Project Details:

Title:
Engineered resistance to soybean cyst nematode via induced gene silencing (RNAi)

Parent Project: Engineered resistance to soybean cyst nematode via induced gene silencing (RNAi)
Checkoff Organization:North Central Soybean Research Program
Categories:Nematodes, Breeding & genetics
Organization Project Code:
Project Year:2013
Lead Principal Investigator:Harold Trick (Kansas State University)
Co-Principal Investigators:
Jiarui Li (Kansas State University)
Tim C. Todd (Kansas State University)
John Finer (The Ohio State University)
Lila Vodkin (University of Illinois at Urbana-Champaign)
Jack Widholm (University of Illinois-Carbondale)
Show more
Keywords: Soybean Bioengineering, Soybean Cyst Nematode - Genetic Resistance

Contributing Organizations

Funding Institutions

Information and Results

Click a section heading to display its contents.

Project Summary

The primary goal of this research project is to establish a new set of biotech traits that have durable resistance to soybean cyst nematode (SCN). Turning off genes by a process known as RNA interference (RNAi) has tremendous potential as a new strategy to increase nematode resistance. Past research with other nematode species has demonstrated the scientific merit of the technique. This project will investigate the opportunities to insert target gene sequences in the SCN that will provide durable genetic materials that will be lethal to SCN populations.

Turning off genes by a process known as RNA interference (RNAi) has tremendous potential as a new strategy to increase nematode resistance. Past research with other nematode species has demonstrated the scientific merit of the technique. This project will investigate opportunities to silence specific SCN gene sequences using RNAi in transgenic soybean providing a durable genetic material that will be lethal to SCN populations.

Project Objectives

1. Engineer stable transgenic soybean plants with traits that can silence specific nematode genes.
2. Evaluate transgenic lines with SCN bioassays to confirm the effectiveness of the level of SCN resistance.
3. Examine the durability of the transgenic traits on single and diverse populations of SCN.

Project Deliverables

Progress of Work

Final Project Results

Six different nematode genes were identified as potential targets for gene silencing. Selection was based on data mined from Caenorhabditis elegans (a free-living nematode) RNAi database and preliminary hairy root assays performed in our laboratory. Each of six genes are required for either the overall fitness of the nematode or required for the proper development of their offspring. Silencing vectors were created last funding cycle and were distributed to the Co-PIs at the four institutions.

The soybean transformations with the six RNAi vectors were initiated and are still underway. Several independent transformation experiments have been performed at each location and are in various stages of selection and regeneration. Some of the transformation experiments were recently initiated and are currently under selection. Other experiments that were initiated earlier in the year have yielded putative events testing positive for the transgene and have been regenerated into plants. To date we have recovered putative transgenic events from all six vectors and identified at least three PCR positive events from each vector. Although plant regeneration is still occurring for these events, we have recovered plants from four of our vector constructions. These plants, a total of nineteen separate events, are being grown to maturity in greenhouses. Once these plants have matured the seeds will be shipped to Kansas State University for evaluations in SCN bioassays.

In addition to the production of transgenic events we have also collected three different populations of nematodes in Kansas and used PCR to amplify the genes used in this study. Specifically, we have sequenced gene regions selected from our vector constructions. Three of the genes have been sequenced for these three populations and we have observed that these genes have high homology (98-100%) in the regions we have searched. This observation is critical as sequence homology among the different populations provides additional data that the constructs have an equal chance to be effective on all of these nematode populations.

Benefit to Soybean Farmers

Performance Metrics

Project Years