Fire-related features of wood anatomy in a sweet chestnut (Castanea sativa) coppice in southern Switzerland

Explorative wood anatomical analysis was conducted on a Castanea sativa stand in southern Switzerland, where a moderate-intensity surface fire burned in April 1997. Cross-sections were sampled at multiple heights from 20 scarred shoots, 20 apparently intact shoots, along with cores taken from 20 reference trees outside the fire area. Thin sections were prepared from uphill and downhill locations on the circumference for up to 5 years preceding and following the event year. The thin sections were visually observed in order to identify a response to the known fire event preserved in the wood anatomical structure. Anatomical features were observed at the uphill and downhill locations on both scarred and intact (unscarred) cross-sections, and they occurred in a subset of the observed samples. The features observed in both scarred and intact cross-sections were an apparent increase in vessel density and a decrease in lumen area of the second row of earlywood vessels, along with tyloses formation in the first row of earlywood vessels. Furthermore, the scarred region exhibited a zone of delayed cambial death following the fire, and the onset of woundwood was often initiated later in the season or the following year. Using the type of features and their location around the circumference, we inferred that the observed features may have formed in response to local heating of the cambium, and likely formed in response to canopy damage.     

Plant species diversity changes in abandoned chestnut (Castanea sativa) groves in southern France

Over the last few decades, marked land use changes have taken place in many Mediterranean ecosystems. For example, many chestnut groves in France are now abandoned and have turned into 'natural' coppice stands while others are now clear-cut every 10 or 15 years for wood. Species composition and life form diversity of the plant communities have changed markedly both in the abandoned groves and in the periodically clear-cut coppice stands. What are the consequences of these changes on biodiversity at local and regional scales, and what are the implications for 'new forestry' management intended to conserve biodiversity at the same time as it optimizes productivity and profitability? To answer these questions we studied plant species diversity in the understorey strata along a successional gradient including cultivated grove; abandoned grove; young (15 years old), medium (40 years), and old (>55 years) coppice stands. The results showed that species richness decreased quickly but not steadily along this gradient, but life-form spectra data highlighted that the high levels of plant species biodiversity in cultivated groves were due primarily to the large number of therophytes (annual plants). At a shorter time scale, we also studied the consequences of experimental clear-cutting on plant species biodiversity. Direct measurements over four years showed that species richness and diversity in the understorey increased in the first two years after clear-cutting, but decreased during the 3rd and 4th years. This decrease corresponded closely with a steady increase in Leaf Area Index of the forest canopy. These results have been used to suggest some possible ways to manage the biodiversity in these ecosystems.     

Influence of mycorrhization on physiological parameters of micropropagated Castanea sativa Mill. plants

 Mycorrhizal micropropagated Castanea sativa plants were studied in terms of growth and physiological parameters following in vitro mycorrhization with Pisolithus tinctorius. Mycorrhization enhanced growth of micropropagated chestnut plants, increased their protein content and photosynthetic rates, decreased the respiratory rates and CO₂ compensation point. RuBisCO activity was not significantly different in mycorrhizal and control plants, although there was an increase in the amount of RuBisCO in the former. Mycorrhization increased plant biomass and improved plants physiological status, thus enhancing the acclimatization process. Accepted: 21 May 1997     

Earlywood vessels of Castanea sativa record temperature before their formation

The aim of this study was to identify the climatic signal contained in the earlywood vessel size of the ring-porous chestnut (Castanea sativa) and the physiological processes involved in the underlying mechanisms. In order to assign the encoded signal to a specific physiological process, bud phenology and vessel formation were monitored along an elevation transect and chronologies of the size of the first row of earlywood vessels were retrospectively correlated with 40 yr of early spring temperatures. The first vessels appeared in late April to early May, after encoding both a negative temperature signal in February-March (during tree quiescence) and a positive temperature signal in early April (at the time of resumption of shoot growth). We hypothesize that February and March temperatures affect cambial sensitivity to auxin, preconditioning tree responses later in the season. Furthermore, April temperature is related to tree activation whereby new hormone production fosters vessel expansion.     

Identification of QTLs affecting adaptive traits in Castanea sativa Mill

A QTL analysis for three different adaptive traits was performed in an F₁ progeny of Castanea sativa Mill. The female and male parents originated from two Turkish chestnut populations adapted to a drought and humid environment, respectively. QTLs for bud flush, growth and carbon isotope discrimination were detected over a 3‐year period. Bud set was also recorded in the last year of measurement. Thirty‐five individual QTLs were detected for phenology, 28 for growth and 17 for carbon isotope discrimination, most of them explaining a low to moderate proportion of the total phenotypic variance. QTLs were distributed throughout the whole genome. Temporally stable QTLs were identified for all the traits analysed, with phenology showing the higher proportion of stable QTLs. Interesting phenotypic correlations and co‐localizations among QTLs for different adaptive traits were observed, allowing the formulation of an hypothesis about the genetic adaptation of the female parent to drought.     

Aspects of sandfly biology in southern Switzerland

Abstract. During three field surveys of phlebotomine sandflies in the region of Sessa, Minor Malcantone, southern Switzerland, annually 1987-89, Phlebotomus pemiciosus and P.mascittii were caught regularly by means of light traps, but only when the traps were in close association with humans. The most productive catching sites were in the middle of Sessa village, in basements or partially closed shelters in the immediate vicinity of human dwellings. All catching sites outside the village were negative. The only other sandfly species caught was Sergentomyia minuta, four specimens only. P.pemiciosus and P.mascittii always occurred together and were found to feed - at least occasionally – on human blood. Glucose and fructose were identified in wild-caught sandflies of both species. A breeding place of P.mascittii was identified in the basement of a house in the middle of Sessa. Analyses of soil samples revealed moderately loamy humus with a slightly acidic pH (5.6).     

Cotyledon storage proteins as markers of the genetic diversity in Castanea sativa Miller

This study has been to analyse the useful nut globulin proteins as a marker of the genetic diversity in Castanea sativa. The evaluated populations were highly polymorphic for the globulins, being detected up to 35 polymorphic bands with a wide distribution among all the evaluated populations. Taken together for populations from all the chestnut regions, about 39.3% of total allelic variation was distributed among the populations. The estimates of genetic similarity between populations were clearly associated with the collecting site. This method of analysis of the nut storage proteins (globulins) could be a useful tool for the evaluation of genetic diversity in this and other species of the Fagaceae.     

Multiple shoot-bud formation and plantlet regeneration on Castanea sativa Mill. seeds in culture

Primordial initiation and development of shoot-buds has been accomplished by using shoots derived from chestnut (Castanea sativa Mill) seedlings cultured with added 6-benzylaminopurine (BAP). Germination of chestnut seeds in the presence of BAP (4 – 40 M) stimulated varying numbers of shoot-buds in those areas of the main axis that were favorably altered. When excised single shoots from these treated seeds were subcultured on a fresh medium containing BAP (4 – 40 M) continual shoot production was observed. Bud growth and shoot elongation were stimulated by transferring cultures to a reduced concentration of BAP (2 M) plus indole-3-butyric acid (IBA 0.4 M). Plant regeneration occurred in the presence of IBA (0.8 M) after a preconditioning treatment in which naphthaleneacetic acid (NAA 50 M) and kinetin (k 2 M) were applied to the tissue culture shoots for 7 days in light.     

Rooting Substances in Water Extracts of Castanea sativa and Salix viminalis

The rooting activity of water extracts of cuttings of chestnut (Castanea sativa Mill.) and willow (Salix viminalis L.) was studied throughout the year by the bean rooting test. The chestnut extract nullified the rooting activity of 10^?5M IAA, but on the whole it did not modify the rooting pattern of the bean cuttings. The willow extract increased rooting and showed a rhythm in the content of endogenous growth substances. IAA was found at the time when the extract showed the strongest rooting activity. The extract from willow contained two synergistic root-promoting fractions with the R_f's of 0.0–0.2 and 0.6–0.8 in isopropanol: ammonia: water. The latter one promoted rooting without added IAA. In this fraction a high amount of catechol was detected. No clear correlation was found between the ability to induce roots and the total phenol content in the two species.     

Proliferation,maturation and germination of Castanea sativa Mill. Somatic embryos originated from leaf explants

Experiments were performed to determine the influence of proliferation medium on the maintenance of embryogenic competence and on repetitive embryogenesis in Castanea sativa Mill. somatic embryos derived from leaf explants. Somatic embryo proliferation was carried out by both direct secondary embryogenesis and by the culture of nodular callus tissue originated from cotyledons of somatic embryos. Both systems led to the production of cotyledonary somatic embryos on Murashige and Skoog proliferation medium supplemented with 0.1 mg l-1 benzyladenine and 0.1 mg l-1 naphthaleneacetic acid. Carbon source and concentration had a marked influence on maturation and subsequent germination ability of chestnut somatic embryos. Plantlet conversion was achieved in embryos matured on media with 6 % sucrose, and on 3 or 6 % maltose, whereas mean shoot length, root length and leaf number of produced plants were not significantly affected by these maturation media. Overall, the best results were obtained with 3 % maltose-matured somatic embryos, giving rise to 6 % plant recovery in addition to 33 % of embryos exhibiting only shoot development. The application of a 2-month cold treatment at 4 degrees C to somatic embryos matured on medium with 3 % maltose was necessary for achieving plant conversion, while partial desiccation did not appear to influence this response. A total of 39 % of embryos eventually produced plants either through conversion to plantlets or indirectly through rooting of shoots. Shoots formed by somatic embryos could be excised, multiplied and rooted following the micropropagation procedures previously developed for chestnut.     

In vitro conservation of chestnut (Castanea sativa) by slow growth

Slow growth storage has been achieved for Castanea sativa (cv. ‘Montemarano’) shoot cultures over a duration of 48 mo at a temperature of 8°C, where 82% of explants survived and were able to resume normal growth after transfer to standard culture conditions at 23°C. The evaluation of the chlorophyll content of leaves also showed no differences between material stored for 48 mo and control material subcultured at 23°C. With a storage temperature of 4°C, the survival of shoots was significantly lower at approximately 56% after 12 mo, and no plants recovered after 24-mo storage. The presence of 6-benzyladenine 0.44 μM in the culture medium proved to be necessary for the recovery of healthy shoots, while pre-treatments with different concentrations of abscisic acid did not significantly influence the survival of shoots following storage conditions. A low level of light during slow growth storage resulted in positive effects on the rate of shoot survival over the longest preservation periods.     

Bacterial expression of an active class Ib chitinase from Castanea sativa cotyledons

Ch3, an endochitinase of 32 kDa present in Castanea sativa cotyledons, showed in vitro antifungal properties when assayed against Trichoderma viride. The characterization of a cDNA clone corresponding to this protein indicated that Ch3 is a class Ib endochitinase that is synthesized as a preprotein with a signal sequence preceding the mature polypeptide. Bacterial expression of mature Ch3 fused to the leader peptide of the periplasmic protein ompT resulted in active Ch3 enzyme. A plate assay was adapted for semi-quantitative determination of chitinase activity secreted from cultured bacteria, which should facilitate the identification of mutants with altered capacity to hydrolyse chitin.     

Effects of fire on forest dynamics in southern Switzerland

Abstract. We analysed the influence of fire on species composition, floristic diversity and indicator values of the vegetation in forests of the Canton Ticino (South Switzerland). Both frequency of fire and time since last fire were taken into account using variance analysis. Results show a strong interaction of these two parameters, i.e. the reaction of the vegetation after a fire depends on the frequency of past fires. A rapid recovery is generally observed after fire, involving both herbaceous and woody species. However, a long-term tendency towards floristic and edaphic impoverishment is observed with increasing frequency of fires. This phenomenon appears to be linked to the progressive dominance of a few fire-tolerant species.     

Study of the temperature effect in three chestnut (Castanea sativa Mill.) cultivars' behaviour

The aim of this work was to analyse the effect of temperature in three chestnut cultivars, Aveleira, Judia and Longal. For this purpose, gas exchange, thylakoid membrane potential, photosynthetic pigment and lipid content data in July, September and October under different temperatures (31, 26 and 18 degrees C) were determined. With respect to gas exchanges, significant changes in photosynthesis rate of Aveleira were observed between July and September (7mumol CO2m(-2)s(-1)). In contrast, Judia and Longal showed a strong increase in this period, 6.1-8.5 and 4.9-6.7 micromol CO2m(-2)s(-1), for Judia and Longal, which represent an increase of about 15% and 43%, respectively. Similar patterns were detected in daylight photosynthesis measurements for Judia and Longal, in which an almost 60% decrease was observed, in contrast to 40% for Aveleira, from morning to midday, when temperatures increased from 27 to 34 degrees C. In addition to high photosynthetic rates in the hottest month, Aveleira was also the sunniest cultivar according its highest value on chlorophyll a/b ratio (3.65). Cultivars also presented maximal thylakoid membrane potential at different temperatures, with their values being 20.8, 17.8 and 17.2 degrees C for Aveleira, Longal and Judia, respectively. These results were also supported by thylakoid fatty acid composition which indicated that the unsaturation index of Aveleira (158) was the lowest in comparison with other two cultivars, 168 and 175, for Longal and Judia, respectively.     

Effect of competitive interactions between ectomycorrhizal and saprotrophic fungi on Castanea sativa performance

In Northeast of Portugal, the macrofungal community associated to chestnut tree (Castanea sativa Mill.) is rich and diversified. Among fungal species, the ectomycorrhizal Pisolithus tinctorius and the saprotroph Hypholoma fasciculare are common in this habitat. The aim of the present work was to assess the effect of the interaction between both fungi on growth, nutritional status, and physiology of C. sativa seedlings. In pot experiments, C. sativa seedlings were inoculated with P. tinctorius and H. fasciculare individually or in combination. Inoculation with P. tinctorius stimulated the plant growth and resulted in increased foliar-N, foliar-P, and photosynthetic pigment contents. These effects were suppressed when H. fasciculare was simultaneously applied with P. tinctorius. This result could be related to the inhibition of ectomycorrhizal fungus root colonization as a result of antagonism or to the competition for nutrient sources. If chestnut seedlings have been previously inoculated with P. tinctorius, the subsequent inoculation of H. fasciculare 30 days later did not affect root colonization, and mycorrhization benefits were observed. This work confirms an antagonistic interaction between ectomycorrhizal and saprotrophic fungi with consequences on the ectomycorrhizal host physiology. Although P. tinctorius is effective in promoting growth of host trees by establishing mycorrhizae, in the presence of other fungi, it may not always be able to interact with host roots due to an inability to compete with certain fungi.     

Effect of ectomycorrhizal fungi on survival and growth of micropropagated plants and seedlings of Castanea sativa mill.

 Four ectomycorrhizal fungi (Amanita muscaria, Laccaria laccata, Piloderma croceum and Pisolithus tinctorius) were used to produce mycorrhiza on seedlings and micropropagated plants of Castanea sativa in vitro. Pisolithus tinctorius was most effective in colonizing roots of both micropropagated plants and seedlings. A. muscaria and L. laccata only colonized a few feeder roots of some plants and Piloderma croceum did not form mycorrhizas. Mycorrhization of micropropagated plants increased survival and growth during weaning. Accepted: 27 February 1996     

Upgrade of Castanea sativa (Mill.) genetic resources by sequencing of barcode markers

Journal of Genetics -