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United States Department of Agriculture

Agricultural Research Service

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Research Project: Fostering Coexistence: Industry-Driven Field and Landscape Research on Pollen Flow in Ge Alfalfa

Location: Forage Seed and Cereal Research

2013 Annual Report


1a.Objectives (from AD-416):
Since 2007, the alfalfa industry has been working to develop coexistence strategies for genetically engineered Round Up Ready alfalfa (RRA). The National Alfalfa and Forage Alliance spearheaded efforts, and more recently, alfalfa seed producers have been working with AOSCA to develop an Alfalfa Seed Stewardship Program (ASSP). With the deregulation of RRA in February 2011, there is an urgent need to complete and implement coexistence strategies to protect the export seed market and other alfalfa markets that are sensitive to the adventitious presence (AP) of transgenic traits. Industry recognizes research is needed at the commercial scale to better understand pollen-mediated RRA transgene flow in the landscape. Extended conversations with alfalfa producers and breeding companies have led to the following objectives of our proposal: (1) to examine how leaf cutter and alkali bees transmit RRA across commercial seed fields and how that will impact proposed harvest strategies that separate seed for non AP- and AP- sensitive markets and to examine the persistence of RRA pollen in honeybee hives; (2) to characterize fitness parameters such as seed production, seed dormancy and viability, longevity in the seed bank, seedling establishment and plant persistence, in feral and feral-RRA hybrid alfalfa to determine how important and to what extent control strategies are required; (3) to track RRA transgene flow from RRA hay and seed production fields planted during the previous deregulation (seed fields were removed in 2007) into feral alfalfa to understand the role feral alfalfa plays as a transgene reservoir and vector for long distance transgene dispersal; (4) to study the transmission of the RR transgene from RRA hay fields to conventional seed fields to refine isolation distances by taking into account landscape variables.


1b.Approach (from AD-416):
This project will focus on the three main alfalfa seed and hay production areas in the Western United States: the Walla Walla valley and Columbia Basin in Washington, the Treasure Valley in Idaho and Oregon, and Fresno County in California. Monsanto has agreed to provide us with the locations of RRA hay and seed fields and will be giving us permission to use their event-specific PCR primers. We will be using greenhouse seedling assays and protein based, commercially available test strips to carry out qualitative tests for RRA presence in leaf, seed and hay sampling. Positive tests will be confirmed using PCR. Landscape genetics is a newly emerging field which provides powerful geostatistical tools to understand how gene flow is influenced by landscape variables. We will be using a landscape genetics approach to analyze transgene movement across the landscape. Since we want to generate information that has immediate relevance to industry’s efforts to establish coexistence strategies, our research will be focused at the commercial field and landscape level.


3.Progress Report:

Funds related to the National Institute of Food and Agriculture grant 60-5348-2-851 “Fostering Coexistence: Industry-Driven Field and Landscape Research on Gene Flow in Genetically Engineered Alfalfa” were distributed in early November 2012. This research relates to Objectives 1 and 2 of the in house project "Improve grass germplasm by developing molecular and biological resources that can be applied to reduce the impact of abiotic stresses on the productivity of grasses used for livestock production, turf and bioenergy", and "Reduce the impact of weeds and stem rust, two biotic stresses that negatively impact seed production and the utilization of grasses for livestock production, turf, and bioenergy". The analysis of samples collected during July, August and September of 2011 and May of 2012 were completed during Jan-April of 2013. A meeting was held in Prosser, Washinton, in February of 2013 to discuss the results of the 2011 studies looking at the presence of the Round Up resistant alfalfa transgene in feral roadside alfalfa and to develop a work plan for 2013. In May of 2013, scientists conducted surveys of alfalfa hay and seed fields in Fresno County, California, Walla Walla County, Washington, and Canyon County, Idaho, to facilitate the identification of seed fields grown for markets sensitive to the presence of the genetically modified trait and conventional seed fields, and their relative location to genetically modified hay/seed fields. Samples were collected and tested for the presence of the Round Up resistant alfalfa transgene using test strips. Positive samples were confirmed by Polymerase Chain Reaction (PCR) using a reliable Deoxyribonucleic acid (DNA) isolation method and protocols/primers developed for this purpose. A new method, using labeled probes was developed to increase the specificity of the real time Polymerase Chain Reaction (PCR) protocol. Maps with the locations of genetically modified and non-genetically modified seed/hay fields were examined to identify source/sink fields for gene flow studies and pollination studies. Additionally, a Deoxyribonucleic acid (DNA)extraction method was developed for alfalfa pollen to facilitate studies related to insect mediated transgene flow studies. Pollinators for insect transmission studies were collected from fields in Fresno County.


Last Modified: 10/23/2014
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