity in vitro. The gene encoding OCI was introduced into poplar (Populus tremula x P. tremuIoides) and transgenic plants expressing OCI at a high level were selected. Feeding tests on these transgenic plants demonstrate the toxicity of OCI-producing poplar leaves against C. tremulae larvae.

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

“…The use of proteinaceous inhibitors in insect control strategies has good potential, because insect digestive proteinases are promising targets in the control of various insects, including lepidopterans such as Manduca sexta [3], Heliothis zea [4], Spodoptera litura [5], and Lucilia cuprina [6], and also various coleopterans [7][8][9][10][11]. Despite several suggested physiological functions in plants [12][13][14], the inhibitors are known for their role in response to abiotic [15,16] biotic stresses, especially in plant defense processes against insect pest attack [9,[17][18][19][20].…”

Tailoring the Specificity of a Plant Cystatin toward Herbivorous Insect Digestive Cysteine Proteases by Single Mutations at Positively Selected Amino Acid Sites

“…Cystatins expressed in transgenic plant lines were shown to alter the development of herbivorous arthropods and root parasitic nematodes relying on Cys proteases for extracellular protein digestion (e.g. Leplé et al, 1995;Atkinson et al, 2003;Outchkourov et al, 2004). These proteins also show potential for microbial control, as suggested by their deleterious effects against a number of fungal and viral pathogens of agronomic significance (GutierrezCampos et al, 1999;Martinez et al, 2005;Yang and Yeh, 2005;Christova et al, 2006).…”

Deleterious effects of plant cystatins against the banana weevil Cosmopolites sordidus

“…Despite some promising developments (Leplé et al, 1995;Kuroda et al, 1996;Koiwa et al, 2000;Liu et al, 2004;Alvarez-Alfageme et al, 2007;Ninkovic et al, 2007), the general usefulness of plant cystatins for the control of coleopteran pests still remains to be established. These insects have developed over time effective strategies to elude the inhibitory effects of plant protease inhibitors, involving the use of complex digestive protease systems with proteases from different mechanistic classes acting in a complementary, coordinated manner (Brunelle et al, 1999;Hernandez et al, 2003;Gruden et al, 2003;Vinokurov et al, 2006a,b;Prabhakar et al, 2007); the over-expression of target proteases following cystatin ingestion to outnumber the inhibitory proteins (Cloutier et al, 2000;Ahn et al, 2004); the constitutive or diet-induced expression of cysteine cathepsins weakly sensitive to the ingested cystatin, the so-called 'cystatin-insensitive proteases' (Michaud et al, 1993(Michaud et al, , 1995aGirard et al, 1998a;Cloutier et al, 1999Cloutier et al, , 2000ZhuSalzman et al, 2003;Brunelle et al, 2004;Gruden et al, 2004;Liu et al, 2004;Koo et al, 2008); the over-expression of proteases from alternative mechanistic classes following cystatin ingestion (Zhu-Salzman et al, 2003;Brunelle et al, 2004;Rivard et al, 2004;Oppert et al, 2005); and the degradation of defensive protease inhibitors using non-target, insensitive proteases (Michaud et al, 1995b;Girard et al, 1998b;Giri et al, 1998;Gruden et al, 2003;Zhu-Salzman et al, 2003).…”

“…OsCYS1 was also expressed as a histidine (His)-tagged protein using the 6x Histidine Tagging System™ (Qiagen, Mississauga ON, Canada), with the vector pAOCI-3 (Leplé et al, 1995) as a DNA template. An OsCYS1-encoding DNA fragment excised from pAOCI-3 by EcoRI/PstI treatment was first cloned between the EcoRI and PstI cloning sites of pBlueScript (Stratagene, La Jolla CA, USA), re-excised as a BamHI/KpnI fragment, and finally introduced in the expression vector pQE31 (Qiagen), in frame with the poly-His tag.…”

“…In this study, we assessed the potential of OC-I and other plant cystatins as possible tools for the production of transgenic banana lines resistant to the banana weevil. Several papers have been published assessing the effectiveness of plant cystatins against herbivorous Coleoptera, with conclusive results in some cases (Leplé et al, 1995;Kuroda et al, 1996;Lecardonnel et al, 1999;Koiwa et al, 2000;Alvarez-Alfageme et al 2007;Ninkovic et al, 2007) but disappointing results in other cases owing to the ability of the target insects to readily elude or even counteract the antidigestive effects of natural or recombinant inhibitors (e.g. Girard et al, 1998a,b;Cloutier et al, 1999Cloutier et al, , 2000Zhu-Salzman et al, 2003;Gruden et al, 2003Gruden et al, , 2004Ahn et al, 2004Ahn et al, , 2007Oppert et al, 2005;Koo et al, 2008).…”