Transgenic soybeans expressing siRNAs specific to a major sperm protein gene suppress Heterodera glycines reproduction
Ryan M. Steeves A , Tim C. Todd B , Juliane S. Essig B and Harold N. Trick B CA Molecular, Cellular, and Developmental Biology Program, Iowa State University, Ames, IA 50011, USA.
B Department of Plant Pathology, Kansas State University, Manhattan, KS 66506, USA.
C Corresponding author. Email: hnt@ksu.edu
Functional Plant Biology 33(11) 991-999 https://doi.org/10.1071/FP06130
Submitted: 24 May 2006 Accepted: 1 August 2006 Published: 1 November 2006
Abstract
The soybean cyst nematode (SCN), Heterodera glycines, is the major disease-causing agent limiting soybean production in the USA. The current management strategy to reduce yield loss by SCN involves the deployment of resistant soybean cultivars and rotation to non-host crops. Although this management scheme has shown some success, continued yearly yield loss estimates demonstrate the limitations of these techniques. As a result, new control strategies are needed to complement the existing methods. Reported here is a novel method of SCN control that utilises RNA interference (RNAi). Transgenic soybeans were generated following transformation with an RNAi expression vector containing inverted repeats of a cDNA clone of the major sperm protein (MSP) gene from H. glycines. The accumulation of MSP-specific short interfering RNA (siRNA) molecules were detected by northern blot analysis of transgenic soybeans. T0 plants displaying MSP siRNA accumulation were deployed in a bioassay to evaluate the effects of MSP interfering molecules on H. glycines reproduction. Bioassay data has shown up to a 68% reduction in eggs g–1 root tissue, demonstrating that MSPi transgenic plants significantly reduced the reproductive potential of H. glycines. An additional bioassay evaluating progeny nematodes for maintenance of reproductive suppression indicated that progeny were also impaired in their ability to successfully reproduce, as demonstrated by a 75% reduction in eggs g–1 root tissue. The results of this study demonstrate the efficacy of an RNAi-based strategy for control of the soybean cyst nematode. In addition, these results may have important implications for the control of other plant parasitic nematodes.
Acknowledgments
The authors thank Dr Marcy Main and Melissa Schapaugh for their technical support with tissue culture and transformation. We thank Tom Oakley for assistance with the SCN bioassays. The ACT2 promoter from Arabidopsis was kindly provided by Dr Richard Meagher, University of Georgia and the SCN J2 library was a kind gift from Dr Eric Davis, North Carolina State University. This research was supported by Kansas State University, the Kansas Soybean Commission and USDA NRICGP Grant # 2004-35607-14970. This article is contribution no. 06-113-J from the Kansas Agricultural Experimental Station, Kansas State University, Manhattan, Kansas.
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