两株地理隔离的桦褐孔菌深层发酵产生抗氧化次生代谢产物的代谢组学分析

采用基于核磁共振代谢组学的方法研究了两株地理隔离的桦褐孔菌中国牡丹江菌株(IOM)和芬兰万塔菌株(IOV)在深层培养过程中主要代谢产物组成的差异。IOM和IOV两菌株在培养过程中积累菌丝体生物量、菌丝体酚类(TMP)以及三萜化合物(TMT)的过程相似,但IOM菌株产生较高水平的TMP。代谢产物组成动力学比较结果表明,IOM在第3天含有较高水平的硬毛素类似物、桦褐孔菌二糖和TMT,但在第5天均有所下降。导致这些差异的代谢产物为fuscoparianolD,21,24-羊毛甾-7,9(11),23-三烯-3β,22,25-三醇,羊毛甾-8,23-二烯-3β,22,25-三醇,23,24-环戊羊毛甾-3β,21,25-三醇-8-烯,桦褐孔菌二糖,phelligridinsC,D及H,甲基地花菌素A和C,地花菌素B,davallialactone及methyl davallialactone。这些代谢产物在IOV的提取物中均参与清除自由基,而在IOM的提取物中主要是酚类化合物参与自由基的清除。两菌株的这些差异反应了环境条件对桦褐孔菌生长及代谢的影响,即不仅影响其次生代谢产物的合成,还影响代谢产物在防止细胞氧化损伤过程中所起的作用。     

大花序桉心边材变异规律和候选基因挖掘研究

为探究大花序桉(Eucalyptus cloeziana)心材比例差异显著的不同家系间心边材变异规律,挖掘心材变异相关的候选基因,为珍贵用材树种高效培育及育种利用提供基因资源。以18 a生的2个心材比例差异显著的大花序桉家系为材料(家系1和2),各制作解析木3株,沿着树干以1 m为区间分段截取圆盘,测量东西和南北2个方向的带皮直径、去皮直径、总年轮数、边材年轮数、边材直径,并开展心材和边材径向和轴向分析。同时利用各解析木胸径处初生木质部样品进行DNA混池测序,发掘等位基因频率差异显著的SNP位点并挖掘相关功能基因。结果表明,大花序桉边材宽度和心材半径的方位变异中家系2大于家系1,平均差值分别为0.7和5.5 cm,在随树高的变异中,家系1和2的心材半径和心材年轮数的下降速率分别为0.40和0.64及0.43和0.36。两家系间基本密度差异显著,家系1为0.80～0.82 g/cm³,家系2为0.75～0.78 g/cm³。基本密度与树高、横截面半径和心材半径呈显著负相关,与顺纹抗拉强度、弦面硬度和部分力学性质呈显著正相关。利用DNA混池测序共...     

杜仲雄花中次生代谢物合成积累的动态变化

对杜仲(Eucommia ulmoides Oliv.)雄株不同花期雄花中次生代谢产物含量进行分析测定,并对杜仲雄花次生代谢的生理基础及不同花期次生代谢产物含量差异进行了探讨.研究结果表明,雄花在不同花期次生代谢产物含量均有差异.总黄酮含量在花蕾期最高(4.010%),始花期最低(2.422%),从盛花期到末花期逐渐上升;桃叶珊瑚苷和绿原酸含量均在花蕾期最高(分别为2.351%和1.075%),盛花期最低(分别为1.463%和0.503%),至末花期含量上升;京尼平苷酸含量在始花期最低(0.217%),从盛花期开始逐渐升高,至末花期高达1.403%;次生代谢产物总量也以花蕾期为最高(7.420%).杜仲雄花的花蕾期和盛花期是兼顾质量和产量的最佳采摘期.     

七种石斛花朵挥发性成分分析

为了解几种石斛属植物鲜花挥发性成分,该文利用固相微萃取(SPME)法结合GC-MS技术分析了7种石斛(含2个品种)花朵挥发性成分及其相对含量。结果表明:(1)从7种石斛(含2个品种)中共鉴定出52种挥发性化合物,包括萜烯类、酯类、芳香族化合物、含氮化合物、烷烃类、醇类和酮类等7类,其中萜烯类总相对含量最高,为83.25%～94.93%,为主要挥发性成分。(2) 7种石斛共同含有(1R)-(+)-α-蒎烯、D-柠檬烯和顺式-β-罗勒烯等3种成分,每个品种的相对含量存在差异。其中,顺式-β-罗勒烯在鼓槌石斛、细叶石斛、流苏石斛、翅梗石斛和春石斛‘H1’等5种石斛中的含量均最高,分别达到46.09%、46.40%、39.02%、65.96%和54.34%;(1R)-(+)-α-蒎烯在鼓槌石斛、流苏石斛、翅梗石斛、春石斛‘H1’和春石斛‘818’等5种石斛中的相对含量相对较高,分别为34.11%、25.61%、15.26%、21.11%和23.21%; D-柠檬烯在翅萼石斛和翅梗石斛中的含量较高,分别为16.02%和6.86%,而在其他5种石斛中的含量均较低。(3)β-蒎烯在流苏石斛和翅萼石斛中的相对含量高达19.39%和45.95%,桧烯仅在春石斛‘818’中能检测到(12.24%)。这些含量较高的成分可能为主要的香气成分或特征性香气成分。综上结果认为,7种石斛花朵挥发性成分既含有相同的成分也含有不同成分,且含量随种类的不同而不同。这些结果可为研究石斛属植物花香代谢以及产品开发等提供参考价值。     

棘孢木霉挥发性次级代谢产物检测及抑菌活性分析

木霉是一类具有重要生防价值的丝状真菌。文中首先对分离自浙江省绍兴市和广东省佛山市共12株棘孢木霉Trichoderma asperellum进行平板拮抗评价,然后采用顶空固相微萃取气质联用法(HS-SPME-GC-MS)检测拮抗性较好的两株菌的挥发性次级代谢产物。结果表明,棘孢木霉ZJSX5003和GDFS1009菌丝生长迅速,对尖孢镰孢菌Fusariumoxysporum抑制率分别达73%和74%。挥发性次级代谢产物主要是醇类和酮类,其中包含异丁醇、异戊醇、3-甲基-3-丁烯-1-醇、3-羟基-2-丁酮、2,3-丁二醇和6-正戊基-2H-吡喃-2-酮(6-PAP)。进一步通过体外抑菌试验,证实6-PAP具有较好的抑制尖孢镰孢菌的效果,为开发以木霉菌代谢产物如6-PAP为主要成分的生防制剂提供指导。     

不同产地大血藤次生代谢产物含量比较

对不同产地大血藤的不同营养器官的总鞣质、总生物碱、皂苷、木质素、绿原酸、总黄酮共6种次生代谢产物的含量进行测定与比较。结果显示：6种次生代谢产物在不同营养器官中的含量具有一定的差异,总含量以叶最高,老茎次之,幼茎最小。6种次生代谢产物除木质素茎含量较高外,其余5种均以叶片的含量最高,差异具有显著性。不同产地大血藤的6种次生代谢产物的含量具有显著性差异。通过逐步回归分析和通径分析,得知大血藤叶片次生代谢产物与土壤生态因子的关系密切。聚类分析显示庆元百山祖及天台的大雷山和天台山的大血藤叶片的次生代谢产物含量较高。     

Changes in log quality at different decay stages in an alpine forest

倒木是高寒森林生态系统重要的碳(C)库和养分库,其不同分解阶段的质量变化,是认识倒木分解过程中C和养分释放的重要基础。以一个分解序列的岷江冷杉(Abies faxoniana)倒木为研究对象,研究了心材、边材和树皮在5个分解阶段的C:N:P化学计量特征,以及木质素和纤维素含量动态。结果显示:I至III分解阶段,随着分解程度加深,树皮C含量升高,而心材和边材C含量降低,从IV分解阶段开始倒木各组分C含量均开始显著降低。除III分解阶段的心材外,倒木各组分N含量总体表现为随着分解程度加深而增加的趋势,除边材N含量在V分解阶段时显著升高外,其余组分均未达到显著性水平。心材和树皮P含量表现为先降后升的变化趋势,最小值分别出现在III和II分解阶段;边材P含量表现为随着分解程度加深而增加。在同一分解阶段,树皮相对于边材和心材均具有最低的C:N:P化学计量比,易分解比例Fm也表明树皮更易于分解。边材在I和II分解阶段的C:N:P化学计量比最高,心材在III到V分解阶段C:N:P化学计量比最高。心材C:P、树皮和边材的C:N和C:P临界值与N和P的初始值成反比。纤维素含量随着倒木分解而降低,不同分解阶段的纤维素含量表现为:心材>边材>树皮;但木质素含量随着分解程度加深而增加,表现为:树皮>边材>心材;倒木3个组分纤维素含量下降均快于木质素,此外,IV和V分解阶段的树皮木质素与纤维素比值显著增高,且一直处于较高水平。统计分析结果表明:倒木N含量显著影响不同分解阶段木质素和纤维素分解。由生态化学计量学理论推测:树皮分解前期易受N限制,整个分解阶段均易受P限制,心材和边材在整个分解阶段均易受N和P限制。     

摇瓶培养条件下雷公藤不定根生长及次生代谢产物含量的研究

以雷公藤(Tripterygium wilfordii Hook. f.)不定根为材料,研究摇瓶悬浮培养条件下接种密度、装液比例、逐级放大、消泡剂、大孔吸附树脂种类及浓度对雷公藤不定根增长量、不定根及培养基中雷公藤内酯醇、雷公藤吉碱、雷公藤次碱含量的影响。结果显示,接种密度在15 g/L (FW)时较适合不定根的继代培养和次生代谢产物的积累。250 m L摇瓶中装入100 m L培养基,即装液量为2/5时,培养基利用率最高。随着摇瓶体积的逐渐放大,不定根增长量和3种次生代谢产物含量略有下降,5 L摇瓶中不定根增长量为对照的91.6%,内酯醇、吉碱和次碱的含量分别为对照的91.8%、91.7%和96.9%。6种大孔吸附树脂中,XAD-7处理对不定根的生长有明显促进作用,培养结束时,3种次生代谢产物产量显著提高,当XAD-7浓度为0.5 g/瓶时不定根增长量为对照的1.2倍,内酯醇、吉碱和次碱产量最高,分别为对照的2.9、2.4和2.2倍。培养基中添加消泡剂后不定根增长量、3种次生代谢产物总产量均不同程度下降,其中,LX-603处理后,虽然不定根增长量为对照的85%,内酯醇、吉碱和次碱产量分别为对照的78%、64%和87%,但明显抑制了培养过程中泡沫的产生。研究结果表明筛选的摇瓶逐级放大培养雷公藤不定根的方法效果较好,可为雷公藤不定根生物反应器放大培养奠定基础。     

高寒森林倒木在不同分解阶段的质量变化

倒木是高寒森林生态系统重要的碳(C)库和养分库, 其不同分解阶段的质量变化, 是认识倒木分解过程中C和养分释放的重要基础。以一个分解序列的岷江冷杉(Abies faxoniana)倒木为研究对象, 研究了心材、边材和树皮在5个分解阶段的C:N:P化学计量特征, 以及木质素和纤维素含量动态。结果显示: I至III分解阶段, 随着分解程度加深, 树皮C含量升高, 而心材和边材C含量降低, 从IV分解阶段开始倒木各组分C含量均开始显著降低。除III分解阶段的心材外, 倒木各组分N含量总体表现为随着分解程度加深而增加的趋势, 除边材N含量在V分解阶段时显著升高外, 其余组分均未达到显著性水平。心材和树皮P含量表现为先降后升的变化趋势, 最小值分别出现在III和II分解阶段; 边材P含量表现为随着分解程度加深而增加。在同一分解阶段, 树皮相对于边材和心材均具有最低的C:N:P化学计量比, 易分解比例F_m也表明树皮更易于分解。边材在I和II分解阶段的C:N:P化学计量比最高, 心材在III到V分解阶段C:N:P化学计量比最高。心材C:P、树皮和边材的C:N和C:P临界值与N和P的初始值成反比。纤维素含量随着倒木分解而降低, 不同分解阶段的纤维素含量表现为: 心材>边材>树皮; 但木质素含量随着分解程度加深而增加, 表现为: 树皮>边材>心材; 倒木3个组分纤维素含量下降均快于木质素, 此外, IV和V分解阶段的树皮木质素与纤维素比值显著增高, 且一直处于较高水平。统计分析结果表明: 倒木N含量显著影响不同分解阶段木质素和纤维素分解。由生态化学计量学理论推测: 树皮分解前期易受N限制, 整个分解阶段均易受P限制, 心材和边材在整个分解阶段均易受N和P限制。     

锐齿栎林非同化器官营养元素含量的分布

锐齿栎林非同化器官营养元素含量的大小为,N：皮＞枝＞根＞边材＞心材,P：根＞枝＞皮＞边材＞心材,K：枝＞皮＞根＞边材＞心材,Ca：皮＞枝＞根＞心材＞边材,Mg：根＞皮＞枝＞边材＞心材。不同枝径和冠层枝营养元素含量存在一定差异。干皮及干木质部营养元素含量与盘高度呈显著的正相关或正相关,随圆盘由基部到稍部高度的增加均呈增加的趋势。干营养元素含量为干皮＞边材＞心材,三者呈极显蓍的正相关或显著的正相关。随林木根径和根系在运动场支中的增加,极系营养元素含量均呈了低的趋势。69年生个体生物量和营养元素积累量分别为1235.782和14.4977kg,其中N、P、K、Ca、Mg的积累量分别占营养元素积累量的20.2%、8.5%、20.4%、53.0%和2.9%;根、干、皮、枝和叶的生物量分别占生物总量的17.05%、49.59%、9.18%、24.30%和0.86%,营养元素积累量分别占总量的14.93%、22.65%、21.19%、36.97%、4.25％。     

Growth rate and ring width variability of teak, Tectona grandis (Verbenaceae) in an unmanaged forest in East Timor

Teak (Tectona grandis) is one of the most valuable timbers in international trade and an important species for tropical forestry. Teak is found on the island of East Timor but no information is available on teak growth from this region. A pure stand planted in 1940-50 in the North of East Timor and left unmanaged was studied. Fifteen trees were sampled in October-November 2003 and stem discs taken at three height levels of its height (1.7m, 9.5m and 18.7m), and cores were collected at DBH. Transverse surfaces of the discs and cores were polished for ring identification. Core cross sections were first digitized and disc cross sections were observed under the microscope. Three randomly selected radii were analyzed in each disc. Ring width measurement and ring counting were done using image analysis software. The distinction between heartwood and sapwood was performed macroscopically by colour difference, and heartwood radius and sapwood width were measured. The relationship between stem and heartwood radius was studied for each disc and heartwood percentage by radius was determined. Radial ring width curves are presented for the different axial positions within the stem, and ring width variability was analyzed. Growth rates were calculated and age-radius relationships were estimated using cumulative growth curves. Growth rings were large and well defined in the juvenile phase, reflecting the specie's fast-growing character. The year-to-year variation of ring width showed a similar pattern among trees. Mean ring width ranged between 4.3-7.3mm for the first 20 years and 3.3-5.1mm for 30 to 45 years. Pith eccentricity was evident in the lower part of the stem and ring wedging occurred. On average, heartwood represented 84% of the radius and sapwood contained 6 to 11 rings. The age-related variation of ring width and the occurrence in the lower part of the tree stems of eccentricity and wedging rings, highlights the importance of appropriate stand management, particularly regarding basal density distribution over time, whenever optimized timber production is envisaged.     

小蓬草精油的提取及GC-MS分析

水蒸馏和水蒸气蒸馏分别提取鲜小蓬草精油的气相色谱-质谱分析结果表明:水蒸馏提取的鲜小蓬草精油挥发性组分较多且柠檬烯质量分数最高(15.36%),香芹酮(10.15%)次之.而水蒸气蒸馏分别提取鲜和干小蓬草精油的气相色谱-质谱分析表明干小蓬草有利于提取更多精油香料成分,且柠檬烯质量分数最高(57.68%),反式-α-佛手...     

Assessment of chemical resistance of wild and cultivated Vigna species to the brown pod-bug Clavigralla tomentosicollis Stal (Hem., Coreidae)

The effects of secondary metabolites in different Vigna species on the development of Clavigralla tomentosicollis were investigated in an artificial seed system using different fractions of crude pod extracts, while the orientation response of this pod‐bug to volatile extracts was studied using a dual‐choice olfactometer. Feeding on the neutral fraction extracts, in contrast to the basic and acidic fractions, resulted in significantly higher mortalities, longer total developmental time, and lower growth index of the insects in comparison with controls. All volatile extracts elicited an avoidance reaction by C. tomentosicollis, except the volatile from the susceptible genotype IT84S‐2246 which generally attracted as many insects as controls. Extracts from wild Vigna species showed higher activity than those from their cultivated relatives. The present study which has established that most secondary metabolites in cowpea pods were localized in the neutral fraction of the crude extract, could facilitate experiments on the separation and characterization of the toxic factors involved.     

Sesquiterpenes and a phenylpropanoid from Cordia trichotoma

Two new secondary metabolites, the phenylpropanoid 3-(2',4',5'-trimethoxyphenyl)propanoic acid (1) and the sesquiterpene (+)-1beta,4beta,6alpha-trihydroxyeudesmane (2) were isolated from the heartwood of Cordia trichotoma Vell., along with the known sesquiterpenes (-)-1beta,4beta,7alpha-trihydroxyeudesmane (3) and (+)-1beta,4beta,11-trihydroxyoppositane (4). Their structures were elucidated by means of spectroscopic data interpretation, mainly 1D and 2D NMR and mass spectrometry.     

Use of direct-infusion electrospray mass spectrometry to guide empirical development of improved conditions for expression of secondary metabolites from actinomycetes

A major barrier in the discovery of new secondary metabolites from microorganisms is the difficulty of distinguishing the minor fraction of productive cultures from the majority of unproductive cultures and growth conditions. In this study, a rapid, direct-infusion electrospray mass spectrometry (ES-MS) technique was used to identify chemical differences that occurred in the expression of secondary metabolites by 44 actinomycetes cultivated under six different fermentation conditions. Samples from actinomycete fermentations were prepared by solid-phase extraction, analyzed by ES-MS, and ranked according to a chemical productivity index based on the total number and relative intensity of ions present in each sample. The actinomycete cultures were tested for chemical productivity following treatments that included nutritional manipulations, autoregulator additions, and different agitation speeds and incubation temperatures. Evaluation of the ES-MS data from submerged and solid-state fermentations by paired t test analyses showed that solid-state growth significantly altered the chemical profiles of extracts from 75% of the actinomycetes evaluated. Parallel analysis of the same extracts by high-performance liquid chromatography-ES-MS-evaporative light scattering showed that the chemical differences detected by the ES-MS method were associated with growth condition-dependent changes in the yield of secondary metabolites. Our results indicate that the high-throughput ES-MS method is useful for identification of fermentation conditions that enhance expression of secondary metabolites from actinomycetes.     

The accumulation pattern of ferruginol in the heartwood-forming Cryptomeria japonica xylem as determined by time-of-flight secondary ion mass spectrometry and quantity analysis

Background and Aims

Heartwood formation is a unique phenomenon of tree species. Although the accumulation of heartwood substances is a well-known feature of the process, the accumulation mechanism remains unclear. The aim of this study was to determine the accumulation process of ferruginol, a predominant heartwood substance of Cryptomeria japonica, in heartwood-forming xylem.

Methods

The radial accumulation pattern of ferruginol was examined from sapwood and through the intermediate wood to the heartwood by direct mapping using time-of-flight secondary ion mass spectrometry (TOF-SIMS). The data were compared with quantitative results obtained from a novel method of gas chromatography analysis using laser microdissection sampling and with water distribution obtained from cryo-scanning electron microscopy.

Key Results

Ferruginol initially accumulated in the middle of the intermediate wood, in the earlywood near the annual ring boundary. It accumulated throughout the entire earlywood in the inner intermediate wood, and in both the earlywood and the latewood in the heartwood. The process of ferruginol accumulation continued for more than eight annual rings. Ferruginol concentration peaked at the border between the intermediate wood and heartwood, while the concentration was less in the latewood compared wiht the earlywood in each annual ring. Ferruginol tended to accumulate around the ray parenchyma cells. In addition, at the border between the intermediate wood and heartwood, the accumulation was higher in areas without water than in areas with water.

Conclusions

TOF-SIMS clearly revealed ferruginol distribution at the cellular level. Ferruginol accumulation begins in the middle of intermediate wood, initially in the earlywood near the annual ring boundary, then throughout the entire earlywood, and finally across to the whole annual ring in the heartwood. The heterogeneous timing of ferruginol accumulation could be related to the distribution of ray parenchyma cells and/or water in the heartwood-forming xylem.     

内生真菌对油樟悬浮细胞培养的影响

该文研究了内生真菌YG42、YG71、YY11和YY26发酵液,对油樟悬浮细胞的生长量及挥发性代谢产物积累量的影响。结果表明：4种内生真菌对油樟悬浮细胞的生长均有抑制作用,抑制强度随发酵液添加量的增加而加强。4种内生真菌对油樟悬浮细胞挥发性代谢产物积累总量及1,8-桉叶油素、γ-叶松油烯和α-油松油醇3种油樟油组分物质积累量的影响多表现为低浓度促进高浓度抑制的趋势。其中,1％添加量的YG42和YY26及0.25％添加量的YY11对悬浮细胞总挥发性代谢产物积累的促进作用相当且最强,其积累量分别是空白组的2.00、1.95、2.01倍;0.25％添加量的YG71对1,8-桉叶油素积累的促进作用最强,其积累量为空白组的11.03倍;0.25％添加量的YG71和YY26对α-松油醇积累的促进作用相当且最强,其积累量分别为空白组的1.72和1.81倍;对于γ-松油烯的积累,在空白组中未检测到其峰值,4种真菌诱导子对γ-松油烯的产生有诱导作用,诱导的最大峰面积为0.19,诱导菌是0.25％添加量的YG71。该研究结果为充实内生菌影响香料植物挥发性代谢产物合成理论奠定了基础,也为生产上内生真菌提高油樟油中有用物质组分含量措施的采用提供了依据。     

Use of solid phase microextraction (SPME) for profiling fungal volatile metabolites

AIMS: The influence of isolation methods: solid phase microextraction (SPME) with different fibres and simultaneous distillation extraction (SDE) on the profile of isolated fungal volatile metabolites was investigated. METHODS AND RESULTS: Four SPME fibre types: Polydimethylsiloxane, Polyacrylate, Carboxen/PDMS and Carboxen/Divinylbenzene/PDMS were evaluated in terms of their efficiency in extracting volatile metabolites emitted by Penicillium roqueforti grown on wheat kernel medium. All fibres showed varied efficiency and selectivity in extracting volatile compounds. Sesquiterpene hydrocarbons were the predominant fraction of volatile compounds isolated by all fibres, and ranged from 55.4 to 93.7% of all volatiles depending on the type of fibre used. Alcohols and ketones ranged from 2.7 to 20.5%, esters from 1.2 to 12.8%, and monoterpene hydrocarbons from 1.2 to 5.4%. Profile of volatile compounds obtained by SDE differed from SPME and the oxygenated sesquiterpenes formed the predominant fraction of volatiles isolated using SDE. SIGNIFICANCE AND IMPACT OF THE STUDY: The data in this study show that analysed profile of volatile compounds emitted by fungi is highly dependent on the extraction method.     

Chemo-profiling of bioactive metabolites from Chaetomium globosum for biocontrol of Sclerotinia rot and plant growth promotion

Chaetomium globosum Kunze ex. Fries has been known to produce diverse bioactive metabolites, attracting researchers to exploit the biocontrol agent for plant disease management. However, distinct research gaps are visible regarding detail characterization of bioactive metabolites. Thus the current study has been planned to characterize volatile and nonvolatile compounds of most potential strain of C. globosum 5157. GC–MS analysis of hexane fraction revealed twenty-six volatile organic compounds, representing 65.5% of total components in which 3-octanone (21.4%) was found to be most abundant. UPLC-QTOF-MS/MS analysis of ethyl acetate and methanolic fractions resulted tentative characterization of fifteen and eleven metabolites, respectively. Among these, nine metabolites were isolated, purified and characterized using ¹H NMR and High resolution mass spectrometric analysis to delineate mass fragmentation pattern for the first time. Antifungal potential of hexane fraction exhibited high inhibitory action against Sclerotium rolfsii (139.2 μg mL^?1) whereas ethyl acetate fraction was highly effective against Sclerotinia sclerotiorum (112.1 μg mL^?1). Comparative assessment of C. globosum 5157 vis a vis Trichoderma harzianum A28 revealed promising effect of C. globosum 5157 with respect to antifungal properties and plant growth promotion of Brassica seedlings.