Seasonal and tissue variation in taxane concentrations of Taxus brevifolia.

Analysis of seven taxanes: taxol, baccatin III, 10-deacetyltaxol, 10-deacetylbaccatin III, 7-xylosyl-10-deacetyltaxol, cephalomannine and brevifoliol in extracts from bark and foliage of pacific yew (Taxus brevifolia) showed a gradient of decreasing concentration from stem base to branch tip. This decrease is attributed to the generally higher concentration of taxanes in the phloem tissue and the decrease in inner bark thickness from base to branch tip. Analysis of taxanes extracted from stem bark and needles sampled over a growing season showed that most taxane concentrations were significantly lower in the needles than in the bark. Typically, taxane concentrations in bark increased from May through August; whereas, in needles, concentrations changed little during that period. Two exceptions were baccatin III, which in the summer reached levels equivalent to bark, and brevifoliol which increased from March to August, reaching levels in needles nine times greater than bark.

Initiation and growth of cell lines of Taxus brevifolia (Pacific yew).

Conditions have been established for the induction and maintenance of callus cultures of Taxus brevifolia (Pacific yew) from bark, stem, and needle tissues. Cultures were established on a modified Gamborg's B5 medium, 1% sucrose, 0.2% casamino acids and 1 mg/L 2,4-D. There was no apparent inhibition of callus induction as a result of taxol concentration in the explant material. Cell lines derived from explants of individual trees were used to investigate growth characteristics. Although none of the cell lines contained taxol, some contained low levels of related taxanes. Variability was observed with each cell line in response to light, and auxin type and concentration. Growth index was most affected by cell line, followed by auxin type and concentration. These culturing methods may be useful for the goal of developing a highproducing cell line applicable for large-scale taxol production.

Influence of drought stress and low irradiance on plant water relations and structural constituents in needles of Pinus ponderosa seedlings.

The influence of low light on tolerance to prolonged drought was tested on unshaded and shaded seedlings of ponderosa pine (Pinus ponderosa var. scopulorum Dougl. ex Laws.). Unshaded seedlings of P. ponderosa var. ponderosa were also drought stressed to compare varietal responses to drought. The maximum irradiance received by shaded seedlings was 10% of full light. Seedlings were progressively drought stressed until predawn water potentials (Psi(x)) were -5.0 MPa. Relative water content (RWC) and the reciprocal of Psi(x) were analyzed by means of an unusual application of the pressure-volume relationship for determination of RWC of the apoplast (RWC(a)), osmotic potential at full turgor (Psi(oft)), and ratio of fully turgid weight to dry weight. Major varietal differences in drought response were in RWC(a) and needle cellulose content. The shaded seedlings showed tissue damage at relative water contents < 60%, and were killed by water deficits from which unshaded seedlings recovered. Correspondingly, shaded plants had significantly higher cell volume/cell mass ratio, Psi(oft), less cellulose in needle tissue, and lower RWC(a) than unshaded plants. These differences suggest that low irradiance restricts drought adaptation in ponderosa pine.

Differences in Proteins Synthesized in Needles of Unshaded and Shaded Pinus ponderosa var Scopulorum Seedlings during Prolonged Drought.

Proteins were radiolabeled and extracted from needles of Pinus ponderosa var scopulorum (Dougl. ex Laws.) seedlings progressively drought-stressed for about 1 month. A set of novel, low molecular weight proteins was detected in fluorographs of two-dimensional gels when relative water content of needles fell below 70%. Their synthesis was undetectable in the fully recovered seedlings within 48 hours after rewatering. In similarly stressed seedlings that were shaded to 10% full light, the low molecular weight polypeptides were not detected or appeared at very low levels. The shaded seedlings, in which drought tolerance was reduced, did not recover upon termination of the drought. The results suggest that protein synthesis induced by water deficit in drought-tolerant seedlings may contribute to resisting the effects of cellular dehydration.