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Basal media including Murashige and Skoog (MS), Woody Plant Medium (WPM), and Driver and Kuniyuki (DKW) media were compared for use in guayule regeneration and propagation. Explants grown on MS basal medium showed large size calli with multiple shoots, but the calli produced on DKW medium were of higher quality, and longer true shoots were produced. Adventitious root formation was best in WPM basal medium. A comparison of macro and micro nutrient levels in MS, DKW, and WPM media revealed significant differences in the calcium levels. Under controlled conditions, guayule plants in vitro incorporated calcium into plant tissues in proportion to the amount available. Moreover, the addition of calcium nitrate tetrahydrate at moderate levels into MS medium significantly improved the emergence of single, elongated true shoots, suggesting a requirement for calcium in guayule tissue culture. We have further explored selection agent (antibiotic) concentration for the identification of transgenic plants using a spectinomycin resistance gene (aadA). Based on the results, we have improved tissue culture protocols for guayule including spectinomycin as the selection agent.

Populus serves as a model tree for biotechnology and molecular biology research due to the availability of the reference genome sequence of Populus trichocarpa (Torr. & Gray) genotype ‘Nisqually-1’. However, ‘Nisqually-1’ has been shown to be very recalcitrant to micropropagation, regeneration and transformation. In this study, a highly efficient micropropagation protocol from greenhouse-grown shoot tips of ‘Nisqually-1’ was established. The optimal micropropagation protocol involves growing in vitro shoots in plant growth regulator-free Murashige and Skoog (MS) basal medium supplemented with 3% sucrose, 0.3% Gelrite® and 5–10 g L−1 of activated charcoal. Plants grown on this medium were significantly longer, and contained significantly higher concentrations of chlorophyll. This highly effective protocol provides a consistent supply of quality leaf and stem materials throughout the year for transformation experiments and other in vitro manipulations, therefore eliminating inconsistency due to seasonal and greenhouse environmental variations and the need for repetitive tissue sterilization.

COBRA proteins have been shown to be involved in both cell wall expansion and/or cellulose deposition. In this paper, we analyzed all 18 COBRA genes (PtCOBRA) from the completely sequenced Populus trichocarpa genome. The 14-member PtCOBRA subfamily I proteins have high similarities to the Arabidopsis (At) COB subfamily, and members with full length sequences were predicted to possess significant potentials for a GPI-anchor site. The 4-member PtCOBRA subfamily II proteins are 45% longer than subfamily I proteins and lack ω-attachment sites at the C terminus, and are more similar to AtCOBL7 subfamily. The expression of the Populus COBRA family genes were regulated in a tissue-specific manner, and were shown to also respond differentially to inductions of hormones and environmental stimuli which affect plant cell expansion. The high levels of expressions, particularly in shoot tip and young root organs, suggests that at least some Populus COBRA genes are likely involved in regulating cell expansion.

Auxin is a plant hormone that regulates many processes of plant growth such as apical dominance, cell growth, adventitious rooting, and fruit and seed development. Expression of a family of YUCCA genes important for auxin biosynthesis has been shown to be spatially and temporally regulated in Arabidopsis. In this study, we report on the identification of 12 YUCCA genes from the completely sequenced Populustrichocarpa genome database and characterized them by comparing them with ArabidopsisYUCCA genes. The PopulusYUCCA genes are distributed on eight chromosomes. All Populus and Arabidopsis YUCCA genes can be divided into two phylogenetic groups, one of which can be divided into two subgroups. Populus YUCCA genes are expressed in the shoot tip, immature and mature leaf, young root, stem, and bark tissues in an tissue-specific manner. Transcript accumulation of the Populus YUCCA genes is reduced by exogenous applications of various plant growth regulators except auxin. Steady-state mRNA levels of different Populus YUCCA genes vary considerably depending on the experimental treatments. These results suggest that, similar to the Arabidopsis and rice YUCCA genes, the Populus YUCCA genes may also have diversified roles in Populus growth and development, and their expression may be regulated in a complex manner by developmental and environmental cues.