1) The ability to produce and study genetic alterations in waterhemp without risking unintentional release into the environment.
2) The ability to safely assess emerging gene drive technology and explore its potential for waterhemp management in North Dakota.

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Research Conducted
Cell suspension cultures, which are living cells suspended in a liquid nutrient solution, are often used in laboratory research. We tested six different solutions for their ability to generate waterhemp protoplasts (cells without cell walls). One solution successfully generated protoplasts, the majority of which were confirmed to be alive and viable. Subsequent cell wall regeneration and cell division, however, was not observed under our tested conditions.

Why the research is important to ND soybean growers
Herbicide-resistant waterhemp represents an important problem in North Dakota soybean production, and new tools are needed to study emerging methods for controlling this weed. When growing herbicide-resistant weeds for research, it is important to minimize the risk of seeds escaping and spreading into fields. This is especially important when researching new genetic methods of weed control, where weed genes may be altered. The cells that make up cell suspension cultures need to be maintained in a laboratory and cannot grow or produce new plants in the field. This makes waterhemp suspension cultures very valuable herbicide resistance research tools. The ability to remove cell walls from these cultured cells, generating protoplasts, adds value to the cultures because of their increased ability to take up genetic material for experimentation compared to intact cells, due to the lack of a cell wall barrier in protoplasts.

Final findings of the research
We found that while most tested solutions did not generate protoplasts from waterhemp cells, a solution containing the enzyme ‘macerozyme R-10’ successfully produced viable waterhemp protoplasts. However, under our tested conditions, waterhemp protoplasts were not observed to subsequently regenerate cell walls or start dividing. This will be an area of future research.

Benefits/Recommendations to North Dakota soybean farmers and industry
Waterhemp plant material in the form of cell suspension cultures are now available for further studies investigating herbicide resistance. Researchers also now have a protocol to generate protoplasts from these cell suspension cultures, facilitating genetic research to investigate new ways for waterhemp control without risk of weed escape into North Dakota fields.