Gene-pool variation in caledonian and European Scots pine (Pinus sylvestris L.) revealed by chloroplast simple-sequence repeats.

We have used polymorphic chloroplast simple-sequence repeats to analyse levels of genetic variation within and between seven native Scottish and eight mainland European populations of Scots pine (Pinus sylvestris L.). Diversity levels for the Scottish populations based on haplotype frequency were far in excess of those previously obtained using monoterpenes and isozymes and confirmed lower levels of genetic variation within the derelict population at Glen Falloch. The diversity levels were higher than those reported in similar studies in other Pinus species. An analysis of molecular variance (AMOVA) showed that small (3.24-8.81%) but significant (p < or = 0.001) portions of the variation existed between the populations and that there was no significant difference between the Scottish and the mainland European populations. Evidence of population substructure was found in the Rannoch population, which exhibited two subgroups. Finally, one of the loci studied exhibited an allele distribution uncharacteristic of the stepwise mutation model of evolution of simple-sequence repeats, and sequencing of the PCR products revealed that this was due to a duplication rather than slippage in the repeat region. An examination of the distribution of this mutation suggests that it may have occurred fairly recently in the Wester Ross region or that it may be evidence of a refugial population.

The effect of previous defoliation of pole-stage lodgepole pine on plant chemistry, and on the growth and survival of pine beauty moth (Panolis flammea) larvae.

A study of the effects of defoliation by insects on the chemistry of lodgepole pine (Pinus contorta), and on the performance of Panolis flammea (Lepidoptera; Noctuidae) larvae, was carried out in a forest in northwest Scotland I year after a severe outbreak of P. flammea had caused extensive defoliation. Larval weight and survival were not significantly different on trees that had experienced different levels of defoliation in 1986. The nitrogen and tannin content of current and previous years' pine needles was not significantly affected by defoliation (although both were slightly greater in the foliage of defoliated trees). Phosphorus content of young pine foliage was lower (but not significantly lower except on one occasion) on heavily defoliated trees. On all sampling occasions, however, the nitrogen: phosphorus ratio was significantly higher on heavily defoliated trees. There were large differences in monoterpene composition of the previous year's shoots associated with defoliation intensity, but these differences had largely disappeared in the new growth. The results are discussed in relation to other studies on the effects of insect damage on plant chemistry and insect performance and in relation to the abundance of P. flammea in Scotland.

CALEDONIAN SCOTS PINE: ORIGINS AND GENETIC STRUCTURE.

Monoterpene and isozyme loci, used as markers to study the genetic structure of Scots pine (Pinus sylvestris L.) native to Scotland, showed that the endemic populations are not genetically impoverished, in spite of severe contraction in range and numbers as a result of both natural and anthropogenic causes. On the contrary, variability in the relict populations is almost the highest: of any plant species studied, with average heterozygosities of 0.33 for monoterpenes (rive loci) and 0.30 for isozymes (16 loci). The overwhelming proportion of this variability (> 95 %) was within populations, even though significant differences in gene frequencies of many individual loci existed among populations. Multiple-locus comparison of gene frequencies among populations, resolved by canonical variate analysis, showed no coherent geographic pattern of differences from population to population or region to region, with one major exception: certain populations in northwestern Scotland (Wester Ross) were distinct from all others and each other. The pattern of variability of the biochemical markers was consistent with that of metrical and physiological traits reported in the literature. These traits, in turn, show relatively little genetic affinity between contemporary Scottish and continental European populations. The genetic evidence, together with the anomalous distribution of pine pollen in the British Isles during the Holocene, suggests that the Caledonian race of Scots pine originated endemically from more than one refugium after the last glaciation.

Effects of light and darkness on polyphenol distribution in the tea plant (Camellia sinensis L.).

1. Flavonoid synthesis was able to proceed in darkness in young shoots and seedlings of the tea plant, but was increased by light. 2. The initial effect of darkness was to inhibit synthesis of the A ring or its linkage to the phenylpropane moiety of the flavonoid, but later the hydroxylation state of the flavanols was affected, leading to smaller proportions of gallocatechins and of complex leucoanthocyanins. 3. The esterification of catechins with gallic acid was less affected, so that the ratio of catechin gallates to simple catechins also increased. 4. The flavylogen content of darkened stems, especially in seedlings, was much less decreased than that of leaves; however, a short subsequent light-treatment caused an increase in polymerization.

The distribution of polyphenols in the tea plant (Camellia sinensis L.).

1. Methods for the separation and determination of the polyphenolic components of the tea plant by thin-layer chromatography and colorimetric reactions have been devised. 2. High concentrations of catechins, flavonols and depsides were found to be restricted to the young vegetative and floral shoots, whereas leucoanthocyanins or flavylogens were characteristic of the more bulky axial tissues of the plant. 3. In the young shoots cell growth was correlated with an increasing degree of flavonoid B-ring hydroxylation. 4. Maximal flavylogen concentrations occurred in the outer protective layers of stem and of seed coat. 5. Mature leaves were shown to contain derivatives of the flavones apigenin and luteolin. 6. Developing seedlings showed a steady increase in polyphenol complexity; flavylogens were concentrated at shoot and root apices and accumulated at the stem base. 7. It is postulated that the flavanols (leucoanthocyanins and catechins), because they can co-polymerize, are of use to the plant for protection of wood and bark against infection and decay.

Studies on the polyphenol metabolism of tissue cultures derived from the tea pant (Camellia sinensis L.).

1. The growth characteristics on various media of solid and liquid suspension cultures derived from the stem of the tea plant are described; chlorophyll and anthocyanin synthesis occurred in the light. 2. Only the simplest catechins and leucoanthocyanins were present in callus tissue, although oligomeric and polymeric leucoanthocyanin fractions were also represented. Light caused an increase in all monomeric components analysed, but inhibited polymerization of the leucoanthocyanins. 3. The polyphenol oxidase activity of cultures was comparable with that of the apical regions of the intact plant, and was inversely correlated with growth rate. 4. Growth was stimulated by hormonal variation, and inhibited by high concentrations of sucrose and by high light-intensity; polyphenol concentrations were generally inversely correlated with growth rate. 5. From the inability of callus tissue and of cultured root apices to synthesize complex catechins, it is inferred that complex catechin formation in intact plants is associated with the process of cell vacuolation.

The separation and distribution of simple and condensed leucoanthocyanins of the tea plant (Camellia sinensis L.).

1. Leucoanthocyanin monomers of high mobilities in aqueous solvents on thinlayer chromatograms, assumed to be structurally simple, were characteristic of mature bulky tissues, whereas members of lower mobility were confined to young vegetative and floral tissues. 2. Flavylogens were separated by gel filtration on Sephadex columns into monomeric, oligomeric and polymeric fractions. 3. The polymeric fraction from young brown stems was heterogeneous, one-half having a molecular weight of about 3400, one-third a molecular weight between 3600 and 17000, and the remainder a molecular weight of over 17000. 4. Leaves had low flavylogen concentrations; only monomers were present. Stem tissues were rich in polymers, which increased with the age of the young stem and decreased inwards through the wood. The maximal flavylogen concentrations were in the phloem and cambium from mature stems, where all three fractions were richly present. The periderm tissue and, to a lesser extent, the seed coat were characterized by a very high polymer/monomer ratio, exhibiting a much higher degree of polymerization than the wood. Root tissues contained high concentrations of monomers. 5. In general, there was an inverse correlation between the extent of polymerization and the complexity of the monomers present. 6. The results are in favour of the thesis that the function of the flavanols is, after polymerization to condensed tannins, to impregnate dead structural tissues and thereby to protect them from infection and decay.