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The Zinnia elegans cell suspension culture is excellent for xylogenesis studies at the cellular and molecular level, due to the high and synchronous in vitro differentiation of tracheary elements (TEs).
The percentage TE differentiation (%TE) in the culture is, however, influenced by a number of factors before and during cell differentiation. One of the factors that is potentially important but has not gotten much attention is the initial osmolarity of the plant material. To examine whether the growth conditions that determine leaf osmolarity (LO) affect the final %TE, we used three light intensities (50, 70 and 100 μmol.m-2s-1) and three electrical conductivity (EC) levels (EC 2, 4 and 6 dS.m-1 ) in hydroponic systems to induce different osmolarities in leaf materials from two cultivars (cvs) of Z. elegans, Envy and Purple Prince. The isolated leaf mesophyll cells were subsequently cultured in a liquid medium (300 mOsm extracellular osmolarity) containing α-naphthalene acetic acid (NAA) (1 mg.l-1) and benzylaminopurine (BA) (1 mg.l-1). The LO increased in both Zinnia cvs with increasing light intensity and increasing EC during growth. Mesophyll cell size correlated negatively with EC, but the correlation was positive with light intensity in both Zinnia cvs. There was an overall positive correlation between %TE and LO although the degree of %TE change versus LO differed between light and EC treatments and also between the two Zinnia cvs. Envy cv is the best known Z. elegans cv for establishing xylogenic cultures. However, it turned out that by subjecting the plants to different growth conditions, the Purple Prince cv produces a higher %TE as compared to the Envy cv. At EC4 the TE differentiation for the Purple Prince cv was 75%, a level that is 25 to 60% higher than those earlier reported. We conclude that light intensity and EC of the root environment affect the LO of Z. elegans which in turn influences the development and therefore dimensions of TEs in an in vitro xylogenic culture. Thus, proper optimization of the growth conditions for the Zinnia plants prior to establishment of xylogenic cultures leads to enhancement of in vitro TE formation.
Keywords: Electrical conductivity, in vitro culture, leaf osmolarity, light intensity, osmotic potential, tracheary element, xylogenesis, Zinnia elegans.