Source: NEW WEST GENETICS INC. submitted to NRP
LAUNCHING AMPLIFY TECHNOLOGY TO DOUBLE HEMP YIELD: A NEW CLIMATE-SMART CASH CROP FOR AMERICAN FARMERS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
SMALL BUSINESS GRANT
Reporting Frequency
Annual
Accession No.
1031805
Grant No.
2024-70031-41899
Cumulative Award Amt.
$174,600.00
Proposal No.
2024-00229
Multistate No.
(N/A)
Project Start Date
Jul 1, 2024
Project End Date
Feb 28, 2025
Grant Year
2024
Program Code
[8.2]- Plant Production and Protection-Biology
Recipient Organization
NEW WEST GENETICS INC.
219 N TIMBERLINE RD UNIT 1
FORT COLLINS,CO 80524
Performing Department
(N/A)
Non Technical Summary
New West Genetics will double industrial hemp grain yields and improve fiber quality by commercializing existing genetics intellectual property. By doing so, hemp will become an economically viable new rotation crop for American farmers that provides manufacturers and consumers with healthy new protein and lipid ingredient options. The intellectual property centers around a genetic factor which, when used as a sire in a cross, skews the proportion of female plants in a hemp population from the traditional dioecious norm of 50%, to a level of 90% or greater in the F1 generation. This inherently increases grain yield since male plants make only pollen. Additional yield increases are expected from heterosis; thus we refer to the F1 seed as Amplify. Further, we have identified specific crosses where the F1 is 100% female plants which paves the way to a scalable hybrid seed production system such as the CMS/Rf system used in other major crops. To make this a reality, we are proposing the following objectives: 1) Demonstrate that the technology can create Amplify consistently above 90% females, 2) Demonstrate the ability to generate Amplify seed which significantly improves grain yield and fiber quality relative to existing check varieties, 3) Identifying new methods for cost-effectively producing Amplify planting seed, and 4) Examining the carbon index of hemp relative to other commodity crops.
Animal Health Component
50%
Research Effort Categories
Basic
20%
Applied
50%
Developmental
30%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2021730108140%
2011730108020%
2061730106030%
6011730301010%
Knowledge Area
601 - Economics of Agricultural Production and Farm Management; 206 - Basic Plant Biology; 202 - Plant Genetic Resources; 201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
1730 - Hemp;

Field Of Science
1060 - Biology (whole systems); 1081 - Breeding; 1080 - Genetics; 3010 - Economics;
Goals / Objectives
a) Demonstrating that the technology can create Amplify consistently above 90% females. We will further test the technology's ability to produce Amplify seed at scale. Based on past results, the percentage of females in a given Amplify population varies dependent upon the genotype used as the female. This is analogous to specific combining ability (SCA) in other hybrid crops. Identification of genotypes more easily skewed towards higher proportions of females will guide the choice of female genotypes used in Phase II commercial scale-up. Genotypes consistently skewed to 100% will be examined further to understand the molecular basis of this tendency. The goal is to identify combinations that reliably create 100% female plants. Several have been identified already. This is analogous to the CMS system used in other hybrid crops and will pave the way for the most cost-effective production of Amplify. We also expect to identify heterotic groups which will facilitate future breeding efforts. Identification of such groups has been critical to modern maize hybrid breedingand will accelerate development of improved Amplify genotypes in our program.b) Demonstrating the ability to generate Amplify seed which significantly improves grain yield and fiber quality relative to existing check varieties. Our results to date provide strong support for the hypothesis that a greater proportion females leads to higher yields. In addition, certain hybrid combinations show substantial better parent heterosis. To fully exploit both genetic improvements, we will expand testing of Amplify genotypes using replicated plot trials grown across multiple locations. Grain yield and fiber quality results will identify the highest performing Amplify genotypes, thus informing which genotypes to produce during Phase II efforts.c) Identifying new methods for cost-effectively producing Amplify planting seed. We will test several agronomic and genetic methods for producing Amplify seed at larger scale in order to provide guidance for practices to be used at a larger scale during Phase II production. Yield and quality data from the plot trials (item b) will be used in models to calculate the extent to which the expected improvements in yield and quality translate to gross returns for farmers. The calculations will include varying price points for the harvested product at the farmgate and how they can lower processor costs because of lower field production costs. Identification of Amplify genotypes with yield and economic parity to soy will confirm hemp's benefit to US farmers.d) Examining the carbon index of hemp relative to other commodity crops. We will work closely with experts in crop Life Cycle Analysis to understand the carbon footprint and potential benefits of hemp production. The lower inputs required to produce hemp along with its large root system and potential to use the stalk byproduct in multitude other applications position it as a climate-smart crop.
Project Methods
Amplify hybrid testcross production in 2021 was managed like small-plot yield trials, with each plot consisting of two rows measuring 5.6 meters in length on a 0.76 m row spacing. Female plots were planted alongside the inducer genotype at a ratio of approximately 80% females and 20% inducer. Males were rogued from the intended female plots every second or third day to eliminate any pollen except that of the inducer. Plots were harvested at physiological seed maturity, which varied considerably due to genetic variation in flowering/maturity times among female genotypes. Seed was threshed and cleaned by hand. In 2022 and 2023, 16 Amplify genotypes were produced using the same general methodology as described above. The only modification is the design was more representative of a commercial seed production scheme in that genotypes were planted in full rows, rather than in a plot format, and harvest was performed using a mechanical swather and combine. The male (pollen producing) genotype was planted at approximately 14% of the total field. The seed produced over the past few years will be used in the following assessments of performance.Demonstrate the ability of the technology to create Amplify consistently above 90% females. Quantifying percentage of females and stability of % females. Amplify seed will be sown into 18-well inserts filled with Pro-Mix HP potting soil. Greenhouse supplemental lighting will be set to 16-hour daylength with temperatures maintained at 20-30 degrees C. Plants will be scored at the initiation of flowering for sex: male, female or hermaphrodite. Further, we will assess the percentage of females for each Amplify genotype as they are trialed across a diverse set of trialing locations.Examination of relationship between lineage and field performance. The current Amplify genotypes were created using dioecious genotypes representing eight pedigrees, including germplasm originating from Canada, China, France and varieties created by New West Genetics (NWG). These genotypes are only a small subset of the diversity in our breeding pool but they generally represent standing genetic diversity. We will compare yield and fiber quality data among Amplify genotypes to look for trends between group membership and performance. Amplify parents will be included in the 2024 trials (explained below) so that heterosis relative to the Amplify parents can be calculated.Demonstrate the yield advantages of Amplify by testing in replicated yield trials across geographically distinct locations. Performance of existing Amplify genotypes will be assessed in trials conducted by NWG in Colorado along with sites managed by cooperators located outside of Colorado. NWG has worked with the proposed cooperators but this will be the most extensive trialing of Amplify thus far. Data will be collected on emergence/stand count, days to flower, percentage of female plants, days to maturity, height, fiber yield and grain yield/quality (fatty acid profile, percentage protein and oil).The experimental design for CO trials will be completely randomized complete blocks with three replications of each genotype. We will also include 5 dioecious genotypes (NWG 2463, NWG 2730, NWG 4000, NWG 4113, X-59) planted from certified seed as checks. Thus, each experimental trial site will consist of 90 plots (30 genotypes x 3 plots). Each plot will measure 11.2 meters in length by 0.76 m row in width. Any rows next to an alleyway will be excluded from data analysis to avoid edge effect. Plots will be swathed at full maturity using a tractor mounted swather and harvested using a plot combine with a pickup header to measure grain yields. A block of dioecious checks and high stalk yielding Amplify plots will be left to ret at two locations so that stalks can be collected to estimate yields and provide material for quality testing with collaborators. Dioecious checks will be excluded from one site in order to confirm that 5 to 10% males provide pollen sufficient for grain fill. Data will be analyzed using mixed model analysis in R (R Core Team, 2021). Replications within trials will be treated as random effects and genotypes will be treated as fixed effects. Comparisons of genotype means will be made from Least Significant Difference estimates when the F-test is considered significant (p < 0.10). Locations with a coefficient of variation greater than 20% will be removed. Collaborator experimental design will be performed similar to that described above but within the constraints of the equipment/facilities available.Identifying new methods for cost-effectively producing Amplify planting seed. We have demonstrated that seed set and yields of F1 seed are good in production scenarios with 14% of the field planted with the male pollinator. While this is similar to other hybrid crops, hemp is anemophilous due to its extraordinarily small pollen grains which have been shown to disperse long distances. We will test the following treatments on production fields to find the lowest planting rate of male genotype with maximal seed yield: 15%, 10%, 5% and 1%. In essence, we will plant larger blocks (consecutive female genotype rows) to see the inflection point at which the plants in the middle blocks begin to show reduced seed set and yield. Based on current seed production cost estimates, reducing male presence from 14% to 1% would lower cost by approximately $0.50/lb, a 10% reduction. We will also expand the use of 100% female populations in hybrid seed production. We have made small-scale quantities of hybrids from 100% female genotypes in greenhouses and small plots. 2023 yield trials have not been harvested at the time of this submission, but observations of hybrids made using genetically 100% female genotypes look promising. Hybrids made using this method will be tested more thoroughly as part of the 2024 trialing plan outlined above. We will also make larger quantities of seed using this method in 2024 using USDA SBIR funding to understand its economic, biologic and agronomic impacts. We will specifically look to answer these questions: 1) Economics: how much less rogueing is required and how does that impact cost of seed production?; 2) Biology: do female populations revert to monoecy as pollen availability declines?; 3) Agronomy: how much does a higher final stand (due to minimal rogueing) impact weed control and, thus, overall quality of the harvested seed?Examining the carbon index of hemp relative to other commodity crops. NWG will provide estimates for agronomy and genetics parameters (yield, fertilizer, fuel, etc.) in Life Cycle analysis models to understand the carbon intensity of hemp production relative to other crops. The higher yields, vigor and extensive root system of hybrid hemp, along with the many uses of the stalk byproduct, make NWG hemp a candidate to stand alone as a climate-smart new crop.

Progress 07/01/24 to 02/28/25

Outputs
Target Audience:The target audience is farmers and processors. We arehighlighting the grain yield and performance advantages of our Amplify hybrid hemp varieties and how they put hemp production economics on par with other major oilseeds.We have identified two main customer profiles, described in the following examples: 1) Growers: Leon is a third generation Iowa grower of organic alfalfa and corn. He is young and wants to reinvigorate his family farm. He has recently added sorghum and rye to his crop rotations and would like to add hemp due to his interest in both grain and fiber. Leon was uncertain where to turn for quality genetics. He reached out to a local hemp industry association, who referred him to NWG. Upon first meeting, NWG asked many questions, then described their seed options, the value of their seed, and explained the agronomic support package. NWG suggests Leon try a small plot, using a dual-purpose variety that would suit his growing region. Leon purchases 20 acres worth of seed. Throughout the season, NWG and Leon held meetings at crucial stages of production to discuss Leon's questions. NWG traveled to his farm to lend additional support, including harvest and post-harvest processing. 2) Manufacturers: A processing company wants to add hemp products to their portfolio. They are feeling consumer pressure to offer products that are US sourced, non-soy, non-GMO, and have a differentiated nutrition/fiber/cannabinoid profile. They are referred to NWG through a hemp industry insiders, Ag Tech VCs, or Ag accelerators they are supporting members of. They are cautious, more cautious than growers, and ask many questions. They frequently request NWG manage the supply chain from genetics to the raw materials. Those outside of the hemp industry would prefer NWG to leverage their network of experience and reliable growers to contract crop production, then deliver it to their door. Changes/Problems:The biggest problem we encountered were the non-significant results at 2 of our trial locations (MT and MB).This is an unfortunate reality of field trialing where the environment often overrides the effect of genetics. Commercial breeding programs of corn, soybean and other major commodities run trials at dozens, hundreds or even thousands of locations because they expect data to be uninformative at a large percentage of them. What opportunities for training and professional development has the project provided?We participated in a hemp Field Day at the University of Illinois on 2 August 2024 (Hemp Research - Lee Laba) to describe our USDA-SBIR funded trial and the technology driving it.We also hosted a tour of our trial conducted at Colorado State (ARDEC) on 7 August 2024 as part of the 2024 Cannabis Research Conference held in Fort Colllins, CO.Agronomists and scientists from many disciplines were in attendance t both events which generated lots of good questions and helped the professional community better understand the benefits and potential offered by gendered skewed hybrid hemp. How have the results been disseminated to communities of interest?We have focused our dissemination efforts on Field Days and a very active social media presence, highlighting the benefits and potential of hybrid hemp. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1. Demonstrating that the technology can create Amplify consistently above 90% females.AMPLIFY hybrid production in 2021-2023 was planted in traditional hybrid design with female (pollen recipient) interspersed with inducer genotype at a ratio of approximately 80% females and 20% inducer. Males were rogued from the intended femals every 2-3 days to eliminate any pollen except that of the inducer. Can we create Amplify seed in the field at larger scale?Yes.AMPLIFY seed lots were grown in greenhouses with supplemental lighting. Plants were scored for sex at flowering.Over 4 years of seed production, we averaged 98.85% female plants across a total of 62 hybrid crosses (Figure). 56% of crosses achieved 100% female plants providing strong evidence that a hybrid seed production system using 100% female populations (akin to CMS or NMS systems in other species) is feasible. Can we predict which genotypes will skew to higher percentages of female plants?Yes, but more research is needed. In crosses with <100% female plants, we suspect the few male plants were due to outcrossing from stray males (e.g. those missed during rogueing dioecious plots). Agronomic practices to facilitate effective rogueing were identified and implemented. Which genetic backgrounds create the highest performing Amplify genotypes?We have identified a heterotic pattern between the current inducer and crosses with female genotypes originating from "high CBD" genetics which averaged 161% grain yield relative to Open-Pollinated (OP) checks. Testcrosses using genotypes derived from our founder population (non "high-CBD") yielded 123% of the OP check, a highly significant result (p = 0.001). Objective 2. Quantifying Amplify grain and fiber yield relative to existing check varieties. Performance of 30 AMPLIFY hybrids were assessed in trials conducted by NWG at the Colorado State University Agriculture Research & Demonstration site, The University of Illinois, Range & Row, LLC (Great Falls, MT) and Ag-Quest, LLC (Minto, MB). These sites represent distinct climates and soils. Due to the large differences in latitude, hybrids were selected which maximized their likelihood for success based on their adaptation. This was essential becauseC. sativais highly responsive to daylength and later flowering genotypes can become too tall for harvest. Thus, we trialed 21 AMPLIFY hybrids at ARDEC and UIL adapted to ~ 40' North. We trialed 5 and 8 AMPLIFY hybrids at Range & Row and Ag-Quest, respectively, adapted to ~ 48' North. Do Amplify genotypes have improved yield and performance, relative to known dioecious varieties?Yes.The average hemp yield was 2,181 pounds/acre at ARDEC and 2,062 pounds/acre at UIL. The Coefficients of Variation (CV) indicate respectable uniformity at 20.0% and 20.9%, respectively. The highest and lowest yielding hybrids differed by a factor of 3X (1,107 lbs/ac versus 3,386 lbs/ac). The 21 hybrids trialed at ARDEC and the University of Illinois demonstrated significant improvements in grain yield relative to OP check varieties, with one hybrid reaching 207% and a maximum estimated yield of 3,386 pounds/acre. Overall, the results across locations were in agreement (r = 0.779). Unfortunately, the trials conducted at Range & Row, LLC and Ag-Quest Holdings, LLC were both inconclusive. Although the trial conducted by Range & Row, LLC was under pivot irrigation, only 7" of water was available which was not sufficient for the full season needs of the crop. The majority of these 7" were applied in July (5.5"). This left only 1.5" during the grain fill in August, an insufficient amount during the most critical period of grain development in a season that only produced 2.4" of rainfall during July and August. This moisture limitation, along with unprecedented bird feeding, led to a high CV (42.9) with no significant differences in yield among genotypes. The trial at and Ag-Quest Holdings, LLC produced no significant differences among genotypes, likely due to seed loss from a large wind storm prior to harvest. Objective 3. Identifying new methods for cost-effectively producing Amplify planting seed. In 2024, we tested the following treatments to explore how low the male inducer planting rate could go while still maximizing seed yield: 12.5%, 10% and 4.5%. We accomplished this by planting increasingly larger bays of the female genotype, thus increasing the distance of females in the middle of the bay from male pollen to 17.5', 22.5' and 55'. Can we produce Amplify cost effectively?Yes.The current method of hybrid seed production is costineffectivedue to the labor required for rogueing male (XY) plants. However, our results from 2024 suggest a path to affordable seed production: Lowering the ratio of a seed production field planted with the male inducer increases the harvested area of female carrying F1 seed.In our experiment testing seed yields of females produced with increasing distance from the male inducer pollen source, we estimated net yields of 2,807 lbs/ac, 3,146 lbs/ac and 3,533 lbs/ac at the 17.5', 22.5' and 55' distance treatments, respectively, and insignificantly different from one another. Thus, we are confident that no more than 4.5% male inducer is required for adequate pollination. This simple increase in seed producing plants translates to a 36% increase in yield per gross acre and a 25% reduction in the seed cost. Using 100% female populations as pollen recipients in hybrid seed production will work.Our production trials using multiple A-line genotypes demonstrate that this approach can work for scaling hybrid seed production. Economics: how much less rogueing is required and how does that impact cost of seed production?Our estimates from 2024 suggest that A-lines can reduce rogueing hours from 389 hours/acre to 20 hours/acre, a reduction of 95%. This results in a 64% reduction in the cost of seed on a per pound basis and, more importantly, makes large-scale production far more realistic. Biology: do female populations revert to monoecy as pollen availability declines?Yes, some genotypes produced male flowers on plants with otherwise female phenotypes. We saw up to 40% plants showing this phenomenon but, in most cases, the number of male flowers and strength of pollen production was low. Agronomy: how much does a higher final stand (due to minimal rogueing) impact weed control and, thus, overall quality of the harvested seed?We did not quantify this, but the anecdotal answer is that the higher populations increased the crop's ability to compete with weeds through better uniformity and agronomy. Objective 4. Examining the carbon intensity of hemp relative to other commodity crops. What is the Carbon Intensity (CI) of hemp?The CI (g/MJ) of hemp is estimated to average 21.84, better than alternative oilseed crops such as soybean, Brassica carinata, canola and camelina. We contracted Trinity Consultants (https://www.trinityconsultants.com/) to perform an analysis of the CI of hemp using the TheGreenhouse Gases, Regulated Emissions, and Energy Use in Transportation (GREET) model, a tool that evaluates the life-cycle impacts of vehicle technologies, fuels, products, and energy systems. It is a transparent system for examining energy and environmental effects of energy and product systems. Since hemp is not a default feedstock in Argonne GREET, Trinity Consultants parameters for soy feedstock based on inputs to represent AMPLIFY hybrid hemp. Soy was chosen as a proxy for modeling because it most closely represents hemp compared to the other GREET feedstock options. The inputs that were updated are compared to soy defaults, including updates to energy used for farming (less herbicide = more mechanical weed control), fertilizer (~10-20X higher because hemp is not a Nitrogen fixing plant), and pesticide use. All other data assumptions within GREET were left as defaults.

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