Limits...
Potential use of a serpin from Arabidopsis for pest control.

Alvarez-Alfageme F, Maharramov J, Carrillo L, Vandenabeele S, Vercammen D, Van Breusegem F, Smagghe G - PLoS ONE (2011)

Bottom Line: Subsequently, the cotton leafworm Spodoptera littoralis Boisduval and the pea aphid Acyrthosiphon pisum (Harris) were fed on artificial diets containing AtSerpin1, and S. littoralis was also fed on transgenic Arabidopsis plants overproducing AtSerpin1.AtSerpin1 supplied in the artificial diet or by transgenic plants reduced the growth of S. littoralis larvae by 65% and 38%, respectively, relative to controls.Nymphs of A. pisum exposed to diets containing AtSerpin1 suffered high mortality levels (LC(50) = 637 µg ml(-1)).

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Gent, Belgium. fernando.alvarez@art.admin.ch

ABSTRACT
Although genetically modified (GM) plants expressing toxins from Bacillus thuringiensis (Bt) protect agricultural crops against lepidopteran and coleopteran pests, field-evolved resistance to Bt toxins has been reported for populations of several lepidopteran species. Moreover, some important agricultural pests, like phloem-feeding insects, are not susceptible to Bt crops. Complementary pest control strategies are therefore necessary to assure that the benefits provided by those insect-resistant transgenic plants are not compromised and to target those pests that are not susceptible. Experimental GM plants producing plant protease inhibitors have been shown to confer resistance against a wide range of agricultural pests. In this study we assessed the potential of AtSerpin1, a serpin from Arabidopsis thaliana (L). Heynh., for pest control. In vitro assays were conducted with a wide range of pests that rely mainly on either serine or cysteine proteases for digestion and also with three non-target organisms occurring in agricultural crops. AtSerpin1 inhibited proteases from all pest and non-target species assayed. Subsequently, the cotton leafworm Spodoptera littoralis Boisduval and the pea aphid Acyrthosiphon pisum (Harris) were fed on artificial diets containing AtSerpin1, and S. littoralis was also fed on transgenic Arabidopsis plants overproducing AtSerpin1. AtSerpin1 supplied in the artificial diet or by transgenic plants reduced the growth of S. littoralis larvae by 65% and 38%, respectively, relative to controls. Nymphs of A. pisum exposed to diets containing AtSerpin1 suffered high mortality levels (LC(50) = 637 µg ml(-1)). The results indicate that AtSerpin1 is a good candidate for exploitation in pest control.

Show MeSH

Related in: MedlinePlus

Western blot immunoassay showing the expression of AtSerpin1 in leaves of the transgenic Arabidopsis lines AtSerpin1OE1, AtSerpin1OE2, and AtSerpin1OE3, and the non-transformed line Col-0.Lanes: (1) Page ruler plus protein standard; (2) 100 ng AtSerpin1; (3) 50 ng AtSerpin1; (4) 25 ng AtSerpin1; (5) 12.5 ng AtSerpin1; (6) 5 ng AtSerpin1; (7) 0 ng AtSerpin1; (8) overproducing line AtSerpinOE3 (6 ng); (9) overproducing line AtSerpinOE2 (6 ng); (10) overproducing line AtSerpinOE1 (6 ng); (11) non-transformed line Col-0. In lanes 7–9, the upper band is the full-length and active form of AtSerpin1, while the lower band is the cleaved form after interaction with a protease.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3104999&req=5

pone-0020278-g003: Western blot immunoassay showing the expression of AtSerpin1 in leaves of the transgenic Arabidopsis lines AtSerpin1OE1, AtSerpin1OE2, and AtSerpin1OE3, and the non-transformed line Col-0.Lanes: (1) Page ruler plus protein standard; (2) 100 ng AtSerpin1; (3) 50 ng AtSerpin1; (4) 25 ng AtSerpin1; (5) 12.5 ng AtSerpin1; (6) 5 ng AtSerpin1; (7) 0 ng AtSerpin1; (8) overproducing line AtSerpinOE3 (6 ng); (9) overproducing line AtSerpinOE2 (6 ng); (10) overproducing line AtSerpinOE1 (6 ng); (11) non-transformed line Col-0. In lanes 7–9, the upper band is the full-length and active form of AtSerpin1, while the lower band is the cleaved form after interaction with a protease.

Mentions: Three transgenic Arabidopsis lines overproducing AtSerpin1 were tested against S. littoralis Expression of AtSerpin1 in leaves of Arabidopsis was confirmed by Western blot using increasing concentrations of purified AtSerpin1 (Figure 3). Differences in the AtSerpin1 expression among the transgenic lines was observed, being higher in AtSerpinOE2 and AtSerpin1OE3. In those lines, about 5 ng AtSerpin ug−1 of total protein content was measured.


Potential use of a serpin from Arabidopsis for pest control.

Alvarez-Alfageme F, Maharramov J, Carrillo L, Vandenabeele S, Vercammen D, Van Breusegem F, Smagghe G - PLoS ONE (2011)

Western blot immunoassay showing the expression of AtSerpin1 in leaves of the transgenic Arabidopsis lines AtSerpin1OE1, AtSerpin1OE2, and AtSerpin1OE3, and the non-transformed line Col-0.Lanes: (1) Page ruler plus protein standard; (2) 100 ng AtSerpin1; (3) 50 ng AtSerpin1; (4) 25 ng AtSerpin1; (5) 12.5 ng AtSerpin1; (6) 5 ng AtSerpin1; (7) 0 ng AtSerpin1; (8) overproducing line AtSerpinOE3 (6 ng); (9) overproducing line AtSerpinOE2 (6 ng); (10) overproducing line AtSerpinOE1 (6 ng); (11) non-transformed line Col-0. In lanes 7–9, the upper band is the full-length and active form of AtSerpin1, while the lower band is the cleaved form after interaction with a protease.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3104999&req=5

pone-0020278-g003: Western blot immunoassay showing the expression of AtSerpin1 in leaves of the transgenic Arabidopsis lines AtSerpin1OE1, AtSerpin1OE2, and AtSerpin1OE3, and the non-transformed line Col-0.Lanes: (1) Page ruler plus protein standard; (2) 100 ng AtSerpin1; (3) 50 ng AtSerpin1; (4) 25 ng AtSerpin1; (5) 12.5 ng AtSerpin1; (6) 5 ng AtSerpin1; (7) 0 ng AtSerpin1; (8) overproducing line AtSerpinOE3 (6 ng); (9) overproducing line AtSerpinOE2 (6 ng); (10) overproducing line AtSerpinOE1 (6 ng); (11) non-transformed line Col-0. In lanes 7–9, the upper band is the full-length and active form of AtSerpin1, while the lower band is the cleaved form after interaction with a protease.
Mentions: Three transgenic Arabidopsis lines overproducing AtSerpin1 were tested against S. littoralis Expression of AtSerpin1 in leaves of Arabidopsis was confirmed by Western blot using increasing concentrations of purified AtSerpin1 (Figure 3). Differences in the AtSerpin1 expression among the transgenic lines was observed, being higher in AtSerpinOE2 and AtSerpin1OE3. In those lines, about 5 ng AtSerpin ug−1 of total protein content was measured.

Bottom Line: Subsequently, the cotton leafworm Spodoptera littoralis Boisduval and the pea aphid Acyrthosiphon pisum (Harris) were fed on artificial diets containing AtSerpin1, and S. littoralis was also fed on transgenic Arabidopsis plants overproducing AtSerpin1.AtSerpin1 supplied in the artificial diet or by transgenic plants reduced the growth of S. littoralis larvae by 65% and 38%, respectively, relative to controls.Nymphs of A. pisum exposed to diets containing AtSerpin1 suffered high mortality levels (LC(50) = 637 µg ml(-1)).

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Gent, Belgium. fernando.alvarez@art.admin.ch

ABSTRACT
Although genetically modified (GM) plants expressing toxins from Bacillus thuringiensis (Bt) protect agricultural crops against lepidopteran and coleopteran pests, field-evolved resistance to Bt toxins has been reported for populations of several lepidopteran species. Moreover, some important agricultural pests, like phloem-feeding insects, are not susceptible to Bt crops. Complementary pest control strategies are therefore necessary to assure that the benefits provided by those insect-resistant transgenic plants are not compromised and to target those pests that are not susceptible. Experimental GM plants producing plant protease inhibitors have been shown to confer resistance against a wide range of agricultural pests. In this study we assessed the potential of AtSerpin1, a serpin from Arabidopsis thaliana (L). Heynh., for pest control. In vitro assays were conducted with a wide range of pests that rely mainly on either serine or cysteine proteases for digestion and also with three non-target organisms occurring in agricultural crops. AtSerpin1 inhibited proteases from all pest and non-target species assayed. Subsequently, the cotton leafworm Spodoptera littoralis Boisduval and the pea aphid Acyrthosiphon pisum (Harris) were fed on artificial diets containing AtSerpin1, and S. littoralis was also fed on transgenic Arabidopsis plants overproducing AtSerpin1. AtSerpin1 supplied in the artificial diet or by transgenic plants reduced the growth of S. littoralis larvae by 65% and 38%, respectively, relative to controls. Nymphs of A. pisum exposed to diets containing AtSerpin1 suffered high mortality levels (LC(50) = 637 µg ml(-1)). The results indicate that AtSerpin1 is a good candidate for exploitation in pest control.

Show MeSH
Related in: MedlinePlus