Effect of sainfoin () variety and harvest maturity on quality, yield, and condensed content

PI and Co-PIs Megan Van Emon, PhD Principal Investigator Extension Beef Specialist at Montana State University 406.874.8286/ [email protected] Kylie Gardhouse Graduate student at Montana State University 818.584.5995/ [email protected] Whit Stewart, PhD Extension Specialist at University of Wyoming 307.766.5374/ [email protected] Harley Naumann, PhD Assistant Professor at University of Missouri 573.882.9896/ [email protected] Patrick Carr, PhD Superintendent and Associate Professor at the MSU Central Ag Research Center 406.423.5421/ [email protected] Jennifer MacAdam, PhD Physiologist at Utah State University Utah State University 435.797.2364/ [email protected]

Abstract Sainfoin is a high quality, non-bloating . The objectives of this study were to determine impact of maturity at harvest and variety on forage quality, yield, condensed tannin content and to determine the economic impacts of production on different varieties.

Introduction Research on sainfoin (Onobrychis viciifolia) production is lacking, often leading to use of recommendations for management. This has potential to create unexpected losses due to unrealized differences between the two forage . Current information regarding best harvest management practices of sainfoin and their impacts on yield, quality, and regrowth is required for growers to optimize production. Sainfoin is a high-quality perennial forage, comparable to other perennial like alfalfa in nutrient quality. Sainfoin is commonly used in place of other perennial forages due to its attractiveness to pollinators, as well as its palatability, non-bloating factors, and its ability to grow in various conditions (Carbonero et al., 2011). One of the biggest attributes of sainfoin is the presence of condensed (CT), making sainfoin a non-bloating forage. In a study conducted by McMahon et al. (1999), a mix of 10% sainfoin and 90% chopped alfalfa decreased the cases of bloat by 93% compared to feeding 100% alfalfa, providing producers options to graze or the forage. Additionally, sainfoin does not have the potential for autotoxicity. This is a major limiting-factor for alfalfa, as stands cannot be interseeded once they begin to decline. However, provided no disease is present, sainfoin can be allowed to reseed itself and improve production, improving producer profitability by lowering and production costs. Another potential benefit of sainfoin is its anthelmintic abilities. Studies have shown that livestock consuming forages with CT have decreased amounts of intestinal parasitic eggs. Arroyo-Lopez (2014), found that in lambs fed sainfoin, total intestinal egg count decreased by 46.9% compared to untreated lambs. Feeding forages containing naturally high levels of CT decreases potential for parasite shedding, leading to decreased need for synthetic anthelmintic use, and potentially even decrease parasite resistance, helping to produce a more sustainable operation. However, it is not yet known if different sainfoin varieties contain significantly different CT levels. Finally, sainfoin is a high-quality forage similar to alfalfa, but different in that sainfoin matures more rapidly. On dryland, the maximum dry matter yield for sainfoin is at 70% bloom, while alfalfa is at 5% bloom, according to Carlton et al. (1968). In this study, sainfoin was higher in nitrogen-free extracts, total digestible nutrients, and phosphorous. The N-free extracts were higher for sainfoin than alfalfa, meaning there is enough energy available in sainfoin for animal production to be similar to alfalfa. Alfalfa was found to be higher in crude protein at all stages of maturity, however the amount of protein in the sainfoin was sufficient to meet beef cattle’s nutritional needs (Carlton et al., 1968). Overall, there is a lot of potential for sainfoin in the Intermountain West and Midwest. The knowledge gained from this study may have the ability to improve ranchers’ productivity, leading to larger profits, and enhance quality of life, due to increased soil health benefits, as well as improved animal performance.

Materials and Methods Three research sites are being used for this study: Moccasin, Montana (CARC; 47.05’, - 109.95; Danvers-Judith clay loam); Bozeman, Montana (BZN; 45.66’, -111.07’; Turner loam and Meadowcreek loam); and Logan, UT (UT; 41.74’, -111.91; Ricks Gravelly loam). In 2017, plots were established in a replicated split-plot design. Each plot measures 9 meters x 4.5 meters, with a 3-meter alley between replications. The main plot includes four different varieties of sainfoin, which were planted in spring 2018. The sainfoin varieties established are ‘AAC Mountainview’ (AAC), ‘Eski’ (ESK), ‘Shoshone’ (SHO), and ‘Delaney’ (DEL). Shaw (SHA) is the alfalfa check variety used in this study at all three locations. Sub-plot treatment includes harvest maturity, with each variety harvested at 10%, 50%, and 100% bloom. Treatments are defined as A (10%), B (50%) and C (100%). At the Moccasin location, we had a second cutting at 10% bloom, which is represented as treatment D. All plots were replicated 4 times. Prior to seeding in 2018, soil samples were taken, and pre-plant fertilizer and herbicide was applied. Plots were seeded at a rate of 33.6 kg PLS/ ha for sainfoin, and 13.5 kg PLS/ ha for alfalfa. In spring of 2019, soil samples were collected and appropriate fertilizer applied. In the summer of 2019, yield and quality samples were taken. When each third of the plot reaches appropriate maturity, two one square meter areas were hand clipped to 5 cm in each subplot to evaluate herbage mass production. Clippings were immediately weighed to obtain wet weights. Subsamples will be collected and weighed to obtain subsample wet weight, then subsamples will be dried at 60C and re-weighed to obtain a dry weight. Subsample wet weights and dry weights were used to estimate dry matter percentage. Plot production was then be estimated by multiplying the sample wet weight by the dry matter percentage and used to estimate production on a kg/ha basis. Subsamples were ground at the MSU Nutrition Center through a 2mm screen using a Wiley Mill (Swedesboro, NJ). These samples were then analyzed for crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), and condensed tannin (CT) content. Statistical analysis was completed utilizing the GLM procedure of SAS (9.4; Cary, NC). Due to environment differences, main effects of variety, treatment, and the interaction were analyzed within location. Statistical differences were determined at P ≤ 0.05. After all quality and production data are obtained, we will begin to work with an agricultural economist in the Ag Economics and Economics Department at MSU. We will be evaluating the tradeoff between yield and quality of the forage entries to determine the ideal recommendation for harvest maturity of sainfoin.

Results and Discussions At the Bozeman location, we saw a treatment by variety interaction for production, NDF, and ADF (P ≤ 0.02; Tables 1, 3, and 4). There was a trend of treatment and variety on condensed tannin content (P = 0.06; Table 2). We observed a treatment effect on crude protein content (P = 0.001; Table 5). Treatment A (10%) and B (50%) had similar crude protein content, while treatment C (100%) had lower crude protein, which is consistent with increased maturity. At the Moccasin location, we saw a treatment, variety interaction for condensed tannin content , NDF , and ADF (P = 0.03; Table 2, 3, and 4). There was a variety by treatment interaction for production (P < 0.0001; Table 1). The shaw alfalfa produced more (P = 0.005) than any of the sainfoin varieties. Treatment B (50%) and C (100%) produced more (P < 0.0001) than treatment A (10%) and D (10% second cutting). Crude protein content was affected by both treatment (P < 0.0001) and variety (P = 0.003; Table 5). Shaw alfalfa had greater (P = 0.003) crude protein content than all varieties of sainfoin and treatment A (10%) had greater (P < 0.0001) crude protein content than the other treatments. At the Utah location, we saw no effect of treatment, variety, or the interaction on production (P ≥ 0.22; Table 1). Both NDF and ADF levels were affected by treatment, P = 0.004 and P = 0.007, respectively (Table 3 and 4). Treatments B and C had greater NDF concentrations than treatment A, which was expected due to increased plant maturity. Crude protein and condensed tannin content were affected by both treatment (P ≤ 0.0004) and variety (P ≤ 0.008; Table 2 and 5). As maturity increased, crude protein and the condensed tannins decreased. The main variety driving the difference in condensed tannins is the shaw alfalfa variety. At the Bozeman location, there were factors that could have influenced results. Wildlife was were a major problem despite the 6-foot deer fence surrounding the plots. Alfalfa weevil was also a concern in the alfalfa portion of the plots. These unforeseen problems could have affected production specifically. Due to the alfalfa weevil we were not able to sample the 10% bloom. We also had to delay the 10% sampling for the sainfoin due to the deer. Due to these issues that is why data were analyzed within location. As seen in other research, shaw alfalfa was more productive than sainfoin varieties. However, this was not seen across all locations. This could be due to environment differences or wildlife factors. It was also seen that shaw alfalfa had much lower condensed tannin content. These higher levels of condensed tannins are what allow sainfoin to be a bloat free forage. As expected with increasing maturity, there was an increase in ADF and NDF with each treatment. It was also noted that the CP level decreased with increasing maturity. Since data were just analyzed we will be sending our results to the economic department at MSU to determine economic impacts of the different varieties and harvest treatments.

Acknowledgements Appreciation is expressed to the Midwest Forage Association for funding this research. A special thanks to all the individuals who helped sample and run quality analysis, Pat Carr, Harley Naumann and Jennifer MacAdam. I would also like to thank Megan Van Emon for helping with statistical analysis.

Table 1. Production at three different locations for four sainfoin varieties and one alfalfa variety. Variety3 P – value Location1 Treatment2 AAC DEL ESK SHA SHO SEM4 Treatment Variety Treatment*Variety U A 4839.7 4749.2 4109.9 6095.4 3618.2 1067.89 0.22 0.43 0.55 U B 6629.4 5064.9 6267.2 ̶ 4117.1 U C 5925.8 3801.7 5034.2 ̶ 5908.3 B A 1662.2a 1807.8a 1576.6a ̶ 2082.3a 398.75 <0.0001 0.03 0.02 b ab ab ab a B B 1927.1 2869.8 2266.2 3064.1 3266.8 a a b a ab B C 3100.3 2934.6 4027.8 2345 3740.5 M A 1966.3 1442.3 1858.8 3771.4 1688.6 612.03 <0.0001 0.005 0.21 M B 2064.9 2270.9 2253.0 3820.7 2602.3 M C 3233.9 2485.9 3793.8 3529.5 3977.4 M D 1339.2 604.7 514.2 1132.3 1126.0 1Location: B = Bozeman, U = Utah, M = Moccasin 2Treatment: A = harvest at 10% bloom, B = harvest at 50% bloom, C = harvest at 100% bloom, D = second cutting at 10% bloom 3Variety: AAC = AAC Mountainview, DEL= Delaney, ESK = Eski, SHA = Shaw, SHO= Shoshone 4Greatest standard error of means presented. a,bMeans within a location and treatment with superscripts are different (P ≤ 0.05).

Table 2. Condensed tannin content at three different locations for four sainfoin varieties and one alfalfa variety. Variety3 P – value Location1 Treatment2 AAC DEL ESK SHA SHO SEM4 Treatment Variety Treatment*Variety U A 46.66 38.77 44.69 1.38 47.07 5.829 0.0004 <0.0001 0.62 U B 34.04 32.16 41.14 1.23 31.94 U C 34.63 32.24 37.42 0.88 31.11 B A 35.07ab 33.75ab 37.12a ̶ 29.90b 3.843 <0.0001 <0.0001 0.06 a b ab c a B B 36.60 29.16 33.49 1.37 31.21 B C 24.77a 23.84a 25.69a 1.12b 26.89a M A 14.31a 11.27a 17.10a 0.98b 15.42a 5.183 <0.0001 <0.0001 <0.0001 ab b a c ab M B 36.38 34.42 42.72 1.27 38.95 a a a b a M C 36.01 32.85 30.88 0.92 35.45 b b a c a M D 29.86 31.38 39.41 1.46 39.96 1Location: B = Bozeman, U = Utah, M = Moccasin 2Treatment: A = harvest at 10% bloom, B = harvest at 50% bloom, C = harvest at 100% bloom, D = second cutting at 10% bloom 3Variety: AAC = AAC Mountainview, DEL= Delaney, ESK = Eski, SHA = Shaw, SHO= Shoshone 4Greatest standard error of means presented. a-cMeans within a location and treatment with superscripts are different (P ≤ 0.05).

Table 3. Acid detergent fiber content at three different locations for four sainfoin varieties and one alfalfa variety. Variety3 P – value Location1 Treatment2 AAC DEL ESK SHA SHO SEM4 Treatment Variety Treatment*Variety U A 26.89 28.98 26.70 28.67 26.15 2.322 0.007 0.27 0.29 U B 33.25 28.09 29.68 31.10 31.86 U C 30.63 29.94 27.68 32.20 30.40 B A 24.52a 26.25a 26.07a 26.64a 26.23a 1.914 <0.0001 0.0003 0.003 a b a c a B B 25.27 31.06 26.06 34.38 27.81 B C 33.18a 31.11ab 31.64ab 35.54c 32.56b M A 25.26 26.89 25.80 27.26 26.65 2.561 <0.0001 0.08 0.03 M B 29.11 26.76 29.80 28.51 29.96 b ab a a ab M C 25.52 27.52 29.11 30.40 28.50 ab c c a bc M D 23.35 19.59 17.15 24.26 19.86 1Location: B = Bozeman, U = Utah, M = Moccasin 2Treatment: A = harvest at 10% bloom, B = harvest at 50% bloom, C = harvest at 100% bloom, D = second cutting at 10% bloom 3Variety: AAC = AAC Mountainview, DEL= Delaney, ESK = Eski, SHA = Shaw, SHO= Shoshone 4Greatest standard error of means presented. a-cMeans within a location and treatment row with superscripts are different (P ≤ 0.05).

Table 4. Neutral detergent fiber content at three different locations for four sainfoin varieties and one alfalfa variety. Variety3 P - value Location1 Treatment2 AAC DEL ESK SHA SHO SEM4 Treatment Variety Treatment*Variety U A 35.00 36.92 34.40 37.93 33.68 2.535 0.004 0.08 0.28 U B 42.24 36.36 37.26 40.93 40.02 U C 39.13 39.16 35.56 41.02 38.93 B A 31.90a 34.09ab 33.76ab 34.73ab 36.27b 2.580 <0.0001 <0.0001 0.002 a b a c b B B 32.58 40.64 33.98 46.01 37.42 B C 42.08a 39.28a 40.22a 46.55b 41.24a M A 32.88b 35.15ab 33.45b 38.43a 34.40ab 3.077 <0.0001 0.001 0.02 M B 37.36 34.51 38.70 37.96 37.96 c bc ab a abc M C 32.70 35.24 37.37 39.58 36.09 a b b a b M D 30.48 26.03 22.61 32.44 25.82 1Location: B = Bozeman, U = Utah, M = Moccasin 2Treatment: A = harvest at 10% bloom, B = harvest at 50% bloom, C = harvest at 100% bloom, D = second cutting at 10% bloom 3Variety: AAC = AAC Mountainview, DEL= Delaney, ESK = Eski, SHA = Shaw, SHO= Shoshone 4Greatest standard error of means presented. a-cMeans within a location and treatment row with superscripts are different (P ≤ 0.05).

Table 5. Crude protein content at three different locations for four sainfoin varieties and one alfalfa variety. Variety3 P - value Location1 Treatment2 AAC DEL ESK SHA SHO SEM4 Treatment Variety Treatment*Variety U A 11.01 10.43 11.41 12.59 11.07 0.729 <0.0001 0.008 0.41 U B 9.16 8.86 9.67 9.28 7.86 U C 7.39 6.82 7.90 9.19 7.54 B A 31.90 34.09 33.76 34.73 36.27 1.158 0.001 0.27 0.07 B B 32.58 40.64 33.98 46.02 37.43 B C 42.08 39.28 40.22 46.55 41.24 M A 13.37 11.97 12.52 15.83 13.15 1.454 <0.0001 0.003 0.09 M B 12.39 11.65 10.98 12.94 10.79 M C 9.82 9.98 8.83 9.51 10.11 M D 8.80 10.41 10.87 12.48 10.13 1Location: B = Bozeman, U = Utah, M = Moccasin 2Treatment: A = harvest at 10% bloom, B = harvest at 50% bloom, C = harvest at 100% bloom, D = second cutting at 10% bloom 3Variety: AAC = AAC Mountainview, DEL= Delaney, ESK = Eski, SHA = Shaw, SHO= Shoshone 4Greatest standard error of means presented. a-cMeans within a location and treatment row with superscripts are different (P ≤ 0.05).

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