Can Aboveground Pest Pressure Disrupt Nitrogen Fixation in Alfalfa?

Morgan N. Thompson & William O. Lamp
University of Maryland, Department of Entomology

Nitrogen is a critical nutrient for forage crop growth and quality. Typically, farmers need to apply additional nitrogen fertilizers to meet the nitrogen demand of crops. Nitrogen-fixing crops, however, do not require nitrogen fertilizer inputs, providing their own nitrogen supply through symbiotic interactions with soil microbes (rhizobia). Rhizobia induce the formation of root nodules in nitrogen-fixing crops, predominantly legumes, and extract inert nitrogen gas from the atmosphere to produce ammonium. In exchange for ammonium, legumes provide the rhizobia carbohydrates to fuel the microbe’s metabolism. Alfalfa is a leguminous forage crop that relies on symbiotic interactions with rhizobia to obtain nitrogen. As a perennial crop, alfalfa stands can last from 3-7 years and typically require no nitrogen fertilizer inputs, making alfalfa a sustainable and high-quality option for forage growers.

Pest pressure can decrease the economic viability of an alfalfa harvest. One particularly devastating pest of alfalfa in Maryland is the potato leafhopper (Empoasca fabae). Potato leafhoppers migrate northward from the southern United States every spring, making the timing of management in the northeast very difficult. Additionally, potato leafhoppers can utilize many alternative host plants, some of which are also of agroeconomic value, such as soybeans and several other fruit and vegetable crops, and leafhoppers can reproduce multiple times during the growing season. To protect alfalfa from potato leafhopper damage (termed ‘hopperburn’), insecticides are often the only option for growers. As a perennial crop, serious pest pressure in one growing season could impact nitrogen fixation in subsequent growing seasons, further accelerating economic losses for growers.

Figure 1. Amount of fixed nitrogen in alfalfa stems and leaves. * represents significant differences between treatments. No Nitrate = No Nitrogen Fertilizer, Moderate Nitrate = Nitrogen Fertilizer Applied; E. fabae- = No Leafhopper Pressure, E. fabae+ = Leafhopper Pressure.

Therefore, in recent field and greenhouse experiments, we sought to determine the effect of potato leafhopper pest pressure on nitrogen fixation in alfalfa. We predicted pest pressure would negatively impact plant growth and carbohydrate production, resulting in reduced nitrogen fixation by rhizobia and uptake of fixed nitrogen by alfalfa. We also predicted losses in nitrogen content of alfalfa due to pest pressure could be offset by nitrogen fertilizer applications. To test our predictions in a field setting, we planted four combinations of small plots: 1) Fixing Cultivar + Nitrogen Fertilizer, 2) Non-Fixing Cultivar + Nitrogen Fertilizer, 3) Fixing Cultivar No Nitrogen Fertilizer, and 4) Non-Fixing Cultivar No Nitrogen Fertilizer. Fixing and non-fixing alfalfa cultivars were utilized to compare plants reliant on both nitrogen fixation and soil nitrogen with plants completely reliant on soil nitrogen. We split each plot in half, applying cages with leafhoppers to one side and cages without leafhoppers to the other. We analyzed the amount of fixed nitrogen in aboveground plant tissue. Results from the field experiment contradicted our predictions, showing nitrogen fertilizer did not increase aboveground nitrogen content of alfalfa under pest pressure. Nitrogen fertilizer (Moderate Nitrate) also decreased aboveground fixed nitrogen content in plants with and without pest pressure (Fig. 1). Unfertilized plants (No Nitrate), in contrast, showed significantly increased amounts of fixed nitrogen content when under pest pressure (Fig. 1). These results contradicted our predictions and suggest alfalfa interactions with rhizobia play a role in helping plants withstand pest damage.

We also examined leafhopper-alfalfa interactions in a greenhouse setting. Here, we analyzed the response of two different cultivars of alfalfa: leafhopper-susceptible (Pioneer 55V50) and leafhopper-resistant (Pioneer 55H94). Nitrogen fertilizer treatments were applied to both cultivars, as well as cages with or without leafhoppers. Results indicate that additional nitrogen fertilizer did not increase the percent nitrogen of plants under pest pressure, regardless of the cultivar (Table 1).

Overall, we conclude leafhopper pest pressure decreases total nitrogen content of alfalfa across all four cultivars tested in both field and greenhouse settings. Amending soils with additional nitrogen fertilizer did not offset losses to leafhopper pressure and we do not recommend this as a management strategy to growers. In our field experiment, however, we found evidence that leafhopper pressure enhances aboveground fixed nitrogen content of alfalfa grown in soils without additional nitrogen. Rhizobia may play an unexamined role in the response of alfalfa to leafhopper pressure. Broader implications of our results highlight how pest damage may increase nitrogen fixation, which may benefit farmers interested in utilizing nitrogen-fixing cover crops.

Acknowledgements: Many thanks to the Western Maryland Research and Education Center staff and greenhouse staff at the University of Maryland aiding in the execution of these experiments, as well as members of the Lamp Lab. This study was funded by Northeastern Sustainable Agriculture Research and Education (Award Number GNE18-187-32231) and the Hatch Project MD-ENTM-1802.

Table 1. Systemic (shoots, crowns, roots) percent nitrogen content of susceptible and resistant alfalfa cultivars in the greenhouse. No Nitrogen Added = No Nitrogen Fertilizer, Nitrogen Added = Nitrogen Fertilizer Applied; Healthy = No Leafhopper Pressure, Injured = Leafhopper Pressure.

Fixing Soybean’s Need For Nitrogen

Article from the American Society of Agronomy

Soybean is rich in protein, which is great for the humans and animals eating it. But this high protein content comes at a cost.

To make protein, soybean plants need a lot of nitrogen. The plants get some of the nitrogen they need by working with specialized bacteria in the soil. These bacteria live in root nodules. They pull nitrogen from the atmosphere and convert it to a form the plants can use.

soybean roots with nodules
A soybean root with nodules. These nodules house bacteria that “fix,” or extract, nitrogen from the atmosphere for the plant’s use. Photo credit Luiz G. Moretti.

But this process–biological nitrogen fixation–may not provide all the nitrogen soybean crops need. Farmers may have to apply nitrogen fertilizer as well.

A new study, however, shows it’s possible to increase the number of soybean root nodules—and the bacteria that live there–to increase crop yields. This could remove the need to apply additional nitrogen fertilizers.

“That opens the possibility of achieving higher yields of soybean based exclusively on biological fixation,” says Mariangela Hungria, a researcher at Embrapa Soja, Brazil.

Hungria, lead author of the study, and her colleagues coated soybean seeds with the bacteria (the usual method used by growers). They supplied additional bacteria by spraying it on the plants during other stages of growth. Soybean plants that received the additional spray inoculation developed more root nodules. And more nodules led to higher yields.

In fact, adding bacteria to seeds increased yields by 27% and 28%. Spraying bacteria on the soy fields during growth pushed up yields even further.

The increase in root nodules after additional spray inoculation surprised Hungria and her colleagues. Previous research indicated that each nodule makes it more difficult for soybean plants to develop subsequent ones. But in this study, soybean plants were able to form new nodules when researchers provided more bacteria.

“To discover that nodules aren’t regulated as strictly as previously thought is an important finding,” says Hungria. “The limitation happens particularly at the beginning of soybean growth when the first nodules appear.” After that initial stage, more nodule growth is possible.

More biological nitrogen fixation, and less nitrogen through fertilizer, can also increase sustainability. First, it reduces carbon emissions. Nitrogen fertilizers are usually produced using fossil fuels. “For every pound of nitrogen fertilizer manufactured, at least 10 pounds of carbon dioxide may be released,” Hungria states.

The second improvement in sustainability is on the field. Excess nitrogen fertilizers from the field can flow into bodies of water. Too much in an aquatic ecosystem can cause algal blooms. These deplete the water of oxygen and lead to “dead zones” devoid of life. Biological fixation using bacteria, however, means more of the nitrogen is used by the crop.

Less fertilizer use also has an economic impact. Nitrogen fertilizer costs can add up quickly, both for farmers and for countries. Brazil imports about 70% of the nitrogen fertilizers used in the country.

Several farms in Brazil began using the study’s strategy in October 2016 (the summer crop in Brazil). Initial results have been promising, says Hungria. The higher soybean yields seen in the study are sustained on these larger scales.

Hungria thinks these results will extend beyond Brazil as well. “But they have to be verified because the genetic background of soybean is different in each country,” she says. Collaborations with Kansas State University, to verify if the results can be extended to the U.S., have just started.

Inoculating soybeans with rhizobia
Inoculating–adding helpful bacteria to soybean seeds–usually occurs at sowing time. However, in this study, soybean crops at various stages of growth were also inoculated by spraying the plants with bacteria. Photo credit Luiz G. Moretti.

Researching bacteria and nitrogen fixation may just be the beginning. “I think microorganisms can be the ‘stars’ of a new era of agriculture, in which we consider not only food security but also sustainability,” she says.

Read more about Hungria’s research in Agronomy Journal. The research in Brazil was funded by Universidade Estadual Paulista, Fundação Agrisus, Embrapa, and Total Biotecnologia.