Toward a Pinus pinaster bacterial artificial chromosome library

Conifers are of great economic and ecological importance, but little is known concerning their genomic organization. This study is an attempt to obtain high-quality high-molecular-weight DNA from Pinus pinaster cotyledons and the construction of a pine BAC library. The preparation incorporates modifications like low centrifugation speeds, increase of EDTA concentration for plug maintenance, use of DNase inhibitors to reduce DNA degradation, use of polyvinylpyrrolidone and ascorbate to avoid secondary metabolites, and a brief electrophoresis of the plugs prior to their use. A total of 72 192 clones with an average insert size of 107 kb, which represents an equivalent of 11X pine haploid genomes, were obtained. The proportions of clones lacking inserts or containing chloroplast DNA are both approximately 1.6%. The library was screened with cDNA probes for seven genes, and two clones containing Fd-GOGAT sequences were found, one of them seemingly functional. Ongoing projects aimed at constructing a pine bacterial artificial chromosome library may benefit from the methods described here.     

Resin tapping in Pinus pinaster: effects on growth and response function to climate

Pinus pinaster is of great ecological and economic importance and has traditionally been subjected to intensive uses such as wood and resin extraction. In the last decade, dendrochronological methods are increasingly being used to analyze the effects of climatic factors on the growth of the maritime pine, although tapped trees were generally avoided because it was thought that their growth was affected by resin extraction. In Spain, however, it is hard to find a long-lived forest of P. pinaster that has not been subjected to tapping for resin. In the present paper, we performed dendrochronological analyses of this species from wood cores and cross sections taken from both resin-tapped trees and resin-untapped trees killed by a fire in 2008 in the central Iberian region. On the one hand, we reconstructed the history of forest management by means of analysis of resin scars in the cross sections of resin-tapped trees. This facet of dendrochronological dating had not heretofore been developed, and little is therefore known about it. We dated 46 scars, which indicate a history of intensive resin extraction in the 1920–1950 period. Moreover, we attempted to answer the question: Have the old resin extractions in P. pinaster precluded the use of their growth rings for dendrochronological and dendroclimatic studies? We found that resin extraction did not alter general short-wavelength variability, and we developed a local chronology with all synchronized series, and the response function with respect to climate is similar to other oldest P. pinaster forests studied in Spain. The information we have recorded can be of use for providing tools to land managers for interpreting forest dynamics in resin-tapped regions.     

Xylogenesis of compression and opposite wood in mountain pine at a Mediterranean treeline

Key message

Comparisons between compression and opposite wood formation in prostrating Pinus mugo indicate that the secondary meristem can produce more tracheids with thicker walls by also increasing the number of contemporaneously differentiating cells, rather than only increasing the duration or the rate of cell formation.

Context

Although cambium tissues within a stem experience the same climatic conditions, the resulting wood structure and properties can strongly differ. Assessing how meristem differently regulates wood formation to achieve different anatomical properties can help understanding the mechanisms of response and their plasticity.

Aims

We monitored the formation of compression (CW) and opposite (OW) wood within the same stems to understand whether achieved differences in wood structure are caused by modifications in the process of cell formation.

Methods

We collected weekly microcores of compression and opposite wood from the curved stem of ten treeline prostrating mountain pines (Pinus mugo Turra ssp. mugo) at the Majella massif in Central Italy.

Results

Results indicate that cambium formed approximately 1.5 times more cells in CW than OW, despite that CW cell differentiation only extended 2 weeks longer and the residence time of CW cells in the wall-thickening phase was only 20% longer. Differences in their formation were thus mainly related to both the rates and the width of the enlarging and wall-thickening zones (i.e., the number of cells simultaneously under differentiation) and less to duration of cell formation.

Conclusion

We conclude that to achieve such a different wood structures, the efficiency of the secondary meristem, in addition of altered rate of cell division and differentiation, can also modify the width of the developing zones. Thus, deciphering what rules this width is important to link environmental conditions with productivity.

     

A Top Height Equation for Pinus pinaster

A number of equations were fitted by means of stepwise regression to data from 171 compartments enumerated for working plan purposes in the southern and south-western Cape Province. Top height (= the mean height of the 100 biggest trees per ha in terms of diameter at breast height) or a function of top height was used as the dependent variable. Mean height, age (A), stems per ha (SPH) and mean diameter at breast height were used as the independent variables. The following equation gave the best results:

Top height = mean height + 0,03 log (SPH—99) A     

Variability of stem and branch maintenance respiration in a Pinus pinaster tree

The relationship between maintenance respiration (Rm) of woody organs and their structural characteristics was explored in adult Pinus pinaster Ait. trees. We measured Rm on 75 stem and branch segments of different ages (from 3 to 24 years) and diameters (from 1 to 35 cm). The temperature response of Rm was derived from field measurements based on a classical exponential function with Q10 = 2.13. Relationships between Rm and the dimensions of the woody organs were analyzed under controlled conditions in the laboratory. The surface area of a woody organ was a better predictor of Rm than volume, but surface area failed to account for the observed within-tree variability of Rm among stems, branches and twigs. Two simple models were proposed to predict the variability of Rm at 15 degrees C in an adult tree. Model 1, a linear function model based on the dry mass and nitrogen concentration of sapwood and phloem tissues, explained most of the variability of Rm in branches and stems (R2 = 0.97). We concluded that the respective contributions of the phloem and sapwood depend on the location and diameter of the woody organ. Model 2, a power-law function model based on the length, diameter and age of the sample, explained the same variance of Rm as Model 1 and is appropriate for scaling Rm to the stand level. Models 1 and 2 appear to explain a larger variability of Rm than models based on stem area or sapwood mass.     

Bursaphelenchus fungivorus from Pinus pinaster bark in Portugal

Bursaphelenchus fungivorus is reported for the first time in Portugal, identified as associated with Pinus pinaster bark and characterized on the basis of morphological and morphometrical characters for this species. Species identification was confirmed using restriction fragment length polymorphism analysis and sequencing of the internal transcribed spacer (ITS) regions of ribosomal DNA. Intraisolate genetic variability was detected among ITS sequences of the Portuguese B. fungivorus isolate. Phylogenetic analysis, obtained from multiple sequence alignment between ITS sequences of Bursaphelenchus species, revealed that the Portuguese B. fungivorus isolate clusters with other B. fungivorus isolates, forming a separate group close to B. seani highlighting a molecular proximity of these two species.     

Improving plantlet yield in Pinus pinaster somatic embryogenesis

Abstract

Somatic embryogenesis is expected to play a significant role in the future forest tree improvement programmes. The main bottleneck of this technique is still the progression from immature embryogenic cultures to mature cotyledonary embryos able to develop properly into well-growing plants. In this work, we present an improved protocol focused on increasing the maturation and conversion rate of Pinus pinaster Ait. embryogenic cultures. Results showed that the optimisation of the nutrient composition in the maturation medium increased the number of mature embryos by 25% (187.8 embryos per gram of fresh mass in average compared to 144.5 embryos in regular medium). It was also shown that 12-month cryostorage did not reduce viability or embryogenic ability of maritime pine cultures. A further increase in the yield of the protocol could be obtained by using benzyladenine in the conversion medium, promoting the bud-break of axillary buds that yielded 5.7 shoots in average per somatic embryo. Rooting of axillary shoots reached 84.3%. This methodology offers an alternative to overcome some problems associated with low somatic embryo production since the plantlet yield could be increased fivefolds.     

2S Arginine-rich proteins from Pinus pinaster seeds

Low molecular weight globulins, which are abundant proteins in the Pinus pinaster Ait. megagametophyte, were purified and characterized. They showed a dimeric structure formed of one large and one small subunit linked by disulfide bridges. They were characterized by a high Arg and Glx content and by a relatively high Cys content. A comparison of their characteristics with those of angiosperm 2S proteins suggests that there is homology between them.     

Physical properties and termite durability of maritime pine Pinus pinaster Ait., heat-treated under vacuum pressure

An original heat treatment performed under vacuum pressure was investigated. Maritime pine samples were treated at six different temperatures: 140, 160, 180, 200, 230 and 260°C. The physical and mechanical consequences, i.e. bending strength (MOR), modulus of elasticity (MOE), hygroscopic behaviour, equilibrium moisture contents and anti-swelling efficiency (ASE) were studied. A no-choice feeding test according to the NF EN 117 standard was achieved. Temperatures up to 200°C had no significant effect on wood properties. However, at 230 and 260°C, the decrease in MOR was severe, reaching 42.5 and 62.5%, respectively. Whatever the treatment conditions, wood samples were still highly degraded by termites, revealing no increase in their durability.     

Improvement of Pinus pinaster wood dimensional stability using cork industry wastes

Summary This article relates to a procedure of preparing solutions for wood treatment by dissolving the condensed waste material from the baking steam of expanded cork board production in organic solvents. These solutions were applied by way of different methods inPinus pinaster wood treatment. The use of these products and methods improved the pine wood dimensional stability giving rise to lower water absorption and swelling. This cork industry waste, is mainly constituted by tannins and waxes, and their solutions are easy to obtain at low cost. A patent application was done.The authors would like to thank the collaboration of our fellow Mr. Paulo Cortiço     

Post-fire production of mushrooms in Pinus pinaster forests using classificatory models

This study was aimed at describing post-fire mushroom production in a Mediterranean ecosystem dominated by Pinus pinaster Ait. in the northwest of Spain and assessing the results by classificatory models. During the autumn periods of 2003–2006, fruit bodies from 115 fungal taxa were collected in burned and unburned areas and were further grouped into the following categories: saprotrophic/mycorrhizal; and edible/non-edible. After wildfires, a significant reduction in the number of fungal species and fruit body biomass production was observed. Based on this relevant information, the first simple classificatory model for this aim is provided. Nine alternative models based on classifications according to combinations of edibility and functional groups were fitted, and four fruiting body biomass production classes were defined as possible responses. As explanatory factors, time after fire and climatic variables significantly related to fruit body production were included. The best predictive results were obtained for edible and edible-mycorrhizal models, for which the correct classification rate of production classes was between 92 and 85 %. Moreover, the models obtained were applied to analyse the effect of time after fire on fungal production. Mycorrhizal and edible fungal production after fire was classified into the lowest class, whereas saprotrophic and non-edible species followed a contrary trend. The classificatory models can be useful to optimise management and harvest of these increasingly appreciated non-timber forest resources.     

引种法国海岸松种子萌发试验研究

通过对从法国引进的海岸松种子进行千粒重测定,认为法国海岸松种质均一,变异系数小;通过对引进种子在试验室及苗圃地不同处理情况下进行萌发试验,可为日后海岸松引种试验提供基础数据。     

Growth and water relations of three geographically separate origins of maritime pine (Pinus pinaster) under saline conditions

Water relations and growth of maritime pine (Pinus pinaster Ait.) were investigated in 2-year-old seedlings of French ('Landes'), Iberian ('Iberian') and Moroccan ('Tamjoute') origin raised for 67 days in a flowing solution culture system containing 0, 50, 150 or 250 mM NaCl. Height growth, and stem, needle and root dry matter were reduced by salinity with minor differences among geographic origins. Predawn needle water potential was decreased by salinity and corresponded approximately to the osmotic potential of the nutrient solution. Stomatal conductance was reduced according to the amount of salinity applied. Whole-plant hydraulic conductance was also reduced, even when expressed on a root dry weight basis. The osmotic potential of xylem sap was five- to sixfold lower than that of the nutrient solution. Seedlings of the most southerly origin (Tamjoute) exhibited a greater ability to decrease osmotic potential under saline conditions than seedlings of more northerly origin (Landes and Iberian) as a result of higher mineral cation transport to the shoot.     

Kinetics and mechanism of acetic acid pulping of detannined Pinus pinaster bark

Summary The observed kinetics of Pinus pinaster bark with acetic acid after alkali treatment, with or without intervening acid prehydrolysis, are satisfactorily explained by a model involving both solubilization and condensation reactions.This study was subsidised by the Xunta de Galicia. J. González is grateful to DGICYT for a research grant     

Recovery of bioactive compounds from Pinus pinaster wood by consecutive extraction stages

Pinus pinaster wood samples, obtained at different positions of three healthy trees, were subjected to two sequential extractions using an Accelerated Solvent Extraction instrument. The first extraction was carried out with hexane (to remove lipophilic extractives) and the second one with acetone/water (95:5 v/v) to recover bioactive phenolic compounds, the target compounds of this study. The extracted fractions were assayed for total yield and composition. The extracts contained a spectrum of phenolic compounds (simple phenolics, phenolic stilbenes, flavonoids and lignans) and non-phenolic components (juvabiones, resin and fatty acids, steryl esters and triglycerides). The fractionation effects achieved by consecutive extractions and the recovery of bioactive phenolics are discussed.     

Modelling the site index of Pinus pinaster plantations in Turkey using ecological variables

The determination of site productivity in forest ecosystems plays a crucial role in resource management.This study was carried out to identify relationships between site characteristics and height growth of Corsican maritime pine(Pinus pinaster Ait.) plantations in Turkey.Sixty-nine sample plots 20 years of age were selected from locations with different inclinations,aspects,elevations,slope positions and site class.Soil samples were taken at various depths.Height and age were measured on a dominant tree after felling in each plot.Physical and chemical properties of the soil were determined.Relationships between site index(SI_(25)) and physiographic factors,climatic attributes as well as soil properties were evaluated using correlation analysis and multiple regression analysis.Site index was significantly related with annual precipitation,mean spring rainfall,rainfall June to September,rainfall of the driest month,length of the dry period,mean maximum temperature,mean temperature of the warmest month,stoniness of the soil,sand,silt,clay,pH,electrical conductivity,and available water capacity.Multiple regression accounted for 57.9% of variations in height growth.The models obtained can be used to determine the site index of potential areas in Turkey for maritime pine.It can be said that the productivity of maritime pine may decline in the future due to global climate change.     

希腊海岸松叶中聚戊烯醇含量与化学结构分析

利用HPLC与聚戊烯醇对照品保留时间对照的定性方法,确定海岸松叶中聚戊烯醇的异戊烯基单元数为14~20;利用1H NMR和13C NMR确定海岸松聚戊烯醇化学结构为ω-trans2-cisn-OH构型。采用Kromasil C18 ODS-1(150 mm×4.6 mm,5μm)色谱柱,以甲醇-异丙醇(体积比9∶16)为流动相,流速为0.50 mL/min,检测波长为210 nm,柱温为25℃,PDA检测器等色谱条件,测定海岸松叶聚戊烯醇在1.10~12.10μg范围内线性良好(r=0.996 3),平均回收率为98.21%,RSD为2.27%(n=6),海岸松干叶中聚戊烯醇含量为0.33%。