3种白腐菌木质纤维素酶活性及酶相关基因的TRAP标记遗传多态性

【目的】木质纤维素是重要的可再生资源,白腐菌对降解木质纤维素具有特殊的优势。研究3种白腐菌的生物学特性、酶学活性以及菌种间的遗传多样性,系统分析3种白腐菌木质纤维素酶的表达与相关基因多态性的关系,为高产木质纤维素酶菌种选育提供理论基础,为白腐菌木材降解分子机制的研究提供科学依据。【方法】以云芝栓孔菌、火木层孔菌和松杉灵芝3种白腐菌为研究材料,测定在不同培养温度下固体培养基中菌落的生长速度和液体培养基中的生物量变化,利用比色法测量白腐菌5种木质纤维素酶活性,运用目标区域扩增多态性( TRAP)分子标记分析3种白腐菌的木质纤维素降解相关酶基因的多态性。【结果】在不同温度下(23,28℃),3种白腐菌在PDA固体培养基上的生长速度均为云芝栓孔菌＞火木层孔菌＞松杉灵芝,云芝栓孔菌和火木层孔菌在28℃的生长速度高于23℃,而松杉灵芝在23℃培养生长更快;液体培养基中云芝栓孔菌的生物量高于火木层孔菌和松杉灵芝。3种白腐菌木质素相关酶活性受木屑诱导显著提高,木质素酶活性最高的是漆酶,其次是木质素过氧化物酶,最低的是锰过氧化物酶;纤维素酶的表达在3种白腐菌中无显著差异,但外切纤维素酶活性明显高于内切纤维素酶活性,玉米秸秆为碳源诱导产生的纤维素酶活性明显高于木屑碳源样品。云芝栓孔菌更偏好于木质素酶的表达,而火木层孔菌和松杉灵芝则更偏好于纤维素酶的表达。TRAP分子标记扩增结果显示,木质素酶相关基因的6对引物共产生109条条带,多态性条带为79条,多态性百分比为72.47%;纤维素酶相关基因的11对引物共产生198条条带,多态性条带为140条,多态性百分率为70.70%。【结论】通过生长速度、生物量、木质纤维素酶活性的检测以及 TRAP 分子标记结果表明3种白腐菌在种间具有较高的遗传差异。3种白腐菌木质素酶活性的大小与其酶基因多态性之间没有明显的对应关系;3种白腐菌纤维素酶活性大致相同,其基因多态性也比较一致。     

毛竹和重组竹腐朽前后的表面颜色及耐腐性能

以毛竹和重组竹为研究对象,用绵腐卧孔菌(PV)、密粘褶菌(GT)两种褐腐菌和彩绒革盖菌(CV)、变色栓菌(TV)两种白腐菌进行腐朽试验,比较分析了毛竹和重组竹腐朽前后表面视觉性质及其质量损失差异。结果发现:毛竹和重组竹颜色均发生了显著变化,主要表现为明度(L~*)降低,色度值(C)、红绿轴色品指数(a~*)和黄蓝轴色品指数(b~*)显著增大,其中绵腐卧孔菌对毛竹及重组竹的色差影响最大。重组竹(耐绵腐卧孔菌)能达到Ⅰ级强耐腐水平;毛竹(耐绵腐卧孔菌)和重组竹(耐密粘褶菌和彩绒革盖菌)达到了Ⅱ级耐腐水平;毛竹(耐密粘褶菌、彩绒革盖菌和变色栓菌)和重组竹(耐彩绒革盖菌)为Ⅲ级稍耐腐水平。整体来看,重组竹对白腐菌和褐腐菌的耐腐效果好于毛竹。绵腐卧孔菌虽然对毛竹和重组竹的降解能力最弱,但对其颜色变化的影响却最大。扫描电镜可观察到彩绒革盖菌通过毛竹导管壁上的纹孔进入薄壁细胞、伴胞等,同时通过分泌木质素酶导致其细胞壁降解。     

6种木材白腐菌对山杨材木质素分解能力的研究

由于不同的木材腐朽菌的生理特性不同 ,所分泌的酶及酶的活性各不相同 ,因此 ,不同的腐朽菌分解木材的各种成分及相对速度就各不相同 ,而且对于木质纤维基质会有不同的中间代谢产物。本项研究选择了火木层孔菌 (Phelliusigniarius)及另外 5种木材分解能力较强的阔叶树上的白腐菌 :粗毛盖菌 (Funaliagallica)、三色革裥菌 (Lenzitestricolor)、冬拟多孔菌 (Polyporellusbrumalis)、偏肿拟栓菌 (Pseudotrametesgibbosa)和血红密孔菌 (Pyc noporussanguineus) ,研究了它们对山杨木材木质素的分解能力 ,测定了经 6种白腐菌分解一定时期的山杨木材木质素的含量 ,作为木材白腐菌对山杨木材木质素生物降解机制的初步研究 ,旨在为山杨木材生物制浆造纸提供应用基础理论研究 ,同时也可为木质素合理的生物转化为有用的化学品、生物漂白、酶处理防止机械浆的返黄、废水治理、纤维素酶解糖化的微生物前处理等提供相关的借鉴研究 ,以期在生产实践中减轻环境污染并充分利用木质素资源。在无菌的条件下 ,将山杨木片样品分别放入以上 6种白腐菌的平板培养基中受菌侵染 ,一定时间后取出 ,去除木片表面的菌丝体 ,然后分别测定未腐朽材和受菌侵染 4 0d、6 0d、80d和 12 0d时木片样品中木质素的含量 ,分析 6种白腐菌对山杨木     

长白落叶松提取物对木材防腐作用的研究

以长白落叶松心材、边材及树皮为原料,分别用乙醚、氯仿、丙酮、甲醇和水5种溶剂对其进行提取,采用滤纸片法研究了不同溶剂提取物分别对白腐菌和褐腐菌的抑菌性能,发现心材的甲醇提取物和丙酮提取物对白腐菌有较好的抑制效果,树皮的水提取物对2种菌都有较好的抑制效果.并研究了这两种提取物在室内对木材的防腐性能,借助扫描电子显微镜(SEM)照片观察了木材腐朽后菌丝生长的情况,与常用木材防腐剂酸性铬酸铜(ACC)进行了对比,结果显示:心材的甲醇提取物对白腐菌的抑制效果较好;而树皮的水提取物对褐腐菌抑制效果较好.     

10种植物醇提物对木腐菌的抑菌活性筛选北大核心

对比了10种植物干叶的50%乙醇提取物对4种常见木材腐朽菌的抑菌活性,筛选出具有广谱杀菌性且效果较好的植物来源,为后续开发植物源木材防腐剂提供依据。以白腐菌采绒革盖菌、黄孢原毛平革菌和褐腐菌密粘褶菌、绵腐卧孔菌为供试真菌,采用生长速率法初步测定香樟、侧柏、闽楠等10种植物醇提物对木腐菌的抑菌效力,比较其不同时间段的(3、5 d和7 d)菌落直径和抑菌率,聚类分析优选出抑菌植物。结果表明:1)在供试浓度为20 mg/mL的条件下,10种植物醇提物对白腐菌和褐腐菌均有一定抑制作用,随着时间延长大多数植物醇提物抑菌活性明显下降,总体抑菌效果褐腐菌比白腐菌更强;2)当菌落培养至7 d时,飞龙掌血、猴樟、圆柏等6种植物对白腐菌和褐腐菌均有抑制作用,其中飞龙掌血和香樟对4种菌均有抑制作用;3)飞龙掌血抑菌率均高于70%,总体抑菌效果较香樟好,而香樟对采绒革盖菌的抑制效果较差,对其他3种菌的抑制效果达到70%以上。聚类分析结果显示,飞龙掌血(1)和香樟(10)的抑菌效果最佳,在不同时间段对4种木材腐朽菌均有较强的抑菌效力,说明这2种植物具有开发植物源木材防腐剂的潜力。     

亚黑管菌母种培养基筛选初步研究

为获得亚黑管菌母种的最佳培养基,以PDA培养基作为基础培养基,根据亚黑管菌母种对营养的需求,添加各类营养元素设计了20种不同的培养基,接种野外采集亚黑管菌腐木菇木分离所获得的母种,研究不同培养基对亚黑管菌菌丝生长的影响,筛选出更适宜于亚黑管菌菌丝生长的最佳培养基,以期获得性状更佳的母种菌丝,也为后续生产提供种植资源。结果表明,最佳母种培养基配方是：南瓜200 g/L、葡萄糖20 g/L、硫酸镁1 g/L、磷酸二氢钾1 g/L、维生素0.01 g/L和琼脂18 g/L,pH自然。     

核桃基腐病菌产生细胞壁降解酶的种类及其活性分析

通过致病力的测定筛选出两株强致病力核桃基腐病菌菌株HA-1和KY-S,通过对酶活性的分析,研究证实核桃基腐病菌均能分泌4种细胞壁降解酶(CWDEs):β-葡萄糖苷酶、多聚半乳糖醛酸酶(PG)、果胶甲基半乳糖醛酸酶(PMG)、纤维素酶(Cx)。在不同碳源培养条件下,核桃基腐病产生细胞壁降解酶的活性存在一定差异,以蔗糖为碳源时,菌株分泌的各种细胞壁降解酶活性较高。在4种培养基中,达到各酶活性高峰的时间仅为3d。在蔗糖培养基中,核桃基腐病菌产生的Cx最大酶活性为1495.5mU/mL,明显高于其他3种酶,且显著高于其他培养基产生的Cx。但在树皮为碳源的培养基中Cx的活性最低。在CMC为碳源的培养基中PG、PMG的活性最高,分别为826.5mU/mL和661.3mU/mL,此外,以果胶为碳源时产生的PMG的活性最高为1068.0mU/mL。     

壳聚糖铜 /锌配合物复配的防白腐效果分析

壳聚糖金属配合物(CMC)是一种环境友好的木材保护剂,根据壳聚糖锌配合物(CZC)对褐腐菌(Poria placenta)的防腐效果优良,而壳聚糖铜配合物(CCC)对白腐菌(Coriolous versicolor)的防腐效果好的特点,本研究将CZC与CCC复配为壳聚糖金属配合物混合液MCMC,进行防白腐试验.结果显示复配后的MCMC,提高了其对木材白腐菌的耐腐能力,防白腐效果与CCA相近.     

怀槐心材提取物的防腐性能及其急性毒性试验研究

以怀槐的心材和树皮为原料,用乙醚、氯仿、丙酮、甲醇和水5种溶剂对其进行浸提,得到10种提取物.这10种提取物对白腐菌和褐腐菌的抑菌性能研究表明,怀槐心材的甲醇和丙酮提取物的抑菌性能良好.将心材的甲醇提取物和无机木材防腐剂酸性铬酸铜(ACC)用于对木材的室内防腐实验,根据腐朽后试样的失重率和扫描电子显微镜(SEM)的照片观测,得出甲醇提取物对白腐菌的防腐作用较好,对褐腐菌的防腐作用很弱,而ACC对两种菌的防腐作用较强.将心材甲醇提取物和ACC用于对斑马鱼的急性毒性试验,测得两者在24、48和 72 h 的半致死浓度,甲醇提取物分别为16.5、12.7、12.0 mg/L,ACC分别为3.2、2.5、2.0 mg/L;两者的安全质量浓度分别为2.26和 0.46 mg/L.     

竹青,竹黄室内耐腐性试验研究

对４种竹材的竹青、竹黄室内耐腐性进行了初步研究。试验结果表明：竹青、竹黄的耐腐性因竹材品种、竹龄及腐配菌菌种的不同而存在差异。竹青的耐腐性高于竹黄，对白腐菌耐腐性高于对褐腐菌耐腐性；不同竹材的竹黄对两种腐配菌的耐腐性差异较大。     

Browsing Damage on Pine (Pinus sylvestris and P. contorta) by a migrating moose (Alces alces) Population in Winter: Relation to Habitat Composition and Road Barriers

In Scandinavia, moose (Alces alces L.) sometimes cause severe browsing damage to economically-important pine. Moose-vehicle accidents have spurred construction of fences along roads, and these may interfere with moose migration between summer and winter ranges, or the road alone may be a barrier. If this happens and moose build up along roads, landowners may suffer economically. Therefore, this study investigated whether roads, fences or other factors influence the use of young pine stands by moose. Eighty stands along roads in northern Sweden were evaluated in which individually-browsed branches were counted on 9972 pines. Moose browsing was not significantly related to birch (Betula pendula Roth, B. pubescens Ehrh.) density, nor did it differ between pines (Pinus contorta Douglas or P. sylvestris L.). However, increased pine density, site productivity and proximity to a highway were associated with increased browsing. Further large-scale studies are needed to understand moose habitat selection and the effects of roads.     

Effects of Root Freezing on the Physiology and Growth of Picea glauca,Picea mariana and Pinus banksiana Seedlings Under Different Soil Moisture Regimes

The root systems of 2-yr-old Picea glauca, Picea mariana and Pinus banksiana seedlings were submitted to various frost temperatures during an artificial frost to induce different levels of root damage. Frost-damaged and control seedlings were placed in a greenhouse under high and low soil moisture regimes. Seedling growth and physiology were evaluated periodically. Seedling survival was reduced when root damage reached levels of 60-80%. Root systems of all three species showed partial to total recovery by the end of the experiment. In general, root freezing damage caused reductions in seedling growth, with these reductions becoming less significant over time. Root damage had little to no effect on black spruce and jack pine seedling physiology, while white spruce CO 2 uptake decreased with increasing root damage. Shoot nitrogen content of all three species decreased slightly with increasing root damage.     

Stand Structure in Pine,Spruce and Deciduous Forests: A Comparison Between Finnish and Russian Karelia

This study investigated the stand structure in pine, spruce and deciduous forests in the border district of Finland and Russia. A total of 46 mature forest stands was selected as pairs, the members of each pair being as similar as possible with respect to their forest site type, age, moisture and topography. The stands were then compared between the two countries by means of basal areas and number of stems. The proportions of dominating tree species were 2-12% lower, and correspondingly the proportions of secondary tree species higher, in Russian forests. The density of the forest stock was also higher in each forest type in Russia. The forests in the two countries differed most radically in terms of the abundance of dead trees. The amount was two to four times higher in Russian deciduous and spruce forests, and in pine forests the difference was 10-fold. The stand structures indicated that Russian coniferous stands, in particular, were more heterogeneous than intensively managed pine and spruce stands in Finland.     

Growth and Terminal Bud Formation in Picea abies Seedlings Grown with Alternating Diurnal Temperature and Different Light Qualities

The root collar diameter and the height:diameter ratio are of particular importance in container-grown seedlings where a high density in the containers may produce spindly seedlings. Temperature regimes and light quality are known to affect plant growth. The aim of this study was to identify responses in Picea abies (L.) Karst. seedlings grown with light providing different red:far-red ratios and under temperature regimes with alternating day (DT) and night temperature (NT) from negative (DT < NT) to positive (DT > NT) difference (DIF) between DT and NT. Experiments were conducted in controlled environment chambers and in a daylight phytotron. Only limited thermoperiodic responses appeared in P . abies seedlings with respect to seedling height and dry weight accumulation. The formation of terminal buds, however, was clearly delayed in seedlings grown at negative DIF. The results indicate a requirement for day extension light that is high in far-red, to prevent terminal bud formation under natural short-day conditions. An extended study should be conducted to clarify the minimum level of light intensity and the optimal light quality needed to prevent terminal bud formation under natural short-day conditions.     

CAD与建筑设计的关系

本文分析了CAD在设计中引起的正面和负面影响,并进行了系统的阐述,从而使设计者在应用中能够保持客观的态度。     

Natural Regeneration in Clear-cuts: Effects of Scarification,Slash Removal and Clear-cut Age

The effects of soil scarification (mounding), slash removal and clear-cut age on the natural regeneration in clear-cuts was evaluated using data from four sites in southern Sweden. The treatments were carried out during a good seed and establishment year for birch ( Betula pubescens Ehrh. and B. pendula Roth). Scarification had the strongest positive effect on the density of naturally regenerated seedlings, especially in birch, but also in pine ( Pinus sylvestris L.) and spruce [ Picea abies (L.) Karst]. Slash removal had a positive effect on birch density. No statistically significant effect of clear-cut age was found. In addition, no statistically significant interactions between clear-cut age and scarification or slash removal were found. The ingrowth of field vegetation was the fastest in areas that were not scarified, less rapid in areas scarified on old clear-cuts, and the slowest after scarification in fresh clear-cuts. In conclusion, it may be possible to control the density of birch during a good establishment year for birch. If birch is desired, the best combination of treatments is to remove the slash and scarify; otherwise, these treatments should be avoided.     

Risk Perception Among Non-industrial Private Forest Owners

Based on an enquiry, risk perception among non-industrial private forest owners is described in relation to climate change and forestry hazards. Of the respondents, 11% took action to remedy the effects of climate change. Out of a given set, hazards were ranked according to each respondent's experience of recent substantial financial loss to the estate and in relation to his or her willingness to make investments aimed at risk reduction. For each hazard, the respondent assessed the risk in four classes ranging from very high to negligible risk. Six hazards were considered most problematic in all three aspects: browsing damage, falling timber prices, damage by wind, spruce bark beetle, root rot and pine weevil. A majority of the respondents claimed to take action to reduce the risk associated with at least one hazard, while 35% did not know whether they did. Excluding climate change, the need for decision support was the largest in relation to damage by wind owing to a combination of perceived high risk and a high level of ignorance in relation to whether risk-reducing measures were taken.     

Interactions in decomposition and N mineralization between tropical legume residue components

In situ produced plant residues contain a mixture of different plant components of varying quality. To assess synergistic or antagonistic interactions occurring during the decomposition and mineralization of such mixtures, individual plant parts (stems, leaves, leaf litter and roots) or the mixture of stems, leaves and leaf litter of the agroforestry species pigeonpea (Cajanus cajan) or of crop residues of peanut (Arachis hypogaea) or of the weed hairy indigo (Indigofera hirsuta) were incubated in pots for 19 weeks. Periodically, remaining plant residues were sieved out (>2 mm), weighed and N content as well as soil mineral N determined. Above- and below-ground residues of peanut decomposed fastest and showed the largest N release in agreement with their high N concentration and low-acid detergent fibre (ADF) : N ratio. Hairy indigo was hypothesized to be of lower quality than pigeonpea because of its high-polyphenol content. However, it decomposed faster than pigeonpea, largely because of the higher N and lower lignin concentration of its components. Ranking of individual plant components for N mineralization resulted in the following pattern, leaves > leaf litter > roots > stems. In mixtures of the different plant components a similar species order in decomposition was obtained, e.g. peanut > hairy indigo > pigeonpea. The amount of N released from the mixture was dominated by stem material that comprised 46–61% of the mixture. The interactions in mixtures were relatively small for peanut (generally high-quality components) as well as for pigeonpea (low proportion of high-quality components, i.e. N rich leaf material). However, a positive interaction occurred during later stages of N mineralization in the mixture of hairy indigo as it had a significant proportion of N rich components and absence of highly reactive polyphenols. Thus, for plants with low to intermediate chemical quality attributes, manipulating plant composition (e.g. by varying harvest age, affecting stem and leaf proportions) will be important to obtain significant interactions during decomposition when its components are mixed.     

Pasture production and tree growth in a young pine plantation fertilized with inorganic fertilizers and milk sewage in northwestern Spain

The use of organic waste materials such as milk sewage as an organic fertilizer could have the dual advantages of organic-waste disposal and reduced dependence on inorganic fertilizers. The effects of fertilization with (1) conventional mineral fertilization, (2) milk sewage sludge at 40 kg N ha⁻¹ target rate and (3) no fertilization on pasture production and tree growth were examined in an experiment consisting of two pasture mixtures under a one-year-old Pinus radiata plantation with a density of 2500 trees ha⁻¹. The two pasture mixtures were: (1) Dactylis glomerata L. var. saborto (25 kg ha⁻¹) + Trifolium repens L. group Ladino (4 kg ha⁻¹) + Trifolium pratense L. var. Marino (1 kg ha⁻¹); (2) Lolium perenne L. var. Tove (25 kg ha⁻¹) + Trifolium repens L. group Ladino (4 kg ha⁻¹) + Trifolium pratense L. var. Marino (1 kg ha⁻¹). The experiment began in the spring of 1995 using a randomized block design with three replicates in Castro Riberas de Lea (Lugo, Galicia, north-western Spain). Plot size was 12 × 8 m², with a 1 m buffer strip between plots. Two-year data showed that fertilization with either material had a positive effect on pasture production, with no significant difference between the two fertilization treatments. Tree growth in the milk sewage sludge plot was significantly higher than in the control plots. Inorganic fertilization increased pasture production, but affected tree growth negatively. The results show that milk sewage sludge could be used as a fertilizer in silvo-pastoral systems. This revised version was published online in June 2006 with corrections to the Cover Date.