小花假泽兰精油超临界CO_2提取工艺及其成分和抑菌活性研究

采用正交试验法研究超临界CO_2萃取法提取小花假泽兰精油的提取条件,采用GC-MS法分析测定精油的化学成分,并初步测定精油的抑菌效果.结果表明:超临界CO_2萃取小花假泽兰精油的较佳工艺条件为:采用先静态后动态的萃取方式,静态萃取时间20 min,动态萃取条件为萃取温度55℃,萃取压力35 Mpa,CO_2体积为40 mL/g,CO_2 流速为0.5～1.0 mL/min;从超临界CO_2萃取的小花假泽兰精油中鉴定了64种成分,占精油总量的79.80%,主要有萜类、醇类、脂肪酸、酯类、甾体类等;生物测定结果表明,超临界CO_2萃取物对小麦赤霉病菌和小麦纹枯病菌菌丝生长的EC_(50)分别为119.55 mg/L和78.27 mg/L. 果.结果表明:超临界CO_2萃取小花假泽兰精油的较佳工艺条件为:采用先静态后动态的萃取方式,静态萃取时间20 min,动态萃取条件为萃取温度55℃,萃取压力35 MPa,CO_2体积为40 mL/g,CO_2 流速为0.5～1.0 mL/min;从超临界CO2萃取的小花假泽兰精油中鉴定了64种成分,占精油总量的79.80%,主要有萜类、醇类、脂肪酸、酯类、甾体类等;生物测定结果表 ,超临界CO_2萃取物对     

小花假泽兰茎叶中杀菌活性成分的研究

以小麦纹枯病原菌为指示菌种,采用活性跟踪法,从小花假泽兰(M ikan ia m icrantha H.B.K.)茎叶中分离出1种对黄瓜霜霉、黄瓜炭疽、小麦赤霉和玉米大斑4种病原菌孢子萌发具有较好抑制作用的物质,在供试浓度为0.50 m g.mL-1时,对黄瓜炭疽、小麦赤霉、玉米大斑3种病原菌孢子萌发抑制率均在70%以上.生长速率法测定其对番茄灰霉病原菌的EC50为0.039 m g.mL-1,组织法测定结果表明该化合物对番茄灰霉病有较好的防效,在供试浓度为2 000 m g.k-g 1时,治疗作用和保护作用防效分别达到了68.6%和48.7%,高于或相当于对照药剂速克灵的防效.经熔点测定、IR、M S1、HNM R1、3CNM R分析,鉴定该化合物为豆甾醇,其为首次从小花假泽兰茎叶中分离得到.     

孜然种子提取物枯茗醛和枯茗酸抑菌作用研究

以番茄早疫病菌、棉花黄萎病菌、小麦纹枯病菌、烟草赤星病菌、小麦全蚀病菌、油菜菌核病菌、辣椒疫霉病菌、水稻纹枯病菌、小麦白粉病菌和番茄灰霉病菌等为供试菌种,采用离体与活体相结合的方法系统地测定了枯茗醛和枯茗酸的抑菌活性。离体抑菌活性测定结果表明,枯茗醛和枯茗酸对多种病原菌具有一定的抑制效果,其中对油菜菌核病菌菌丝生长的抑制效果高于其它供试病原菌,有效中浓(EC50)分别为2.1和7.3 mg/L;枯茗醛和枯茗酸对小麦白粉病的防治实验结果表明,供试浓度为1000 mg/L时,两种药剂的保护防效均高于50%;相同处理浓度下,枯茗酸对油菜菌核病的保护防效与速克灵处理相当,达到57.52%。     

20种植物提取物抑制植物病原菌活性研究

以小麦纹枯病菌(Rhizoctonia cerealis)、番茄灰霉病菌(Alternaria solani)这两种植物病原真菌为供试菌,对采自江西省吉安市的20种植物提取物的抑制菌丝生长活性及孢子萌发进行测定.结果表明,在供试质量浓度为1 mg/mL时,黄花草木樨乙醇提取物的乙酸乙酯萃取部分抑菌效果最好,对小麦纹枯病菌和番茄灰霉病菌两种菌的抑制率均达100%;窃衣、小白酒草、羊蹄和车前草的抑制作用次之,对两种菌的抑制率均大于80%;单从对一种病原菌的抑制作用看,还有黄荆对小麦纹枯病菌率为100%,空心莲子草、窃衣、小白酒草和鬼针草对番茄灰霉病菌的抑制率为100%,但是,这20种供试植物的石油醚相和水相萃取物对两种病原真菌的抑制效果均不强.抑制病原菌孢子萌发亦得到类似结果.以上结果提示植物抑菌活性成分主要存在于乙酸乙酯萃取部分,黄花草木樨、窃衣、小白酒草等的提取物作为植物源杀菌剂值得进一步研究.     

大花金挖耳内生真菌H-18菌株的鉴定及其抑菌活性研究

对大花金挖耳内生真菌H-18菌株的菌丝提取物进行了抑菌活性研究并对菌株进行了鉴定.离体试验结果表明,H-18菌株菌丝体丙酮提取物在500mg/L浓度下对玉米大斑病菌、番茄灰霉病菌和辣椒疫霉病菌的菌丝生长抑制率均大于80%;组织法和盆栽试验结果表明,H-18菌株菌丝体丙酮提取物在2000mg/L浓度下对番茄灰霉病和辣椒疫霉病的保护效果和治疗效果均在50%以上.结合形态分类学和ITS-5.8S rDNA序列分析结果鉴定菌株H-18为同色镰孢菌(Fusarium concolor).     

灰色链霉菌YD3309菌丝体抗菌活性成分研究

采用生物活性跟踪法,从一株灰色链霉菌菌丝体中分离得到2个具有抑菌活性的化合物,通过核磁共振波谱和质谱等技术鉴定2个化合物的结构为新刺孢霉素A(1)和N-乙酰基色氨醇(2);化合物1首次从放线菌中分离得到。抑菌活性测定结果表明:化合物1和2对番茄灰霉病菌(Botrytis cinerea)、茄子黄萎病菌(Verticillium dahliae)和辣椒枯萎病菌(Fusarium oxysporum)等多种蔬菜病原真菌有抑制作用,其中化合物1对番茄灰霉病菌和番茄早疫病菌菌丝具有强烈的抑制作用,化合物2对茄子黄萎病菌的菌丝具有强烈的抑制作用;化合物1对番茄灰霉病菌和番茄早疫病菌菌丝生长的半抑菌浓度(IC_(50))分别为30.6和28.8 mg/L,化合物2对茄子黄萎病菌菌丝生长的半抑菌浓度(IC50)为34.3 mg/L。     

大花金挖耳花蕾中精油的化学组成及其杀菌活性研究

采用GC-MS联用技术检测了阴干的大花金挖耳花蕾中精油的化学组成及相对含量,并测定了该精油对14种常见作物病原菌的抗菌活性.结果表明,大花金挖耳花蕾精油得率为2.9%,从中鉴定出53种成分,占色谱峰总面积的86.01%,主要化学成分为（E,E）-3,7,11,15-四甲基-1,6,10,14-十六烷四烯（15.33%）、甲苯（11.32%）、（3α,5α）-3,5,14,19-四羟基强心甾-20（22）-烯（6.00%）、2-乙氧基四氢呋喃（5.44%）、亚油酸（3.51%）等.杀菌实验结果表明,该精油对小麦全蚀病菌（Gaeumannomyces gramims）和小麦纹枯病菌（Rhizoctonia cerealis）抑制作用最强,在0.5 mg/mL浓度下,5 d后对这2种病菌菌丝生长的抑制率均大于97%;对小麦赤霉病菌（Fusarium graminearum）和番茄叶霉病菌（Fulvia fulva）等也有较强的抑制效果.大花金挖耳花蕾精油的化学组成及其杀菌活性研究均为首次报道.     

大花金挖耳内生菌的分离及抑菌活性筛选

从菊科植物大花金挖耳（Carpesium macrocephalum）的花、茎、叶、果实和根中分离出92株内生菌,其中真菌68株,细菌16株,放线菌8株;对峙法生物活性测定结果表明,其中22株内生菌分别对小麦赤霉病菌、黄瓜灰霉病菌、辣椒疫霉病菌、玉米大斑病菌和苹果炭疽病菌均有较好的拮抗作用;采用生长速率法测定22株内生菌发酵产物的抑菌活性,结果表明,内生细菌H-32的发酵液稀释10倍和内生真菌H-18、Y-12、G-4、G-6的菌丝丙酮提取物在0.5 mg·mL^-1（菌丝干重）的剂量下,均对番茄灰霉病菌和玉米大斑病菌菌丝生长具75%以上的抑制效果;活体组织法测定结果,H-32的发酵液和H-18、J-1、Y-17、G-6的菌丝丙酮提取物在2.0 mg·mL^-1（菌丝干重）的剂量下,对番茄灰霉病的防治效果均在50%以上.综合分析认为,H-32、H-18和G-6等内生菌的抑菌作用值得进一步研究.     

蜡质芽胞杆菌XF的发酵条件优化及对番茄灰霉病的防治效果

由灰葡萄孢菌(Botrytis cinerea)引起的番茄灰霉病是一种土传病害,近年来施用生防菌被认为是一种有效的防治手段。以灰葡萄孢菌为指示菌,生物量和抑菌率为指标,采用单因素试验、正交优化试验相结合的方法,优化蜡质芽胞杆菌XF株的发酵培养基和发酵条件,从而提高菌株XF对番茄灰霉病菌的抑制效果,并于室内盆栽条件下探讨其对番茄灰霉病的防治效果,为今后该菌剂生产工艺条件的进一步确定及其防治应用的进一步开展奠定基础。结果表明,蜡质芽胞杆菌XF株发酵最适培养基(质量分数):葡萄糖1.00%,蛋白胨1.00%,酵母粉0.50%,NaCl 1.00%,MgSO_4·7H_2O 0.10%,色氨酸100 mg/L;最佳发酵条件:初始pH 6.0,温度30℃,发酵周期60 h,摇床转速200r/min,接种量5%。菌株XF稀释10倍发酵液对番茄灰霉病菌的抑制率为98.19%,对番茄离体叶片、盆栽的防效分别可达66.58%、83.97%。菌株XF具有很强的生防潜力。     

植物精油对草菇双额岩小粪蝇的产卵驱避效果及毒力测定

在室内采用选择性试验测定了14种植物精油单组分化合物对草菇双额岩小粪蝇Bifronsina bifrons(Stenhammar)的产卵驱避效果及在室外采用非选择试验测定了其中3种化合物的产卵驱避效果及毒力测定。结果表明:各精油浓度为10 000 mg/L时,α-松油醇、丁香酚和茴香醛表现出明显的产卵驱避效果,其选择性产卵驱避率分别是60.17%±9.57%、51.07%±2.63%和47.66%±7.10%,产卵量均显著低于对照;月桂烯、α-蒎烯、β-蒎烯和异松油烯产卵驱避作用效果不明显。α-松油醇、丁香酚和茴香醛对双额岩小粪蝇幼虫均具有一定的毒性,对幼虫的校正死亡率分别是5.11%±3.54%、42.68%±3.94%和14.47%±6.92%;丁香酚对双额岩小粪蝇幼虫的毒力最大,LC_(50)为16 965.159 mg/L。其次是茴香醛,LC_(50)为311 941.332 mg/L。α-松油醇毒力最差,LC_(50)为40 676 080.550 mg/L。进一步非选择性试验结果表明α-松油醇(1 250 mg/L～20 000 mg/L)、丁香酚(625 mg/L～20 000 mg/L)和茴香醛(2 500 mg/L～20 000 mg/L)对双额岩小粪蝇具有明显的产卵驱避效果,驱避效果随施用浓度的增加而显著增加。在浓度为20 000 mg/L时α-松油醇、丁香酚和茴香醛的非选择性产卵驱避率分别为69.75%±3.98%、60.75%±1.94%和57.23%±3.38%。本研究为草菇生产中双额岩小粪蝇的无公害防治及高效环保的植物源驱避剂的开发提供参考。     

Chemical composition and antifungal activity of essential oil and various extract of Silene armeria L

The aims of this study were to examine the chemical composition of the essential oil isolated from the floral parts of Silene armeria L. by hydrodistillation, and to test the efficacy of essential oil and various leaf extracts (n-hexane, chloroform, ethyl acetate and methanol) as an antifungal potential. The GC-MS analysis determined that 28 compounds, which represented 89.03% of total oil, were present in the oil containing mainly 1-butene, methylcyclopropane, 2-butene and caryophyllene oxide. The oil (1000ppm/disc) and the leaf extracts (1500ppm/disc) revealed remarkable antifungal effect against Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Colletotrichum capsici, Sclerotinia sclerotiorum, Botrytis cinerea and Rhizoctonia solani, in the growth inhibition range of 39.6-67.6% and 9.3-61.3%, respectively, along with their respective MIC values ranging from 62.5 to 1000mug/ml and 125 to 2000mug/ml. The essential oil had also a strong detrimental effect on spore germination of all the tested plant pathogens along with concentration as well as time-dependent kinetic inhibition of B. cinerea. Thus, the results obtained in this study demonstrate that S. armeria essential oil and various organic extracts possess a wide range spectrum of fungicidal activity and could become an alternative to synthetic fungicides for controlling certain important plant fungal diseases.     

Fumigant activity of essential oils and components of Illicium verum and Schizonepeta tenuifolia against Botrytis cinerea and Colletotrichum gloeosporioides

To develop a natural fungicide against Botrytis cinerea and Colletotrichum gloeosporioides, a total of 25 essential oils were tested for their fumigant activity against post-harvest pathogens. The vaporous phases of oils were treated to each fungus on potato dextrose agar medium in half-plate separated Petri plates at 10 microg per plate. The essential oil of Illicium verum strongly inhibited the mycelial growth of both B. cinerea and C. gloeosporioides by over 90%. On the other hand, the essential oil of Schizonepeta tenuifolia showed inhibitory activity against mycelial growth of only B. cinerea by over 90%. Gas chromatography-mass spectrometry and bioassay indicated trans-anethole in I. verum and menthone in S. tenuifolia as a major antifungal constituent. The essential oils of I. verum and S. tenuifolia and their major constituents could be used to manage post-harvest diseases caused by B. cinerea and C. gloeosporioides.     

Chemical characterization and antifungal activity of essential oil of capitula from wild Indian Tagetes patula L.

Summary. The essential oil extracted by steam distillation from the capitula of Indian Tagetes patula, Asteraceae, was evaluated for its antifungal properties and analyzed by gas chromatography and gas chromatography–mass spectrometry. Thirty compounds were identified, representing 89.1% of the total detected. The main components were piperitone (24.74%), piperitenone (22.93%), terpinolene (7.8%), dihydro tagetone (4.91%), cis-tagetone (4.62%), limonene (4.52%), and allo-ocimene (3.66%). The oil exerted a good antifungal activity against two phytopathogenic fungi, Botrytis cinerea and Penicillium digitatum, providing complete growth inhibition at 10 μl/ml and 1.25 μl/ml, respectively. The contribution of the two main compounds, piperitone and piperitenone, to the antifungal efficacy was also evaluated and ultrastructural modifications in mycelia were observed via electron microscopy, evidencing large alterations in hyphal morphology and a multisite mechanism of action. Correspondence and reprints: Dipartimento delle Risorse Naturali e Culturali, Università di Ferrara, Corso Porta di Mare 2, 44100 Ferrara, Italy.     

番茄内生菌St24的鉴定及其对灰霉病的生防作用

从番茄植株根茎部分离到1株有抑菌活性的植物内生放线菌菌株St24,对其分类地位以及对灰霉病菌的防治效果进行研究.结果表明: 菌株St24为酒红链霉菌.St24发酵液经石油醚萃取得到的粗提物对多种病原菌有抑制作用,其中对灰霉病菌的抑制作用最强,抑制菌丝生长的EC₅₀为11.78 mg·L^-1.粗提物处理灰霉菌后,菌丝量减少,菌丝体皱缩、断裂、原生质外渗,菌体细胞表面有许多瘤状畸形.处理后的灰霉病菌培养液在260 nm处比对照多出现一吸收峰,说明粗提物对病原菌的细胞膜透性有影响.经盆栽试验, St24发酵液对番茄灰霉病有保护和治疗的作用,100 mg·L^-1粗提物叶面喷雾的保护作用效果最好,24 h后防效达到94.3%,120 h后为85.4%.     

11种植物精油对6种植物病原真菌的抑菌活性研究

为筛选有效的植物杀菌成分,采用菌丝生长法,测定了香茅油、薰衣草油、菊花油、月桂油、柠檬油、广藿香油、肉桂油、天竺葵油、迷迭香油、茶树油、薄荷油对6种植物病原真菌的抑菌活性。发现在2 g/L的浓度下,上述11种精油对6种供试病菌均有明显的抑制作用,其中香茅油、肉桂油、天竺葵油、月桂油、茶树油和薄荷油对6种病原真菌的抑制率均为100%。剂量效应试验表明,肉桂油对灰葡萄孢（Botrytis cinerea）和禾谷镰孢菌（Fusariumgraminearum）的EC50值分别为29.05μg/mL和42.96μg/mL,而天竺葵油对灰葡萄孢（Botrytis cinerea）和禾谷镰孢菌（Fusarium graminearum）的EC50值分别为34.02μg/mL和68.48μg/mL。     

酶法制备壳寡糖对番茄灰霉病病菌抑制机理初探

目的：研究酶降解法制备的壳寡糖对番茄灰霉病病菌的抑制机理。方法：测定壳寡糖对番茄灰霉病菌菌丝发育、孢子萌发的抑制作用以及对菌丝形态、细胞膜透性和对菌丝体可溶性蛋白质含量的影响。结果：当壳寡糖处理浓度为1.5mg/mL以上时,对菌丝生长的抑制率达70%以上,EC50为0.72mg/mL。当处理浓度为6mg/mL时,对分生孢子的抑制率显著优于其他浓度水平,达87.4%,EC50为1.48mg/mL。显微观察,浓度为0.8mg/mL的壳寡糖处理可诱导菌丝分枝增多、菌体分隔增加,分生孢子形成受到抑制。此外壳寡糖能够引起菌丝细胞膜透性发生变化,造成菌丝体内含物的渗漏。壳寡糖处理66 h后,菌丝体可溶性蛋白含量比对照降低了36.7%。结论：壳寡糖对番茄灰霉病病菌抑制机理的探究为研究壳寡糖这一新型生物农药的作用机制提供依据。     

Study of antifungal activities of seven essential oils from some Iranian medicinal plants against various postharvest phytopathogenic fungi

The aim of this study was to find the antifungal activities of seven essential oils from some Iranian medicinal plants that have maximum (100%) inhibition effect on the mycelium growth of postharvest phytopathogenic fungi. Among 20 examined species belonging to three families, only 7 species could stop the mycelium growth of phytopathogenic fungi. The selected plants include Trachyspermum ammi, Zataria multiflora Boiss., Satureia hortensis, Caryophillum aromaticus, Menthe piperita, Cuminum cyminum L. and Carum carvi, and fungi include Aspergillus flavus, Botrytis cinerea, Penicillium italicum, Penicillium expansum, Penicillium commune, Rhizopus stolonifer and Rhizopus lyococcus. The results showed that the essential oil of these plants could stop the mycelium growth at 500 ppm, but could not completely inhibit the spore germination, however reduced the spore germination to 80–90%. Among the fungi Rhizopus stolonifer and Rhizopus lyococcus are more resistant to the inhibition effects of essential oils. Among the plants, Cuminum cyminum L. and Carum carvi were slightly weaker than other plants. Also except for Cuminum cyminum L. and Carum carvi, the essential oils of other plants had fungicide effect while these two plants in most cases had fugistatic effect. The results showed that these essential oils can be used as an effective alternative control method.     

黄花草木樨提取物抑菌活性初探

采用离体和活体试验方法分别测定了黄花草木樨不同溶剂提取物对12种植物病原真菌的抑菌活性.结果表明:各溶剂提取物对12种植物病原真菌均具有不同程度的抑菌活性,其中以乙酸乙酯提取物的抑菌活性最高,对油菜菌核病菌、玉米大斑病菌和白菜黑斑病菌抑制菌丝生长的EC50分别为0.62、0.83、0.64g/L,对稻瘟病菌和玉米大斑病菌抑制孢子萌发的EC50分别为0.67、0.97g/L.离体组织法测定表明其乙酸乙酯提取物对番茄灰霉病菌具有较高的保护和治疗作用,在浓度为5.0g/L时,防治效果分别为75.41%和59.18%(6d).活体试验表明乙酸乙酯提取物对小麦白粉病和小麦条锈病也有一定的保护作用,在浓度为10.0g/L时,防治效果分别为73.39%和63.27%.