Source: OHIO STATE UNIVERSITY submitted to NRP
SUSTAINABLE PRODUCTION OF ALTERNATIVE POLYMERS, FILLERS AND PRODUCTS FROM DOMESTIC CROPS AND WASTE STREAMS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1014257
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2017
Project End Date
Sep 30, 2022
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691
Performing Department
Horticulture and Crop Science
Non Technical Summary
In May 2017, the Secretary general of the Association of Natural Rubber Producing Countries (ANRPC) announced that, based on preliminary estimates, the world supply of natural rubber during January to May 2017 was nearly 600,000 tonnes in deficit and the shortfall is anticipated to widen to around 700,000 tonnes by the end of June 2017. World supply during 2017 is anticipated at 12.76 million tonnes. The United States imports 1.2 million tonnes per year so this shortfall puts the US and other importing countries at serious risk of high prices and even unavailability of this critical agricultural raw material. A systems approach will be taken to develop a novel natural rubber and industrial sugar crop (Taraxacum kok-saghyz, TK) in the northern US for bioenergy, biofuels and bioproducts. A TK crop that generates 1.2 million metric tons (mt) of natural rubber/yr to meet US demand also produces 3.2 mt of sugars for conversion into 1 billion gallons of biofuel. Current TK value of $2,946/ha is conservatively based on 6 g dry root/plant and 100 mg rubber/g dry root. This compares to $2,321/ha/yr for ethanol and DDGS from the average yield (188 bu/acre; 464 bu/ha) of Iowa corn (USDA, 2016). The OSU already has TK genotypes capable of producing >12 g dry root/plant and >150 mg rubber/g dry root, a crop value of $8,838/ha/yr. Objectives include: 1. Integrate TK into existing cropping systems and determine ecological (including honey bees) and water quality benefits of this bioenergy crop. 2. Optimize a scalable water-based process that recovers >95% of rubber, inulin and sugars in TK roots at purity levels that meet industry specifications for TK rubber and sugar yield and quality. 3. Educate/Extension to identify/mitigate risks, develop crop name, and train workforce. 4. Develop and assess waste-derived fillers as complete or partial replacements of carbon black and silica reinforcing fillers.
Animal Health Component
40%
Research Effort Categories
Basic
40%
Applied
40%
Developmental
20%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2050199106020%
1020199106020%
2010199108020%
2130199114020%
4020199202020%
Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships; 205 - Plant Management Systems; 201 - Plant Genome, Genetics, and Genetic Mechanisms; 213 - Weeds Affecting Plants; 402 - Engineering Systems and Equipment;

Subject Of Investigation
0199 - Soil and land, general;

Field Of Science
2020 - Engineering; 1060 - Biology (whole systems); 1080 - Genetics; 1140 - Weed science;
Goals / Objectives
Accelerate development and commercialization of domestic natural rubber crops to address global shortfalls of this critical raw material.Integrate TK into existing cropping systems, based on recent successes in TK agronomy, genetic tools and germplasm improvement, and determine ecological impacts of this regional industrial crop.Develop a scalable water-based process that recovers >95% of rubber, inulin and sugars from TK roots at purity levels that meet industry specifications.Optimize and model the partial and/or complete replacement of carbon black or silica reinforcing fillers with waste-derived fillers
Project Methods
Germplasm Development: A wide range of enabling technologies for TK were developed in ourprevious Hatch project. These technologies include the draft TK genome sequence, initial SNPmarkers[17], CRISPR/Cas9 gene editing[18], Agrobacterium tumafaciens transformation, A. rhizogenestransformation[19], colchicine induction of polyploidy, seed priming and pelletization, and high throughputphenotyping. Germplasm, and hybrid lines, are needed that maximize rubber, inulin, and sugarcontent in large rooted plants adapted to cultivation in the soils and environments across the northernUS. These plants must combine yield traits with additional domestication traits such as seedling vigorand herbicide tolerance. Methods to restrict seed dispersal are needed, such as plants that do notflower in their first year and seed heads with a dysfunctional pappus, which would have the additionaladvantage of enhanced seed collection. Plastidic engineering will capitalize on the ability of plastids toproduce 50x the amount of product/cell (substrates and end?products) produced by the nucleargenome[20,21].The projected yield is based on expansion of our current best lines and so should be attainable, i.e. plantmeans of 150 mg rubber, at least 200 mg inulin and 200 mg sugar/g root dry wt. Increased root size,including through tetraploid induction and propagation, will increase bagasse weight by at least 4 g/rootand thus, concomitantly, the C5/C6 mixed sugar stream. Research will continue to further improvegermplasm and increase the complement of stacked domestication and yield traits. The existing diploidTK genome assembly has been completed using Illumina reads produced by BGI. Optimization has beenpursued in house, but artifacts due to the heterozygous nature and prevalence of repetitive sequencewithin the TK genome have prevented a cytologically related (i.e. with resemblance to the chromosomicstructure of the genome) assembly, i.e. the current assembly is highly fragmented. We will performSingle Molecule, Real?Time (SMRT) Sequencing (a.k.a. PacBio sequencing) of the same genotype used forthe short Illumina reads, pursue a hybrid assembly, then consolidate the existing Illumina reads with askeleton guided by the scaffolding of the longer reads by PacBio. Thus, PacBio sequencing will augmentthe current assembly, not replace it, which will allow the assembly of a less fragmented genomesequence also allowing a much more detailed and complete gene annotation, overall, considering thatannotated genome sequences have been recently released for related crops, such as lettuce andsunflower. Linkage mapping will be pursued in parallel. The comparative genomics analyses we willpursue must include distinct levels: (1) Compare genomes as entities within the Asteraceae family(lettuce, sunflower, guayule, horseweed and globe artichoke), in which some species have welldeveloped resources; (2) Compare rubber?related genes (transferases, rubber particle ontogeny, laticiferbiogenesis, etc) across rubber?bearing species.Since TK is cross?pollinated and ensures heterozygosity through self?incompatibility and male sterility,inbred lines cannot be developed in the short term, and this development will continue, and, oncesuccessful, will rapidly expand with the direct involvement of seed companies, some of which havealready expressed interest.Tetraploids will be produced from the seed of progeny of high x high rubber content plants by treatmentof the seeds with 0.2% colchicine for 24h using a previously established method (unpublished).Seedlings are screened phenotypically and tetraploids confirmed with flow cytometry. Matureflowering tetraploids will be interbred and the F1 generation grown up and tested for yield and fitness.The highest performing tetraploids will be rapidly scaled up by interbreeding. Multilocation, replicatedfield trials will assess improvement in seedling vigor, and mature plant root size and rubber yield.ALS herbicide resistance will be produced by targeting known sites in the ALS gene for amino acidsubstitution using CRISPR/Cas9 using established methods[18.19]. Transgenic plants will be interbredwith high yielding genotypes and the progeny screened for the mutated genes and for herbicideresistance. Genotypes containing the mutation genes will be interbred to generate plants homozygousfor mutated ALS. Seed will be generated from resistant lines homozygous for mutated ALS. ALSherbicide resistance and plant performance will be tested in replicated field trials. In addition, seedfrom these lines will be used to generate tetraploids in order to enhance vigor and yield whilemaintaining strong herbicide resistance.TK genes homologous to Gerbera pappus development genes, previously identified in the draft TKgenome, will be mutated using CRISPR to disrupt pappus development and reduce wind mediated seeddispersal. The protocol will essentially follow the protocol of ALS gene mutation, except that pappusphenotypic disfunctionality will be assessed instead of herbicide resistance.In addition, transgenic and edited genotypes previously produced will be bred to homozygosity andevaluated in greenhouse and field trials. Lines with improved yield will be scaled up and tetraploidsproduced. These genotypes target assimilate partitioning (carbon to rubber instead of inulin) andrubber substrate synthesis. All generations will be evaluated for growth rate, rubber and inulin yieldusing established methods[11]. Plastidic engineering will be pursued using both ballistic and protoplasttransformation methods{21].

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Industry, researchers, general public, program managers, elementary, middle and high school students and teachers, technology transfer personnel, international and national alternative rubber scientists. Changes/Problems:Our goals have remained unchanged but the program was significantly negatively impacted by (i) COVID 19 research shutdowns which led to extensive delays in research field trials and breeding, (iv) crop failure of our planned one acre chemical weed control demonstration due to a too late planting date, (ii) reduction in previously stable sources of funding, (iii) student graduations reducing available worker hours. Unfortunately, these research delays also led to outright loss or indefinite postponment of several sponsored programs which were under negotiation before the shut down. What opportunities for training and professional development has the project provided?Cornish served on a Rubber and Plastics News Latex Livestream Panel(one of four experts) held on 8/6/2020 to cover sustainability, issues regarding the natural rubber trade, ongoing work with alternative forms of natural rubber, and any impact from the pandemic.This was attended by 141 international registrants. One visiting scientist (Gabor Kaszas, PolyFiberMatrix LLC), three graduate students, three Post-doctoral fellows and two visiting professors (Marek Kowalczuk from Poland, Lingyun Liu from Akron) have worked on the project in the reporting period and received technical and grants preparation and managment training. How have the results been disseminated to communities of interest?In addition to the activities and conferences detailed in earlier sections of this report, we have shared results at monthly meetings of the academic members of the Program of Excellence In Natural Rubber Alternatives Consortium, and of our multi-univierstiy BRDI funded team. We have also shared relevant results at weekly meetings of two DOD-DARPA Phase I funded proposals. We also have shared results with several different collaborativeteams working on new grants proposals, to NSF, DOE and USDA-NIFA, and which led to eight new proposals being submitted. COVID19 shutdowns led to extensions being required for the Phase I grants and direclty led to our failure to acheive Phase II for these projects. However, anearlier USDA PHASE II application was funded to develop the guayule radiation attenuation medical glove. What do you plan to do during the next reporting period to accomplish the goals?In field selections will continue to develop regionally adapted rubber dandelion populations. Data will be collected for future submission of Plant Variety Protections prior to the release of germplasm. High through put selections for improved rubber concentration and vigor will be performed in hydroponics using partial root harvests. Best performers will be identified and interbred. Seed will be tested in hydroponics and in the field to determine if this selection method is relevant to field perfomance as well as to hydroponic performance. Gene edited and genetically transformed plants will be grown to maturity and analyzed, including the relative amount of latex and solid rubber produced and any changes in plant biomass and total rubber concentration under APHIS approvals where required. One acre field demonstrations will be attempted in Ohio, Nebraska and Oregon, conditions permitting. The naturally-selected herbicide resistant population will be included in these field trials in Ohio, Oregon and Nebraska. Plants from the fourth generation selections of protoporphyrinogen oxidase inhibitor herbicides will be planted and tested/selected for tolerance again. Progeny of crosses among colchicine-induced tetrapolids and among colchine induced triploids (which do not appear to be stable apomicts) will be tested for chromosome numbers. and then interbred again for a seed increase. Latex will be extracted from greenhouse grown guayule transgenics of multiple ages and rubber molecualr configuration characterized. Latex samples with similar configurations will be pooled to determine the effect of conformational changes on rheological and mechanical performance. Processing research will continue to maximize yield and purity of dandelion rubber. Further TEA and LCA is covered in a different Hatch project (led by Dr. Ajay Shah)

Impacts
What was accomplished under these goals? Agronomy Research has continued to improve rubber root dandelion stand establishment and to control weeds. The optimum planting-time window was between 500 and 700 Growing Degree Days (GDD), corresponding to May 25 to June 8, when 10-cm soil temperature were over 17C.The Jang seeder resulted in 88% establishment compared with 70% for broadcast seeds. Root weight was higher for plants grown on raised beds. For weed control, the most consistent treatments were preemergence applications of imazethapyr or sulfentrazone at standard rates, which provided 60 to 88% weed control and 5-15% crop injury. Emergence differentiation experiments indicated that plants that emerge quickly do not have higher yield at maturity and so are not going to incorporated into our breeding schemes. Population Development Trial results from field-grown plants harvested in December 2019 indicated a 206% rubber yield increase in the F generation when compared to source B generation material. All gains were made in root biomass as rubber concentrations remained constant. Large root size is a primary selection target for robust growth in soil and in hydroponic systems. The fourth generation of seed is being developed from selected crosses with tolerance to the Protoporphyrinogen oxidase inhibitor herbicides. These will be planted and tested for tolerance again. 1,814,000 seed was collected from, plants with ALS herbidice resistance (from natural slections). Development of a population capable of germination significantly below 0.25 inches has continued. We collected seed and replanted a depth study. Seed will be collected from a vernalized collection of plants, all of whichestablished after being planted at 1" or greater depth. currently about to flower in a greenhouse. The plants will be interbred and clean seed will be sent to our collaborators at the University of Nebraska, Scottsbluff, for field trials in 2021. This location requires deeper planting than Ohio for stand establishment because their soil surface dries out too quickly to permit germination of shallowly planted seed adn plant establishment. Selections are ongoing to create populations which will not flower without vernalization. F3 plants are vernalizing. Also, we have proven that an 80% flowering rate was achieved with only 3 weeks of vernalization. Golf generation (sixth generation of selections from an alpha generation selected for large size and high rubber content in field-grown transplants of seed from the USDA germplasm collection) were produced for field trials in Oregon, Nebraska and Ohio to select plants best adapted to these different growing regions. Unfortunately, the plantings failed in Oregon and Ohio due to too late planting windows caused by Covid19 shutdowns. However, Nebraska successfully harvested and shipped live plants back to Ohio for selection and crossing. Additionally, top performing Alpha generation plants were selected and crossed to produce a new breeding line called Washington. Seed Our project produces and supplies rubber root dandelion seed to several collaborators for both field and indoor controlled environment agriculture production trials. Current inventory is over 8 million seed from different populations and selections. 2.5 million were distributed to collaborators/partners for spring 2020 planting. Despite shutdowns and restrictions, 2.5 kg of seed (Foxtrot generation, selected fr high yield from golf field-grown plants) were produced and collected in this reporting period. This seed (4.9 million) will be cleaned, pelletized and distributed among growing partners. Molecular Enhancement: We have been taking multiple strategies to accelerate carbon routed to rubber biosynthesis. In one approach, a specific gene has been targeted to alter the flow of carbon from storage carbohydrates to rubber. The CRISPR/cas9 expression vector to knock out this gene was transformed into competent wild type A. rhizogenes K599 strain and A. tumefaciens EHA105 strain through electroporation. Rubber dandelion Fox Trot plants (Fox Trot is an advanced selected population with twice the root size of the initial population), were used for CRISPR/cas9 driven mutagenesis. Root fragments from 12 weeks old plants and leaves from 6-10 weeks old plants were used as explants for Agrobacterium-mediated plant transformation. Root fragments were transformed with A. rhizogenes harboring the CRISPR/cas9 construct and leaves were transformed with both A. rhizogenes and A. tumefaciens harboring the construct. The regenerated plantlets are being maintained in shoot elongation medium. In a second approach, site directed mutagenesis was carried out to generate the mutations A378V, S328L and V189I in the large subunit of RubisCO to reduce the ability of oxygen to inhibit carbon fixation from atmospheric carbon dioxide. More carbon captured should lead to more carbon being routed to plant growth or rubber biosynthesis (a terminal carbon sink). Mutations were confirmed with sequencing. Plasmid constructs were bombarded into TK wild type plant leaves. Three replicates were done for each construct. Non transformed plants were maintained as the control. The regenerated plantlets are maintained in root induction medium supplemented with 5 mg/l spectinomycin for selection. In a complementary project, we are targeting an enzyme implicated in in vivo latex coagulation. If we can reduce or eliminate latex coagulation, hydroponically grown rubber root dandelion could be used to produce latex for the manufacture of gloves - a vital component of personal protective equipment during the current COVID19 pandemic. CRISPR/Cas9 target sequences were selected using the OSU and Chinese rubber dandelion draft whole genomes and RNA seq data from six discrete rubber dandelion genotypes. The two gRNAs were modified to specifically target two adjacent sites in the rubber dandelion PPO-1 gene using PCR, utilizing four short primers, among which two forward and two reverse primers are partially overlapping. The Cas9 expression vector pKSE401 was purchased from Addgene (Addgene plasmid # 62202). The final expression vector was successfully constructed by ligating the modified two gRNA expression cassette with the pKSE401 vector through a restriction-ligation reaction, using Bsa1 and T4 ligase. Polymer Characterization For the first time, polymer macromolecular structural analysis of latex rubber extracted from transgenic guayule was attempted. Preliminary results support the hyothesis that designer rubber lines can be developed, expanding potentialmarkets for guayule and dandlelion rubber by producing stable rubber variants. Processing technologies Processing improvements for the extraction of industrial grade dandelion rubber included optimization of milling media (size and composition), mill rotational velocity, and enzyme composition and loading. Above 95% yields were achieved with 97.8% purity while reducing processing time from 48 h to 3 h - a highly significant improvement. Leveraging of Results The data from this project and from related grants provided the preliminary information needed to support several submitted grant applications, including •NSF FMG •USDA-NIFA-AFRI (3), •NSF-ERC preproposal, •DOE-BOTTLE (2) •DOE NTEL •USDA-NIFA SBIR (Phase i) (4) •OSU-CFAES SEED Note: Progress in TEA and LCA is reported in a different Hatch project (led by Dr. Ajay Shah)

Publications

  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Ren, X., Barrera, C.S., Tardiff, J.L., Gil A., Cornish, K., Liquid guayule natural rubber, a renewable and multifunctional rubber additive. Journal of Cleaner Production 2020, 122933.
  • Type: Book Chapters Status: Published Year Published: 2020 Citation: 5. Abdul Ghaffar, M.A., Cornish, K. New Developments in rubber particles biogenesis of rubber-producing species. Chapter 10: 153-168, In: The Rubber Tree Genome, Compendium of Plant Genomes, Eds: Minami Matsui and Keng-See Chow, Springer Nature Switzerland AG, 2020.
  • Type: Book Chapters Status: Published Year Published: 2020 Citation: Cornish, K. Perspectives and on-going challenges. Chapter 11: 169-175. In: The Rubber Tree Genome, Compendium of Plant Genomes, Eds: Minami Matsui and Keng-See Chow, Springer Nature Switzerland AG, 2020.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: 7. Okonkwo, C.C., Ujor, V., Cornish., K., Ezeji, T.C., Inactivation of the levansucrase gene in Paenibacillus polymyxa DSM 365 diminishes exopolysaccharide biosynthesis during 2,3-butanediol fermentation. Applied and Environmental Microbiology 86:e00196-20. 2020.
  • Type: Journal Articles Status: Published Year Published: 2020 Citation: Zhao, X., Cornish, K., Vodovotz., Narrowing the gap for bioplastic use in food packaging: an update. Environmental Science and Technology, 2020. Pages a-u. https://dx.doi.org/10.1021/acs.est.9b03755
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Cornish, K. Valorization and scalability of alternative natural rubber through tailoring properties, Proceedings of the Tire Technology International Conference, Hannover, Germany, February 25-27, 2020.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Cornish, K. Minimizing risks around NR through alternative crops. Proceedings of the Tire Technology International Conference, Hannover, Germany, February 25-27, 2020
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: 11. Zhao, X., Ji, K., Koelling, K., Cornish, K., Vodovotz, Y. Optimal mechanical properties of biodegradable natural rubber-toughened PHBV bioplastics intended for food packaging applications. Food Packaging and Shelf Life 21: 100348, (11 pages) 2019.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Jara, F.M., Cornish, K., Carmona, M. Potential applications of guayulins to improve feasibility of guayule cultivation, Agronomy 9: 804, 2019.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Barrera, C.S., Cornish, K., Review: Characterization of agricultural and food processing residues for potential rubber filler applications. Journal of Composites Science 3: 102-122, 2019. doi:10.3390/jcs3040102
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 15. Cornish, K., Valorization and scalability of alternative rubber through properties and production processes. Proceedings of the International Conference on Development of Alternatives Sources of Natural Rubber, Montpellier, France, November 4-6, 2019.
  • Type: Other Status: Published Year Published: 2019 Citation: Cornish, K., Rubber dandelions. Natural History Magazine. October 2019, 15-19, 2019.
  • Type: Other Status: Published Year Published: 2019 Citation: Cornish, K., Kopicky, S.E., Madden, T., Hydroponic cultivation of rubber dandelion has high annual rubber yield potential. Rubber and Plastics News, Technical Note Oct 7: 23-25, 2019.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 18. Zhao, X., Storts, S., Cornish, K., Vodovotz., Y. Natural Rubber-Toughened PHBV/PLA Bioplastic Films for Quieter Packaging Applications. Proceedings of the Fall 196th Technical Meeting of Rubber Division, ACS, Cleveland, OH, October 8?10, 2019, ISSN: 1547?1977
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ifeyinwa Ijeoma Obianyo, Azikiwe Peter Onwualu, Alfred B. O. Soboyejo, (2019), Mechanical Behaviour of Lateritic Soil Stabilized with Bone Ash and Hydrated Lime for Sustainable Building Applications, Case Studies in Construction Materials, https://doi.org/10.1016/j.cscm.2020.e00331
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ijeoma, Ifeyinwa. Anosike-francis, Esther Nneka. Odochi, Gina. Geng, Yang. Jin, Ruoyu. Peter, Azikiwe. Soboyejo, Alfred B O. 2020. Multivariate Regression Models for Predicting the Compressive Strength of Bone Ash Stabilized Lateritic Soil for Sustainable Building. Construction and Building Materials 263: 120677, 2019. https://doi.org/10.1016/j.conbuildmat.2020.120677.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Cornish, K., Natural Rubber Security and High-Performance Biobased Materials, ACS Rubber Division, International Elastomer Conference (Virtual), October 19-23, 2020
  • Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: Nikita Amstutz, Brandon Wheeler, and Katrina Cornish Emergence Differentiation and Yield in Taraxacum kok-saghyz OSU-CFAES Research Conference, 2020
  • Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: David Lankitus, Y. Zhang, K. Benzle, M. Liu, J. Fresnedo-Ramirez, & K. Cornish Enhancing rubber dandelion germplasm by increasing mevalonate pathway activity OSU-CFAES Research Conference, 2020
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Cornish, K. Convergent Rubber Science and Engineering, NSF Convergent Accelerator Workshop, Sept 30, 2020
  • Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: M. DORGET, S. SAKR, K. CORNISH, Towards an European Dandelion Rubber Supply Chain! RUBBERCON 2020, Paris, France, June 4-55, 2020
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Salgado, A.; Mancl, K.M.; Puskas, J.E. Tire Consumption and Recycling in South-East Asia. Chemistry and Materials Engineering for Industry Symposium, December 6, Detroit, Michigan (2019).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: de Freitas, M.A.; Mancl, K.M.; Puskas, J.E. Natural Rubber from Hevea brasiliensis: Economic Progress or Environmental Regress? Chemistry and Materials Engineering for Industry Symposium, December 6, Detroit, Michigan (2019)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Puskas, J.E. Sustainable Polymers: Closing the Loop. Chemistry and Materials Engineering for Industry Symposium, Dec. 6, Detroit, MI (2019).
  • Type: Other Status: Published Year Published: 2019 Citation: Dandelions as a cash crop? by Kamie Stephen, Start-Herald, Scottsbluff, NE https://wahoo-ashland-waverly.com/agriculture/latest/dandelions-as-a-cash-crop/article_de48bfc3-484e-5d97-b60e-df001ba2f95c.html This was distributed through a state-wide group of newspapers and at least 25,000 subscribers received it.


Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Industry, researchers, general public, program managers, middle and high school students and teachers, technology transfer personnel, international alternative rubber scientists Changes/Problems:No major changes were made. With respect to field work, the 2019 spring was very rainy, so some anticipated planting dates were delayed, and subsequent planting times were adjusted accordingly. What opportunities for training and professional development has the project provided?Project personnel continue to keep up with pesticide training. Training in advanced Excel was broadly provided. How have the results been disseminated to communities of interest?Industry executives, other researchers What do you plan to do during the next reporting period to accomplish the goals?Herbicide experiments will be repeated and a one acre production field will be attempted with chemical weed control. Polyploids will be identificed and characterized. Population selections and advancement will continue. Polymer macromolecular structure will be characterized. Hydoponionically grown plahnts will be evaluated. Extraction and purificationprocesses will be improved.

Impacts
What was accomplished under these goals? Field experiments were conducted to evaluate the interaction between seedbed preparation and planting method; evaluate field performance of most 'advanced' TK lines available; determine optimum time of planting for successful TK establishment; evaluate treatments that suppress weeds without injury to the TK crop; and evaluate postemergence herbicide treatments that suppress weeds without injury to the TK crop. Establishment success was about 40% of viable seeds sown; subsequent survival ranged from 65 to 81%. Weed control can be up to 86% depending on weeds present. Crop injury varied greatly with herbicide and application conditions. Low injury was usually balanced by marginal weed suppression. Planting on a raised soil bed resulted in larger roots; surface-applied compost increased initial seedling survival. The 2018 TK field trial/ generational comparison (A,B,C,D,E) was harvested in December. Rubber yield results were as expected and each successive generation outperformed the previous with E generation exhibiting a 47% increase from the source material B. Selections were made from the E field population based on root size, 50 plants were selected for advancement and moved to the greenhouse for crossing and propagation of the next generation of seed, F. Screened F1 Potential Biennials were also harvested from the field in December and brought into the greenhouse for crossing and propagation of the F2 generation. Greenhouse screening of the PB F2s began in March 2019 and will continue for one year. Four TK lines were planted in May for the 2019 field nursery (A for examination and selection of more wildtype germplasm, B for control and comparison, F for selection, and PB for screening and selection). Greenhouse experiments included the completion of a mycorrhizae amendment experiment, a vernalization experiment to determine an optimal period, an emergence differentiation experiment to determine if selecting for early emergence has an effect on yield (ongoing), and propagation of inbred lines based on high rubber concentration (ongoing). In the production greenhouses, two complete cycles, and a partial cycle, were harvested generating over 2 kg of E seed. A post-harvest storage experiment was conducted using 25 diploid (2n) and 25 triploid (3n) TK plants. Two-month-old plants were transplanted into raised beds in a Madison Greenhouse (MGH) and were harvested after 6 months (February, 2019). Plants were stored at5-8°C.Latex quantification occurred at days 0, 20, 33, 49 and 62 on five 2n and five 3n plants (selected randomly) at each time point.Root bagasse was dried, ground and scanned with NIRS to predict rubber concentration. Rubber content per rootwas stable during the stroage period.Collected 5,000 seed from 4nG0 and 3nG1 plants. Proprietary CRISPR/Cas9 gene editing and site directed mutatgeneiss was performed to generate improved germplasm. Processing oils are rubber additives commonly used to improve the processability of a compound. Most processing oils are petroleum-based, like naphthenic oil (NO), and negatively impact the carbon footprint of the rubber industry. Moreover, these aids act as diluents that lower the mechanical performance of the resultant rubber products. In this research, liquid guayule natural rubber (LGNR) was produced by thermal degradation of guayule natural rubber and tested as a sustainable alternative to NO in natural rubber (Hevea and guayule rubber) and synthetic (styrene-butadiene) rubber composites filled with carbon black (CB). LGNR effectively facilitated rubber compounding. Moreover, unlike NO, LGNR acted as an active additive in the vulcanization reaction. Mechanical properties, durability and dynamic mechanical properties increased when NO was replaced with LGNR in the compounds. The higher mechanical properties may be explained by additional crosslinking networks formed between LGNR and the rubber matrices and strong LGNR-CB interactions. LGNR, as a renewable processing aid, can address rising performance goals, and reduce carbon footprint for rubber products. Processing oils are rubber additives commonly used to improve the processability of a compound. Most processing oils are petroleum-based, like naphthenic oil (NO), and negatively impact the carbon footprint of the rubber industry. Moreover, these aids act as diluents that lower the mechanical performance of the resultant rubber products. In this research, liquid guayule natural rubber (LGNR) was produced by thermal degradation of guayule natural rubber and tested as a sustainable alternative to NO in natural rubber (Hevea and guayule rubber) and synthetic (styrene-butadiene) rubber composites filled with carbon black (CB). LGNR effectively facilitated rubber compounding. Moreover, unlike NO, LGNR acted as an active additive in the vulcanization reaction. Mechanical properties, durability and dynamic mechanical properties increased when NO was replaced with LGNR in the compounds. The higher mechanical properties may be explained by additional crosslinking networks formed between LGNR and the rubber matrices and strong LGNR-CB interactions. LGNR, as a renewable processing aid, can address rising performance goals, and reduce carbon footprint for rubber products. Production of petroleum-based synthetic rubbers (SR) causes an enormous carbon footprint for the rubber industry. Carbon footprint would be reduced if natural rubber (NR) could take a larger market share and replace significant quantities of SR. However, SR has higher oil resistance and stiffness than NR and in applications where these properties chemically-modified NR will be required. Epoxidation is a chemical modification of NR which partially converts unsaturated bonds on the backbone of NR to epoxy groups. In this research, epoxidized guayule natural rubber (EGNR)/ guayule natural rubber (GNR) blends, and GNR were used to make carbon black (CB) filled composites. The processability, mechanical properties, swelling behaviors and dynamic mechanical properties were characterized at various epoxide fractions. Composites made with EGNR/GNR blends had higher oil resistance, wet traction and stiffness than GNR composites, although tensile strength and elongation at break were reduced by epoxidation. EGNR is expected to lead to development of new NR products with similar properties to SR, eroding SR markets and increasing the sustainability of the rubber industry. A new lab is being set up for Puskas. Most important is the Size Exclusion Chromatography/Field Flow Fractionation equipment (~$400,000 value) that will be used for molecular weight measurements. A new mill, DSC, TGA, GC-MS and other ancillary equipment is available.

Publications

  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ren, X., Cornish, K., Eggshell improves dynamic properties of durable guayule rubber composites co-reinforced with silanized silica. Industrial Crops and Products 138: 111440 https://doi.org/10.1016/j.indcrop.2019.06.003
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Cornish, K., Bates, G.M., Slutzky, J.L., Meleshchuk, A., Xie, W., Sellers, K., Mathias, R., Boyd, M., Caste�eda, R., Wright, M., Borel L. Extractable protein levels in latex products, and their associated risks, emphasizing American dentistry. Biology and Medicine 11:2 (7 pages) 2019. DOI: 10.4172/0974-8369.1000456
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2019 Citation: Ren, X., Geng, Y., Soboyejo, A.B.O., Cornish K., Reinforced mechanical properties of functionalized silica and eggshell filled guayule natural rubber composites, Rubber Chemistry and Technology, (accepted 11/12/2018) 2019.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Junkong, P., Matsushima, Y., Phakkeeree T., Cornish, K., Ikeda, Y. Influence of strain-induced crystallization on stress softening of sulfur cross-linked unfilled guayule and dandelion natural rubbers. Rubber and Chemistry Technology 92: 388-398, 2019.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Dhanania, S., Mahata, D., Prabhavale, O., Cornish, K., Nando, G.B., Phosphorylated cardanol prepolymer grafted guayule natural rubber: an advantageous green natural rubber, Iranian Polymer Journal 27: 307-318, 2018.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Junkong, P., Takami, S., Miyajib,K., Cornish, K., Ikeda, Y., Origin of unique features for guayule natural rubber. Proceedings of the International Conference on Advanced and Applied Petroleum, Petrochemicals, Polymers, Chulalongkorn University, Thailand, Dec 18-20, 2018.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Bates, G.M., Cornish, K., Rapid and complete removal of guayule (Parthenium argentatum) leaves by cryodefoliation, and freeze and thaw induction of rubber particle coagulation. Industrial Crops and Products 125: 491-495, 2018.
  • Type: Other Status: Published Year Published: 2018 Citation: Cornish. K., What is the chance of herbicide resistance in Buckeye Gold  the rubber dandelion industrial crop  jumping over to common dandelion? Ohios County Journal April 19, 2018. http://ocj.com/2018/04/what-is-the-chance-of-herbicide-resistance-in-buckeye-gold-the-rubber-dandelion-industrial-crop-jumping-over-to-common-dandelion/
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Junkong, P., Matsumishima, Y., Cornish, K., Ikeda, Y. Study on Guayule and Dandelion Natural Rubbers for Sustainability. Proceedings of the International Rubber Conference, Kuala Lumpur, Malaysia, Sept 4-6, 2018.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Cherian, S., Ryu, SB, Cornish, K., Natural rubber biosynthesis in plants, the rubber transferase complex, and metabolic engineering progress and prospects. Plant Biotechnology Journal (2019), pp. 121 doi: 10.1111/pbi.13181
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Bates, G.M., McNulty, S.K., Amstutz, N.D., Pool, V.K., Cornish, K., Planting density and harvest season effects on actual and potential latex and rubber yields in Taraxacum kok-saghyz. Hort Science, 54: 1338-1344, 2019.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Ramirez-Cadavid, D.A., Cornish, K., Hathwaik, U., Kozak, R., McMahan. C.M., Michel, F.C.,Jr. Development of novel processes for the aqueous extraction of natural rubber from Taraxacum kok-saghyz (TK). J Chem Technol Biotechnol, 2019, DOI 10.1002/jctb.6027
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: 8. Zhao, X., Ji, K., Koelling, K., Cornish, K., Vodovotz, Y. Optimal mechanical properties of biodegradable natural rubber-toughened PHBV bioplastics intended for food packaging applications. Food Packaging and Shelf Life 21: 100348, (11 pages) 2019.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Zhao, X., Cornish, K., Vodovotz, Y. Synergistic mechanisms underlie the peroxide and coagent improvement of natural rubber toughened poly (3-hydroxybuty-rate-co-3-hydroxyvalerate) mechanical performance. Polymers I Special Issue: Recent Advances in Bioplastics: 26;11(3). pii: E565. doi: 10.3390/polym11030565.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: 15. Zhao, X., Venoor, V., Koelling, K., Cornish, K., Vodovotz, Y., Bio-based blends from poly (3-hydroxybutyrate-co-3-hydroxyvalerate) and natural rubber for packaging applications, Journal of Applied Polymer Science, 136, 47334-47348, 2019. DOI: 10.1002/APP.47334
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Cornish, K., Crosby, P., Ogajiaku, C., Slutzky, J.L., Li, Z. Guayule latex strippable paints and coatings for particulate removal. Proceedings of the International Latex Conference, Fairlawn, Ohio, August 6th and 7th, 2019.


Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Industry, researchers, general public, program managers, middle and high school students and teachers, technology transfer personnel Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We have provided training and professional development to four graduate students and one post-doctoral researcher, a capstone team of senior undergraduates. These opportunities have included presentations at OSU and national and international conferences. Specialized training on specific equipment has also been provided. How have the results been disseminated to communities of interest?Many posters and oral presentations have been made to academic and industrial audiences, growers and general public audiences, including at the Wayne County Fair. Also, a variety of press reports have extended our reach. for example: http://ocj.com/2018/04/what-is-the-chance-of-herbicide-resistance-in-buckeye-gold-the-rubber-dandelion-industrial-crop-jumping-over-to-common-dandelion/ What do you plan to do during the next reporting period to accomplish the goals?Research and development will continue under the logical progression of the work decribed in this report.

Impacts
What was accomplished under these goals? We have continued to improve our seed production capacity and currently have 5 million clean viable seeds stored. We demonstrated that rubber dandelion may be grown on three contrasting Ohio soil types and similar root and rubber yields were obtained. Selections are ongoing to develop improved populations, included a true biennial line (no issue with seed dispersal when grown as an annual), larger root size, and higher rubber yield. CRIPST/Cas 9 gene editing is being used to improve carbon capture, yield and herbicide tolerance and progeny of edited parents are being evaluated. A major accomplishment this year was our successful submittal and award of a BRDI grant for $2 million lead by OSU, in collaboration with Oregon State University and the University of Nebraska. The funds will primarily focus on the optimization of agronomic production of rubber dandelion across these states, as well as further processing improvements. Rubber can now be extracted from roots at >99% purity. Our sustainability goals also are addressed by waste derived fillers. Our egg-shell derived reinforcing filler has been proven in natural rubber composites as a partial and full replacement of bifunctionalized silica filler. The eggshell particles improve the dispersion of the silica particles for a synergistic improvement of mechanical performance over either filler alone. This material is now being manufactured in 500 kg lots and our eggshell industrial partner (a major egg processor) is moving toward commercial scale production. Ford Motor Company, one of our rubber consortium (PENRA) partners, has developed elastomeric product applications including eggshell fillers, which meet performance specifications, to address their company's sustainability goals. With additional support from a grant from the NSF I/UCRC Center for Applied Processing and Packaging, we have further improved the properties of rubber-toughened biodegradable bioplastics by dynamic vulcanization of, using bioplastic (PHBV), natural rubber, food grade organic peroxide and coagent. Freezer to microwave trials have demonstrated satisfactory mechanical performance with below limits migration of food or color from the food into the package. This bioplastic rubber blend could be scaled up to commercially-meaningful volumes and a major international food company is exploring this opportunity.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Li, Z., Cornish, K. Guayule medical radiation attenuation glove. Rubber & Plastics News, Technical Note, October 2, 20-25, 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Junkong, P., Cornish, K., Ikeda, Y., Characteristics of mechanical properties of sulphur cross-linked guayule and dandelion natural rubbers. RSC Advances 7: 50739-50752, 2017.
  • Type: Journal Articles Status: Published Year Published: 2107 Citation: Barrera, C.S., Cornish, K., Alternate Reinforcing Fillers can Reduce Power Consumption during Compounding. Rubber & Plastics News, Technical Note, October 30, 16-18, 2017.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Luo, Z., Iaffaldano, B.J., Cornish, K., Colchicine-induced polyploidy has the potential to improve rubber yield in Taraxacum kok-saghyz. Industrial Crops and Products 112: 75-81, 2018.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Debat, H., Luo, Z., Iaffaldano, B.J., Zhuang, X., Cornish, K. Molecular Identification and Characterization of Two Rubber Dandelion Amalgaviruses, Annals of Applied Biology (on line 12/6/2017) doi: http:?/dx.doi.org/10.1101/229443
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Barrera, C.S, Soboyejo, A.B.O., Cornish. K., Quantification of the contribution of filler characteristics on natural rubber reinforcement using principal component analysis. Rubber Chemistry and Technology 91:79-96, 2018. DOI: 10.5254/rct.82.83716
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Cornish, K., Young, C.Y., Challenges and solutions to scale up of unconventional but viable bioenergy feedstocks. The Digest 2018.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Adams, R.P., TeBeest, A.K., McNulty, S.K., Holmes, W., Bartel, J.A., Cornish, K., Geographic variation in natural rubber yields in natural populations of Helianthus annuus (Asteraceae, Sunflowers). Phytologia 62:100(1), 62-70, 2018.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Ramirez-Cadavid, D.A., Valles-Ramirez, S., Cornish, K., Michel Jr., F.C. Simultaneous quantification of rubber, inulin, and resins in Taraxacum kok-saghyz (TK) roots by sequential solvent extraction. Industrial Crops and Products 122: 647656, 2018.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Cornish, K., Scott, D.J., Xie, W., Mau, C.J.D., Zheng, Y.F., Liu, X., Prestwich, G.D. Unusual subunits are directly involved in binding substrates for natural rubber biosynthesis in multiple plant species. Phytochemistry 156:55-72, 2018.
  • Type: Book Chapters Status: Published Year Published: 2018 Citation: Ren, X., Cornish, K., Waste conversion into sustainable and reinforcing fillers for rubber composites. Encyclopedia of Renewable and Sustainable Materials. Chapter 10547, 2018. https://doi.org/10.1016/B978-0-12-803581-8.10547-8
  • Type: Book Chapters Status: Published Year Published: 2018 Citation: Sikandar, S., Afzal, I., Ali, N., Cornish, K., Lignocellulose degrading thermophilic fungi and their prospects in natural rubber extraction from plants. In: Fungi in Extreme Environments: Ecological Role and Biotechnological Significance. Eds: Sonia M. Tiquia-Arashiro, Martin Grube, Pub. Springer. 2018.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Luo, Z., Iaffaldano, B.J., Zhuang, X., Fresnedo-Ramirez, J., Cornish, K. Variance, inter-trait correlation, heritability and marker-trait association of rubber yield related characteristics in Taraxacum kok-saghyz. Plant Molecular Biology Reporter 36(4), 576-587. 2018. doi.org/10.1007/s11105-018-1097-8,
  • Type: Other Status: Published Year Published: 2018 Citation: Cornish. K., What is the chance of herbicide resistance in Buckeye Gold  the rubber dandelion industrial crop  jumping over to common dandelion? Ohios County Journal April 19, 2018. http://ocj.com/2018/04/what-is-the-chance-of-herbicide-resistance-in-buckeye-gold-the-rubber-dandelion-industrial-crop-jumping-over-to-common-dandelion/