Abscisic acid from Pinus densiflora pollen

(+)-Abscisic acid was isolated as the methyl ester from Pinus densiflora pollen and identified spectroscopically.     

Endogenous Inhibitors for Pollen Germination and Pollen Tube Elongation in Pinus densiflora SIEB,et Zucc.

Sulfur metabolism in Cephalosporium acremonium was investigated using a mutant, 8650⁺/ OAH^?/SeMe^R, which could not convert cysteine or inorganic sulfur to methionine. The production of cephalosporin by the mutant depended on the amount of S-sulfocysteine in a chemically defined medium supplemented with a low level of methionine sufficient to support optimal growth. S-Sulfocysteine was detected in an extract of cells grown in the presence of sodium thiosulfate and l-serine. Furthermore, an NADPH-linked reduction of S-sulfocysteine to cysteine was demonstrated in a cell-free extract. These facts suggest that S-sulfocysteine is a direct precursor in cysteine biosynthesis in C. acremonium and an alternative pathway involving the compound is one of the most important ones in cephalosporin C production by this fungus.     

Determination of Abscisic Acid in Pinus densiflora by Selected Ion Monitoring

Abscisic acid (ABA) in the stem of akamatsu (Pinus densiflora) was identified and quantified by gas chromatography-mass spectrometry-selected ion monitoring using hexadeuterated ABA as an internal standard. tert-Butyldimethylsilyl ester was used as a derivative of ABA. This derivative had high sensitivity and selectivity for ABA determination. ABA concentrations in cambial region scrapings were independent of the cessation of cambial activity.     

Purification and Some Properties of Malate Synthase from the Pollen of Pinus densiflora Sieb,et Zucc.

Malate synthase (EC 4.1.3.2), termed MS_H (mol. wt. 630,000), was purified from the pollen of Pinus densiflora Sieb, et Zucc. to apparent homogeneity as judged by SDS-PAGE. Part of MS_H was converted to the low molecular weight form of MS, termed MS_L (mol. wt. 62,000), by incubation with 5 mm ATP. Both forms of MS were maximally active at pH 7.6 in the presence of 10 mm MgCl₂ and 0.01 mm EDTA. MS_L was completely inactivated by heating at 50°C for 3 min, but MS_H activity was retained about 65%. MS_H preincubated with 5mm ATP showed unstability similar to that of MS_l. MS_h activity was more strongly inhibited than that of MS_L by phosphoenolpyruvate or ATP. Km values of MS_H for acetyl-CoA and glyoxylate were about one half of those of MS_L. The F_max of MS_h was about 5 times as high as that of MS_L. ATP was a noncompetitive inhibitor with respect to acetyl-CoA and Ki values of ATP for MS_H and MS_L were 1.0 × 10^–3 m and 2.0 × 10^–2 m, respectively. From these results we suggested that MS_H and MS_L are the active and low active forms of MS, respectively.     

Novel p-coumaric acid esters from Pinus densiflora pollen

Solvent-extractable lipids in Pinus densiflora pollen were investigated. The cis- and trans-isomers of 1,16-dioxo-, 1-hydroxy-16-oxo- and 1, 16-dihydroxyhexadecan-7-yl p-coumarates were identified.     

Pollen tube reuses intracellular components of nucellar cells undergoing programmed cell death in Pinus densiflora

Through the process known as programmed cell death (PCD), nucelli of Pinus densiflora serve as the transmitting tissue for growth of the pollen tube. We sought to clarify the processes of degradation of nucellar cell components and their transport to the pollen tube during PCD in response to pollen tube penetration of such nucelli. Stimulated by pollination, synthesis of large amounts of starch grains occurred in cells in a wide region of the nucellus, but as the pollen tube penetrated the nucellus, starch grains were degraded in amyloplasts of nucellar cells. In cells undergoing PCD, electron-dense vacuoles with high membrane contrast appeared, assumed a variety of autophagic structures, expanded, and ultimately collapsed and disappeared. Vesicles and electron-dense amorphous materials were released inside the thickened walls of cells undergoing PCD, and those vesicles and materials reaching the pollen tube after passing through the extracellular matrix were taken into the tube by endocytosis. These results show that in PCD of nucellar cells, intracellular materials are degraded in amyloplasts and vacuoles, and some of the degraded material is supplied to the pollen tube by vesicular transport to support tube growth.     

Two new triterpenoids from the roots of Pinus densiflora

Chemical investigation of the roots of Pinus densiflora led to the isolation of two new triterpenoids, (24S)-3β-methoxy-24,25-epoxy-lanost-9(11)-ene (1) and 29-acetoxy-3α-methoxyserrat-14-en-21α-ol (2), together with three known serratene-type triterpenoids (3–5) and four known diterpenoids (6–9). Their structures were determined by spectroscopic analyses.     

天然赤松个体生物量的研究

对延边地区不同密度天然赤松林中不同生长势的赤松个体地上部生物量进行了研究。结果表明,不同生长势赤松生物量变化受密度影响的顺序为：优势木〈平均木〈被压木;各器官生物量分配比例受密度影响的顺序为：干〉枝〉叶〉皮。不同生长势赤松的生物量垂直分布虽然各具特点,但都表现为：树干和树皮生物量主要分布在６ｍ树高范围内,树枝则集中在６～１０ｍ范围,针叶在上、中、下３层林冠中均分,上层林冠的枝叶量受密度影响最小。     

Competition between ectomycorrhizal fungi colonizing Pinus densiflora

 Interactive competition of Pisolithus tinctorius (Pers.) Coker et Couch with an unidentified species Tanashi 01 and Suillus luteus (L.: Fr.) S. F. Gray was investigated using a rhizobox. Pinus densiflora Sieb. et Zucc. was used as the host plant and mycelia were distinguished by hyphal color. The speed of mycelial spread differed between the fungi;P. tinctorius and Tanashi 01 grew faster than S. luteus. A P. tinctorius mycorrhizal seedling and a Tanashi 01 mycorrhizal seedling were transplanted on opposite sides of the rhizobox. The mycelia and mycorrhizae of P. tinctorius were overgrown by Tanashi 01 hyphae and development of P. tinctorius was gradually inhibited. The areas occupied by mycelia and mycorrhiza of P. tinctorius decreased by 52% and 37%, respectively, 154 days after transplantation relative to that at 91 days. In the overlap area of P. tinctorius and Tanashi 01, the latter fungus infected new root tips emerging from P. tinctorius mycorrhiza, which lacked a mantle of P. tinctorius hyphae, and formed a composite mycorrhizal structure. P. tinctorius mycorrhizae were progressively replaced by Tanashi 01 mycorrhizae. Mycelial spread of P. tinctorius and S. luteus were naturally inhibited but there was no interaction in mycorrhizal formation. Accepted: 18 June 1999     

Factors Affecting the Emission of Monoterpenes from Red Pine (Pinus densiflora)

The mechanism of monoterpene emission from Pinus densiflora was studied using an environmentally controlled gas cabinet. It was found that monoterpene emission rate increases exponentially with temperature and is also influenced by light. These observations were explained reasonably by a mechanism whereby monoterpene emission rate depends on the monoterpene amount in the leaf oil and its saturated vapor pressure.     

山东赤松种群的数量动态

通过静态生命表分析赤松（Ｐｉｎｕｓ ｄｅｎｓｉｆｌｏｒａ Ｓｉｅｂ．ｅｔ Ｚｕｃｃ．）种群的生命结构与数量动态。结果表明,赤松种群具有不同年龄等级结构,死亡高峰出现在５～１５年,此时正是幼龄期向成年期的过渡阶段,度过此阶段的赤松个体大多能达到生理寿命。由此看出,赤松种群静态生命表能较精确地反映赤松种群的数量动态规律。     

云南松花粉中钙调素结合蛋白的分离和鉴定

通过ＣａＭ－Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析方法从云南松花粉中提取出１０种ＣａＭ结合蛋白。它们均能抑制ＣａＭ对ＰＤＥ的激活,但这种抑制可被随后加入的过量的ＣａＭ所消除。酶活测定表明ＣａＭ结合蛋白中有Ｃａ２＋－依赖的ＡＴＰａｓｅ活力,但无植酸酶、过氧化物酶、酸性磷酸酶和磷脂酶Ｄ活性。     

Biological pretreatment of softwood Pinus densiflora by three white rot fungi

The effects of biological pretreatment on the Japanese red pine Pinus densiflora, was evaluated after exposure to three white rot fungi Ceriporia lacerata, Stereum hirsutum, and Polyporus brumalis. Change in chemical composition, structural modification, and their susceptibility to enzymatic saccharification in the degraded wood were analyzed. Of the three white rot fungi tested, S. hirsutum selectively degraded the lignin of this sortwood rather than the holocellulose component. After eight weeks of pretreatment with S. hirsutum, total weight loss was 10.7%, while lignin loss was the highest at 14.52% among the tested samples. However, holocellulose loss was lower at 7.81% compared to those of C. lacerata and P. brumalis. Extracelluar enzymes from S. hirsutum showed higher activity of ligninase and lower activity of cellulase than those from other white rot fungi. Thus, total weight loss and changes in chemical composition of the Japanese red pine was well correlated with the enzyme activities related with lignin- and cellulose degradation in these fungi. Based on the data obtained from analysis of physical characterization of degraded wood by X-ray Diffractometry (XRD) and pore size distribution, S. hirsutum was considered as an effective potential fungus for biological pretreatment. In particular, the increase of available pore size of over 120 nm in pretreated wood powder with S. hirsutum made enzymes accessible for further enzymatic saccharification. When Japanese red pine chips treated with S. hirsutum were enzymatically saccharified using commercial enzymes (Cellulclast 1.5 L and Novozyme 188), sugar yield was greatly increased (21.01%) compared to non-pretreated control samples, indicating that white rot fungus S. hirsutum provides an effective process in increasing sugar yield from woody biomass.     

Spatiotemporal transfer of carbon-14-labelled photosynthate from ectomycorrhizal Pinus densiflora seedlings to extraradical mycelia

Seedlings of Pinus densiflora colonized by an unidentified ectomycorrhizal fungus (T01) were labelled photosynthetically with 14C. Movement of 14C-labelled photosynthates within the underground part of the seedlings was investigated by temporal autoradiography using an imaging plate. Within 1 day, 14C was transferred from the shoot to the underground part that included roots, mycorrhizae, and the extraradical mycelium; within 3 days, the 14C in the underground part reached its maximum density. Mycorrhizae and actively growing root tips were large C sinks. Three days after 14C labelling, counts of 14C radioactivity in the underground part of the mycorrhizal seedlings were 2.6 times those of nonmycorrhizal seedlings. The mycorrhizae of mycorrhizal plants accumulated 5.2 times the 14C counts in the short-root tips of nonmycorrhizal plants. 14C counts in various areas of the extraradical mycelium demonstrated that all 14C-photosynthate transfer from the host root to the extraradical mycelium occurred within 3 days after 14C labelling, and that there was only a short lag of < 1 day between 14C accumulation in the basal and distal parts of the mycelium. Although more 14C accumulated in the distal than in the basal parts, 14C counts per unit hyphal biomass were similar between the two. These results suggest that 14C spread rapidly throughout the entire mycelium. Thirteen days after 14C labelling, we estimated 14C allocation to extraradical mycelia by taking autoradiographs after removing host roots. About 24% of 14C counts in the underground part of the mycorrhizal seedlings had been allocated to extraradical mycelia in this system, indicating that the fugal mycelium is an important sink for photosynthates.     

Seedling regeneration and vegetative resprouting after fires in Pinus densiflora forests

Modes of recruitment of each regenerated species after fire were investigated quantitatively in a Pinus densiflora forest in central Japan. There was a continuum of regeneration modes from species with abundant seedling populations (more than one individual m^–2) to sprouters. The former included Erigeron canadensis, Eupatorium chinense var. simplicifolium, Rubus crataegifolius, Lespedeza bicolor forma acutifolia, Mallotus japonicus, Rhus javanica, Eurya japonica, Aralia elata and Weigela decora, whose seedlings accounted for 70% of total recruitment on the forest floor. Sprouts from stem bases which comprised much of the canopy prior to fire (e.g. Quercus serrata, Castanea crenata, Fraxinus sieboldiana and Prunus spp.), were vigorous and grew rapidly. Plants on both ends of the continuum dominated post-fire vegetation in the study sites.     

Paenibacillus pinihumi sp. nov., a cellulolytic bacterium isolated from the rhizosphere of Pinus densiflora

A novel cellulolytic bacterium, strain S23^T, was isolated from the rhizosphere of the pine trees in Daejeon, Republic of Korea. This isolate was Gram-positive, strictly aerobic, rod-shaped, catalase-negative, oxidase-positive, motile by means of peritrichous flagella, and tested positive for alkaline phosphatase, esterase lipase, leucine arylamidase, α-galactosidase, and β-galactosidase activities. The DNA G+C content was 49.5 mol%. The main cellular fatty acids were anteiso-C_15:0 (51.9%), iso-C_16:0 (14.7%), and iso-C_15:0 (13.2%). The major isoprenoid quinone was menaquinone 7 (MK-7). Diagnostic diamino acid in the cell-wall pepti-doglycan was meso-diaminopimelic acid. Comparative 16S rRNA gene sequence analysis showed that this strain clustered with Paenibacillus species. The 16S rRNA gene sequence similarity values between S23^T and other Paenibacillus species were between 89.9% and 95.9%, and S23^T was most closely related to Paenibacillus tarimensis SA-7-6^T. On the basis of phylogenetic and phenotypic properties of strain S23^T, the isolate is considered as a novel species belonging to the genus Paenibacillus. Therefore, the name, Paenibacillus pinihumi sp. nov., is proposed for the rhizosphere isolate; the type strain is S23^T (=KCTC 13695^T =KACC 14199^T =JCM 16419^T)     

An allelopathic substance in red pine needles (Pinus densiflora)

Aqueous methanol extracts of red pine (Pinus densiflora) needles inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), alfalfa (Medicago sativa), ryegrass (Lolium multiflorum), timothy (Pheleum pratense), Digitaria sanguinalis and Echinochloa crus-galli. Increasing the extract concentration increased inhibition, suggesting that the pine needles may have growth inhibitory substances and possess allelopathic potential. The aqueous methanol extract of the pine needles was purified, and a main inhibitory substance was isolated and determined by spectral data as 9alpha,13beta-epidioxyabeit-8(14)en-18-oic acid. This substance inhibited root and shoot growth of cress and Echinochloa crus-galli seedlings at concentrations greater than 0.1 mM. The endogenous concentration of the substance was 0.13 mmol/kg pine needle. These results suggest that 9alpha,13beta-epidioxyabeit-8(14)en-18-oic acid may contribute to the growth inhibitory effect of the pine needles and may play an important role in the allelopathy of red pine.     

马尾松花粉中植物甾醇成分分析

采用正己烷超声浸提法提取破壁马尾松花粉中的植物甾醇成分,L-B比色法(Liebermann-Burchard)测定马尾松花粉中总甾醇含量,并用GC-MS(气相色谱-质谱联用法)首次对马尾松花粉中的植物甾醇进行详细定性分析。测得马尾松花粉中总甾醇含量为1.680 mg/g,并鉴定出8种植物甾醇成分,分别为:Δ5-菜油甾醇,豆甾醇,Δ7-菜油甾醇,β-谷甾醇,Δ5-燕麦甾醇,禾本甾醇,环阿屯醇和24-亚甲基环木菠萝烷醇。