Free amino acid and protein levels,and γ‐glutamyltransferase activity in Pinus sylvestris apical buds and shoots during the growing season

Free amino acid and protein levels, and γ‐glutamyltransferase activity in apical buds and shoots of Scots pine during the growing season. The aim of the study was to obtain the basic information about nitrogen mobilization needed in carrying out studies on the optimal nitrate and ammonium ratios in the metabolism of Scots pine (Pinus sylvestris L.). Considerable seasonal changes in the concentrations of free amino acids and other ninhydrin‐positive low molecular‐weight compounds were observed in the buds and shoots of Scots pine. 43 different amino compounds were identified, the concentrations of arginine, glutamine, glutamic acid, γ‐aminobutyric acid, alanine and aspartic acid being highest at the break of dormancy. The amounts of certain amino compounds decreased during the growing season, those of arginine, ethanolamine and various ammonium compounds in particular. The amount of glutamic and aspartic acids, glycine, alanine and γ‐aminobutyric acid, however, remained relatively constant. The protein concentration and the specific activity of γ‐glutamyltransferase increased towards the end of the growing period. Slight differences were found between the fertilized trees and the control trees.     

Effects of root temperature on growth and photosynthesis in conifer seedlings during shoot elongation

Growth and gas exchange characteristics were studied in pine (Pinus sylvestris L.) and spruce (Picea abies Karst.) seedlings grown in hydroponic culture in the presence of N (50 mg l(-1)) and transferred at the start of their second growing season to tap water at 5, 8, 12, 16 or 20 degrees C (air temperature between 18-20 degrees C) for 3 weeks (pine) or 5 weeks (spruce). Root growth of both species was completely inhibited at root temperatures of 5 and 8 degrees C, but increased almost exponentially as root temperature increased. Shoot growth was maximal at 12 degrees C in both pine and spruce and decreased at low root temperatures. In both species, CO(2) uptake was decreased at low root temperatures and appeared to be influenced by the pattern of nitrogen retranslocation. In pine seedlings, as root temperature increased, an increasing proportion of the total nitrogen pool was retranslocated to the new shoot, whereas in spruce seedlings nitrogen was retranslocated to the roots. Differences in the retranslocation of nitrogen in the two species were reflected in the amount of soluble protein in needles, which at the end of the experiment increased with increasing root temperature in pine, but decreased in spruce. Our data suggest that in spruce, but not pine, CO(2) uptake was limited by the amount of Rubisco.     

Characterization of glutathione S-transferase from dwarf pine needles (Pinus mugo Turra)

Glutathione S-transferase activity conjugating xenobiotics with glutathione (GSH) was found in extracts from needles of dwarf pine (Pinus mugo Turra). In vivo incubation of needle segments with the herbicide fluorodifen at 25 degrees C resulted in conversion of the xenobiotic to water-soluble products at initial rates of 0.7 nmol h(-1) g(fw) (-1). At 15 degrees C, the initial rate of product formation was decreased to 0.1 nmol h(-1) g(fw) (-1). In vitro conjugation studies with chloro-2,4-dinitrobenzene (CDNB) and 1,2-dichloro-4-nitrobenzene (DCNB) as model substrates gave apparent K(m) values of 0.5 mM GSH and 1.14 mM CDNB in the GSH/CDNB system and 0.3 mM GSH and 0.44 mM DCNB in the GSH/DCNB system. The pH optimum was between 7.7 and 7.9 for both the GSH/CDNB and the GSH/DCNB systems. The temperature optimum for these model substrates was between 30 and 35 degrees C, and only minute amounts of enzyme activity were detected at 15 degrees C. The activation energy in the temperature range of 15 to 30 degrees C was 46 kJ mol(-1). Dwarf pine glutathione S-transferase exhibited an approximate molecular weight of 52 kD.     

不同松树感染松材线虫后NO和核酸酶变化与其抗病性的关系

以黑松、马尾松和火炬松为研究对象,对其接种松材线虫后4,12,24,48,72,96和120 h的NO和核酸酶的活性进行测定.3种松树接种松材线虫后,针叶内NO平均含量的积累为火炬松＜马尾松＜黑松,这种差异在12 ～48 h尤为明显;48h后较其割伤处理(CK1)的增加幅度为火炬松＞马尾松＞黑松.同时3种松树接种后的核酸酶变化趋势存在差异,单链核酸酶活性在4～48 h为火炬松＞马尾松＞黑松,而火炬松双链核酸酶活性在4～96h时却一直低于马尾松和黑松,这表明不同松树体内单链或双链核酸酶的活性变化与其抗病性有关.且3种松树在接种松材线虫后,表现出症状的时间顺序为黑松最早,马尾松次之,火炬松最晚.这表明NO和核酸酶的变化与松树感染松材线虫后的病程发展密切相关.     

Accumulation of free amino acids in the shoots and roots of three northern conifers during drought

The effects of drought on the free amino acid pools in 21- to 23-week-old seedlings of black spruce (Picea mariana (Mill.) Britt.), white spruce (Picea glauca (Moench.) Voss) and jack pine (Pinus banksiana Lamb.) were followed during soil drying. Although water and pressure potentials were sensitive to water deficits, large changes in osmotic potential were not recorded until after the development of severe drought. Total soluble amino nitrogen in the shoots and roots of the three species rose as turgor declined, with peak concentrations attained late in the drought period when the pressure potentials of the shoots approached zero. All white spruce seedlings were alive at zero turgor and showed large decrements in osmotic potential, but concentrations of free amino nitrogen in the roots and shoots showed only modest increases, reaching 125 to 150% of their control values. In contrast, large numbers of black spruce and jack pine were dead or severely damaged at zero turgor, and only small changes in osmotic potential were detected during soil drying. Nevertheless, concentrations of soluble amino nitrogen in both species reached 150 to 200% of control values a few days before the seedlings died. Alanine, arginine, aspartic acid/asparagine, glutamic acid/glutamine, glycine, hydroxyproline and proline were the major components of the free amino acid pools under both water-stressed and non-stressed conditions, with the largest and most consistent increases observed in the roots of all three conifers. Although proline was an important and dynamic component of the free pools, absolute concentrations were commony equalled or exceeded by other free amino acids in the roots and shoots and nearly always exceeded by the concentration of aspartic acid/asparagine in both tissues. Differences in drought resistance among the three conifers were not reflected by unique patterns of amino acid accumulation or by large differences in absolute concentrations of the free amino acid pools.     

Seasonal changes in amino acids, protein and total nitrogen in needles of fertilized Scots pine trees

Seasonal changes in amino acids, protein and total nitrogen in needles of 30-year-old, fertilized Scots pine (Pinus sylvestris L.) trees growing in Northern Sweden were investigated over two years in field experiments. The studied plots had been fertilized annually for 17 years with (i) a high level of N, (ii) a medium level of N, or (iii) a medium level of N, P and K. Trees growing on unfertilized plots served as controls. In control trees, glutamine, glutamic acid, gamma-aminobutyric acid, aspartic acid and proline represented 50-70% of the total free amino acids determined. Arginine was present only in low concentrations in control trees throughout the year, but it was usually the most abundant amino acid in fertilized trees. Glutamine concentrations were high during the spring and summer in both years of study, whereas proline concentrations were high in the spring but otherwise low throughout the year. In the first year of study, glutamic acid concentrations were high during the spring and summer, whereas gamma-aminobutyric acid was present in high concentrations during the winter months. This pattern was less pronounced in the second year of investigation. The concentrations of most amino acids, except glutamic acid, increased in response to fertilization. Nitrogen fertilization increased the foliar concentration of arginine from < 1 micromol g(dw) (-1) in control trees to a maximum of 110 micromol g(dw) (-1). Trees fertilized with nitrogen, phosphorus and potassium had significantly lower arginine concentrations than trees fertilized with the same amount of nitrogen only. Protein concentrations were similar in all fertilized trees but higher than those in control trees. For all treatments, protein concentrations were high in winter and at a minimum in early spring. In summer, the protein concentration remained almost constant except for a temporary decrease which coincided with the expansion of new shoots. Apart from arginine, the amino acid composition of proteins was similar in all treatments.     

Responses of Scots pine (Pinus sylvestris) seedlings grown in different nutrient regimes to changing root zone temperature in spring

We examined effects of nutrient availability and changing root zone temperature (RZT) on growth, gas exchange and plasma membrane H(+)-ATPase (PM-ATPase) activity of roots of 1-year-old Scots pine (Pinus sylvestris L.) seedlings during spring flushing. The 6-week growth-chamber experiment was carried out in hydroponic cultures that supplied the seedlings with low (0.5 mM N) or high (3 mM N) nutrient concentration and two rates of increase in RZT were simulated: slow warming (SW-treatment) and fast warming (FW-treatment). Air temperature, humidity, and light conditions were similar in all treatments. Growth of roots and shoots was retarded at low RZT, and fresh mass increment of roots was closely correlated with RZT sum. High nutrient availability increased nitrogen concentrations of needles and stems, but only at RZTs >13 degrees C. Low RZT and low availability of nutrients suppressed gas exchange of the seedlings. Real PM-ATPase activity was highly dependent on RZT. At high RZTs, real PM-ATPase activity was affected by nutrient availability but this effect was related to root growth. We conclude that, under conditions of high nutrient availability, Scots pine seedlings can compensate for the suppressive effects of long-term exposure to low RZT by rapidly accelerating growth, gas exchange and root metabolism, but only when RZT has increased above a threshold value, which was 13 degrees C in this study.     

Free amino acids and protein in Scots pine seedlings cultivated at different nutrient availabilities

Scots pine (Pinus sylvestris L.) seedlings of a provenance from northern Sweden were cultivated hydroponically for 7 weeks in a climate chamber. The nutrient solution contained either 2.5 (low-N) or 50 (high-N) mg N l(-1) with other essential elements added in a fixed optimal proportion to the nitrogen. After 5 and 7 weeks, the seedlings were analyzed for growth, total nitrogen and other essential nutrients, protein and free amino acids. Low-N seedlings grew more slowly and had higher root/shoot ratios than high-N seedlings. With respect to total nitrogen, the effect of the lower nutrient supply was mainly on the nitrogen content of the whole plant and the allocation of nitrogen among tissues, not on tissue nitrogen concentration. This was also the case for potassium, phosphorus, calcium and magnesium. The proportions by weight among these macronutrients in the whole seedlings were similar in both nutrient regimes. The proportion and concentration of sulfur were significantly lower in low-N seedlings than in high-N seedlings, because of a lower net uptake of sulfur than of other macronutrients. The shoot, needles and stem of low-N seedlings had higher concentrations of free amino acids and lower concentrations of protein than the shoot, needles and stem of high-N seedlings. Arginine dominated the pool of free amino acids in the low-N seedlings, whereas glutamine predominated in the high-N seedlings. We conclude that Scots pine seedlings accumulated soluble nitrogen as arginine when net protein synthesis was limited by factors other than nitrogen availability. Nutritional imbalance, as revealed by growth characteristics and a suboptimal proportion and concentration of sulfur in the seedlings, probably affected synthesis of S-amino acids, resulting in the diversion of assimilated nitrogen to arginine instead of protein.     

Characterization and application of recombinant β-glucosidase (BglH) from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> KCTC 1918

β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC.3.2.1.21) catalyzes the hydrolysis of β-glucosidic bonds between saccharides and aryl or alkyl groups. A gene encoding β-glucosidase from Bacillus licheniformis KCTC 1918, an anaerobic spore-forming soil bacterium, was cloned and characterized. The structural gene for the β-glucosidase consists of 1410 bp encoding 469 amino acid residues, and has a molecular weight of 53.4 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis with 12% separating gel. The enzyme activity was determined against pNPG as a substrate. The enzyme was optimally active at pH 6.0 (citrate-phosphate buffer) and 47°C. β-Glucosidase retained 100% of its original activity for 24 h. The activity of the enzyme was stimulated by glycerol and urea and was decreased by Ca²⁺, Cu²⁺, Hg²⁺, Mg²⁺, and Mn²⁺. In particular, Cu²⁺ had the strongest negative effect on β-glucosidase activity. The purified β-glucosidase was active against pNPG and cellobiose. When the β-glucosidase was tested for cellulose hydrolysis, the supplement of β-glucosidase with cellulose increased the glucose yield from pine wood powder by 139.8%.     

Changes in freezing tolerance, plasma membrane H+-ATPase activity and fatty acid composition in Pinus resinosa needles during cold acclimation and de-acclimation

It has previously been suggested that plasma membrane ATPase (PM H+-ATPase, EC 3.6.1.3.) is a site of incipient freezing injury because activity increases following cold acclimation and there are published data indicating that activity of PM H+-ATPase is modulated by changes in lipids associated with the enzyme. To test and extend these findings in a tree species, we analyzed PM H+-ATPase activity and the fatty acid (FA) composition of glycerolipids in purified plasma membranes (PMs) prepared by the two-phase partition method from current-year needles of adult red pine (Pinus resinosa Ait.) trees. Freezing tolerance of the needles decreased from -56 degrees C in March to -9 degrees C in May, and increased from -15 degrees C in September to -148 degrees C in January. Specific activity of vanadate-sensitive PM H+-ATPase increased more than two-fold following cold acclimation, despite a concurrent increase in protein concentration. During de-acclimation, decreases in PM H+-ATPase activity and freezing tolerance were accompanied by decreases in the proportions of oleic (18:1) and linoleic (18:2) acids and increases in the proportions of palmitic (16:0) and linolenic (18:3) acids in total glycerolipids extracted from the plasma membrane fraction. This pattern of changes in PM H+-ATPase activity and the 18:1, 18:2 and 18:3 fatty acids was reversed during cold acclimation. In the PM fractions, changes in FA unsaturation, expressed as the double bond index (1 x 18:1 + 2 x 18:2 + 3 x 18:3), were closely correlated with changes in H+-ATPase specific activity (r2 = 0.995). Changes in freezing tolerance were well correlated with DBI (r2 = 0.877) and ATPase specific activity (r2 = 0.833) in the PM fraction. Total ATPase activity in microsomal fractions also closely followed changes in freezing tolerance (r2 = 0.969). We conclude that, as in herbaceous plants, simultaneous seasonal changes in PM H+-ATPase activity and fatty acid composition occur during cold acclimation and de-acclimation in an extremely winter hardy tree species under natural conditions, lending support to the hypothesis that FA-regulated PM H+-ATPase activity is involved in the cellular response underlying cold acclimation and de-acclimation.     

Characterization of nitrogenous solutes in tissues and xylem sap of Leucaena leucocephala

Amino acid profiles of leaf, stem, and root tissues from nodulated and nonnodulated Leucaena leucocephala (Lam.) de Wit plants were determined by gas chromatography-mass spectrometry. High concentrations of mimosine and several other potentially toxic nonprotein amino acids, including pipecolic acid and two isomers of hydroxypipecolic acid, were identified in the tissues. Five metabolites remain unidentified. Of the foliar free amino acid nitrogen, 57-66% was associated with the potentially toxic amino acids. Major constituents in the leaf tissues of nonnodulated plants were mimosine and hydroxypipecolic acid (isomer 1). Mimosine was recovered in both the neutral plus basic and acidic amino acid fractions. Major differences between amino acid profiles of nodulated and nonnodulated roots were the low percentages of asparagine + aspartate (3.6% of the total pool compared to 33% in nodulated plants) and pipecolic acid in nonnodulated roots (1% of the total compared to 12.5% in nodulated plants). A novel plant betaine (dihydroxypipecolic acid betaine) was identified by fast-atom-bombardment mass spectrometry in leaf tissues, albeit at relatively low concentrations (< 1 micro mol per gram fresh weight). Analyses of the xylem sap collected from nodulated plants confirmed that Leucaena is an asparagine transporter, as suggested by the high concentrations of asparagine and the low concentrations of ureides in its root nodules. Amino acid profiles of xylem sap from nonnodulated plants showed extremely low concentrations of asparagine + aspartate (0.12 micro mol ml(-1)), whereas asparagine + aspartate was the major constituent (4.38 micro mol ml(-1)) in the xylem sap of nodulated plants. Two nonprotein amino acids, pipecolic acid and hydroxypipecolic acid, were major constituents of the xylem sap of nodulated and nonnodulated plants, respectively. Three unidentified compounds detected in xylem sap samples from both nodulated and nonnodulated plants did not correspond with any of the peaks characterized from tissue samples.     

Influence of ozone and nitrogen deposition on bark beetle activity under drought conditions

Four years of severe drought from 1999 through 2003 led to unprecedented bark beetle activity in ponderosa and Jeffrey pine in the San Bernardino and San Jacinto Mountains of southern California. Pines in the San Bernardino Mountains also were heavily impacted by ozone and nitrogenous pollutants originating from urban and agricultural areas in the Los Angeles basin. We studied bark beetle activity and bark beetle associated tree mortality in pines at two drought-impacted sites in the San Bernardino Mountains, one receiving high levels of atmospheric pollutants, and one with more moderate atmospheric input. We also investigated the effects of nitrogen addition treatments of 0, 50 and 150 kg N ha⁻¹ year⁻¹ at each site. Tree mortality and beetle activity were significantly higher at the high pollution site. Differences in beetle activity between sites were significantly associated with ozone injury to pines, while differences in tree mortality between sites were significantly associated with both ozone injury and fertilization level. Tree mortality was 9% higher and beetle activity 50% higher for unfertilized trees at the high pollution site compared to the low pollution site. Tree mortality increased 8% and beetle activity increased 20% under the highest rates of nitrogen additions at the low pollution site. The strong response in beetle activity to nitrogen additions at the low pollution site suggests that atmospheric nitrogen deposition increased tree susceptibility to beetle attack at the high deposition site. While drought conditions throughout the region were a major factor in decreased tree resistance, it appears that both ozone exposure and atmospheric nitrogen deposition further increased pine susceptibility to beetle attack.     

Pistagremic acid,a glucosidase inhibitor from Pistacia integerrima

Pistacia integerrima Stewart in traditionally used as folk remedy for various pathological conditions including diabetes. In order to identify the bioactive compound responsible for its folk use in diabetes, a phytochemical and biological study was conducted. Pistagremic acid (PA) was isolated from the dried galls extract of P. integerrima. Strong α-glucosidase inhibitory potential of PA was predicted using its molecular docking simulations against yeast α-glucosidase as a therapeutic target. Significant experimental α-glucosidase inhibitory activity of PA confirmed the computational predictions. PA showed potent enzyme inhibitory activity both against yeast (IC₅₀: 89.12 ± 0.12 μM) and rat intestinal (IC₅₀: 62.47 ± 0.09 μM) α-glucosidases. Interestingly, acarbose was found to be more than 12 times more potent an inhibitor against mammalian (rat intestinal) enzyme (having IC₅₀ value 62.47 ± 0.09 μM), as compared to the microbial (yeast) enzyme (with IC₅₀ value 780.21 μM). Molecular binding mode was explored via molecular docking simulations, which revealed hydrogen bonding interactions between PA and important amino acid residues (Asp60, Arg69 and Asp 70 (3.11 Å)), surrounding the catalytic site of the α-glucosidase. These interactions could be mainly responsible for their role in potent inhibitory activity of PA. PA has a strong potential to be further investigated as a new lead compound for better management of diabetes.     

Freezing tolerance of conifer seeds and germinants

Survival after freezing was measured for seeds and germinants of four seedlots each of interior spruce (Picea glauca x engelmannii complex), lodgepole pine (Pinus contorta Dougl. ex Loud.), Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata Donn ex D. Donn). Effects of eight seed treatments on post-freezing survival of seeds and germinants were tested: dry, imbibed and stratified seed, and seed placed in a growth chamber for 2, 5, 10, 15, 20 or 30 days in a 16-h photoperiod and a 22/17 degrees C thermoperiod. Survival was related to the water content of seeds and germinants, germination rate and seedlot origin. After freezing for 3 h at -196 degrees C, dry seed of most seedlots of interior spruce, Douglas-fir and western red cedar had 84-96% germination, whereas lodgepole pine seedlots had 53-82% germination. Freezing tolerance declined significantly after imbibition in lodgepole pine, Douglas-fir and interior spruce seed (western red cedar was not tested), and mean LT50 of imbibed seed of these species was -30, -24.5 and -20 degrees C, respectively. Freezing tolerance continued to decline to a minimum LT50 of -4 to -7 degrees C after 10 days in a growth chamber for interior spruce, Douglas-fir and lodgepole pine, or after 15 days for western red cedar. Minimum freezing tolerance was reached at the stage of rapid hypocotyl elongation. In all species, a slight increase in freezing tolerance of germinants was observed once cotyledons emerged from the seed coat. The decrease in freezing tolerance during the transition from dry to germinating seed correlated with increases in seed water content. Changes in freezing tolerance between 10 and 30 days in the growth chamber were not correlated with seedling water content. Within a species, seedlots differed significantly in freezing tolerance after 2 or 5 days in the growth chamber. Because all seedlots of interior spruce and lodgepole pine germinated quickly, there was no correlation between seedlot hardiness and rate of germination. Germination rate and freezing tolerance of Douglas-fir and western red cedar seedlots was negatively correlated. There was a significant correlation between LT50 after 10 days in the growth chamber and minimum spring temperature at the location of seedlot origin for interior spruce and three seedlots of western red cedar, but no relationship was apparent for lodgepole pine and Douglas-fir.     

Acquired thermotolerance of jack pine, white spruce and black spruce seedlings

The acquired thermotolerance of first-year seedlings of jack pine (Pinus banksiana Lamb.) hardened at 36, 38, 40 or 42 degrees C for 90, 180 or 360 minutes and of black spruce (Picea mariana (Mill.) B.S.P.) hardened at 34, 36, 38 or 40 degrees C for 30, 90, 180 or 360 minutes was determined by comparison of needle damage to that of non-hardened seedlings (25 degrees C) following exposure to temperatures of 49 and 47.5 degrees C, respectively. Compared to seedlings kept at 25 degrees C, heat injury sustained from exposure to high temperatures was markedly reduced following hardening for 180 minutes at 36 and 38 degrees C in jack pine and black spruce, respectively. Increasing the exposure time at 36 degrees C in jack pine, and at 36 to 40 degrees C in black spruce, also reduced needle damage. The duration of increased thermotolerance was investigated in jack pine, black spruce and white spruce (Picea glauca (Moench) Voss) by comparing heat injury from high temperatures in non-hardened seedlings and in seedlings hardened at 38 degrees C for 180 minutes a day for either 1, 3 or 6 days. In all three species, the duration of acquired thermotolerance increased with the number of days of heat hardening. For jack pine and white spruce seedlings hardened at 38 degrees C for 6 days, increased thermotolerance persisted for at least 14 and 10 days, respectively, after the end of the hardening treatment. In contrast, the thermotolerance of black spruce seedlings hardened at 38 degrees C for 6 days remained elevated for only 4 days.     

Responses of loblolly pine seedlings to elevated CO(2) and fluctuating water supply

Osmotic adjustment of loblolly pine (Pinus taeda L.) seedlings to fluctuating water supply in elevated CO(2) was investigated. Seedlings were grown in controlled-environment chambers in either 350 or 700 micro l l(-1) CO(2) with weekly watering for four months, after which they were either watered weekly (well-watered treatment) or every two weeks (water-stress treatment) for 59 days. Osmotic adjustment was assessed by pressure-volume analysis of shoots and by analysis of soluble carbohydrates and free amino acids in roots during the last drying cycle. In well-watered seedlings, elevated CO(2) increased the concentration of soluble sugars in roots by 68%. Water stress reduced the soluble sugar concentration in roots of seedling growing in ambient CO(2) to 26% of that in roots of well-watered seedlings. Elevated CO(2) mitigated the water stress-induced decrease in the concentration of soluble sugars in roots. However, this was probably due, in part, to carbohydrate loading during the first four months when all seedlings were grown in the presence of a high water supply, rather than to osmotic adjustment to water stress. Water stress caused a doubling in the concentration of free primary amino acids in roots, whereas elevated CO(2) reduced primary amino acid and nitrogen concentrations to 32 and 74%, respectively, of those in roots of seedlings grown in ambient CO(2). There was no indication of large-scale osmotic adjustment to water stress or that elevated CO(2) enhanced osmotic adjustment in loblolly pine.     

Castilleja and Pedicularis confirmed as telial hosts for Cronartium ribicola in whitebark pine ecosystems of Oregon and Washington

The primary objective of this research was to determine whether native species of Castilleja and Pedicularis are naturally infected by Cronartium ribicola in whitebark pine ecosystems of the Oregon and Washington Cascade Range, USA. Secondary objectives were to monitor the phenology of aecial and telial hosts to determine whether there is sufficient time for C. ribicola to complete its life cycle within high‐elevation stands and to evaluate the variety of susceptible native host species within these genera through field and growth chamber inoculation. These objectives were approached through fieldwork in 2008 and 2009 in whitebark pine ecosystems at Mt. Rainier, Mt. Adams, Mt. Hood, Mt. Bachelor, Tumalo Mtn. and Crater Lake. Forty‐nine observational study plots were established and monitored. Natural C. ribicola infection was detected on 84 Pedicularis racemosa plants and five Castilleja plants (C. applegatei, C. miniata and C. parviflora). Field observation provided evidence that there is sufficient time for C. ribicola to complete its life cycle on hosts within high‐elevation whitebark pine stands. In 2009, 18 field inoculation plots were established at Mt. Rainier and Crater Lake. Field inoculation confirmed the susceptibility of two additional species within these genera, C. arachnoidea and P. bracteosa. All four Castilleja species inoculated in the growth chamber developed infection, with an overall infection incidence of 62% (167 out of 270 plants). The identity of the rust species on field specimens as C. ribicola was verified through PCR and sequencing of the ITS1‐5.8S‐ITS2 region of DNA. Improved understanding of the role of these newly recognized hosts in white pine blister rust epidemiology should be used to prioritize sites for the restoration of ecologically valuable whitebark pine.     

Analysis of needle proteins and N-terminal amino acid sequences of two photosystem II proteins of western white pine (Pinus monticola D. Don)

A method is described for extracting needle proteins from mature western white pine (Pinus monticola D. Don) trees. Extracted proteins were separated into 26 components by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The predominant protein component (32-39%) had a molecular weight of 57 kDa. The second most predominant protein component (14-25%) had a molecular weight of 16 kDa. The amino acid composition of the twenty-six protein components was determined and a few selected proteins were subjected to further amino acid sequence analysis. Of these, two proteins, designated as Pin m I and Pin m II, were identified by sequence homology with other Photosystem II proteins. The Pin m I and Pin m II proteins had molecular weights of 22.6 kDa and 23.4 kDa, respectively. Pin m I had a much higher proline content than Pin m II.     

Effects of elevated nitrogen deposition on soil microbial biomass carbon in major subtropical forests of southern China

2003

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Wang X F.Li S Y.Bai K J.Kuang T Y Influence of doubled CO2 on plant growth and soil microbial biomass C and N 1998(12)

Xue J H.Mo J M.Li J.Li D J The short-term response of soil microorganism number to simulated nitrogen deposition 2007(02)

Yao W H.Yu Z Y The nutrient content of throughfall inside the artificial forests on downland 1995

Yi Z G.Yi W M.Zhou L X.Wang X M Soil microbial biomass of the main forests in Dinghushan Biosphere Reserve 2005(05)

Zhou G Y.Yan J H The influence of region atmospheric precipitation characteristics and its element inputs on the existence and development of Dinghushan forest ecosystems [其它论文] -Acta Ecologica Sinica2001

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