CHARACTERIZATION OF MOLECULAR MASS OF SIX WATER-SOLUBLE POLYSACCHARIDE -PROTEIN COMPLEXES FROM GANODERMA TSUGAE MYCELIUM

Six water-soluble polysaccharide-protein complexes coded as GM1, GM2, GM3, GM4, GM5 and GM6 wereisolated from the mycelium of Ganoderma tsugae by extracting with 0.2 mol/L phosphate buffer solution at 25, 40 and80℃, water at 120℃, 0.5 mol/L aqueous NaOH solution at 25 and 65℃, consecutively. Their chemical components wereanalyzed by using IR, GC, HPLC and ^(13)C-NMR, and some new results were obtained. The four samples GM1, GM2, GM3and GM4 are heteropolysaccharide-prote in complexes, in which, α- (1→3) linked D-glucose is the major monosaccharidewhile galactose, mannose and ribose are the secondary ones. GM5 and GM6 are β-(1→3)-D-glucan-protein complexes. Theprotein content increased from 32% to 69% with the progress of isolation. Weight-average molecu1ar mass M_w and theintrinsic viscosity [η] of the GM samples in 0.5 mol/L aqueous NaCl solution at 25℃ were measured systematically by laserlight scartering (LLS), size exclusion chromatography (SEC) combined with LLS, and viscometry. The M_w of GM1 to GM6are 35.5, 46.8, 58.9, 41.6, 3.3 and 22.0×10~4, respectively. The conformation and molecular mass of the two fractions of sample GM5 were characterized satisfactorily by SEC-LLS without further fractionation.

Antitumor Active Protein-containing Glycans from the Body of Ganoderma tsugae

To explore the effects of traditional herbal medicine Ganoderrna tsugae（G, tsugae） on immunomodulatory and antitumor activities, the crude polysaccharides of G. tsugae were purified by filtration, diethylaminoethyl（DEAE） sepharose-fast flow chromatography and sephadex G-100 size-exclusion chromatography. Two main fractions, pro- tein-containing glyeans CSSLP-1 and CSSLP-2, were obtained via the gradient elution. The protein content, molecu- lar weight, and monosaccharide composition of the two fractions were analyzed. Furthermore, the influence of the protein-containing glycans from G. tsugae on the activation of human acute monocytic leukemia cell line（THP-l） and their antitumor activities to the human hepatocellular liver carcinoma cell（HepG-2） in vitro were evaluated. The re- sults indicate that CSSLP-1 and CSSLP-2 could increase the pinocytie activity of THP-1 cells and induce THP-1 cells to produce the eytokines of TNFa and IL-2, significantly. CSSLP-1 and CSSLP-2 also played an inhibiting effect on the cancer cell（HepG-2）. Moreover, the anti-proliferation activity of CSSLP-1 and CSSLP-2 increased with the par- ticipation of TNFa and IL-2 or other antitumor factors induced from THP-1 cells by G. tsugae protein-containing glycan fractions.

Fitness and physiology of Adelges tsugae （Hemiptera： Adelgidae） in relation to the health of the eastern hemlock

The hemlock woolly adelgid, Adelges tsugae Annand is an invasive insect that frequently causes hemlock （Tsuga spp.） mortality in the eastern United States. Studies have shown that once healthy hemlocks become infested by the adelgid, nutrients are depleted from the tree, leading to both tree decline and a reduction of the adelgid population. Since A. tsugae is dependent on hemlock for nutrients, feeding on trees in poor health may affect the ability of the insect to obtain necessary nutrients and may consequently affect their physiological and population health. Trees were categorized as lightly or moderately impacted by A. tsugae based on quantitative and qualitative tree health measurements. Population health ofA. tsugae on each tree was determined by measuring insect density and peak mean fecundity; A. tsugae physiological health was determined by measuring insect biomass, total carbon, carbohydrate, total nitrogen, and amino nitrogen levels. Adelges tsugae from moderately impacted trees exhibited significantly greater fecundity than from lightly impacted trees. However, A. tsugae from lightly impacted hemlocks contained significantly greater levels of carbohydrates, total nitrogen, and amino nitrogen. While the results of the physiological analysis generally support our hypothesis that A. tsugae on lightly impacted trees are healthier than those on moderately impacted trees, this was not reflected in the population health measurements. Adelges tsugae egg health in response to tree health should be verified. This study provides the first examination of A. tsugae physiological health in relation to standard A. tsugae population health measures on hemlocks of different health levels.

铁杉属的起源、扩散与传播——基于化石证据

铁杉属是松科的重要组成部分,具有典型的北美—东亚间断分布特征。关于该属的起源、扩散与传播,目前存在较多的争议。本次研究收集了亚洲铁杉属大化石记录,整合了铁杉属花粉记录,获取地质历史时期该属在中国的分布,推测该属的起源和传播路径。该属大约起源于晚白垩世的西欧地区,于马特斯里赫特期—达宁期通过欧亚大陆传播到中国,而在古新世—始新世时期通过北大西洋陆桥传播到北美。早始新世以后北美与东亚的铁杉属植物可以通过白令陆桥进行物种交流,当图尔盖海峡在渐新世干涸后,铁杉属能够通过西西伯利亚和中亚哈萨克斯坦地区在欧亚大陆上交流。在晚中新世铁杉属可以通过北大西洋陆桥进行交流。依据化石记录推断的铁杉属起源和传播的结果与分子学研究存在一定差异,需要更多的铁杉属化石记录及分子学研究提供更可靠的证据。

Spatial Distribution of Hemlock Woolly Adelgid Induced Hemlock Mortality in the Southern Appalachians

Hemlock woolly adelgid (Adelges Tsugae Annand, HWA) outbreaks are posing a major threat to eastern hemlock (Tsuga canadensis L. Carr.) and Carolina hemlock (Tsuga caroliniana Engelm.) forest landscapes in the eastern USA. As foundation species, hemlocks play a variety of functional roles in forest landscapes. These species usually occur as isolated canopies and mixed species in landscapes where variation in topography is extreme. Spatially explicit inventory information on HWA induced hemlock mortality at landscape scale does not exist. High resolution aerial imageries enable landscape scale assessment even at the individual tree level. Accordingly, our goal was to investigate spatial pattern and distribution of HWA induced hemlock mortality using a high resolution aerial image mosaic in the Linville River Gorge, Southern Appalachians, western North Carolina. Our study objectives were: 1) to detect dead trees within the Lower Linville River watershed;2) to estimate the area occupied by dead trees in the forest canopy surface;3) to investigate the relationship of dead hemlocks and topography;and 4) to define the spatial pattern of the dead trees. We found ca. 10,000 dead trees within the study area, occupying over 7 ha of the canopy surface with an average area of 36 m2 per dead tree. The density of the dead trees was higher in proximity to the Linville River, at higher elevations, and on northern and northwestern aspects. Spatial pattern of the dead trees was generally clustered at all spatial scales. We suggest that although the reduction in plant biomass resulting from herbivory within the landscapes is modest, impact of the clustered distribution of hemlock mortality, especially in the riparian zones, is noteworthy. Our analysis of the pattern of hemlock decline provides new means for projecting future impacts of HWA on the range of hemlock distribution in eastern North America.

A New Species of Tsuga(Pinaceae) based on Lignified Wood from the Late Miocene of Central Yunnan, China, and Its Paleoenvironmental Implications

A new species, Tsuga nanfengensis sp.nov.（Pinaceae）, is described on the basis of lignified fossil wood from the late Miocene of the Xianfeng Basin, central Yunnan, southwestern China.Detailed observation of the fossil wood specimens show the following characteristics： distinct growth rings, absence of resin canals, uniseriate bordered pits in the radial wall of tracheids, ray tracheids and piceoid and cupressoid cross-field pits.These features indicate similarities to the wood of extant Tsuga canadensis, T.chinensis, and T.dumosa.According to the fossil record, Tsuga was present in Xundian County during the Miocene.Today Tsuga is drought intolerant, preferring wet conditions with no extant species growing naturally in Xundian County.The presence of Tsuga in the Miocene of Xundian County indicates a humid climate consistent with previous palaeoclimatic reconstructions showing a wetter and probably shorter dry season in the Miocene, relative to the present day.Therefore, the change in the local climate such as increasing aridity through the Miocene might explain the local extinction of Tsuga from central Yunnan.

Volatile Constituents of Juniperus communis L., Taxus canadensis Marshall. and Tsuga canadensis （L.） Carr. from Canada

The essential oil composition leaves of Juniperus communis L., Taxus canadensis Marshall. and Tsuga canadensis （L.） Carr. from Canada were investigated by head space solid phase microextraction （HS-SPME） and gas chromatography/mass spectrometry （GC-MS）. Thirty-three, thirty and thirty-one components were identified representing 95.78%, 93.89%, 96.14% of the oil, respectively. Limonene （26.12%）, benzene （15.62%）, 13-mrycene （9.08%） and β-pinene （7.30%） were found to be the main constituents of J. communis; 1-propanone （36.38%）, morpholine （10.95%）, methylamine （9.10%） and methanone （8.14%） were detected main components of Taxus canadensis; bornylacetate （26.84%）, α-pinene （23.74%）, camphene （11.93%） and limonene （6.02%） were determined as major constituents of Tsuga canadensis. The chemical distributions of the essential oil compounds in the genus pattern were discussed in means of chemotaxonomy and natural products.

Ecological factors influencing the beneficial endosymbionts of the hemlock woolly adelgid (Hemiptera:Adelgidae)

Bacterial endosymbionts of sap-sucking insects provide their host with a num- ber of beneficial qualities,including the supply of nutrition,defense against parasitoids, and protection from heat stress.Damage to these bacterial associates can therefore have a negative impact on the fitness of their insect host.We evaluated observational and experi- mental factors regarding the normative hemlock woolly adelgid (Adelges tsugae Annand) (Hemiptera:Adelgidae)to help understand the roles of its three recently identified symbionts,including under heat stress conditions.The prevalence of A.tsugae's facultafive symbiont (Serratia symbiotica)was examined at different spatial scales to determine how variable infection rates are for this symbiont.There was no significant difference found in infection rates between adelgids on a tree,within a plot,or within a state.However, significantly more adelgids in Georgia (95%)had S.symbiotica compared to those in New York (68%).Microsatellite genotyping of the adelgids found that this difference was most likely not the result of a second introduction ofA.tsugae into eastern North America.Comparison orS.symbiotica proportions between first and fourth instars showed that symbiont absence did not affect the ability ofA.tsugae to survive aestivation.Evaluations of sym- biont densities within each adelgid found that when S.symbiotica was absent,the density of obligate symbionts was significantly higher.Exposure to heat stress (32.5℃)was not consistently correlated with changes in symbiont densities over a 4-d period.Overall,we have shown that symbiont prevalence and densities vary within the broad population of A.tsugae in eastern North America,with potentially significant effects upon the ecology of this important pest.

The importance of large-diameter trees to the creation of snag and deadwood biomass

Background:Baseline levels of tree mortality can,over time,contribute to high snag densities and high levels of deadwood(down woody debris)if fire is infrequent and decomposition is slow.Deadwood can be important for tree recruitment,and it plays a major role in terrestrial carbon cycling,but deadwood is rarely examined in a spatially explicit context.Methods:Between 2011 and 2019,we annually tracked all trees and snags≥1 cm in diameter and mapped all pieces of deadwood≥10 cm diameter and≥1 m in length in 25.6 ha of Tsuga heterophylla/Pseudotsuga menziesii forest.We analyzed the amount,biomass,and spatial distribution of deadwood,and we assessed how various causes of mortality that contributed uniquely to deadwood creation.Results:Compared to aboveground woody live biomass of 481 Mg ha^(−1)(from trees≥10 cm diameter),snag biomass was 74 Mg ha^(−1) and deadwood biomass was 109 Mg ha^(−1)(from boles≥10 cm diameter).Biomass from large-diameter trees(≥60 cm)accounted for 85%,88%,and 58%,of trees,snags,and deadwood,respectively.Total aboveground woody live and dead biomass was 668 Mg ha^(−1).The annual production of downed wood(≥10 cm diameter)from tree boles averaged 4 Mg ha^(−1) yr^(−1).Woody debris was spatially heterogeneous,varying more than two orders of magnitude from 4 to 587 Mg ha^(−1) at the scale of 20 m×20 m quadrats.Almost all causes of deadwood creation varied in importance between large-diameter trees and small-diameter trees.Biomass of standing stems and deadwood had weak inverse distributions,reflecting the long period of time required for trees to reach large diameters following antecedent tree mortalities and the centennial scale time required for deadwood decomposition.Conclusion:Old-growth forests contain large stores of biomass in living trees,as well as in snag and deadwood biomass pools that are stable long after tree death.Ignoring biomass(or carbon)in deadwood pools can lead to substantial underestimations of sequestration and stability.

Effect of Gap Size on Seedling Establishment of Tsuga longibracteata

Effects of three gaps which are large (118 m2), medium (86 m2) and small (20 m2), respectively, and under canopy of Tsuga longibracteata forest on the seedling establishment of T. longibracteata were studied through seed burial experiments from December 2003 to January 2005 in Tianbaoyan National Nature Reserve of Fujian, China. The results showed that the area of gap had an evident effect on the seedling establishment of T. longibracteata. The seedling emergence rates of T. longibracteata in plots of large gap, medium gap, small gap and under canopy were 10%, 10%, 4% and 6%, representing an increasing trend along with the gap size increasing without a significant difference. Rain eroding and insects feeding were two main factors leading to seedling death. The larger the gap size was, the more seedlings were killed by rain erosion and the fewer seedlings were killed by insects feeding. The emergence time of seedlings was almost same in all plots while their death time was different respectively. The gap size had a significant impact on seedling survival rate. The seedling survival rate was highest in the medium gap plot (27.0%) and next to the highest in large gap plot (7.3%), and seedling in small gap plot and under canopy plot died out after one growing season. Increased light supply in gaps was favorable for the seedlings growth and survival. Increased light supply in the large gap could enhance the growth of seedling leaf and root of T. longibracteata, and the seedling in turn allocated more dry mass to root and leaf, but it has little impact on the growth of stem. This research indicates that T. longibracteata is a pioneer species and its seedling establishment need a medium or large gap (>50 m2).