采用ARDRA研究双孢蘑菇培养料后发酵过程中的细菌群落结构(Ⅰ)——细菌16S rDNA全长文库的建立

采用化学裂解和酶解相结合的方法,以加入PVPP的高盐缓冲液作为细胞裂解反应体系,用PEG-8000进行DNA沉淀,从双孢蘑菇堆肥后发酵的4个代表时期培养料样品中提取微生物总DNA,再以总DNA为模板,以专用引物F27和R1498进行PCR扩增,获得16SrDNA片段,经纯化后构建4个时期样品的细菌16SrDNA文库。试验结果表明,本试验获得的总DNA质量较好,采用PCR扩增可获得多个细菌、放线菌和真菌特异片段;细菌16SrDNA文库的目的片段插入效率在90％以上。     

外源DNA片段导入草菇原生质体的研究

以平菇DNA为供体,草菇原生质体为受体,用PEG,CaCl_2作诱导剂,将平菇DNA导入草菇原生质体,选育出V_(157-1)菌株,该菌株在菇型、温型、酶活力及生物转化率上与亲本V_(157)有显著差异。本研究为食用菌育种提供了一条新的途径。     

广东省微生物所近年食用菌研究成果简介

广东省微生物研究所(前身中南真菌研究室)成立于1963年,是一个以研究真菌为主的研究所,食用真菌是其中一个研究方面,现有30个科技人员从事这方面工作,主要任务是改革提高已生产的食用菌栽培技术;通过细胞融合、杂交育种、不同菌株栽培评比等方法,培育或筛选出优良高产适应当地的菌株或新的品种;开发新的食用菌资源,驯化有重要经济价值的野生食用菌。1979年以前,主要研究香菇和草菇,在选育优良品种,引进国外先进     

不同食用菌菌糠中微生物的群落结构

食用菌菌糠是食用菌栽培过程中收获子实体后剩下的栽培基质，随着食用菌产业的发展，食用菌菌糠数量越来越多，菌糠中微生物群落结构的研究未见相关报道。本试验中以不同食用菌菌糠为材料，采用高通量测序的方法对平菇、香菇、金针菇、杏鲍菇菌糠中的细菌、真菌群落进行了分析。结果表明，所有菌糠样品细菌群落在门水平上的优势群落为变形菌门和厚壁菌门，从属水平来看，未分类的细菌群落占优势；所有菌糠样品真菌群落在门水平上的优势群落为子囊菌门和担子菌门，从属水平来看，优势属为食用菌自身所在属。     

灵芝制种过程多糖含量变化研究

灵芝是一类具有较高药用价值的腐生型真菌。本研究选取4株灵芝真菌,进行人工制种,并测定其不同阶段的多糖含量。结果表明,Ganoderma atrum Hz2、Ganoderma lucidum Hz5、Ganoderma sinense Hz1三株灵芝真菌在制种过程中多糖含量呈递减的趋势,而菌株Ganoderma neojaponicum Wzs1二级种在制种过程中多糖含量最多为1.75%;4株菌株中Hz2制种完成后多糖含量最大为0.88%。以上结果反应了灵芝多糖的含量随着制种的进行而逐渐减少,此外黑芝Ganoderma atrum的人工栽培品种相比其它3种灵芝具有较高的药用价值。     

大杯伞柄多糖与菌丝体粗多糖的免疫识别方法初探

采用热水(80℃)提取和75%乙醇沉淀、Sepharose CL-4B凝胶过滤及DEAE Sepharose CL-4B阴离子交换等方法分离纯化大杯伞(Clitocybe maxima)柄粗多糖,获得伞柄多糖纯品,制备多糖抗体;采用热水法提取大杯伞菌丝体粗多糖。用碳二亚胺法将纯化伞柄多糖与牛血清白蛋白进行共价化学偶联,并将偶联产物(拟糖蛋白)免疫兔子以制备抗血清,以酶联免疫法检测抗体滴度。依据该抗体与来自相同菌株的菌丝体粗多糖免疫反应差异对这两种多糖进行免疫识别,结果表明:经过亲和层析纯化后,第六次免疫抗血清对伞柄多糖的抗体滴度可达64K,而且对菌丝体粗多糖反应呈阴性,显示该抗体可识别这两种多糖之间的结构差异。     

过氧化物酶同工酶在真菌分类上的研究

自同工酶被发现以来,对它的研究已相当广泛和深入.近年来,由于对植物、动物及人类的各种同工酶的研究,加深了人们对生物形态、功能以及起源的认识.关于食用菌同工酶的研究目前虽有开展,但报道甚少,尤其是我们所研究的若干菌种尚未见报道.本实验旨在探求不同酶谱进行品种间鉴定,方法简便、快速、准确,对解决当前国内菌种混乱具有现实意义.同时,采用此法将有助于提高真菌间的细胞杂交的主动性,为真菌细胞工程提供必要的材料.本实验采用聚丙烯酰胺凝胶圆盘电泳方法,对平菇八个菌株的过氧化物酶同工酶进行了分析,现将结果报告如下.     

浙江益圣菌物发展有限公司

本公司是一家专业从事食药用真菌多糖提取及保健食品生产，致力于人类的健康事业和食用菌产业发展的商科技企业，公司由浙江省林科院及著名食用菌专家吴学谦等高层次科技人员发起组建。公司设在世界香菇发原地和全国著名食用菌生产基地的丽水市水阁经济开发区内，厂区占地面积1400m^2，已建成了年提取加工100吨食用菌多糖产品的先进生产线和食用菌单组分纯化多糖中试车间，     

三种食用菌多糖及其复合多糖功能性评价研究

以大球盖菇、金针菇、香菇为原料,分别经热水浸提,分离纯化得到真菌多糖,并通过复配后形成复合多糖,研究3种食用菌多糖及复合多糖的DPPH·清除能力、·OH清除能力、还原能力、α-糖苷酶抑制率、α-淀粉酶抑制率等抗氧化及降血糖活性指标。结果显示,大球盖菇、金针菇、香菇三种食用菌,真菌多糖的提取率分别为8.58%、4.71%、6.26%。三种真菌多糖均具有抗氧化和降血糖的作用,且复合多糖的抗氧化能力与降血糖能力均优于单一品种多糖。三种食用菌多糖浓度均为2 mg/mL时,复合多糖的DPPH清除率、羟自由基清除率、还原力分别达到60.4%、37.26%与0.31。同时以α-糖苷酶抑制率和α-淀粉酶抑制率来表征多糖的体外降血糖作用,结果显示复合多糖的降血糖能力也优于单一品种多糖,复合食用菌多糖浓度为2 mg/mL时,α-糖苷酶抑制率和α-淀粉酶抑制率分别为42.26%和50.01%。     

平菇DNA导入草菇原生质体选育优良品种的研究

以湘平992DNA为供体,浏阳麻菇原生质体为受体,用PEG400、CaCl2作诱导剂,将平菇DNA导入浏阳麻菇原生质体,选育出V1568新菌株,该菌株在菇型、温型、酶活力及生物转化率上与亲本浏阳麻菇有显著差异。该研究为食用真菌外源DNA导入育种增添了新的内容。     

Ergebnisse der Leistungsprüfungen der Süßkirschensorte ‚Stella‘ auf Gisela- und Weiroot-Unterlagen in Bulgarien

Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden.     

Relationship between renal hypertension and cardiac α1-adrenoceptor autoantibodies in rats

AIM: To examine the autoantibody against α₁-adrenoceptor and its biologic activities during the development of renal hypertension. METHODS: Renal hypertension of rat was achieved by clipped renal artery, the titre of autoantibody to α₁-adrenoceptor was detected using ELISA immunoassay. Furthermore, the biological offects of these autoantibodies on cultured cardiomyocytes were also examined. RESULTS: After two weeks of clipping renal arteries, both the frequency of occurrence and the titre of autoantibodies to cardiac α₁-adrenergic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 12 weeks. The biological effects of these autoantibodies displayed an "agonistic-like" activities on the beating frequency of cultured neonatal cardiomyocytes. CONCLUSION: Autoantibodies against α₁-adrenoceptor may play a role in the elevation of peripheral vascular resistance and in the development of cardiac hypertrophy in rats with renal hypertension.     

Effects of radiation from 188Re on smooth muscle cell proliferation

AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β^-particles emission from ¹⁸⁸Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by ¹⁸⁸Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [³H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of ¹⁸⁸Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from ¹⁸⁸ Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G₀/G₁ arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.     

Influence of Mineral Nutrients on Strawberry Fruit Quality and Their Accumulation in Plant Organs

Abstract

This review is based partly on complete articles and partly on abstracts. Three of the 60 articles deal with the total uptake of elements in strawberry plant organs in two different strawberry production systems, both considered as optimal concerning amount and balance of elements. The effect on fruit quality may be dramatic if the level of a particular element is outside this range, but there may also be effects initiated by differences within the optimal range of elements. Most articles refer to product oriented quality, but some focus on consumer oriented quality, as discussed by Shewfelt (1999). The discussion here is on a general basis, so one should keep in mind that there are cultivar differences and that specification of nutrition ideally should mirror the needs of a single cultivar, or a group of cultivars with similar requirements. Also, to get a complete understanding of the subject future reviews should embrace a broader access of information including the effect on plant development of individual elements, such as the role of calcium in fruit firmness and its importance in cell wall structure. However, the intention here is to narrow the information to results that suggest a direct connection between nutrient uptake and fruit quality.     

Effect of puerarin on pulmonary arterioles wall collagen metabolism of chronic hypoxia hypercapnia rats

AIM:To investigate the effect of puerarin on pulmonary vessel collagen metabolism in pulmonary hypertension rats induced by chronic hypoxia and hypercapnia.METHODS:Collagen Ⅰ, Ⅲ and their mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization.RESULTS:① Light microscopy showed media thickness of pulmonary arterioles was much higher in HH(hypoxic-hypercapnia) group than that of NC(normal control) group, and, vessel cavity turned more straiter in HH group than that of NC group.However, the damage of pulmonary arterioles in HP(hypoxic-pueratin) group was much slighter than that of HH group. ② The levels of plasma ET-1 and lung homogenates Hyr were much higher in HH group than those of NC group(P＜0.01), and lower in HP group than HH groups(P＜0.01).Plasma NO content in group HH was lower than that of group NC(P＜0.01), it was higher in group HP than that of group HH(P＜0.01).③Expression of collagen Ⅰ and collagen Ⅰ mRNA in pulmonary arterioles were significantly higher in HH groups than those of NC group (P＜0.01), and they were lower in HP group than those of HH group (P＜0.01).Expression of collagen Ⅲ and collagen Ⅲ mRNA showed no difference among three groups(P＞0.05).CONCLUSION:Puerarin inhibited the deposition of collagen and improved pulmonary vessel remodeling.     

Mixed-severity fire regime in a high-elevation forest of Grand Canyon,Arizona, USA

Fire regime characteristics of high-elevation forests on the North Rim of the Grand Canyon, Arizona, were reconstructed from fire scar analysis, remote sensing, tree age, and forest structure measurements, a first attempt at detailed reconstruction of the transition from surface to stand-replacing fire patterns in the Southwest. Tree densities and fire-/non-fire-initiated groups were highly mixed over the landscape, so distinct fire-created stands could not be delineated from satellite imagery or the oldest available aerial photos. Surface fires were common from 1700 to 1879 in the 4,400 ha site, especially on S and W aspects. Fire dates frequently coincided with fire dates measured at study sites at lower elevation, suggesting that pre-1880 fire sizes may have been very large. Large fires, those scarring 25% or more of the sample trees, were relatively infrequent, averaging 31 years between burns. Four of the five major regional fire years occurred in the 1700s, followed by a 94-year gap until 1879. Fires typically occurred in significantly dry years (Palmer Drought Stress Index), with severe drought in major regional fire years. Currently the forest is predominantly spruce-fir, mixed conifer, and aspen. In contrast, dendroecological reconstruction of past forest structure showed that the forest in 1880 was very open, corresponding closely with historical (1910) accounts of severe fires leaving partially denuded landscapes. Age structure and species composition were used to classify sampling points into fire-initiated and non-fire-initiated groups. Tree groups on nearly 60% of the plots were fire-initiated; the oldest such groups appeared to have originated after severe fires in 1782 or 1785. In 1880, all fire-initiated groups were less than 100 years old and nearly 25% of the groups were less than 20 years old. Non-fire-initiated groups were significantly older (oldest 262 years in 1880), dominated by ponderosa pine, Douglas-fir, or white fir, and occurred preferentially on S and W slopes. The mixed-severity fire regime, transitioning from lower-elevation surface fires to mixed surface and stand-replacing fire at higher elevations, appeared not to have been stable over the temporal and spatial scales of this study. Information about historical fire regime and forest structure is valuable for managers but the information is probably less specific and stable for high-elevation forests than for low-elevation ponderosa pine forests.This revised version was published online in May 2005 with corrections to the Cover Date.     

瓜叶绿素降解关键酶基因PPH和PAO cDNA片段的克隆与表达初步分析

 以西双版纳黄瓜和‘华黄5号’黄瓜叶片为试验材料,研究了衰老过程中两种黄瓜叶片叶绿素含量变化及脱镁叶绿素脱镁叶绿酸水解酶基因PPH和脱镁叶绿酸a加氧酶基因PAO的表达。结果表明：随着叶片衰老,叶绿素a和叶绿素b含量显著降低,叶绿素a与叶绿素b的比值在0.5左右,推测叶绿素b的含量可能对黄瓜叶片保绿起到重要作用。荧光定量RT-PCR分析结果显示：PPH和PAO在西双版纳黄瓜叶片生长35 d时表达量最高,分别为5.50和1.86;PPH在‘华黄5号’叶片35 d时表达量最高,为7.14,而PAO在35和45 d表达量都很高,分别为3.90和3.91;PPH和PAO在西双版纳黄瓜叶片中的表达量低于在‘华黄5号’中。     

Effects of simvastatin on PDGF-BB and serum-induced proliferation of vascular smooth muscle cells and on the expression of tumor suppressor gene PTEN

AIM:To observe the effect of simvastatin on the proliferation of vascular smooth muscle cells(VSMCs) induced by serum and growth factor PDGF-BB and the effect of simvastatin on the expression of PTEN,a important regulator of G₁/S cell cycle transition. METHODS:The DNA synthesis was determined by [³H]-TdR incorporation, cell cycle was examined with flow cytometry, the protein level of PTEN was measured by Western blot method. RESULTS: (1)Simvastatin inhibited [³H]-TdR incorporation in a dose dependent manner. (2) Flow cytometric DNA analysis revealed that simvastatin induced significantly enhancement of G₀/G₁ phase and decrease in S phase VSMCs.(3)Simvastatin increased protein level of PTEN and mevalonate, a metabolite of HMG-COA, reversed the effect of simvastatin on PTEN protein expression. CONCLUSION:Simvastatin may inhibit proliferation of VSMCs and retarded cell cycle in G₀/G₁ phase by increasing PTEN expression through inhibiting synthesis of mevalonate.     

Multi-scale landscape and seascape patterns associated with marbled murrelet nesting areas on the U.S. west coast

Habitat for wide-ranging species should be addressed at multiple scales to fully understand factors that limit populations. The marbled murrelet (Brachyramphus marmoratus), a threatened seabird, forages on the ocean and nests inland in large trees. We developed statistical relationships between murrelet use (occupancy and abundance) and habitat variables quantified across many spatial scales (statewide to local) and two time periods in California and southern Oregon, USA. We also addressed (1) if old-growth forest fragmentation was negatively associated with murrelet use, and (2) if some nesting areas are more important than others due to their proximity to high quality marine habitat. Most landscapes used for nesting were restricted to low elevation areas with frequent fog. Birds were most abundant in unfragmented old-growth forests located within a matrix of mature second-growth forest. Murrelets were less likely to occupy old-growth habitat if it was isolated (> 5 km) from other nesting murrelets. We found a time lag in response to fragmentation, where at least a few years were required before birds abandoned fragmented forests. Compared to landscapes with little tono murrelet use, landscapes with many murrelets were closer to the ocean's bays, river mouths, sandy shores, submarine canyons, and marine waters with consistently high primary productivity. Within local landscapes (≤ 800ha), inland factors limited bird abundance, but at the broadest landscape scale studied (3200 ha), proximity to marine habitat was most limiting. Management should focus on protecting or creating large, contiguous old-growth forest stands, especially in low-elevation areas near productive marine habitat. This revised version was published online in July 2006 with corrections to the Cover Date.     

Relationships of landscape,prey and agronomic variables to the abundance of generalist predators in cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis>) fields

A two-year field study investigated the possible effects of grain sorghum (Sorghum bicolor [L.] Moench) and uncultivated areas on the abundance of generalist predators in commercially-managed cotton (Gossypium hirsutum L.) fields in Texas, USA. From 63 to 70 fields were sampled for pests and predators over nine consecutive weeks during early stages of cotton development. Additional data on agronomic practices and landscape composition at three spatial scales were also collected for each field. Stepwise regression analyses were used to determine the relationships of landscape, agronomic and prey variables to the abundance of generalist predators. Because the variables most closely linked to predator levels could vary over time, separate regressions were conducted for three time periods corresponding to stages of grain sorghum growth (half-bloom, hard-dough, maturity) in each year. Significant relationships between predator abundance and agricultural landscape composition appear in both years and in all three time periods, but the specific relationships of landscape variables to cotton predator levels differed between and within years. At maturity in 2001, predator levels rose as the amount of uncultivated land from 1.6 to 3.2 km distant and the perimeter shared with grain sorghum increased. During 2002, the area of grain sorghum (half-bloom) and uncultivated land (hard-dough) within 1.6 km of cotton fields were both positively related to predator numbers. Cotton planting dates and the abundance of cotton fleahoppers (Pseudatomoscelis seriatus [Reuter]) were also strongly linked to predator numbers during both years. Results suggest that the total amount of grain sorghum or uncultivated land in an area is more important than the presence of these habitats adjacent to cotton fields, and that landscape composition may sometimes be the most important factor in determining predator abundance.