Abstract
BackgroundCry8-like from Bacillus thuringiensis (Bt) encodes an insecticidal crystal (Cry) protein. Holotrichia parallela (Coleoptera: Scarabaeoidae), commonly known as the dark black chafer, is a troublesome pest of soybean (Glycine max). To test whether cry8-like can confer resistance against H. parallela to soybean, we introduced cry8-like from the Bt strain HBF-18 into soybean cultivar Jinong 28.ResultsQuantitative reverse transcription-PCR analysis demonstrated that cry8-like was expressed most highly in soybean leaves. In addition, Southern blot assays revealed that one copy of the integrated fragment was present in the transformed plants. Eight independent cry8-like transgenic lines were subsequently fed on by H. parallela. Under H. parallela feeding stress, the survival rates of the non-transgenic plants were 92% lower than those of the transgenic plants. The mortality rate of H. parallela increased when the larvae fed on the roots of T1 transgenic soybean plants. Moreover, the surviving larvae were deformed, and their growth was inhibited.ConclusionsCollectively, our data suggest that transgenic soybean plants expressing the cry8-like gene are more resistant to H. parallela than non-transgenic plants and that transgenic expression of the cry8-like gene may represent a promising strategy for engineering pest tolerance. The events generated in this study could thus be utilized in soybean breeding programs.
Highlights
Cry8-like from Bacillus thuringiensis (Bt) encodes an insecticidal crystal (Cry) protein
Production of transgenic soybean plants expressing the cry8-like gene The recombinant plasmid designated pCAMBIA3300cry8-like was introduced into Agrobacterium tumefaciens strain (Fig. 2a)
Compared with cry8Ga, Cry8-like exhibits 3-fold greater toxicity [23], confirming that cry8like would be the best choice for expression in transgenic soybeans. These findings suggest that the insertion of an exogenous cry8-like gene into soybean is a feasible approach to produce insect resistance
Summary
Cry8-like from Bacillus thuringiensis (Bt) encodes an insecticidal crystal (Cry) protein. Holotrichia parallela (Coleoptera: Scarabaeoidae), commonly known as the dark black chafer, is a troublesome pest of soybean (Glycine max). Merr.) is a globally important crop species that is grown for its oil and protein; its seeds are used for human consumption, animal feed, and industrial raw materials [1]. Soybean is self-pollinated, and soybean seeds are rich in protein (38–40%), in addition, soybean crops are utilized in crop rotations because of the ability of the plants to increase the amount of nitrate in the soil via symbiotic nitrogen fixation. Holotrichia parallela (Coleoptera: Scarabaeoidea), commonly known as the dark black chafer, is one of the most important pests in agriculture and forestry in China [3]. H. parallela has a three-stage life cycle: the egg, larva and adult (Fig. 1).
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