Genome-Wide Characterization and Functional Validation of the ACS Gene Family in the Chestnut Reveals Its Regulatory Role in Ovule Development.

Ovule abortion significantly contributes to a reduction in chestnut yield. Therefore, an examination of the mechanisms underlying ovule abortion is crucial for increasing chestnut yield. In our previous study, we conducted a comprehensive multiomic analysis of fertile and abortive ovules and found that ACS genes in chestnuts (CmACS) play a crucial role in ovule development. Therefore, to further study the function of ACS genes, a total of seven CmACS members were identified, their gene structures, conserved structural domains, evolutionary trees, chromosomal localization, and promoter cis-acting elements were analyzed, and their subcellular localization was predicted and verified. The spatiotemporal specificity of the expression of the seven CmACS genes was confirmed via qRT-PCR analysis. Notably, CmACS7 was exclusively expressed in the floral organs, and its expression peaked during fertilization and decreased after fertilization. The ACC levels remained consistently greater in fertile ovules than in abortive ovules. The ACSase activity of CmACS7 was identified using the genetic transformation of chestnut healing tissue. Micro Solanum lycopersicum plants overexpressing CmACS7 had a significantly greater rate of seed failure than did wild-type plants. Our results suggest that ovule fertilization activates CmACS7 and increases ACC levels, whereas an overexpression of CmACS7 leads to an increase in ACC content in the ovule prior to fertilization, which can lead to abortion. In conclusion, the present study demonstrated that chestnut ovule abortion is caused by poor fertilization and not by nutritional competition. Optimization of the pollination and fertilization of female flowers is essential for increasing chestnut yield and reducing ovule abortion.

Auxin and CmAP1 regulate the reproductive development of axillary buds in Chinese chestnut (Castanea mollissima).

KEY MESSAGE: Auxin accumulation upregulates the expression of APETALA1 (CmAP1) and subsequently activates inflorescence primordium development in axillary buds of chestnut. The architecture of fruiting branches is a key determinant of chestnut yield. Normally, axillary buds at the top of mother fruiting branches develop into flowering shoots and bear fruits, and the lower axillary buds develop into vegetative shoots. Decapitation of the upper axillary buds induces the lower buds to develop into flowering shoots. How decapitation modulates the tradeoff between vegetative and reproductive development is unclear. We detected inflorescence primordia within both upper and lower axillary buds on mother fruiting branches. The level of the phytohormones 3-indoleacetic acid (IAA) and trans-zeatin (tZ) increased in the lower axillary buds in response to decapitation. Exogenous application of the synthetic analogues 1-naphthylacetic acid (NAA) or 6-benzyladenine (6-BA) blocked or promoted, respectively, the development of the inflorescence primordia in axillary buds. The transcript levels of the floral identity gene CmAP1 increased in axillary buds following decapitation. An auxin response element TGA-box is present in the CmAP1 promoter and influenced the CmAP1 promoter-driven expression of ß-glucuronidase (GUS) in floral organs in Arabidopsis, suggesting that CmAP1 is induced by auxin. We propose that decapitation releases axillary bud outgrowth from inhibition caused by apical dominance. During this process, decapitation-induced accumulation of auxin induces CmAP1 expression, subsequently promoting the reproductive development of axillary buds.

Impact of 'brown rot' caused by Gnomoniopsis castanea on chestnut fruits during the post-harvest process: critical phases and proposed solutions.

BACKGROUND: The brown rot fungus, Gnomoniopsis castanea, is the main organism responsible for the outbreak of chestnut postharvest decay that is threatening the sustainability of the chestnut market in Europe. Currently, no specific strategy is available to mitigate the impact and remediate the high losses of fruits in postharvest storage. In the present study, the different phases of chestnut handling in a standard facility plant were analyzed by evaluating the amount of fruit rot and infection by G. castanea at each phase. RESULTS: The warm bath (48 °C) was identified as the critical phase, requiring strict parametrization to effectively inactivate G. castanea in fruits. Laboratory tests indicated that maintaining fruits at 50 °C for a maximum of 45 min provided optimal conditions to completely inactivate G. castanea inoculum during postharvest handling. However, the warm bath at 50 °C and over was not effective in inactivating the complex of fungal taxa responsible for contamination and development of molds. Higher temperatures and extended treatment times caused significant losses in fruit quality, as indicated by taste panel evaluation. Upscaling of postharvest facilities is discussed and critically evaluated. CONCLUSION: The warm bath (50 °C for 45 min) is effective in completely inactivating G. castanea in fruits but did not reduce the impacts of the complex of molds responsible for external contamination and mycotoxin production. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Friendly neighbours: Hydraulic redistribution accounts for one quarter of water used by neighbouring drought stressed tree saplings.

Hydraulic redistribution (HR) can buffer drought events of tree individuals, however, its relevance for neighbouring trees remains unclear. Here, we quantified HR to neighbouring trees in single- and mixed-species combinations. We hypothesized that uptake of HR water positively correlates with root length, number of root tips and root xylem hydraulic conductivity and that neighbours in single-species combinations receive more HR water than in phylogenetic distant mixed-species combinations. In a split-root experiment, a sapling with its roots split between two pots redistributed deuterium labelled water from a moist to a dry pot with an additional tree each. We quantified HR water received by the sapling in the dry pot for six temperate tree species. After 7 days, one quarter of the water in roots (2.1 ± 0.4 ml), stems (0.8 ± 0.2 ml) and transpiration (1.0 ± 0.3 ml) of the drought stressed sapling originated from HR. The amount of HR water transpired by the receiving plant stayed constant throughout the experiment. While the uptake of HR water increased with root length, species identity did not affect HR as saplings of Picea abies ((L.) Karst) and Fagus sylvatica (L.) in single- and mixed-species combinations received the same amount of HR water.

Fruit development of Lithocarpus (Fagaceae) and the role of heterochrony in their evolution.

Stone oaks, or Lithocarpus species of Fagaceae are ecologically important canopy trees in the tropical and subtropical forests over East Asia, and the fruits of which are important food sources for insects and vertebrates there. The great fruit morphological variation of this genus represents two fruit types, acorn and enclosed receptacle fruit types. However, the evolutionary mechanisms of differentiation into these two fruit types with contrasting morphology remain a puzzle. To reveal the morphogenetic properties of two fruit types, we observed tissue differentiation and development among 20 Lithocarpus species from fruit set to maturity. Unlike in fruits of Quercus, the endocarp differentiation in Lithocarpus fruits occurred later than exocarp and mesocarp. Cupules provided further protection of developing seeds, particularly of acorn-type fruits. Fruits of Lithocarpus and Quercus acorns share similar insect predators. At fruit set, both acorn and enclosed receptacle types were largely identical, with similar tissue morphology and the sequence of differentiation. The distinct difference between two fruit types at maturity came from varied rates and degrees of development between the pericarp and receptacle tissues. We found that heterochrony between two tissues could create substantially divergent ecological strategies for protection and dispersal of their seeds, which is essential for the evolution of two fruit types.

Growth Features of Castanea sativa Mill. in Forest Stands Infected by Cryphoneciria parasitica (Murrill) М. Е. Barr. in Abkhazia.

Currently, the forest plantations of Castanea sativa Mill. are largely damaged by phytopathogenic organisms, of which Cryphoneciria parasitica (Murrill) М. Е. Barr. has the most negative impact. The spread of this type of disease reduces the vital state and significantly reduces the life expectancy of C. sativa trees. For the first time, using dendrochronological analysis, the specificity of distribution and degree of phytopathogenic damage to C. sativa trunk tissues were assessed. It was revealed that in its lower part the dieback of the peripheral layers of wood occurs fairly evenly with a total degradation of cellular structures in the amount of 18-20%. In the middle segments of the trunk this figure increases almost twice. And the greatest degree of wood dieback is observed in the upper part of damaged trees. It was found that climatic changes of the last decades determine the strengthening of destructive processes in the woodland of C. sativa of the Caucasus.

Roles of the GA-mediated SPL Gene Family and miR156 in the Floral Development of Chinese Chestnut (Castanea mollissima).

Chestnut (Castanea mollissima) is a deciduous tree species with major economic and ecological value that is widely used in the study of floral development in woody plants due its monoecious and out-of-proportion characteristics. Squamosa promoter-binding protein-like (SPL) is a plant-specific transcription factor that plays an important role in floral development. In this study, a total of 18 SPL genes were identified in the chestnut genome, of which 10 SPL genes have complementary regions of CmmiR156. An analysis of the phylogenetic tree of the squamosa promoter-binding protein (SBP) domains of the SPL genes of Arabidopsis thaliana, Populus trichocarpa, and C. mollissima divided these SPL genes into eight groups. The evolutionary relationship between poplar and chestnut in the same group was similar. A structural analysis of the protein-coding regions (CDSs) showed that the domains have the main function of SBP domains and that other domains also play an important role in determining gene function. The expression patterns of CmmiR156 and CmSPLs in different floral organs of chestnut were analyzed by real-time quantitative PCR. Some CmSPLs with similar structural patterns showed similar expression patterns, indicating that the gene structures determine the synergy of the gene functions. The application of gibberellin (GA) and its inhibitor (Paclobutrazol, PP333) to chestnut trees revealed that these exert a significant effect on the number and length of the male and female chestnut flowers. GA treatment significantly increased CmmiR156 expression and thus significantly decreased the expression of its target gene, CmSPL6/CmSPL9/CmSPL16, during floral bud development. This finding indicates that GA might indirectly affect the expression of some of the SPL target genes through miR156. In addition, RNA ligase-mediated rapid amplification of the 5' cDNA ends (RLM-RACE) experiments revealed that CmmiR156 cleaves CmSPL9 and CmSPL16 at the 10th and 12th bases of the complementary region. These results laid an important foundation for further study of the biological function of CmSPLs in the floral development of C. mollissima.

Forecasting effects of tree species reintroduction strategies on carbon stocks in a future without historical analog.

American chestnut (Castanea dentata) was once an important component forests in the central Appalachians (USA), but it was functionally extirpated nearly a century ago. Attempts are underway to reintroduce blight-resistant chestnut to its former range, but it is uncertain how current forest composition, climate, and atmospheric changes and disturbance regimes will interact to determine future forest dynamics and ecosystem services. The combination of novel environmental conditions (e.g. climate change), a reintroduced tree species and new disturbance regimes (e.g. exotic insect pests, fire suppression) have no analog in the past that can be used to parameterize phenomenological models. We therefore used a mechanistic approach within the LANDIS-II forest landscape model that relies on physiological first principles to project forest dynamics as the outcome of competition of tree cohorts for light and water as a function of temperature, precipitation, CO2 concentration, and life history traits. We conducted a factorial landscape simulation experiment to evaluate specific hypotheses about future forest dynamics in two study sites in the center of the former range of chestnut. Our results supported the hypotheses that climate change would favor chestnut because of its optimal temperature range and relative drought resistance, and that chestnut would be less competitive in the more mesic Appalachian Plateau province because competitors will be less stressed. The hypothesis that chestnut will increase carbon stocks was supported, although the increase was modest. Our results confirm that aggressive restoration is needed regardless of climate and soils, and that increased aggressiveness of chestnut restoration increased biomass accumulation. The hypothesis that chestnut restoration will increase both compositional and structural richness was not supported because chestnut displaced some species and age cohorts. Although chestnut restoration did not markedly enhance carbon stocks, our findings provide hope that this formerly important species can be successfully reintroduced and associated ecosystem services recovered.

Functional lifespans of xylem vessels: Development, hydraulic function, and post-function of vessels in several species of woody plants.

PREMISE OF THE STUDY: Xylem vessels transition through different stages during their functional lifespan, including expansion and development of vessel elements, transition to vessel hydraulic functionality, and eventual transition to post-functionality. We used information on vessel development and function to develop a model of vessel lifespan for woody plants. METHODS: We examined vessel functional lifespan using repeated anatomical sampling throughout the growing season, combined with active-xylem staining to evaluate vessel hydraulic transport functionality. These data were combined with a literature review. The transitions between vessel functional lifespans for several species are illustrated, including grapevine (Vitis vinifera L., Vitaceae), English oak (Quercus robur L., Fagaceae), American chestnut [Castanea dentata (Marshall) Borkh.; Fagaceae], and several arid and semi-arid shrub species. KEY RESULTS: In intact woody plants, development and maturation of vessel elements may be gradual. Once hydraulically functional, vessel elements connect to form a vessel network that is responsible for bulk hydraulic flow through the xylem. Vessels become nonfunctional due to the formation of gas emboli. In some species and under some conditions, vessel functionality of embolized conduits may be restored through refilling. Blockages, such as tyloses, gels, or gums, indicate permanent losses in hydraulic functional capacity; however, there may be some interesting exceptions to permanent loss of functionality for gel-based blockages. CONCLUSIONS: The gradual development and maturation of vessel elements in woody plants, variation in the onset of functionality between different populations of vessels throughout the growing season, and differences in the timing of vessel transitions to post-functionality are important aspects of plant hydraulic function.

Multi-cropping edible truffles and sweet chestnuts: production of high-quality Castanea sativa seedlings inoculated with Tuber aestivum, its ecotype T. uncinatum, T. brumale, and T. macrosporum.

The plantation and management of sweet chestnut (Castanea sativa Mill.) orchards is a common and traditional land use system in many areas of Europe that offers the advantage of simultaneous production of nuts and timber. During the last decades, sweet chestnut has declined dramatically in many regions because of the profound social changes in rural areas coupled with pathogen attacks. Truffles, the hypogeous ascocarps of the ectomycorrhizal genus Tuber, are currently cultivated using host trees inoculated with these fungi for improving production in truffle orchards. The production of good forestry quality chestnut seedlings inoculated with European truffles in nurseries is essential for multi-cropping plantation establishment, but so far, it has not been implemented in agroforestry practices. Moreover, it is necessary to assess the physiological condition of the seedlings due to the high calcium amendment needed for the growth of Tuber spp. mycelium that can become toxic for the host plants. In this study, seedlings of C. sativa were inoculated with Tuber aestivum and its ecotypes T. uncinatum, T. brumale, and T. macrosporum and were grown in a greenhouse using culture conditions favorable for the production of high-quality plants for forestry purposes. At the end of the assay, levels of root colonization and morphological and physiological parameters of the seedlings were measured. The colonization of C. sativa with T. aestivum, its ecotype T. uncinatum, and T. brumale was successful, and the seedlings showed normal growth. Inoculation protocols with T. macrosporum need to be improved. Tuber species formed well-developed ectomycorrhizae on C. sativa in nursery conditions.

Quantification and identification of microorganisms found on shell and kernel of fresh edible chestnuts in Michigan.

BACKGROUND: Chestnut is a relatively new cultivated crop for Michigan, and postharvest loss due to decay has been problematic as production has increased each year. In 2007, more than 25% of chestnuts were lost to postharvest decay, equivalent to approximately 5300 kg of fresh product. To determine the organisms responsible for decay, a microbiological survey was performed in 2006 and 2007 to identify microorganisms involved in postharvest shell (external surface) mold and internal kernel (edible portion) decay of chestnuts. RESULTS: Filamentous fungi including Penicillium expansum, Penicillium griseofulvum, Penicillium chrysogenum, Coniophora puteana, Acrospeira mirabilis, Botryosphaeria ribis, Sclerotinia sclerotiorum, Botryotinia fuckeliana (anamorph Botrytis cinerea) and Gibberella sp. (anamorph Fusarium sp.) were the predominant microorganisms that negatively impacted fresh chestnuts. Populations of microorganisms varied between farms, harvesting methods and chestnut parts. CONCLUSION: Chestnuts harvested from the orchard floor were significantly (P < 0.05) more contaminated than chestnuts harvested directly from the tree, by more than 2 log colony-forming units (CFU) g(-1) . In addition, a significant difference (P < 0.05) in the microbial population was seen between chestnuts submitted by different growers, with average count ranges of fungi, mesophilic aerobic bacteria (MAB) and yeasts equal to 4.75, 4.59 and 4.75 log CFU g(-1) respectively. © 2016 Society of Chemical Industry.

Experimental evidence for an intraspecific Janzen-Connell effect mediated by soil biota.

The negative effect of soil pathogens on seedling survival varies considerably among conspecific individuals, but the underlying mechanisms are largely unknown. For variation between heterospecifics, a common explanation is the Janzen-Connell effect: negative density dependence in survival due to specialized pathogens aggregating on common hosts. We test whether an intraspecific Janzen-Connell effect exists, i.e., whether the survival chances of one population's seedlings surrounded by a different conspecific population increase with genetic difference, spatial distance, and trait dissimilarity between them. In a shade-house experiment, we grew seedlings of five populations of each of two subtropical tree species (Castanopsis fissa and Canarium album) for which we measured genetic distance using intersimple sequence repeat (ISSR) analysis and eight common traits/characters, and we treated them with soil material or soil biota filtrate collected from different populations. We found that the relative survival rate increased with increasing dissimilarity measured by spatial distance, genetic distance, and trait differences between the seedling and the population around which the soil was collected. This effect disappeared after soil sterilization. Our results provide evidence that genetic variation, trait similarity, and spatial distance can explain intraspecific variation in plant-soil biotic interactions and suggest that limiting similarity also occurs at the intraspecific level.

Oviposition preference and offspring performance of Mechoris ursulus (Coleoptera: Attelabidae) in Cyclobalanopsis glaucoides (Fagales: Fagaceae) and Quercus franchetii (Fagales: Fagaceae) in Central Yunnan, China.

Mechoris ursulus (Roelofs) (Coleoptera: Attelabidae) is a pest weevil of Fagaceae oak trees in eastern Asia. The female has a distinct branch-cutting behavior in conjunction with oviposition in the acorns of its host plant. This study analyzed the factors influencing oviposition preference by carrying out continuous field surveys over the course of 2009-2010 and through laboratory rearing. The field survey showed that for both of the hosts, Cyclobalanopsis glaucoides Schottky (Fagales: Fagaceae) and Quercus franchetii Skan, M. ursulus preferred branches with fewer acorns and larger acorns on the same branch for oviposition. Laboratory rearing experiments showed offspring performance (i.e., survival rate and fresh weight of larvae) was significantly and positively correlated with acorn size. Preference for larger acorns could maximize the fitness of offspring by providing sufficient food source and space.

Ectomycorrhizal inoculum potential of northeastern US forest soils for American chestnut restoration: results from field and laboratory bioassays.

American chestnut (Castanea dentata) was once a dominant overstory tree in eastern USA but was decimated by chestnut blight (Cryphonectria parasitica). Blight-resistant chestnut is being developed as part of a concerted restoration effort to bring this heritage tree back. Here, we evaluate the potential of field soils in the northern portion of the chestnut's former range to provide ectomycorrhizal (EM) fungus inoculum for American chestnut. In our first study, chestnut seedlings were grown in a growth chamber using soil collected from three sites dominated by red oak (Quercus rubra) as inoculum and harvested after 5 months. Of the 14 EM fungi recovered on these seedlings, four species dominated in soils from all three sites: Laccaria laccata, a Tuber sp., Cenococcum geophilum, and a thelephoroid type. Seedlings grown in the nonsterilized soils were smaller than those growing in sterilized soils. In the second study, chestnut seedlings were grown from seed planted directly into soils at the same three sites. Seedlings with intermingling roots of established trees of various species were harvested after 5 months. Seventy-one EM fungi were found on the root tips of the hosts, with 38 occurring on chestnut seedlings. Multiple versus single host EM fungi were significantly more abundant and frequently encountered. The fungi observed dominating on seedlings in the laboratory bioassay were not frequently encountered in the field bioassay, suggesting that they may not have been active in mycelial networks in the field setting but were in the soils as resistant propagules that became active in the bioassay. These results show that soil from red oak stands can be used to inoculate American chestnut with locally adapted ectomycorrhizal fungi prior to outplanting, a relatively cost effective approach for restoration efforts.

Effects of roads on Castanopsis carlesii seedlings and their leaf herbivory in a subtropical forest in China.

The effects of a forest road on Castanopsis carlesii (Hemsley) Hayata (Fagales: Fagaceae) seedlings and their leaf herbivory were investigated in a subtropical forest at Jiulianshan National Nature Reserve, Jiangxi, China. A total of 1124 seedlings, 33949 leaves, 468 leaf mines, and 205 leaf galls were found. Generally, individual numbers, tree heights, and leaf numbers of C. carlesii seedlings became lower with increasing distances from the road. These results might indicate that old seedlings were fewer and survival rate of seedlings was lower in forest interiors. Leaf miners preferred the seedlings close to the forest road, while leaf gallers preferred the seedlings about 2 m from the road. Species diversity of leaf miners was higher in the forest interior area, while species diversity of leaf gallers was higher near the road. However, both leaf miners and leaf gallers decreased in general from the road to the interior forest. There were interspecific differences in the effects of roads on leaf miner species and leaf galler species. The effects of the road on seedlings and insects could be explained by varying microhabitat conditions and different ecological strategies.

[Short-term nitrogen addition reduces soil microbial nitrogen fixation rate in subtropical Pinus taiwanensis and Castanopsis faberi forests]. / çŸ­æœŸæ°®æ·»åŠ é™ä½Žäºšçƒ­å¸¦é»„å±±æ¾æž—å’Œç½—æµ®æ ²æž—åœŸå£¤å¾®ç”Ÿç‰©å›ºæ°®é€ŸçŽ‡.

Biological nitrogen (N) fixation is an important source of N in terrestrial ecosystems, but the response of soil microbial N fixation rate to N deposition in different forest ecosystems still remains uncertain. We conducted a field N addition experiment to simulate atmosphere N deposition in subtropical Pinus taiwanensis and Castanopsis faberi forests. We set up three levels of nitrogen addition using urea as the N source: 0 (control), 40 (low N), and 80 g N·hm-2·a-1(high N) to examine the chemical properties, microbial biomass C, enzyme activities, and nifH gene copies of top soils (0-10 cm). We also measured the microbial N fixation rate using the 15N labeling method. Results showed that N addition significantly reduced the soil microbial N fixation rate in the P. taiwanensis and C. faberi forests by 29%-33% and 10%-18%, respectively. Nitrogen addition significantly reduced N-acquiring enzyme (i.e., ß-1, 4-N-acetylglucosaminidase) activity and nifH gene copies in both forest soils. There was a significant positive correlation between the microbial N fixation rate and soil dissolved organic C content in the P. taiwanensis forest, but a significant negative relationship between the rate of soil microbial nitrogen fixation and NH4+-N content in the C. faberi forest. Overall, soil microbial N fixation function in the P. taiwanensis forest was more sensitive to N addition than that in the C. faberi forest, and the factors affecting microbial N fixation varied between the two forest soils. The study could provide insights into the effects of N addition on biological N fixation in forest ecosystems, and a theoretical basis for forest management.

[Characterizations of soil enzyme activities and stoichiometry in three subtropical forest stands]. / äºšçƒ­å¸¦ä¸‰ç§æž—åˆ†åœŸå£¤é ¶æ´»æ€§å’Œé ¶åŒ–å­¦è®¡é‡æ¯”ç‰¹å¾.

We conducted in a common garden experiment to explore the differences in soil enzyme activity, stoichiometry, and their influencing factors among a secondary Castanopsis carlesii forest, 10-year-old C. carlesii plantation, and Cunninghamia lanceolata plantation. The results showed that compared to the secondary forest, the soil organic carbon, total nitrogen, and dissolved organic carbon significantly decreased by 42.6%, 47.4%, and 60.9% in C. carlesii plantation, and by 42.9%, 36.7%, and 61.1% in C. lanceolata plantation. Soil microbial biomass C, microbial biomass N (MBN), and microbial biomass phosphorus decreased significantly by 40.6%, 35.5%, and 45.9% in C. carlesii plantation, and by 53.7%、56.4%, and 61.7% in C. lanceolata plantation. Compared to the secondary forest, soil enzymes activities in C. carlesii plantation did not change significantly, but in C. lanceolata plantation, the activities of ß-1,4-glucosidase and cellobiohydrolase significantly decreased by 51.2% and 59.8%, ß-N-acetyl glucosaminidase and acid phosphatase decreased significantly by 41.0% and 29.8%, and enzymatic C:N acquisition ratio and enzymatic C:P acquisition ratio significantly decreased by 11.3% and 7.7%, respectively. Results of redundancy analysis indicated that MBN and NO3--N were the primary factors influencing soil enzyme activity and enzymic stoichiometry. Collectively, there were significant differences in soil enzyme activity and microbial nutrient demands among different forest stands. Compared to secondary forests, the establishment of C. lanceolata plantations would intensify nutrient competition between plants and microbes, and exacerbate the N and P limitations for microbes.

Short-term responses of soil enzyme activities and stoichiometry to litter input in Castanopsis carlesii and Cunninghamia lanceolata plantations.

Litter input triggers the secretion of soil extracellular enzymes and facilitates the release of carbon (C), nitrogen (N), and phosphorus (P) from decomposing litter. However, how soil extracellular enzyme activities were controlled by litter input with various substrates is not fully understood. We examined the activities and stoichiometry of five enzymes including ß-1,4-glucosidase, ß-D-cellobiosidase, ß-1,4-N-acetyl-glucosaminidase, leucine aminopeptidase and acidic phosphatase (AP) with and without litter input in 10-year-old Castanopsis carlesii and Cunninghamia lanceolata plantations monthly during April to August, in October, and in December 2021 by using an in situ microcosm experiment. The results showed that: 1) There was no significant effect of short-term litter input on soil enzyme activity, stoichiometry, and vector properties in C. carlesii plantation. In contrast, short-term litter input significantly increased the AP activity by 1.7% in May and decreased the enzymatic C/N ratio by 3.8% in August, and decreased enzymatic C/P and N/P ratios by 11.7% and 10.3%, respectively, in October in C. lanceolata plantation. Meanwhile, litter input increased the soil enzymatic vector angle to 53.8° in October in C. lanceolata plantations, suggesting a significant P limitation for soil microorganisms. 2) Results from partial least squares regression analyses showed that soil dissolved organic matter and microbial biomass C and N were the primary factors in explaining the responses of soil enzymatic activity to short-term litter input in both plantations. Overall, input of low-quality (high C/N) litter stimulates the secretion of soil extracellular enzymes and accelerates litter decomposition. There is a P limitation for soil microorganisms in the study area.

Clonal growth and fine-scale genetic structure in tanoak (Notholithocarpus densiflorus: Fagaceae).

The combination of sprouting and reproduction by seed can have important consequences on fine-scale spatial distribution of genetic structure (SGS). SGS is an important consideration for species' restoration because it determines the minimum distance among seed trees to maximize genetic diversity while not prejudicing locally adapted genotypes. Local environmental conditions can be expected to influence levels of clonal spread and SGS, particularly in the case of disturbance regimes such as fire. Here, we characterize fine-scale genetic structure and clonal spread in tanoak from drier upland sites and more mesic lowland woodlands. Clonal spread was a significant mode of stand development, but spread was limited on average to about 5-6 m. Gene dispersal was decomposed into clonal and sexual components. The latter varied according to whether it was estimated from all ramets with the clonal component removed or for a single ramet per genet. We used the difference in these 2 estimates of gene dispersal as a measure of the effect of clonality on effective population size in this species. Although upland sites had a greater number of ramets per genet, most of the other indices computed were not significantly different. However, they tended to show greater heterozygote excess and shorter gene dispersal distances than the lowland sites. The average distance among inferred sibships on upland sites was approximately at the scale of maximum clonal range. This was not the case on lowland sites, where sibs were more dispersed. We recommend minimum distances among seed trees to avoid selecting clones and to maximize genetic diversity for restoration.

Nutritional evaluation and health promoting activities of nuts and seeds cultivated in Greece.

Available data suggest that genetic as well as environmental factors may influence nuts and seeds nutrients content. In this context nuts and seeds cultivated in Greece were studied. Macronutrients content was in agreement with that from other areas. Total phenolics content was in the range of 43.0 ± 2.1-1512.7 ± 60.7 mg GAE/100 g for chestnut and walnut, respectively. Thirteen to 22 individual phenolics were identified in the studied species. Oleanolic acid was in the range of 0.10-9.03 mg/100 g. Pumpkin seeds contained the higher squalene content (71.6 mg/100 g). ß-Sitosterol predominated in all samples except pumpkin seeds. Tocopherols ranged from 8.9 mg/100 g (chestnut) to 29.3 mg/100 g (almond). Nuts and seeds hydrophilic extracts at quantities corresponding to the estimated daily consumption by the Greeks succeeded in inhibiting LDL oxidation in vitro by increasing lag time 1.1-14.1 times. One serving of nuts or seeds may cover a significant fraction of health promoting microconstituents daily intake.