地形对赤霞珠干红葡萄酒香气成分的影响

采用液液萃取法提取3种地形下赤霞珠干红葡萄酒的香气成分,用气相色谱-质谱进行分离测定,结合计算机检索技术对分离化合物进行鉴定,并应用色谱蜂面积归-法测定各成分的相对含量.结果表明:海拔较低的山地葡萄酒主要香气成分的种类和相对含量高于海拔高的山地葡萄酒,其中吲哚-3-乙醇、2-乙基己醇、己酸乙酯仅在前者中被检出;平地葡萄酒主要香气成分的种类和相对含量仅次于地势较低的山地葡萄酒,其中酯类化合物含量明显高于山地葡萄酒;苯乙醇、乙酸异戊酯等构成葡萄酒特征香气的组分在高海拔山地葡萄酒中未被检出.     

云南切梢小蠹对云南松树干和枝梢挥发性物质趋向反应的差异分析

利用GC-MS联用仪和计算机信息检索法,通过对云南松Pinus yunnanensis树干韧皮部和枝梢的挥发性物质成分的分析发现,二部位所含挥发性物质在组成成分和含量上存在明显差异。云南松树干挥发物以α-蒎烯为主,含量达81.01%,枝梢为挥发物以α-蒎烯和β-水芹烯为主,含量分别达29.20%和30.52%。与云南松枝梢挥发物相比,在云南松树干的挥发性物质中含有较多的α-蒎烯和3-蒈烯;而在云南松枝梢挥发物中,则含有较多的莰烯、桧烯、柠檬烯、β-水芹烯、石竹烯和异松油烯。室内趋向试验表明,蛀梢期云南切梢小蠹Tomicus yunnanensis对云南松枝梢挥发物表现出一定的正趋向反应,而对云南松树干挥发物则表现出一定的忌避反应。     

绿盲蝽对七种锦葵科植物挥发物的EAG和趋向行为反应

研究绿盲蝽Apolygus lucorum(Meyer-Dür)对锦葵科植物挥发物的EAG和趋向行为反应,为研发绿盲蝽引诱剂或驱避剂策略提供科学依据。田间采用动态顶空采样方法收集了7种锦葵科植物的挥发物,通过室内气相-质谱联用系统(GC-MS)进行物质鉴定及定量分析,最终测定了雌、雄绿盲蝽成虫对11种候选特征挥发物的触角电位(EAG)反应和嗅觉趋向选择行为。结果表明,7种锦葵科植物的挥发物种类和含量有较大差异,而且4种棉花品种之间的挥发物差异也很显著。棉花石远321的挥发物总量最大,其中4,8-二甲基-1,3,7-壬三烯(DMNT)含量最高,壬醛和罗勒烯其次。蜀葵的挥发物种类和含量均最少。11种候选特征挥发物均能够引起绿盲蝽雌、雄成虫EAG反应,且雄性成虫的反应值大都高于雌性成虫。其中,绿盲蝽雌、雄成虫触角对反-2-己烯酸丁酯EAG反应值最大,芳樟醇、乙酸叶醇酯、甲基庚烯酮及DMNT也能够激发绿盲蝽较大的EAG反应。在趋向行为检测中发现,DMNT及柠檬烯等对雄性成虫具有吸引作用。对雌性成虫有吸引作用的物质有γ-萜品烯、DMNT、月桂烯、α-石竹烯、罗勒烯及乙酸叶醇酯等,其中γ-萜品烯和乙酸叶醇酯对雌性成虫具有极显著吸引效果。锦葵科候选植物挥发物对绿盲蝽雌、雄虫具有不同EAG和趋向行为反应影响,候选气味化合物可以作为设计绿盲蝽引诱剂组分进行害虫防控田间实践。     

斑翅果蝇对寄主果实及其挥发物嗅觉趋性的研究

斑翅果蝇是一种主要为害软皮水果的害虫, 诱捕剂诱捕是防治该害虫的重要手段。为明确斑翅果蝇对寄主果实及其挥发性物质的趋性, 进而研发植物源引诱剂, 本研究利用四臂嗅觉仪测定了斑翅果蝇成虫对不同种类（葡萄、树莓、杨梅、蓝莓、樱桃和草莓）和不同成熟程度鲜果的趋性, 并分析了寄主果实挥发物成分。结果表明:1)斑翅果蝇成虫对过熟鲜果的趋性最强, 其次为成熟鲜果和未熟的鲜果; 2)成虫对葡萄的趋性最强, 其次分别为树莓、杨梅、蓝莓、樱桃和草莓; 3)葡萄挥发物中含量最高的成分为反式-2-己烯醛, 樱桃和树莓中为苯甲醇, 蓝莓中为乙醇。研究结果为进一步开发高效的斑翅果蝇植物源引诱剂奠定了基础。     

固相微萃取和同时蒸馏萃取法分析臭椿叶挥发性组分的初步研究

本实验采用同时蒸馏萃取法(simultaneous distillation extraction,SDE)和固相微萃取法(solidphase micro-extraction,SPME)提取臭椿(Ailanthus altissima)叶片的挥发性物质,经气相色谱/质谱联用仪分析,共检测出8类42种挥发性化合物,挥发物中烯类最多,22种,其次是醇类6种、烷烃类6种,另外酮类、酯类、芳香烃类、酸类和酚类分别检测出2、2、2、1、1种。采用SPME法提取的挥发性化合物中醇类、酮类和烷烃类的种类多于SDE法,烯类少于SDE法。酚类物质仅在SPME法检出,而芳香烃类仅在SDE法检出。检测到的挥发物组分中有对昆虫起驱避作用的成分。     

白三叶草挥发物的化感作用及其化学成分分析

采用培养皿生物测定法,研究了白三叶草花、叶挥发物对稗草、苘麻种子萌发及幼苗生长的影响,并对其挥发物化学成分进行了分析,以明确白三叶草挥发物对稗草和苘麻的化感作用。结果显示:花挥发物的抑制作用大于叶挥发物,其中,花挥发物处理苘麻萌发率和芽长的化感效应指数分别为-0.27和-0.29,稗草根长和鲜重的化感效应指数分别为-0.30和-0.28。花挥发物中共检测出23种化合物,4-甲基-1,3戊二烯、乙酸叶醇酯相对含量较大;叶挥发物中共检测出24种化合物,乙烯基乙醚、乙酸叶醇酯、丙酮相对含量较大。研究表明,白三叶草挥发物对稗草和苘麻均有化感作用,其挥发物的化学成分以烯和酯类化合物为主。     

棉籽壳中除草活性物质提取及鉴定

为了开发利用棉籽壳中的除草活性物质，分别以无水乙醇、正丁醇、石油醚和乙酸乙酯为溶剂，采用索氏提取法对棉籽壳中的活性物质进行了提取，并对各溶剂粗提物进行了除草活性测定。结果发现:用无水乙醇提取的粗提物对稗草生长的抑制活性最高，经气相色谱-质谱(GC-MS)联用分析，发现该粗提物中主要含有甘油、三环己基3-烯-6-辛酮、4-乙烯基-2-甲氧基苯酚、(邻甲基苯酚)-2-溴-2氯-乙酰酯、十四酸、十四酸乙酯、十六烷酸、辛酸异戊酯和亚油酸9种化合物。进一步的除草活性测定结果表明，亚油酸、辛酸异戊酯、4-乙烯基-2-甲氧基苯酚和(邻甲基苯酚)-2-溴-2氯-乙酰酯4种化合物对稗草表现出一定的除草活性，其中亚油酸活性最强，其IC₅₀值为14.5 mg/L。     

一种缓释型双条杉天牛引诱剂缓释效果的研究

本试验旨在了解缓释型双条杉天牛引诱剂主要挥发性物质在使用过程中的变化,为缓释型双条杉天牛引诱剂在防治森林害虫的应用提供依据.采用SPME-GC-MS测定了自主研制的缓释型和普通型双条杉天牛引诱剂在林间使用0d、10d和30d后的主要挥发性物质的种类和含量,检测结果表明,两种双条杉天牛引诱剂共检测出主要挥发性物质31种,其中对双条杉天牛的引诱具有重要作用的烯类化合物21种.在林间使用0d、10 d和30 d后,缓释型双条杉天牛引诱剂挥发的烯类化合物相对含量分别为88.87％、84.79％和82.53％,0～10d和10～30 d的相对含量下降率分别为4.6％和2.7％,而普通型双条杉天牛引诱剂在林间使用0d、10d和30 d后,挥发的烯类化合物相对含量分别为79.03％、50.74％和49.35％,0～10d和10～30d相对含量下降率分别为35.8％和2.7％.普通型双条杉天牛引诱剂挥发的烯类化合物相对含量在林间使用10～30 d期间明显减少,而缓释型双条杉天牛引诱剂仍保持80％以上的较高相对含量,在实际使用中能够实现长效缓释的目的.     

花椒窄吉丁气味结合蛋白AzanOBP3与其寄主挥发物的分子对接

为明确花椒窄吉丁Agrilus zanthoxylumi气味结合蛋白AzanOBP3与其寄主花椒Zanthoxylum bungeanum主要气味挥发物的结合模式及结合能力,利用Signal P 5.0和ProParam Tool-ExPASy软件对AzanOBP3的基本理化性质进行生物信息学分析,采用Swiss-Model对AzanOBP3序列进行同源建模,通过SAVES 5.0软件中Verify-3D、ERRAT以及ProCheck模块评价模型,应用AutoDock Tool 1.5.6软件对AzanOBP3模型和46种主要寄主挥发物进行分子对接分析。结果显示,花椒窄吉丁气味结合蛋白AzanOBP3同源建模所得模型质量较好,其Verify-3D、ERRAT以及ProCheck得分均符合同源建模要求。AzanOBP3与46种花椒气味挥发物主要以氢键、疏水作用和范德华力相结合,其中, 13种链状配体与AzanOBP3的结合能力总体上弱于33种环状配体,链状配体中的芳樟醇和环状配体中的荜澄茄油烯与AzanOBP3表现出较好的结合能力,自由结合能分别为-5.2 kJ/mol和-6.6 kJ/mol。最终筛选出4种链状配体（乙酸芳樟酯、芳樟醇、乙酸香叶酯和别罗勒烯）和10种环状配体（乙酸松油酯、β-石竹烯、葎草烯、3-蒈烯、荜澄茄油烯、右旋大根香叶烯、β-榄香烯、乙酸叶醇酯、可巴烯和倍半水芹烯）可与AzanOBP3稳定结合。表明花椒窄吉丁气味结合蛋白AzanOBP3能与寄主植物花椒的多种挥发性物质结合,推测AzanOBP3可能在其寻找寄主与取食行为中发挥着重要作用。     

外源褪黑素对番茄果实品质和挥发性物质的影响

为探究外源褪黑素对番茄果实品质及挥发性物质的影响,以番茄（Solanum lycopersicum）‘航粉高糖’品种为试材,以果实表面喷施蒸馏水为对照（CK）,设4个不同浓度褪黑素喷施果实处理,分别为T1（50 μmol·L^-1）、T2（100 μmol·L^-1）、T3（150 μmol·L^-1）、T4（200 μmol·L^-1）。结果表明：（1）100 μmol·L^-1外源褪黑素处理能显著增加番茄果实中果糖、葡萄糖和可溶性总糖的含量,降低有机酸含量,从而显著提高番茄果实糖酸比。（2）5个处理中共检测出86种挥发性物质,包括12种酮类、32种醛类、14种醇类、4种酯类、13种烃类、7种酚类和4种其他类物质。各处理挥发性物质总数量和总含量依次为：CK（55种,1 279.18 μg·kg^-1）、T1（69种,1 960.95 μg·kg^-1）、T2（66种,2 258.70 μg·kg^-1）、T3（62种,1 818.84 μg·kg^-1）、T4（59种,1 695.04 μg·kg^-1）,均为醛类含量最高,酚类含量最低,其中100 μmol·L^-1处理可提高番茄果实中酮类、醛类、醇类和酯类挥发性物质总含量。检测出的挥发性物质共包含10种番茄特征香气成分,100 μmol·L^-1处理的特征挥发性物质含量最高,为1 307.34 μg·kg^-1。（3）与对照相比,喷施褪黑素提高了番茄果实单果质量以及可溶性固形物、可溶性蛋白、维生素C、番茄红素含量,促进果实对N、K、Mg、Cu、Fe、Zn等矿质元素的吸收。（4）主成分分析表明,各处理的综合得分表现为T2>T1>T3>T4>CK。综上,外源喷施褪黑素可显著提高番茄果实的营养和风味品质,且100 μmol·L^-1浓度效果最佳。     

Role of strawberry volatile organic compounds in the development of Botrytis cinerea infection

Botrytis cinerea, the main pathogen of strawberry, has the ability to remain quiescent in unripe tissue and develop disease symptoms in ripe fruit. As strawberry ripening is characterized by an increase of aroma compounds, the role of volatile emission in the development of infection was investigated. Thirty‐five strawberry volatile organic compounds (VOCs) were tested on B. cinerea in vitro and volatile emission was analysed in strawberry harvested at four ripening stages by headspace solid‐phase microextraction/gas chromatography–mass spectrometry and proton transfer reaction–time of flight–mass spectrometry. The coupling of such data sets made it possible to conclude that key strawberry aroma compounds stimulate B. cinerea conidial germination and some typical wound‐volatiles stimulate pathogen conidial germination or mycelial growth. This study is the first report of fungal stimulation by some VOCs naturally occurring in strawberry: the esters ethyl butanoate, cis‐3‐hexenyl acetate, trans‐2‐hexenyl acetate, methyl butanoate and hexyl butanoate, the furanones furaneol and mesifurane, and the alcohol trans‐2‐hexenol. The results of this work provide advances in understanding the functional role of fruit VOCs and suggest, for the first time, that fruit VOCs may influence the development of B. cinerea from the latent phase and that they could favour the invasive growth of B. cinerea after wounding. In particular, ethyl butanoate and furaneol could signal strawberry ripening, and the green leaf volatiles trans‐2‐hexenol, trans‐2‐hexenyl acetate and cis‐3‐hexenyl acetate could signal the presence of damaged tissues that are easier sites for penetration by B. cinerea.     

不同杀菌剂对小麦纹枯病和赤霉病的防治效果

为明确不同杀菌剂对小麦纹枯病、赤霉病的防治效果,在大田条件下以‘开麦21’为试验材料研究了12种杀菌剂对小麦纹枯病、11种杀菌剂对赤霉病的防治效果及对小麦产量的影响。结果表明:30%己唑醇悬浮剂76.5g/hm2、75%肟菌·戊唑醇水分散粒剂168.75g/hm2处理对小麦纹枯病的防效较好,拔节期病指防效(乳熟期枯白穗防效)分别为62.77%(56.21%)和60.58%(53.45%),小麦产量较对照分别增加27.65%和21.30%;48%氰烯·戊唑醇悬浮剂432g/hm2、25%氰烯菌酯悬浮剂750g/hm2处理对小麦赤霉病的防效较好,病指防效(病穗防效)分别为70.78%(56.57%)和68.84%(46.86%),小麦产量较对照分别增加25.44%和24.40%。综合分析,以上药剂可做田间大面积推广应用。     

Early Events During Quiescent Infection Development by Colletotrichum gloeosporioides in Unripe Avocado Fruits

Inoculation of avocado pericarp tissue with Colletotrichum gloeospori-oides and treatment of avocado cell cultures with the cell wall elicitor of C. gloeosporioidesboth increased the production of reactive oxygen species (ROS). However, whereas the production of ROS could be detected within minutes in avocado cell suspensions, it was detected only after 2 h following inoculation of pericarp tissue. Protein kinase inhibitors such as K-252a and staurosporine and the phosphatase inhibitor microcystin-LR inhibited the release of H(2)O(2) from avocado cell suspensions. When 1 mM H(2)O(2) was exogenously applied to pericarp tissue, it enhanced ROS, phenyl-alanine ammonia lyase (PAL) activity, and epicatechin levels. But, when H(2)O(2) treatment was applied following staurosporine treatment, PAL activity was no longer induced. The uninduced ROS production in pericarp tissue of freshly harvested, unripe, resistant fruit was twice as high as in ripe, susceptible fruit. Challenge inoculation of resistant fruit further increased the ROS level; however, this increase did not occur in susceptible fruits. The current findings are consistent with the hypothesis that production of ROS is induced by fungal infection of unripe fruits and, consequently, may modulate resistance, resulting in the inhibition of fungal development and quiescence.     

拮抗内生芽孢杆菌BEB17分离鉴定及其挥发性物质抑菌活性分析

 项目组从健康香蕉植株根部中分离得到一株对多种植物病原真菌具有抑菌活性的内生细菌。结合形态特征观察、生理生化分析及分子鉴定,鉴定该菌株为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),命名为BEB17。双皿对峙实验结果表明,该菌株产生的挥发性有机物(Volatile organic compounds, VOCs)可抑制多种病原真菌生长,其中对香蕉枯萎病菌(Fusarium oxysporum f. sp. cubense race 4,Foc4)抑制效果最强,抑制率达66.86%±0.90%,并能抑制其孢子萌发。用BEB17产生的挥发性物质做熏蒸处理后,Foc4菌丝发生缢缩膨大,细胞膜被破坏。采用顶空法固相微萃取结合气质联用(HS-SPME-GC-MS)对BEB17产生的挥发性有机物(VOCs)进行收集鉴定,得到10类33种单体有机物组分。     

内生解淀粉芽孢杆菌BEB17脂肽类和聚酮类化合物的抑菌活性分析

BEB17是本实验室从健康香蕉植株根部中分离得到的一株具有较强抑制真菌活性的内生解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。在本研究中,我们通过PCR检测多个脂肽类和聚酮类化合物的合成相关基因,并结合基质协助激光解吸附离子化-飞行时间质谱(MALDI-TOF-MS)和液相色谱-质谱联用(LC-MS)分析,发现该菌株能产生表面活性素Surfactin、泛革素Fengycin和伊枯草菌素Iturin三类脂肽类化合物以及Bacillibactin、Difficidin和Bacillaene三类聚酮类化合物。通过比较两种发酵培养基(M9和Landy培养基)和两种提取方法(酸沉淀法和大孔树脂抽提法),结果发现酸沉淀法提取较为高效,Landy培养基更易于BEB17脂肽类和聚酮类化合物的发酵。活性检测表明,两种提取方法提取的粗提物对香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense race 4,Foc4)都具有较好的抑制作用,其中大孔树脂抽提法提取的物质抑制效果最好,当其浓度为300μg·m L-1时抑制率可达80.75%,说明该方法制备的粗提物的活性物质含量相对较高。进一步研究表明,该提取物能引起Foc4菌丝膨大和畸形,并抑制其孢子萌发。分析表明,该提取物对Foc4细胞膜具有破坏作用。     

Antifungal activity, acid and sugar content in the wood apple (Limonia acidissima) and their relation to fungal development

Unripe wood apple fruit is generally free from visible fungal growth before and at harvest but a succession of fungi appears on the fruit shell, and sometimes in the pulp, during ripening. A TLC- Cladosporium bioassay of the chloroform extract taken from unripe fruit shell demonstrated three inhibition areas. Similar extracts from stem-bark and root-bark produced these three, and one additional, inhibition areas. The four compounds responsible for inhibition were identified as psoralene, xanthotoxin, 2,6-dimethoxybenzoquinone and osthenol.
Concentrations of the three antifungal compounds on unripe fruit shell increased during the first 4 days after harvest and then declined. They remained much below those required to inhibit the development of three fungi on TLC plates. Titratable acidity of the unripe fruit pulp was high but decreased by about 50% during ripening. Levels of reducing sugars were very low in the unripe fruit pulp but increased by about five times during ripening. Levels also increased in the fruit shell and its washings. The possible role of these factors in restricting fungal growth in unripe fruits is discussed.     

Cacao resistance to Phytophthora: Effect of pathogen species, inoculation depths and pod maturity

Two species of Phytophthora (P. palmivora and P. capsici) and inoculations at two depths (3 mm and 9 mm) were tested each on 10 clones of Theobroma cacao to determine their effects on pod resistance. Ripe and unripe pods were also assessed to determine the influence of physiological status of the pod on the expression of resistance. The two pathogens tested (P. palmivora and P. capsici) differed significantly in their reactions on pods, with P. palmivora being more aggressive than P. capsici. However, the lack of interaction between clones and pathogen species and the similarity in the ranking of clones based on lesion size suggested that selection for resistant clones can be based on one of the two pathogens, preferably the more aggressive one. Pod reactions differed between inoculation depths (3 mm and 9 mm), and between pod maturity stages (ripe and unripe pods) with relatively larger lesions being recorded at 9 mm depth and on unripe pods as compared to those observed at 3 mm depth and on unripe pods, respectively. The magnitude of increase in lesion sizes, however, varied with genotypes, indicating that inoculation depth and pod maturity stage should be standardized in screening cacao germplasm for resistance to Phytophthora.     

Tracing Latent Infection of Colletotrichum acutatum on Strawberry by PCR

Colletotrichum acutatum, a quarantine organism on strawberries in the EU, was found in Finland for the first time in 2000. Concern about rapid, unnoticeable spread of this pathogen has necessitated studies to find methods with which the quiescent fungus infection can be detected in imported, cold-stored strawberry plant material. Successful detection of C. acutatum in strawberry tissues by polymerase chain reaction (PCR) is dependent on the method of DNA extraction used. Good-quality nucleic acid, free of PCR inhibitors, was successfully prepared by slightly modifying the DNA extraction method of a commercially available kit. Species-specific primers, previously described in the literature, were successfully used in the PCR reaction. C. acutatum was detected by PCR both on symptomatic and asymptomatic plant parts and in artificially and naturally infected strawberry tissues. Positive PCR results were obtained from ripe and unripe berries, runners, petioles and different parts of crowns. The data demonstrate that the PCR technique can be used to detect C. acutatum in strawberry tissue even in plant parts that do not show visible symptoms.