Source: TEXAS A&M UNIVERSITY submitted to NRP
COMPLIANT HEMP ACCELERATED IMPROVEMENT NETWORK (CHAIN)
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1029301
Grant No.
2022-38624-38369
Cumulative Award Amt.
$201,174.00
Proposal No.
2022-06905
Multistate No.
(N/A)
Project Start Date
Sep 1, 2022
Project End Date
Aug 31, 2025
Grant Year
2022
Program Code
[HW]- Supplemental and Alternative Crops
Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001
Performing Department
(N/A)
Non Technical Summary
Despite global industrial hemp markets exceeding USD $5 billion annually, expansion throughout the U.S. has been severely limited. The majority of current industrial hemp cultivars were developed in Europe or northern latitudes and have strong photoperiod responses that induce reproductive growth as days shorten. This leads to premature flowering at southerly U.S. latitudes (37°N and lower) and crops with little to no value. The 60+ year U.S. ban on C. sativa has until recently further prevented any formal improvement of the crop, and the newly formed NPGS hemp germplasm collection currently includes only a handful of accessions--none of which are fiber or grain types. The goal of the Compliant Hemp Accelerated Improvement Network (CHAIN) is to publicly release and evaluate a collection of novel accessions produced by Texas A&M University's Hemp Conversion Program (HCP) alongside other candidate materials in order to identify germplasm well adapted to the southern U.S. The project establishes a north-south transect of three locations from 26.2°N to 33.5°N. Twelve fiber and six grain cultivars plus selected HCP accessions will be tested at four planting dates per location. The state extension hemp specialist will develop a team of extension colleagues across southern states to provide information on management of hemp photoperiod responses. The diverse, public HCP germplasm collection incorporates the majority of genetic variation across all Cannabis spp. (C. sativa, C. indica, C. ruderalis) and will benefit farmers with high yielding cultivars, refiners with reliable feedstocks, customers with quality end-products, and society with beneficial ecosystem services.
Animal Health Component
80%
Research Effort Categories
Basic
10%
Applied
80%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2020199108190%
2050199303010%
Goals / Objectives
Objectives1. Southern U.S. Public Germplasm Collection--Develop and distribute a significant array of southern adapted hemp accessions from the HCP program including fiber, grain, cannabinoid, and multi-use ideotypes across universities, extension services, and private organizations.2. Germplasm Testing--Evaluate the initiation of reproductive growth in fiber and grain industrial hemp cultivars from commercial sources and the companion southern U.S. breeding program at three coordinated Texas A&M AgriLife test sites which will develop into a self-supporting fee-based testing program after two years:A. A range of southern latitudes (26.2 to 33.5ºN--a north-south transect of ~600 miles.B. Test four planting dates (28-day intervals) from early spring to early summer.C. Determine THC concentrations at designated pre-harvest conditions to further eliminate unsuitable cultivars.3. Extension--Make ongoing results quickly available online after data collection and summary for immediate Extension reporting to stakeholders. A preliminary guide for cultivar responses will be prepared with other southern state hemp colleagues, followed by a scientific review of prior southern testing results and the photoperiod response impact.4. Type-V Cannabinoid-free hemp-Develop novel Type-V (nil cannabinoid) hemp via traditional breeding (and E-beam mutagenesis as a contingency), followed by entry into the HCP to generate THC-compliant, southern adapted Type-V accessions.
Project Methods
Objective 1 - Development and distribution of southern U.S. adapted hempEstablished HCP protocols and plant materials will be utilized. Female plants of Type IV hemp derived from grain x fiber hybridizations at Texas A&M University will be propagated and placed under floral induction light regimes at the 4-node stage. Single plants will be placed inside pollen exclusion tents prior to flowering. Using sterilized fine-tipped brushes, donor pollen will be lightly applied to at least 5 female flowers in an isolation room at both week 2 and week 3 flowering stages. Each female plant will be maintained in the pollen tent until seed maturity, and seed will be hand harvested and cleaned. Twenty F1 seed from each hybridization will be germinated, placed under a floral induction light regime at the 4-node stage, and classified as male or female plants. Selected plants from each cross will be placed and maintained in pollen exclusion tents and induced into hermaphrodites via plant growth regulator treatments. Bulked F2 seed will be randomly selected from each line, germinated, and propagated through maturity. During late flowering stages, each will be assayed for THC utilizing a Light Lab 3 (Orange Photonics) HPLC in PI Jessup's lab in order to realize cost efficiencies ($5 per sample for THC and cannabinoid profiling). All plants (anticipated ~75%) at or above 0.3% THC will be destroyed. The remaining compliant plants (below 0.3% THC) will be vegetatively propagated from axillary buds using standard protocols and then isolated as a polycross under a larger pollen tent. Flowering time will be monitored to ensure at least two F2 hybrids are synchronous, and the number of parents in each polycross will be documented. All 'Syn1' hemp seed will be harvested at maturity, cleaned, packaged, and made available for distribution in the CHAIN program.Objective 2 - Germplasm TestingThe north-south transect of Texas A&M AgriLife experiment sites (Table 3) for field testing had no restrictions for pollen flow to cannabinoid trials or commercial CBD production. Yield trials and other agronomic work commence in the future after photoperiod adaptation is established. Working with hemp seed companies and hemp contractors 20 prospective fiber (majority) and grain cultivars will be chosen for a range of photoperiod conditions/latitudes we believe will be productive at southern latitudes. In addition, observation tests will be conducted on all experimental releases from PI Jessup's TAMU HCR breeding program. Commercial cultivars will be checked in Kentucky Dept. of Agriculture's comprehensive field test report of THC sample results (KDA, 2022). Cultivar selections will include Europe, newer Canadian lines, Australia, China, and the U.S. Two controls for fiber (e.g., 'Eletta Campana,' Italy; 'Anka,' Canada) and one grain ('Helena,' Italy) represent premature flowering cultivars. Where sufficient seed is available, seeds will be assessed for percent germination (200 seed) in advance using Association of Seed Analysts (AOSA) procedures for hemp. Test weight and thousand seed weight will be determined. Pure Live Seed results will adjust planted seed from 200 (base number) per each of two randomized single-row plots (4-m length on ~40" rows) for cultivars relative to 80% germination (PLS80). Experience finds a combination of heat, soil crusting (broken by hand when needed) from irrigation, and reduced seed germination/seedling vigor may result in <50 plants per row. (At a minimum this is an observational test to assess a larger number of cultivars for photoperiod and relative estimated performance.) Seeding will occur on 28-day intervals from mid-February at Weslaco to late March at Lubbock. Relative emergence (%) and seedling vigor (scale 0 to 5) is recorded. Plants will be observed weekly for initial onset for each of male and female reproductive structures and the corresponding height recorded. Fiber harvest timing will be noted when an ~20% of plants demonstrate female reproductive growth (Kostiuk and Williams, 2019). We will project pre-harvest THC sampling within 30 days of theoretical harvest. THC analysis on each row will be analyzed in the Jessup lab via HPLC. Data is checked for trends in %THC vs. planting date. In addition to the commercial or advanced company cultivars tested, industry will be invited to submit additional fee-based entries $400/location (four planting dates). The two-year research program will seek to establish recognized baseline testing so these trials for photoperiod response will become a self-supporting, fee-based program of Texas A&M AgriLife's official Crop Testing Program. Results will serve all southern U.S. states. Qualitative yield estimates will be recorded for standing fiber and grain lines. Means of Analysis--Statistical analysis for reproductive growth stages and plant height will be conducted using ANOVA then regression to gauge varietal response over time.Objective 3 - Extension CommunicationsThis project will issue cumulative in-season reports near the first of the month, April to July. Likewise, seed companies (even if not participating) will see how to best deploy their seed stock and where. There will be an incentive to avoid marketing their seed too far south. We will directly distribute to clientele and university Extension colleagues in the 12 southern states. The Texas A&M AgriLife Crop Testing hemp page will host project results once available. Results will be distributed through commodity organizations, the national S-1084 hemp collaboration, and southern state variety trial cooperators. Early reporting from March and April plantings will be available soon enough that growers with planting seed on hand of a cultivar in this trial will know whether to plant it. PI Trostle has frequent contact with traditional agriculture and hemp media providing another ready outlet for results. Finally, an annual field meeting (including Extension change-in-knowledge survey work) for stakeholders at each site will highlight of our overall Texas Extension education programs. A virtual field tour will be held from Lubbock for distant clientele.Objective 4 - CHAIN will produce a Type-V Cannabinoid free Hemp lineIn order to develop a Type-V line the nil cannabinoid phenotype exhibited among some members of USO31 must first be stabilized in order to create consistent Type-V breeding stock. A minimum of 2,000 USO31 plants will be cultivated and screened for Type-V identification (via cannabinoid profiling with the LightLab 3 HPLC platform) and self-pollinated to produce a large number of seed for breeding material. This material will then be interbred with heat-tolerant TAMU lines in order to create a Type-V line optimized for growth in southern latitudes. From the breeding stock markers for null cannabinoid production can be discovered which can greatly speed up the interbreeding process. These markers will include variants of a type III PKS and olivetolic acid cyclase, which function together in order to generate olivetolic acid. Olivetolic acid is the precursor to all cannabinoids, so defects in its metabolic pathway are useful markers to identify potential Type-V Hemp. If Type-V individuals are not identified within USO31 plants, an additional 10,000 USO31 seed will be placed in high-density polyethylene (HDPE) bags (1000 seed per bag) and irradiated at an electron beam (eBeam) dose of 500 Gy.

Progress 09/01/23 to 08/31/24

Outputs
Target Audience:Relevant stakeholders for the project include industrial hemp: seed companies, private and public plant breeders, germplasm collection curators, agronomists, and producers. Changes/Problems:A lack of appropriate plant grow room space has persisted and significantly impacted (slowed) progress. While we have achieved more than the proposed number of donations (100) and initialhybridizations, advancement of plant materials through all stages has been delayed. Progresswill be continued with available facilities. What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?The 7 accessions submitted to the NPGS will be publicly released following their internal review, evaluation, and seed increase. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? A total of 7 accessions have completed all CHAIN stages (parental hybridization, F1 inbreeding, F2 population compliance screening, and Syn1 population seed increase. These 7 accessions have been submitted to the NPGS collection for industrial hemp.

Publications


    Progress 09/01/22 to 08/31/23

    Outputs
    Target Audience:Relevant stakeholders for the project include industrial hemp: seed companies, private and public plant breeders, germplasm collection curators, agronomists, and producers. Changes/Problems:Our field lab facility suffered a security breach in July, 2023, that was reported to USDA-NIFA-SAC program managers. Numerous plants were lost, which has required use of reserve seed and slowed progress advancing accessions to completion. We're making every effort to maximize use of available space and resources, but a request for a no-cost extension may be necessary before Aug. 2024. What opportunities for training and professional development has the project provided?One graduate student and one undergraduate student have been trained on hemp breeding (monoecy induction, pollen isolation, controlled hybridization, seed harvest/cleaning), HPLC chemotyping, and mutagenesis methods. How have the results been disseminated to communities of interest?A running status of the program is maintained on LinkedIn (Hemp Conversion Program) as a group. What do you plan to do during the next reporting period to accomplish the goals?Additional pollen donations will be sought to reach a minimum total of 100 accessions. Field trials will be performed in 2024. All accessions advanced through the entire program will be submitted for public release--small lots through Texas A&M in 2024 and once seed is further increased submission into the NPGS hemp collection. At least one publication on the program and one extension document on ecoregion adaptation across a subset of the accessions are also planned.

    Impacts
    What was accomplished under these goals? A total of 61 pollen donations were received and entered into the compliance conversion program. Twelve were advanced sufficiently for field evaluations at the three latitudes planned. Field trials for 2023--3 locations with4 planting dates--and data collection are ongoing. An additional 49 accessions have been initiated in the program and advanced to varied P, F1, F2, and seed increase stages. Type V development activities have begun, with both physical (eBeam) and chemical (EMS) mutagenesis underway. Dose response curves have been completed, larger seed lots have been treated, and M0 plant cultivation/ screening is underway.

    Publications