THE FATE OF THE DWARF SHOOT APEX IN BRISTLECONE PINE (PINUS LONGAEVA)

The fate of the pine dwarf shoot (DS) apex after needle initiation has been controversial. Dwarf shoot primordia of Pinus longaeva were examined to determine the developmental basis for DS with and without interfoliar buds. Interfoliar buds are microscopic buds derived from the original terminal apex of the DS. In October, all the DS primordia are similar in size and appearance. However, as the needles elongate in the following June the apices of more proximal DS decrease in size, such that by July there is a clear diminishing size gradient of apical domes in going from the most distal to the most proximal positions. The distal DSs start to form bud scales in July and have fully formed interfoliar buds by mid-August. In contrast, those DS apices lacking protective bud scales at needle maturity become suberized and can never proliferate into long shoots. The distal placement of interfoliar buds may be due to a group effect, where each developing DS inhibits the more proximal DSs in the long shoot terminal bud.     

PINUS TRIPHYLLA AND PINUS QUINQUEFOLIA FROM THE UPPER CRETACEOUS OF MASSACHUSETTS

Dwarf shoots and needle leaves of Pinus triphylla Hollick and Jeffrey and P. quinquefolia Hollick and Jeffrey have been discovered recently in a Late Cretaceous age clay deposit on Martha's Vineyard Island, Massachusetts. Detailed study of these fossils provides further information on the internal construction of the two taxa. This new information permits both species to be compared easily with similar fossil forms from the Upper Cretaceous of Japan as well as with dwarf shoots and needles of modern species of Pinus. As a result, the relationships between Cretaceous pines and the extinct genus Prepinus are found to be less than previously believed. Moreover, the dwarf shoots and needles of Cretaceous members of the genus Pinus appear to be most like those of present-day species included in the subgenus Pinus.     

SECONDARY GROWTH IN NEEDLE LEAVES OF PINUS LONGAEVA (BRISTLECONE PINE) AND OTHER CONIFERS: QUANTITATIVE DATA

Needle leaves of Pinus longaeva can be accurately dated and remain alive on branches for 30 or more yrs, making this species ideal to study secondary growth in leaves. Field observations and regression analysis of needle age versus mean needle length both indicate primary (elongation) growth of needles is completed in the first year. Statistical analysis of cell counts for one- to 33-yr-old needles indicate production along the entire length of needles of 1.0-1.7 cell layers per year of secondary phloem, but no secondary xylem. Microscopic measurements and cell counts reveal that with advancing needle age phloem increases radially and transfusion tissue buckles, but the number of endodermal cells and xylem width do not change. Living phloem remains constant in amount (ca. 9 layers) with advancing needle age, indicating yearly replacement of old by new phloem. For comparison to P. longaeva, needle leaves were also studied for ten other conifer taxa with maximum needle longevities ranging from 3 to 19 yrs. In needles of each taxon no secondary xylem is produced, but secondary phloem production occurs throughout the post-elongation lifespan of the needles regardless of maximum needle longevity.     

Direct somatic embryogenesis and shoot regeneration from leaves and internodes of Pluchea lanceolata (DC.) C.B. Clarke

Pluchea lanceolata (DC.) C.B. Clarke is a threatened native medicinal plant. Increasing the propagation of this plant will preserve the wild population and provide material for medicinal use. In vitro and field-collected shoots and leaves were tested for response to 2,4-dichlorophenoxyacetic acid (2,4-D) and thidiazuron (TDZ), for initiation of direct shoot regeneration (DSR), or direct somatic embryogenesis (DSE). Leaves and internodes collected from field-grown plants produced only callus, while in vitro–raised shoots exhibited DSR and DSE on Murashige and Skoog (MS) medium with 2,4-D and TDZ. Direct shoot regeneration occurred on medium with TDZ from internode and leaf segments obtained from in vitro–developed shoots. In vitro–grown shoots were rooted on half-strength MS medium with 2 mg L⁻¹ indole-3-butyric acid and acclimatized. Survival in natural conditions was 62.5% for DSE and 79% for DSR plantlets.

     

SILICIFIED PINUS REMAINS FROM THE MIOCENE OF WASHINGTON

Silicified leaves, dwarf shoots, pollen cones, and seed cones of Pinus from a Late Miocene chert bed within the Yakima Basalt Formation near Yakima, Washington are interpreted as coming from a single new species, P. foisyi. The needles and dwarf shoots are those of a three-needle pine. The needles contain two to four medial resin canals, a biform hypodermis, and endodermal cells with uniformly thickened walls. The pollen cones are ellipsoidal and about 1 cm long, and many contain bisaccate pollen grains. The seed cones are at least 6 cm long and are slightly asymmetrical. The cone axis has a broad sclerotic outer cortex, and the seed wing extends from a thick parenchymatous base. The scale apex bears a conspicuously swollen projection. The foliage and seed cones are identifiable with the Subgenus Pinus, Section Pinus, Subsection Oocarpae independently of one another, and together indicate a fossil species related to the modem Californian closed cone pines. Pinus foisyi represents one of the earliest occurrences of cone asymmetry associated with this group. However, cone serotiny characteristic of the modem species appears to have evolved after the Late Miocene.     

Osservazioni sulla lunghezza e sulla longevità degli aghi di pino domestico (Pinus pinea L.)

Abstract

Observations on needle length and longevity of Pinus pinea L. - We investigated needle length and longevity of Pinus pinea L. to understand their ecological significance. Samplings regarded five coastal pinewoods (four in the province of Grosseto, one in the province of Lucca) and isolated trees in different sites of Tuscany. We found a significant variation in needle length among different positions of the crown (central, intermediate and peripheral), among individuals of the same site and among different sites; isolated trees showed longer needles and a longer retention than pines in stands. We also obtained a highly significant linear regression between length of different-aged needles (from 1988 to 1992) and rainfall of the period march-august of the same years. Our results suggest that, despite its high variability, needle ength is influenced by climatic conditions (overall by rainfalls) and by the water availability of the site where pines grow. Needle retention depends largely on the position of needles within the crown and on the drought effect: at the upper position, three or, seldom, four-year old needle can persist if the summer drought period is not too long and the site conditions are favourable; at the lower level only two-year old needles are retained. Needle longevity decreases sharply in years with severe drought, therefore it can be used as an index of climatic and edaphic stress.     

The effect of crown position and date of sampling on biomass,nutrient concentrations and contents of needles and shoots in European larch

 The nutrient concentrations and contents of needles and shoots of 22-year-old European larch (Larix decidua Mill.) were evaluated with respect to crown position, age of tissues and sampling date during a complete growing season. Concentrations of N, P, K, Ca, Mg and Zn in the needles and of N, P and K in the shoots differed significantly among the dates of sampling. The concentrations of N and Mn in the needles and all nutrients in the shoots (except Mg) also differed significantly with crown position. Maximum needle biomass was observed in the middle crown position (55% of the total) and maximum shoot biomass, in the lower crown position (52% of the total). Maximum needle and shoot nutrient contents were observed in the middle position of the living crown for long shoot, short shoot-1, short shoot-2, short shoot-3 and, short shoot-4 age classes while highest contents for short shoot-5 and short shoot-6 age classes were observed in the lower crown position. Biases up to 42% for Mg in the needles and 200% for K in the shoots were obtained when only long shoot tissues are used for content evaluation. For needles and shoots, Mg and K are more difficult nutrients to evaluate. A sampling methodology is proposed for evaluating nutrient contents of the living crown. Accepted: 10 August 1995     

Population history of European mountain pines Pinus mugo and Pinus uncinata revealed by mitochondrial DNA markers

The dwarf mountain pine (Pinus mugo) and the Pyrenean pine (P. uncinata) constitute a pair of closely related coniferous taxa of poorly resolved evolutionary history and affinity, which inhabit numerous stands scattered over subalpine environments of European mountain ranges. The aim of the study was to investigate their phylogeography and mutual relationships, shedding new light on their taxonomy and the past of the alpine flora. Previous evolutionary reconstructions of the mountain pines relied mainly on bi‐parentally or paternally inherited markers that quickly homogenize between populations, showing rather shallow and recent differentiation of gene pools. Therefore, to contrast these pictures, we analyzed diversity and differentiation within a large set of new mitochondrial loci, inherited in maternal line and distributed by seeds at short geographical distances. Samples of the taxa were taken from 27 natural populations representing their range‐wide distributions—17 populations of P. mugo and 10 of P. uncinata. All markers appeared polymorphic, providing a total of 31 multilocus haplotypes. Two of the loci proved to be species‐diagnostic and nearly fixed between analyzed samples. Distribution of mitotypes indicate that allopatric populations of the taxa constitute separate mitochondrial haplogroups, and the two mountain pines have independent evolutionary history. However, introgression of P. mugo mitotypes by P. uncinata specimens revealed in the species contact zone in Western Alps shows that their speciation is not fully completed.     

Spiral grain in bristlecone pines (Pinus longaeva) exhibits no correlation with environmental factors

Six hundred and eleven Great Basin bristlecone pines (Pinus longaeva) were surveyed in two separate groves in California’s White Mountains. The presence and direction of spiral grain were recorded for each tree as well as elevation above sea level, horizon angles, latitude and longitude, trunk diameter, whether the tree was dead, and whether the trunk was broken. The proportions of left-handed, right-handed and straight trees were similar in every part of both groves, although the groves lie at different elevations. No significant correlation was found between the direction of spiral grain and any environmental factor. The hypothesis that spiral grain is an adaptation to distribute sap evenly between the roots and the crown in Pinus longaeva is not strongly supported, since spiral grain is not correlated with asymmetric environments and most trees exhibit <90° rotation through the main stem. The data also do not support the idea that spiral grain makes the tree more resistant to breaking in strong winds. Right-handed spiral grain is predicted by this hypothesis, but most bristlecone pines are either left-handed or exhibit no spiral grain. Bristlecone pines are often uprooted from thin soils by strong winds, but rarely are the main stems broken by this mechanism. Spiral grain in Pinus longaeva growing in California’s White Mountains does not appear to be under environmental control.     

Somatic embryogenesis and shoot regeneration from intact seedlings of Phaseolus acutifolius A., P. aureus (L.) Wilczek,P. coccineus L., and P. wrightii L.

A rapid, one-step procedure has been developed for inducing direct organogenesis and somatic embryogenesis in cultures of Phaseolus coccineus L., P. acutifolius A., P. aureus L. [Vigna radiata L. Wilczek] and P. wrightii L. Development of somatic embryos and shoot buds occurred within 6–8 weeks of culture from intact seedlings raised on MS (Murashige and Skoog 1962) medium supplemented with N⁶-benzylaminopurine (BAP). Shoot buds or embryoids originated from subepidermal tissue of the regions adjacent to the shoot apex, hypocotyl and cotyledonary axils. While P. acutifolius and P. aureus were regenerated via shoot formation and P. wrightii by somatic embryogenesis, both embryogenesis and shoot regeneration were observed in P. coccineus. Relatively higher levels of BAP, 50–80 M, were found to be optimal for inducing regeneration while lower concentrations were ineffective. About 40–70 shoots and 70–250 somatic embryos were produced per responding seedling. Regenerated shoots and somatic embryos developed into whole plants on a basal medium or the one supplemented with 1 M naphthaleneacetic acid.     

Breeding performance and longevity of Tomicus destruens on Mediterranean and continental pine species

The pine shoot beetle Tomicus destruens (Wollaston) (Coleoptera: Curculionidae: Scolytinae) is one of the main pests of Mediterranean forests, where it is oligophagous on Mediterranean pines. However, possible global warming may make the insect move to higher latitudes and altitudes, allowing it to attack new pine species. In this respect, the aim of the present article was to assess both the acceptance and performance of T. destruens offered host and non‐host pine species. A no‐choice breeding experiment was set up under laboratory conditions, using logs of three Mediterranean (Pinus pinea L., Pinus pinaster Miller, and Pinus halepensis Aiton) and two continental (Pinus nigra Arnold and Pinus sylvestris L.) pine species. Log debarking at the end of adult emergence assessed parent fecundity, egg, and larval mortality. The quality of callow adults emerging from each tested pine was evaluated on the basis of their longevity on a semiartificial diet. Tomicus destruens colonised all tested pine species, but did not reproduce in Scots pine, taking about 79 days to complete development with no differences among pines tested. The best breeding performance, evaluated as female fecundity and adult production, was observed in P. halepensis, and the lowest in P. pinaster. On average, adults emerging from P. pinea survived longer (83 days) than from other pines, and adult longevity was the lowest in males emerging from P. nigra. Austrian pine, which under natural conditions is usually not a host of T. destruens, allowed insect development and adult production similar to P. pinea and P. pinaster.     

Release of phylogenetic constraints through low resource heterogeneity: the case of gall-inducing sawflies

Abstract. 1. A group of six unusual sawfly species, which do not conform to the phylogenetic constraints hypothesis as it has been applied to sawflies, was examined in natural populations. All species were in the genus Pontania (Hymenoptera: Tenthredinidae), which induce galls on leaves of willow species (Salicaceae). An understanding of these non‐conformist species was important as a test of the validity of the general hypothesis. 2. The six species of sawfly, Pontania mandshurica, P. cf. arcticornis, P. aestiva, P. arcticornis, P. pacifica, and P. nr. pacifica, showed no oviposition preference for long, vigorous shoots, in contrast to 37 documented tenthredinid species that have demonstrated such a preference. Rather, the non‐conformist species attacked the shortest shoot length classes more frequently and larval establishment in galls was successful. 3. The evident escape from the phylogenetic constraint, which commonly limits sawfly attack to the most vigorous shoots in a willow population, resulted from low apparent heterogeneity of the resources exploited by these Pontania species. At the time of female oviposition, shoots and leaves were too uniform to allow discrimination by females among shoot length classes, resulting in random, or near random attack of shoots. 4. The unusual relative uniformity of resources to which sawflies were exposed resulted from several characteristics. (1) Females emerged early relative to shoot growth phenology, making discrimination among shoot length and vigour difficult or impossible. (2) Low heterogeneity in leaf length resulted in resource similarity independent of shoot length. (3) Abscission of leaves occurred after emergence of larvae from leaf galls so that differential abscission of leaves in relation to shoot length became irrelevant. (4) In some cases, low variance in shoot lengths was evident in old ramets lacking long, vigorous shoots. Probably as a result of low resource heterogeneity, larvae survived well across all shoot length classes, revealing no ovipositional preference and larval performance linkage related to the exploitation of the longest shoot length classes in a population of willows, as in the conformist species. Therefore, larval survival did not provide positive feedback on female preferential behaviour for long shoots, as in the conformist species studied.     

Cones structure and seed traits of four species of large‐seeded pines: Adaptation to animal‐mediated dispersal

Seed dispersal selection pressures may cause morphological differences in cone structure and seed traits of large‐seeded pine trees. We investigated the cone, seed, and scale traits of four species of animal‐dispersed pine trees to explore the adaptations of morphological structures to different dispersers. The four focal pines analyzed in this study were Chinese white pine (Pinus armandi), Korean pine (P. koraiensis), Siberian dwarf pine (P. pumila), and Dabieshan white pine (P. dabeshanensis). There are significant differences in the traits of the cones and seeds of these four animal‐dispersed pines. The scales of Korean pine and Siberian dwarf pine are somewhat opened after cone maturity, the seeds are closely combined with scales, and the seed coat and scales are thick. The cones of Chinese white pine and Dabieshan white pine are open after ripening, the seeds fall easily from the cones, and the seed coat and seed scales are relatively thin. The results showed that the cone structure of Chinese white pine is similar to that of Dabieshan white pine, whereas Korean pine and Siberian dwarf pine are significantly different from the other two pines and vary significantly from each other. This suggests that species with similar seed dispersal strategies exhibit similar morphological adaptions. Accordingly, we predicted three possible seed dispersal paradigms for animal‐dispersed pines: the first, as represented by Chinese white pine and Dabieshan white pine, relies upon small forest rodents for seed dispersal; the second, represented by Korean pine, relies primarily on birds and squirrels to disperse the seeds; and the third, represented by Siberian dwarf pine, relies primarily on birds for seed dispersal. Our study highlights the significance of animal seed dispersal in shaping cone morphology, and our predictions provide a theoretical framework for research investigating the coevolution of large‐seeded pines and their seed dispersers.     

Morphological and histological differences in the development of dwarf mutants of sexual and somatic origin in diverse woody taxa

Morphological and histological observations were made on eight dwarf mutants arising either as seedlings from sexual reproduction or from somatic bud mutations (witches'-brooms in the crowns of normal trees). The most predominate morphological trait contributing to the expression of dwarfism in all taxa was the reduction of final internode lengths along the shoot axis. In taxa of sexual origin, with the exception of Prunus, there was a consistent reduction in the number of preformed leaves contained in the winter buds. In addition, in two taxa (Liquidambar and Tsuga) there was an almost complete absence of neoformed leaves and sylleptic branches on current year shoots. Conversely, in mutants of somatic origin there was no apparent reduction in the number of preformed leaves. Genetic dwarfness in this group resulted solely from decreases in final internode length. Significant differences in the cellular basis of dwarfism between mutants of different genetic origins are clearly evident. In dwarf trees arising from sexual reproduction, reduction in final internode length is attributed predominately to inhibition of mitotic activity in developing internodes resulting in highly significant decreases in final cell number, and not cell length. In mutants of somatic origin, the reduction in length of mature internodes results from a decrease in final cell length, rather than a decrease in cell number. Physiological mechanisms associated with the genetic expression of these morphogenetic differences are suggested.     

Needle area measurement by the cut method and estimation of specific leaf area inCryptomeria japonica

Needle surface area inCryptomeria japonica was measured using a newly proposed cut method. Sample needles of various lengths were taken from foliage shoots belonging to various height layers of nine trees growing at three sites. Needles were cut into small pieces with a hand-made cutter made of razor blades and washers. By measuring the circumference and thickness of each piece, its lateral area was calculated and summed to give the total surface area of the needle. For estimating the surface area of a needle (s), two linear parameters of needle size termedy _n* and /were proposed:y _n* was the distance between the needle tip and the uppermost point of attachment of the needle to the shoot, whilel was the distance between the needle tip and the lowermost point of attachment. The power-form relationship betweens andl was superior to thes-y _n* relationship, since the former did not differ significantly among shoots of different diameter. Based on thes-l relationship, the total surface area of a shoot was estimated from thel-census of the shoot. Specific leaf area of a shoot (SLA), defined as half of the shoot surface area divided by the dry weight of the shoot, decreased from 90 to 3 [cm²g(dry wt)⁻¹] with the diameter of the woody tissue of the foliage shoot.     

Shoot and leaf demography of Carex curvula ssp. Curvula and Carex curvula ssp. rosae in the Central Alps

Abstract. Populations of Carex curvula ssp. curvula, C. curvula ssp. rosae and their intermediate form were investigated in the Central Alps over a three-year period. The closely related taxa showed a different dominance behaviour in their respective communities. This may be caused by different growth strategies and a different reproduction biology. Therefore, the main aim of the study was to compare the demography of the three taxa. Shoot and leaf turnover, flower and seed production and population half-lives were determined. Differences in growth dynamics were less pronounced between the two species of Carex curvula than between these taxa and their intermediate form which showed the highest shoot turnover, highest number of fertile inflorescences and highest number of seeds per inflorescence. Carex curvula ssp. rosae showed a slightly higher shoot and leaf production but a lower reproduction capacity than Carex curvula ssp. curvula. Recruitment of populations of the three Carex-forms was only by vegetative shoots. Flowering had a striking effect on the shoot dynamics of the species grown in the grassland sites: up to about 70 % of all dead shoots could be identified as dead flowering shoots from the previous year. The different dominance behaviour of the three taxa could not be explained by their demographical features. Interspecific interactions and the occurrence of microniches might affect the growth and reproduction processes of the taxa.     

Variability in the essential oils of wood and foliage of Pinus aristata and Pinus longaeva

Variations in the monoterpenoid composition of the essential oil of wood, and of the mono- and sesquiterpenoids of foliage collected from more than 200 specimens of pines of subsection Balfourianae (P. aristata, P. longaeva and P. balfouriana) substantiate the previously reported morphological and chemical separation of P. aristata and P. longaeva and suggest a restricted intermediacy in northern Arizona and eastern California. On the basis of wood essential oil the limited population of P. aristata in northern Arizona, while exhibiting to a slight degree morphological intermediacy, is found to be practically identical to the main populations of P. aristata in Colorado and New Mexico. On the basis of foliage oil, however, the same population is clearly intermediate, but closer to P. aristata. The detailed chemical data suggest a distinct chemical status for the Arizona population, with selection for the present-day chemophenotypes being the dominant factor in their evolution. Analysis of monoterpenoids from P. balfouriana × longaeva hybrids indicates that incomplete selective elimination of 3-carene-producing P. longaeva chemophenotypes rather than gene exchange with P. balfouriana is responsible for the presence of abnormal P. aristata-like 3-carene-producing trees in eastern California. The latter trees could have resulted from limited gene exchange between P. aristata and P. longaeva by routes no longer available to them lying to the south of the Grand Canyon.     

In vitro phyllomorphic regeneration of shoot buds and shoots in Picea abies

Needles (10–15 mm) of frost-hardened 20–22-week-old (physiological age equivalent to 1 year) plants of Picea abies L. excised just after flushing, were induced to form adventitious shoot buds and shoots on media supplemented with BAP (6-benzylaminopurine) and NAA (1-napthaleneacetic acid). The addition of nanomolar concentrations (0.5–50) of NAA combined with 1–10 μM BAP considerably stimulated formation of pseudobulbils on the basal to mid-part of the needle axis, as well as their subsequent development into shoot buds and shoots. On a medium containing 10 μM BAP, pseudobulbils that formed at the needle base did not develop further, but became necrotic and died with the omission of NAA. With 5 μM BAP + 50 nM NAA the initial phase of development was slow, but later showed good response and up to 22% of the needles produced shoot buds. Two to three shoots per needle could be excised and subcultured individually onto fresh media. It is concluded that the level of endogenous auxin decreases progressively from the needle's base to its tip, so that that concentration of exogenous auxin (50 nM NAA) which promotes pseudobulbil and shoot-bud formation part-way along the needle axis, simultaneously inhibits their induction at the needle base.     

Testing a new hypothesis: plant vigor and phylloxera distribution on wild grape in Arizona

Summary Longer, meaning more vigorous, shoots of a wild grape clone (Vitis arizonica) were more susceptible to attack by second and third generations of leaf-galling grape phylloxera,Daktulopsphaira vitifoliae, as the growing season progressed. Although there was no significant difference in mean shoot length between attacked and unattacked shoots within a clone at the beginning of shoot elongation, attacked shoots were significantly longer than unattacked shoots when elongation had ceased (P<0.01). Also, long attacked shoots had a significantly greater population of phylloxera galls than short attacked shoots (P<0.01) as the season progressed. The phylloxera population on long shoots increased rapidly while the population on short shoots remained the same. Longer shoots also produced significantly more axillary shoots than shorter shoots as the season progressed (P<0.001), and the number of axillary shoots accounted for 66 percent of the variance in number of attacked leaves on a shoot. Experimental evidence showed that there was a significantly greater percentage of available leaves attacked on long shoots than on short shoots (P<0.05) and the leaves on long shoots generally had a greater number of galls per leaf. The relationship between shoot length and probability of attack was also tested by comparing shoots lengths of 10 attacked clones and 10 unattacked clones at a second location. Mean shoot lengths of attacked clones were significantly longer than mean shoot lengths of unattacked clones (P<0.05), and mean shoot lengths of attacked shoots within a clone were significantly longer than unattacked shoots (P<0.001). Longer shoot length accounted for 81 percent of the variance in probability of attack. The reason for this pattern of attack was that long shoots produced newly expanding leaves over a longer time during the growing season and multivoltine phylloxera require undifferentiated tissue to initiate gall formation. Patterns of attack within a shoot were characterized by an uneven distribution of galls among leaves. This was due to development time between generations and the current availability of undifferentiated tissue at times of colonization. This study supports the hypothesis that some herbivore species are favored more by vigorous plants than by stressed plants.