Colleen M. McMahan

We report functional genomics studies of a CYP74 rubber particle protein from Parthenium argentatum, commonly called guayule. Previously identified as an allene oxide synthase (AOS), this CYP74 constitutes the most abundant protein found in guayule rubber particles. Transgenic guayule lines with AOS gene expression down-regulated by RNAi (AOSi) exhibited strong phenotypes that included agricultural traits conducive to enhancing rubber yield. AOSi lines had higher leaf and stem biomass, thicker stembark tissues, increased stem branching and improved net photosynthetic rate. Importantly, the rubber content was significantly increased in AOSi lines compared to the wild-type (WT), vector control and AOS overexpressing (AOSoe) lines, when grown in controlled environments both in tissue-culture media and in greenhouse/growth chambers. Rubber particles from AOSi plants consistently had less AOS particle-associated protein, and lower activity (for conversion of 13-HPOT to allene oxide). Yet plants with downregulated AOS showed higher rubber transferase enzyme activity. The increase in biomass in AOSi lines was associated with not only increases in the rate of photosynthesis and non-photochemical quenching (NPQ), in the cold, but also in the content of the phytohormone SA, along with a decrease in JA, GAs, and ABA. The increase in biosynthetic activity and rubber content could further result from the negative regulation of AOS expression by high levels of salicylic acid in AOSi lines and when introduced exogenously. It is apparent that AOS in guayule plays a pivotal role in rubber production and plant growth.

Guayule (Parthenium argentatum A. Gray) is a rubber-producing desert shrub native to Mexico and the United States. Guayule represents an alternative to Hevea brasiliensis as a source for commercial natural rubber. The efficient application of modern molecular/genetic tools to guayule improvement requires characterization of its genome. The 1.6 Gb guayule genome was sequenced, assembled and annotated. The final 1.5 Gb assembly, while fragmented (N50 = 22 kb), maps > 95% of the shotgun reads and is essentially complete. Approximately 40,000 transcribed, protein encoding genes were annotated on the assembly. Further characterization of this genome revealed 15 families of small, microsatellite-associated, transposable elements (TEs) with unexpected chromosomal distribution profiles. These SaTar (Satellite Targeted) elements, which are non-autonomous Mu-like elements (MULEs), were frequently observed in multimeric linear arrays of unrelated individual elements within which no individual element is interrupted by another. This uniformly non-nested TE multimer architecture has not been previously described in either eukaryotic or prokaryotic genomes. Five families of similarly distributed non-autonomous MULEs (microsatellite associated, modularly assembled) were characterized in the rice genome. Families of TEs with similar structures and distribution profiles were identified in sorghum and citrus. The sequencing and assembly of the guayule genome provides a foundation for application of current crop improvement technologies to this plant. In addition, characterization of this genome revealed SaTar elements with distribution profiles unique among TEs. Satar targeting appears based on an alternative MULE recombination mechanism with the potential to impact gene evolution.

Parthenium argentatum, commonly known as guayule, is a desert shrub cultivated as a domestic source of natural rubber in the semi-arid southwestern United States. Guayule natural rubber (GR) may be used to replace petroleum-based rubber or in place of Hevea natural rubber (NR), but substitution must take into consideration differences in physical and chemical properties. Currently, Hevea NR is required in tire applications, especially aircraft and truck tires, because of its high oxidative resistance, rapid cure rate, and exceptional stress–strain response. These outstanding features are attributed to the presence of nonrubber constituents, mainly proteins and lipids, which cause the rubber to gel, and they contribute to strain-induced crystallization. In contrast, GR is low in proteins and is thus deprived of some attributes of Hevea. Addition of amino acids and proteins to guayule could potentially improve performance and thereby widen the range of applications for use. In a previous study, amino acids blended with GR latex improved thermo-oxidative stability, served as plasticizers and cure accelerators, and enhanced green strength slightly, but tensile strength was not improved. Here, a series of bio-based commercial proteins (gelatin, soy, albumin, casein, zein, gliadin, and gluten) were added to GR as a latex blend. In general, protein addition reduced bulk viscosity and improved thermo-oxidative stability. The gel content and green strength of the polymer–protein blends were increased, with the exception of gliadin, but not to levels observed for Hevea. Effects on vulcanization and mechanical properties in compounds were surprisingly influenced by the choice of antioxidants used. Our results demonstrate the potential of proteins as bio-based rubber compounding additives.

Guayule (Parthenium argentatum) is under cultivation in the southwestern United States as an alternative source of natural rubber free from proteins that cause Type I latex allergies. However, since guayule lacks the protein‐polymer interactions present in Hevea latex, its physical and chemical properties may differ. The solvent‐soluble (Sol) and insoluble (Gel) fractions from guayule and Hevea natural rubbers were isolated through a solubilization/centrifugation deproteinization process. Protein could be reduced or removed by centrifugation, or concentrated in the gel fraction for both Hevea and guayule rubber. Separation of the sol fraction of Hevea rubber reduced the overall protein level, in some cases to below detection limits, without impacting rubber thermo‐oxidative stability. Notably, no detectable cross reactions took place between guayule protein antibodies and Hevea‐based materials, nor vice‐versa. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42051.

Background: Parthenium argentatum (guayule) is an industrial crop that produces latex, which was recently commercialized as a source of latex rubber safe for people with Type I latex allergy. The complete plastid genome of P. argentatum was sequenced. The sequence provides important information useful for genetic engineering strategies. Comparison to the sequences of plastid genomes from three other members of the Asteraceae, Lactuca sativa, Guitozia abyssinica and Helianthus annuus revealed details of the evolution of the four genomes. Chloroplast-specific DNA barcodes were developed for identification of Parthenium species and lines. Results: The complete plastid genome of P. argentatum is 152,803 bp. Based on the overall comparison of individual protein coding genes with those in L. sativa, G. abyssinica and H. annuus, we demonstrate that the P. argentatum chloroplast genome sequence is most closely related to that of H. annuus. Similar to chloroplast genomes in G. abyssinica, L. sativa and H. annuus, the plastid genome of P. argentatum has a large 23 kb inversion with a smaller 3.4 kb inversion, within the large inversion. Using the matK and psbA-trnH spacer chloroplast DNA barcodes, three of the four Parthenium species tested, P. tomentosum, P. hysterophorus and P. schottii, can be differentiated from P. argentatum. In addition, we identified lines within P. argentatum. Conclusion: The genome sequence of the P. argentatum chloroplast will enrich the sequence resources of plastid genomes in commercial crops. The availability of the complete plastid genome sequence may facilitate transformation efficiency by using the precise sequence of endogenous flanking sequences and regulatory elements in chloroplast transformation vectors. The DNA barcoding study forms the foundation for genetic identification of commercially significant lines of P. argentatum that are important for producing latex.