微波辅助法提取印楝素的研究

介绍了一种有效利用微波辅助从印楝(Azadirachta indica A.Juss)种仁中提取印楝素的方法。通过考察辐射时间、溶剂、物料比以及辐射功率等因素对提取率的影响，得出结论为：预处理过的印楝种仁粉末6.65g(相当于印楝种仁10g)，以甲醇做溶剂、微波功率210w、物料比1：3(g／mL)、辐射时间为3min*3次，提取率最高。与传统的磁搅拌法相比，该方法不仅工艺简单，而且可以提高产率，缩短提取时间。用高压液相法定量地测定了所提取印楝素的含量，其提取率可达0．519％．     

印楝素A无细胞合成体系构建及其合成前体

【目的】印楝素是一种植物源杀虫剂,印楝素A是印楝素的主要活性成分。本研究通过对印楝素A无细胞合成体系制备条件和反应条件的优化,以及印楝素A合成前体的筛选,探讨印楝素A的离体生物合成途径,以为指导构建印楝素异源生物合成平台,实现印楝素的异源生物合成奠定基础。【方法】通过对缓冲液类型、浓度、起始pH、提取时间及料液比进行单因素试验及正交试验优化,构建印楝素A无细胞合成体系;从反应终止试剂及稳定剂、反应温度、反应时间、底物浓度以及辅助因子5个方面优化无细胞合成体系的反应条件。利用印楝素A无细胞合成体系分别以2,3-环氧鲨烯、羊毛甾醇、大戟二烯醇、丁酰鲸鱼醇、nimbin、茄碱苷和脱乙酰茄碱苷为底物,以印楝素A相对产量为评价标准,探讨印楝素A合成前体。【结果】印楝素A无细胞合成体系的构建:以pH7.0的200 mmol·L~(-1) Tris-HCl缓冲液,按1∶20(g·mL~(-1))料液比加入印楝叶片,提取1 h。优化的印楝素A无细胞合成条件为:反应总体积800μL,其中含印楝无细胞提取物300μL(相当于印楝叶0.015 g),100μL 250μmol·L~(-1)的底物(鲨烯)和400μL含有1 mmol·L~(-1) Mg~(2+)、1 mmol·L~(-1) Mn~(2+)、0.1 mmol·L~(-1) ATP、0.1 mmol·L~(-1) NADPH~+和5 mmol·L~(-1)抗坏血酸的缓冲液,在30℃下反应60 min,迅速加入200μL乙酸终止反应。分别以2,3-环氧鲨烯、丁酰鲸鱼醇、大戟二烯醇、nimbin、茄碱苷和脱乙酰茄碱苷为底物,反应完成后,印楝素A含量均有不同程度的增加;羊毛甾醇抑制体系中印楝素A的合成。【结论】羊毛甾醇不是印楝素A的合成前体,2,3-环氧鲨烯和丁酰鲸鱼醇是印楝素A的前体,nimbin、大戟二烯醇、脱乙酰茄碱苷和茄碱苷极可能是印楝素A的合成前体,且茄碱苷位于合成途径较下游的位置。     

印楝种子品质性状的遗传多样性及稳定性分析

在云南印楝资源全面调查的基础上,以实生株系的种子为研究对象,运用HPLC技术分析种子印楝素组分含量,采用描述统计、方差分析、系统聚类和重复力估计,定性、定量分析印楝实生株系种子品质的遗传多样性及稳定性。结果表明:云南引种栽培印楝实生株系的种子品质存在广泛变异,种子印楝素A、B、AB含量和印楝素A与印楝素B含量比的变异幅度分别为0.28% 0.85%、0.04% 0.39%、0.37% 1.15%和1.698.25;种子印楝素A、B、AB含量分为3个类型:高含量型(azA ≥ 0.69%、azB ≥ 0.25%、azAB ≥ 0.92%)、低含量型(azA ≤ 0.44%、azB ≤ 0.10%、azAB ≤ 0.51%)和中等含量型(azA=0.68% 0.45%、azB=0.24% 0.11%、azAB=0.91% 0.52%);按此标准将高印楝素AB含量型分为"印楝素AB优异型(azAB ≥ 1.15%)"、"印楝素A优、B劣型"(azA ≥ 0.85%、azB ≤ 0.21%)、"印楝素A劣、B优型"(azA ≤ 0.67%、azB ≥ 0.39%)和"印楝素AB含量高型"(azAB=1.05% 0.92%);反映在重复力上,印楝实生株系种子印楝素A、AB的重复力分别为0.2063、0.3252,都属于"中"重复力性状。"印楝素AB优异型"即具有优异种子品质性状的植株是印楝药用原料林培育的繁殖材料;"印楝素A优、B劣型"和"印楝素A劣、B优型"即种子品质性状优缺点互补的植株是药用印楝品质育种的杂交亲本。     

影响印楝素稳定性及其含量的主要因素

印楝素的稳定性及其含量的高低受许多因素的影响。印楝素对光敏感，暴露在光下会逐渐失去活性；室温及低于20℃的贮藏温度有利于印楝素的稳定，而高于室内温度会加速印楝素的降解；印楝素在中性条件下比较稳定，室温条件下，在pH值为4～6的酸性水溶液中也稳定，但在碱性和强酸溶液中不稳定；中性有机溶剂对其稳定性有利。微生物的活动会加快印楝素的降解，不同的地理种源和种植土壤对印楝种实中印楝素的含量高低影响很大，在干季所结果实的种子中印楝素含量高于在雨季种植所结果实的种子。而不同组培条件对组培体的印楝素含量也会产生影响。     

印楝素制剂防治松毛虫及松小蠹室内试验初报

应用印楝素乳油进行了松毛虫、松小蠹的室内防治效果试验。试验结果表明用0．3％印楝素乳油400倍“触杀 胃毒”处理松毛虫，施药后20天，防治效果达到88．5％；用0．3％印楝素乳油100倍“触杀 胃毒 内吸”处理松小蠢，施药后15天，防治效果达到95．6％。观察发现，施用印楝素后，使松毛虫2龄幼虫产生较强的拒食作用，并能抑制其生理发育。     

印楝无性系当代种子农药品质性状的遗传变异及农药型印楝优株评价

[目的]建立一套适合印楝药用优树无性系测定的分析评价方法,并进行农药型印楝优株评价。[方法]以印楝药用优树和平均木无性系为试验材料,通过不完全谱系设计建立无性系品比试验,采用重复力估算法在表型水平上对无性系当代种子农药品质性状进行遗传变异分析,提出"农药型印楝"的概念和标准,根据该标准进行农药型印楝优株评价。[结果]无性系种子印楝素组分含量随优树种子印楝素组分含量的变化而变化,但在无性系内和无性系间都存在不同程度变异:同一无性系各分株间的印楝素B变异(5.26%37.55%)较印楝素A变异(6.06%15.80%)和印楝素AB变异(2.95%16.14%)大,印楝素B和印楝素AB几乎呈退化和/或趋中遗传倾向,而印楝素A则既有退化和/或趋中遗传倾向又有超母遗传倾向;无性系间种子印楝素组分含量性状具有异质性(P0.01)。无性系印楝素A重复力(0.78)、印楝素B重复力(0.81)和印楝素AB重复力(0.83)均较高,表明无性系间印楝素组分含量的差异主要是由遗传效应和一般环境效应造成的,受特殊环境效应影响较小。按农药型印楝标准,评定LD0505、WW0401、AT0515三个无性系为"农药型印楝品种",LD0505和WW0421是"农药型印楝亲本"。[结论]重复力估算法适用于印楝药用优树无性系测定的分析评价,优树种子农药品质性状的遗传传递力强,无性系能保持优树农药品质的优良特性;评定LD0505、WW0401、AT0515为农药型印楝品种,LD0505和WW0421还可作为农药型印楝亲本,其余优树无性系不宜选择。     

印楝素和苦楝素对植物病原菌的抑制作用

印楝(Azadirachta indica．A．juss)系楝科楝属乔木，被美国农业部誉为“可解决全球问题之树”。从印楝植株中发现了以印楝素(Azadirachtin)为主的80余种活性物质，可防治10目400余种农林、仓库和卫生害虫，是理想的生物杀虫剂。其杀真菌活性的研究在国内未见报道。本文选用4种植物病原菌，测定了其抑菌活性；并与另一种     

印楝生物学特性及引种栽培

报道了印楝的形态特征和3个来自不同水分条件的印楝种源在引种地的适应性及经济性状的表现。初步认为3个印楝种源;（1）物候期不同;（2）保存率,生长量,结果枝数,结果枝比等生长指标也都存在明显差异。（3）种子的印楝素量＞0．4％,高于原产地的平均水平。从适应性和经济性状比较,来自干旱,半干旱地区的2种源优于来自湿润地区种源。     

快速搅拌法室温浸提印楝油和印楝素

在室温下用间歇式快速搅拌法从印楝种仁中浸提印楝油和印楝素。讨论了料液比以及浸提时间对得率的影响。结果表明:印楝种仁300g,以750mL石油醚做溶剂,在快速搅拌条件下浸提21min时印楝油得率最高;将300g去油后的印楝种仁以900mL甲醇做溶剂,在快速搅拌下浸提21min时印楝素得率最高。由此得出,快速搅拌法浸提印楝油的最佳条件是:溶剂石油醚、料液比1∶2.5(g∶mL,下同)、时间每次7min、浸提3次,出油率30.4%;快速搅拌法浸提印楝素的最佳条件是:溶剂为甲醇、料液比1∶3、时间每次7min、浸提3次,印楝素A的得率为0.438%,印楝素B的得率为0.142%。     

中国林业专利选摘

☆印楝素植物性杀虫剂制剂专利号 :0 0 1 1 4 2 2 7 5　公告号 :ＣＮ1 2 761 61Ａ专利权人 :华南农业大学 (广州市 )摘　要 :以印楝素和印楝油为主要杀虫活性成分 ,通过加入一定比例的多种配料配制成印楝素乳油 ,可湿性粉剂、悬浮剂三种剂型。是一类高效、低毒、无残毒 ,对人畜安全的印楝素植物性杀虫剂制剂。对小菜蛾、黄曲条跳甲、菜青虫等多种害虫具有优异的防效 ,使用该农药尤其适于防治已产生抗性的各类害虫。☆一种银杏茶专利号 :0 0 1 1 4 364 6　公告号 :ＣＮ1 2 761 64Ａ专利权人 :石延中 (河南省鲁山县 )摘　要 :由银杏叶提取物干…     

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.