<i>Bacillus subtilis Strains</i>with Antifungal Activity against the Phytopathogenic Fungi

Bacillus strains isolated from the rhizosphere soil of potato roots were evaluated for the potential antagonistic activity against fungal pathogens in vitro and in vivo. Two bacterial isolates were identified as new Bacillus subtilis strains by 16S rRNA and GyrB gene sequencing and were designated GM2 and GM5, respectively. Strains were characterized by their ability to inhibit growth of a number of phytopathogenic fungi. It was shown that GM5 strain inhibited growth of phytopathogenic fungi more effectively than GM2 strain. Both strains were capable of producing a number of hydrolytic enzymes as well as antimicrobial metabolites (ammonia and HCN). In addition, GM2 strain also produced siderophores. Four genes encoding antimicrobial peptides were identified in the genome of GM2 strain: ituC, bmyB, bacA and srfA. Genome of GM5 contained two genes encoding for antimicrobial peptides, srfA and fenD. Purified lipopeptide fraction from GM5 but not from GM2 strain was able to control Fusarium solani spread in the plate assay. Furthermore, Bacillus subtilis strain GM2 promoted growth of wheat but only GM5 strain was able to protect wheat seedlings from Fusarium oxysporum infection.

PCR-Mediated Detection of Endophytic and Phytopathogenic Fungi from Needles of the Japanese Black Pine, <i>Pinus thunbergii</i>

A specific and sensitive polymerase chain reaction (PCR) assay based on the internal transcribed spacer (ITS) region of rDNA sequences was developed to detect endophytic and phytopathogenic fungi from needles of the Japanese black pine, Pinus thunbergii. Sequences of the ITS regions of Lophodermium conigenum, Lecanosticta acicola, Pestalotiopsis neglecta, Rhizosphaera kalkhoffii, and Septorioides pini-thunbergii were compared, and each specific primer pair for these species was designed. First, the designed primer pairs were tested for their specificity to detect each species. A PCR product was amplified only each combination of species and its specific primer pair, confirming the specificity of the designed primer pairs. These primer pairs were also tested on DNA extracted from the needles of P. thunbergii. The PCR products were amplified not only in needles with lesions but also in healthy needles without symptoms. Furthermore, several endophytic and phytopathogenic fungi could be simultaneously detected from the same region in a needle. The PCR-mediated detection method developed in this study will be a valuable tool for the detection of the endophytic and phytopathogenic fungi, not only as a rapid diagnostic tool for early detection but also for monitoring variations in both the quality and quantity of the endophytic and phytopathogenic fungi in needles in Japanese black pines.

Isolation and Screening of Silicate Bacteria from Various Habitats for Biological Control of Phytopathogenic Fungi

Silicate solubilizing bacteria (SSB) can play an efficient role in soil by solubilizing insoluble forms of silicates. In addition to this some SSB can also solubilize potassium and phosphates, hence increasing soil fertility and enhancing plant defense mechanisms. A total of 111 bacterial strains were isolated from various habitats of Pakistan and screened for solubilization of silicate, phosphate and potassium on respective media. Out of these, 35 bacterial isolates were capable of solubilizing either silicate, phosphate or potassium. Amongst these 7 bacterial isolates were capable of solubilizing all three minerals tested. The highest silicate (zone diameter 54 mm) and phosphate solubilization (zone diameter 55 mm) was observed for bacterial isolate NR-2 while the highest potassium solubilization was observed for NE-4b (zone diameter 11 mm). Dual culture antagonistic assays were carried out by using these bacterial isolates against four plant pathogenic fungi Magnaporthae grisae, Rhizoctonia solani, Altarnaria alternata and Macrophomina pheasolina. Mean zone of inhibition of these bacterial isolates against the four pathogenic fungi ranged between 4 mm to 39 mm. The largest zone of inhibition against all four bacterial strains was recorded for bacterial isolate NR-2 followed by NE-4b. These strains will be further investigated for their plant growth promoting activities in the future.

Antiviral Effect of Ribonuclease from <i>Bacillus pumilus</i>against Phytopathogenic Rna-Viruses

Background: Viruses can cause different diseases in plants. To prevent viral infections, plants are treated with chemical compounds and antiviral agents. Chemical antiviral agents usually have narrow specificity, which limits their wide application. Alternative antiviral strategy is associated with the use of microbial enzymes, which are less toxic and are readily decomposed without accumulation of harmful substances. The aim of this work is to study the effect of Bacillus pumilus ribonuclease on various phytopathogenic viruses with specific focus on the ability of enzyme to eliminate them from plant explants in vitro. Materials and methods: Extracellular ribonuclease of B. pumilus is tested as an antiviral agent. To study the antiviral effect of RNase, depending on concentration and the time of application several plant-virus model systems are used. Virus detection is conducted by serological testing and RT-PCR. Results: Bacillus pumilus ribonuclease possesses antiviral activity against plant Rna-viruses RCMV (red clover mottle virus), PVX (Potato Virus X) and AMV (Alfalfa Mosaic Virus). The maximum inhibitory effect against actively replicating viruses is observed when plants are treated with the enzyme in the concentration of 100 ug/ml prior to infection. In case of local necrosis ribonuclease in the concentration of 1 ug/ml completely inhibits the development of RCMV virus on bean plants. The enzyme is able to penetrate plants and inhibit the development of viral infection, inhibiting effect for untreated surfaces decreased on average for 20%. It is also found that B. pumilus ribonuclease protects apical explants of sprouts of potato tubers from PVM and PVS viruses. Conclusion: B. pumilus ribonuclease possesses antiviral activity against plant Rna-viruses and produces viruses-free plants in the apical meristem culture.

Antifungal Activity of Oleoresin and Fractions of Pinus elliottii Engelm and Pinus tropicalis against Phytopathogens

Microorganism resistance to the existing products is yet another difficulty that agriculturalists have to deal with. In this context, the search for new agricultural products that can fight phytopathogens has become increasingly important. Plants have played an important role in this process, because they can serve as a source of new compounds for drug discovery. Plants belonging to the genus Pinus produce an oleoresin that protects the plant against herbivores and pathogens. With a view to developing products that can combat fungal pathogens without harming the environment, this work aimed to evaluate the antifungal activity of the oleoresins and fractions of Pinus elliottii Engelm and Pinus tropicalis against phytopathogens. The methodology based on NCCLS M38-A standards aided antifungal activity assessment. The microdilution method helped to determine the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC). The oleoresins of P. elliottii and P. tropicalis afforded the most significant results—they displayed fungicidal activity against all the tested species. MIC values were promising, especially the MIC of the oleoresin of P. elliottii against S. rolfsii (1.95 μg·mL-1). The MIC values of the oleoresins of P. elliottii and P. tropicalis ranged from 1.95 to 1000 μg·mL-1 and from 31.25 to 250 μg·mL-1, respectively. Fraction PT2 of P. tropicalis furnished the best results among all the assayed fractions: MIC values lay between 125 and 500 μg·mL-11. In conclusion, the oleoresin of P. tropicalis is a promising source of new antifungal agents for application in the treatment of phytopathogenic infections.

Discovery of cryptolepine derivatives as novel promising agents against phytopathogenic bacteria

To ensure the production of food crops,a series of cryptolepine derivatives were synthesised,after which their antibacterial activities and mechanism of action against three plant pathogens were investigated.Our bioassay results indicated that most of the target compounds displayed potent inhibitory effects against Xanthomonas oryzae(X.oryzae)and Xanthomonas axonopodis pv.citri(X.axonopodis pv.c.).Remarkably,compound 9 exhibited the best in vitro antibacterial activity against X.oryzae,with a minimum inhibitory concentration(MIC)value of 0.78μg·mL^(-1).Compound 2 exhibited the best in vitro antibacterial activity against X.axonopodis pv.c.,with an MIC value of 0.39μg·mL^(-1).These activities were superior to those of copper quinolate(MIC=6.25,25μg·mL^(-1))and thiodiazole copper(MIC=100,200μg·mL^(-1))against X.oryzae and X.axonopodis pv.c.In vivo experiments demonstrated the promising applicability of compound 9 for the control of rice bacterial infections.Furthermore,compound 9 was selected as a candidate to conduct preliminary analyses of the antibacterial mechanisms of cryptolepine derivatives.Scanning electron microscopy and transmission electron microscopy observations,extracellular polysaccharide production,biofilm formation,transcriptomic,quantitative reverse transcription-polymerase chain reaction analyses,and molecular docking assays were performed.Collectively,our findings demonstrated that compound 9 might act via multifarious mechanisms to down-regulate virulence factors and cause cell death.

石榕树中补骨脂素分离鉴定及其抗真菌活性

[目的]分离出石榕树中能够有效抑制植物病原真菌的活性单体化合物并探索其抗真菌活性。[方法]采用生物活性追踪和化学分离相结合的方法对石榕树甲醇提取物进行分离,运用质谱、核磁共振氢谱和碳谱并结合相关文献数据鉴定活性单体化合物。采用菌丝生长速率法测定活性单体化合物对7种植物病原真菌的抑菌活性。[结果]石榕树(叶)甲醇提取物对甘蔗凤梨病菌具有良好抑菌活性,在10 mg·mL^(-1)浓度下,抑菌率为100%。在浓度为1.5 mg·mL^(-1)时,石榕树(叶)石油醚、乙酸乙酯、正丁醇萃取物和水层萃余物对甘蔗凤梨病菌的抑菌率分别为58.73%、100.00%、39.49%和6.63%。表明石榕树(叶)乙酸乙酯层萃取物对甘蔗凤梨病菌的抑制作用最高,其EC_(50)值为0.430 7 mg·mL^(-1)。石榕树(叶)乙酸乙酯层萃取物进行柱色谱分离后得到对甘蔗凤梨病菌有较高抑菌活性的是LMNO1bc1a活性组分,0.1 mg·mL^(-1)的LMNO1bc1a活性组分对甘蔗凤梨病菌的抑菌率为100.00%。LMNO1bc1a组分经结晶、重结晶纯化后命名为SRS-1,SRS-1经NMR和MS技术鉴定为补骨脂素(psoralen)。SRS-1对甘蔗凤梨病菌、甘蓝黑斑病菌、玉米大斑病菌、罗汉果白绢病菌、柑橘砂皮病菌、茶轮斑病菌、烟草黑胫病菌这7种植物病原真菌都有一定的抑菌活性,其EC_(50)值分别为0.018 2、0.038 8、0.040 9、0.051 2、0.080 7、0.092 7、0.115 4 mg·mL^(-1)。[结论]从石榕树中分离出抑菌活性单体化合物SRS-1,鉴定为补骨脂素,其对甘蔗凤梨病菌、甘蓝黑斑病菌、玉米大斑病菌、罗汉果白绢病菌、柑橘砂皮病菌、茶轮斑病菌、烟草黑胫病菌这7种植物病原真菌都有一定的抑菌活性。

Release of active oxygen from phytopathogenic bacteria and their regulation

Some phytopathogenic bacteria including Xanthomonas oryzae pv. oryzae （a rice leaf bacterial blight pathogen） and its virulent gene mutant XooM3105,Xanthomonas campestris pv. campestris, Erwinia carotovora subsp, carotovora and so on, could release active oxygen such as O2 and H2O2 by themselves.Ca2+, NADPH and O2 concentration regulated the response. The response is sensitive to heat and proteinase. These results show that the component releasing active oxygen in the bacterial plant pathogen may involve special protein and enzyme and be secreted to culture liquid. Various components of rice tissue can significantly induce the release of active oxygen in Xanthomonas oryzae pv. oryzae.

以粪污水热资源化为核心的高效种养结合系统分析与展望

种养结合系统是实现养殖业和种植业可持续发展的重要途径。水热技术可以将畜禽粪污快速直接转化生产生物原油、水热炭、水相产物和气体产物,生成物分别具有燃料、土壤改良、农用杀菌剂和气体肥料应用潜力,以粪污水热资源化为核心构建的高效种养结合系统有助于提升畜禽粪污的无害化资源化水平和消纳能力、构建生态可持续的绿色循环农业,实现畜禽粪污资源化利用、种植业和养殖的协同高效发展。该研究基于国内外近十余年的研究,系统综述了粪污水热资源化产物在种养结合生态循环农业模式中的作用和应用潜力,论述了畜禽粪污水热转化生物原油在农业内燃机中作为燃料的研究现状,阐述了水相产物作为潜在的农用杀菌剂在作物病害防控中的作用,梳理了畜禽粪污源水热炭在转化机理、理化特性和还田应用中的研究进展,分析了气相产物中的组分和用于温室种植的潜力。在此基础上总结了以粪污水热资源化为核心的高效种养结合系统面临的主要挑战,并对水热资源化产物农业循环应用的研究方向进行展望。研究为畜禽粪污水热资源化和高效种养结合系统研究提供理论参考。

菌毛在生防细菌定殖植物病原线虫中功能的研究进展

植物病原线虫对林业生产危害严重,缺乏有效的防控措施,已造成巨大的经济损失。生防细菌已被用于林木病原线虫的绿色防控,并已取得良好的防治效果。然而,生防细菌防控林木线虫病害的作用机制尚不明确,严重阻碍了防线虫微生物制剂的研发与高效应用。目前,已有大量的研究结果证实,生防细菌在线虫上的定殖能力与其杀线防病效果密切相关,且细菌菌毛在生防细菌于线虫宿主的定殖过程中发挥着重要功能。本文综述了革兰氏阴性细菌菌毛的种类以及菌毛在生防细菌定殖线虫中的功能,讨论了生防细菌菌毛对运动性、黏附性、生物膜形成等定殖相关生物学性状的影响,并对菌毛在细菌防控林木线虫病害中功能的研究与应用进行了展望。

Reprogramming of phytopathogen transcriptome by a non-bactericidal pesticide residue alleviates its virulence in rice

Bacteria equipped with virulence systems based on highly bioactive small molecules can circumvent their host's defense mechanisms.Pathogens employing this strategy are currently threatening global rice production.In the present study,variations in the virulence of the highly destructive Barkholderiaplantarii were observed in different rice-producing regions.The environment-linked variation was not attributable to any known host-related or external factors.Co-occurrence analyses indicated a connection between reduced virulence and 5-Amino-l,3,4-thiadiazole-2-thiol(ATT),a non-bactericidal organic compound.ATT,which accumulates in rice plants during metabolization of specific agrochemicals,was found to reduce virulence factor secretion by B.plantarii up to 88.8%and inhibit pathogen virulence by hijacking an upstream signaling cascade.Detailed assessment of the newly discovered virulence inhibitor resulted in mechanistic insights into positive effects of ATT accumulation in plant tissues.Mechanisms of virulence alleviation were deciphered by integrating high-throughput data,gene knockout mutants,and molecular interaction assays.TroK,a histidine protein kinase in a two-component system that regulates virulence factor secretion,is likely the molecular target antagonized by ATT.Our findings provide novel insights into virulence modulation in an important plant-pathogen system that relies on the host's metabolic activity and subsequent signaling interference.

Design, synthesis and SAR study of novel sulfonylurea derivatives containing arylpyrimidine moieties as potential anti-phytopathogenic fungal agents

Acetohydroxyacid synthase(AHAS) was considered as a promising target for antifungal agents.Herein,three series of novel sulfonylureas(SUs) 9-11 containing aromatic-substituted pyrimidines were designed and synthesized according to pharmacophore-combination and bioisosterism strategy.The in vitro fungicidal activities against ten phytopathogenic fungi indicated that most of the title compounds exhibited broad-spectrum and excellent fungicidal activities.Based on the preliminary fungicidal activities,a CoMFA model was constructed and the 3 D-QSAR analysis indicated that either a bulky group around the 5-position of the pyrimidine ring or electropositive group around the 2-position of the benzene ring would be favour to fungicidal activities.In order to study interaction mechanism,10 k was automatically docked into yeast AHAS and it further indicated that bearing bulky groups-aryl at the pyrimidine ring was critical to enhance antifungal activities.It revealed that the antifungal activity of derivatives 9-11 probably results from the inhibition of fungal AHAS.Thus,the present results strongly showed that SUs should be considered as lead compounds or model molecules to develop novel antiphyt o pathogenic fungal agents.

微生物除草剂的研究进展

杂草对农业生产构成了巨大的威胁,对农业经济带来了深远的影响。过度依赖化学除草剂的现象引发了大量的抗药性杂草种群出现,这不仅增加了除草剂的使用频率和用量,还会引发恶性循环。通过对现有文献和研究数据的分析,阐述了微生物除草剂的分类及其对目标杂草物种的生物除草活性和特异性。微生物除草剂凭借其延缓杂草产生耐药性、对生态环境影响较小、安全性高和资源充足等独特优势,有望满足可持续农业发展的需求。本文进一步分析了国内外微生物除草剂的发展现状及面临的限制因素,并展望其未来的发展方向和利用趋势。

新型吡唑乙酰胺类琥珀酸脱氢酶抑制剂的设计、合成及抑菌活性

为开发出新型杀菌剂候选分子,通过柔性改造吡唑甲酰胺杀菌剂结构中的二元酰胺键得到了一系列潜在靶向真菌琥珀酸脱氢酶(SDH)的新型吡唑乙酰胺分子.借助菌丝生长速率法发现了其中具有广谱抑菌特性的二苯醚联吡唑乙酰胺分子(6l),其在药剂质量浓度为50μg/mL时对水稻纹枯病菌、小麦赤霉病菌和草莓灰霉病菌的抑制效果均优于对照药剂噁霉灵.化合物6l对水稻纹枯病菌的半最大效应浓度(EC_(50)值)为19.92μg/mL,抑菌活性明显优于对照药剂噁霉灵和氟吡菌酰胺(EC_(50)分别为76.74和40.36μg/mL).SDH酶活力测试结果表明,真菌体内的SDH是化合物6l抑制水稻纹枯病菌生长的潜在作用靶标.分子对接研究结果显示,化合物6l分子结构中的二苯醚单元能以多种方式与靶标口袋的关键残基结合,对分子发挥抑菌活性起到了关键作用.研究结果表明,二苯醚联吡唑乙酰胺分子对植物病原真菌具有较显著的抑制作用,在其结构基础上进行深度的优化和改造有望得到可有效防控植物病原真菌的新型杀菌剂候选分子.

根癌农杆菌中DapB突变株功能的研究

在对根癌农杆菌中二氢吡啶二羧酸还原酶DapB蛋白序列分析的基础上,利用转座子诱变敲除dapB基因.结果发现,根癌农杆菌中的DapB序列具有典型的保守的二核苷酸结合域和底物结合域.dapB基因突变后,突变株的生长明显延迟,外加二氨基庚二酸可恢复突变株的生长;突变株可能通过在细胞膜中合成更多的蛋白质和提高细胞壁碳水化合物含量来提高细胞的韧性以对抗外界环境;与此同时,dapB基因突变影响到农杆菌对GABA的响应.以上结果说明根癌农杆菌中DapB蛋白是一个有功能活性的二氢吡啶二羧酸还原酶,与细胞的生长、细胞壁的合成以及细胞对GABA的响应有关.本研究为寻求新的抗生素靶位点以控制根癌农杆菌中冠瘿瘤奠定了基础.

葱莲地上部和地下部总生物碱提取物对植物病原菌的体外抑制效应

为了探究葱莲(Zephyranthes candida(Lindl.)Herb.)在生物农药上的应用价值,采用水浴加热浸提法提取野生葱莲地上部和地下部的总生物碱,并比较质量浓度500μg·mL^(-1)地上部和地下部总生物碱提取物对常见植物病原菌(包括12种植物病原真菌和4种植物病原细菌)的抑制率。结果表明:葱莲地上部和地下部总生物碱的提取率分别为18.241%和9.725%。葱莲地上部和地下部总生物碱提取物对稻瘟病菌(Magnaporthe oryzae)的抑制率均较高,分别为75.29%和82.62%;2种提取物对其余植物病原真菌和植物病原细菌的抑制率均较低,多在20%以下。并且,葱莲地上部和地下部总生物碱提取物对多数植物病原菌的抑制率无显著差异。研究结果显示:葱莲地上部和地下部总生物碱提取物具有防治稻瘟病的潜力,且地上部总生物碱提取物的使用成本更低。

植物病原菌Nep1-Like蛋白研究进展

NLPs[necrosis-and ethylene-inducing peptide 1(Nep1)-like proteins]是一类质外体分泌蛋白,存在于多种植物病原菌中,在病原菌侵染植物的过程中发挥着重要的作用。根据结构特点,NLP蛋白分为Ⅰ、Ⅱ和Ⅲ3种类型,存在有细胞毒性和无细胞毒性两种形态。具有细胞毒性的NLP蛋白与植物的NTCD4蛋白结合后促进自身寡聚化,引起双子叶植物细胞死亡和感病,而对单子叶植物无影响。无细胞毒性的NLP蛋白功能仍不清楚。NLP蛋白还能作为MAMP分子激发植物免疫反应,许多植物已经进化出特异性识别NLP蛋白的免疫系统,例如拟南芥通过受体蛋白RLP23识别类型Ⅰ的NLP蛋白保守多肽nlp20/24,从而激发免疫反应。本文综述了近年来国内外关于NLP蛋白结构、表达模式、致病机理及被寄主识别机制的研究进展,以期为病原菌寄主互作机制及病害绿色防控研究提供理论依据和参考。

西沙海绵共附生放线菌ITBB-ZK-a5的分离鉴定及抗菌活性研究

海洋放线菌因其所处环境特殊,在生理性状和遗传背景方面均具有独特的性质,因此海洋放线菌具有产生特殊代谢产物的巨大潜力。本研究以西沙海绵为研究对象,挖掘其具有抑菌活性的共附生放线菌资源。首先,利用平板分离法从海绵样品中分离纯化出一株活性放线菌菌株ITBB-ZK-a5,对其进行16S rRNA基因测序,结合构建的系统发育树和菌落形态特征,确定该菌株为链霉菌。其次,采用平板对峙法研究菌株ITBB-ZK-a5对植物病原真菌的抑菌广谱性和持效性。结果表明:该菌株对供试的16株病原真菌均有较好的抑菌活性,抑菌谱较广,且对峙14、30 d仍具有活性,抑菌持效性较好;为了进一步验证该菌株平板通过产生活性物质抑制病原菌生长,对菌落边缘的无菌琼脂块进行抑菌活性研究,结果表明抑菌率与菌落距离呈负相关,说明放线菌可能通过向平板中分泌活性物质以抑制病原菌的生长。该菌株连续转接5代后,仍具有较好的世代稳定性。再次,利用大米固体培养基对菌株ITBB-ZK-a5进行发酵,乙酸乙酯萃取浓缩得到提取物,通过滤纸片法对发酵粗提物进行光稳定性和热稳定性研究,结果表明,随着水浴温度升高和紫外照射时间的增加,菌株ITBB-ZK-a5对香蕉炭疽病病原菌的抑制效果呈下降趋势,但仍具有较强的抑菌活性。最后,测定菌株ITBB-ZK-a5对香蕉炭疽病病原菌的半最大效应浓度(EC_(50))为38.88μg/mL,对木瓜炭疽病病原菌的EC_(50)值为11.70μg/mL。本研究对南海海绵共附生放线菌进行了分离鉴定,筛选出具有抑制多种热带作物病原真菌活性的生防菌株ITBB-ZK-a5,并对其进行抑菌活性研究,为后续生防菌株的开发利用奠定基础。

芳基噻唑胺类化合物合成及其抑菌活性研究

为了获得低毒、高效、环境友好的小分子植物病原真菌抑制剂,本文合成了22个芳基噻唑胺类衍生物,并采用抑制菌丝生长速率法测试了化合物对九种常见植物病原真菌的抑制作用。活性测试结果表明,目标化合物2p、2q和2s具有优异的广谱性抑菌活性,其抑制率为41.1~99.5%,显著优于商品化抑菌剂噁霉灵(25.4~86.5%)和百菌清(24.7~71.4%)。构效关系研究表明:在噻唑胺结构中引入萘环和含有卤素取代的芳环时有利于其抑菌活性的提高。此研究为后期基于芳基噻唑胺骨架的农用抑菌剂开发提供了理论指导。

山慈菇活性成分微波提取工艺优化、指纹图谱与抗植物病菌活性分析

本文综合运用单因素和Box-Behnken响应面试验,以山慈菇提取率和其中3种主要化学成分:双[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯-2-葡萄糖苷(dactylorhin A)、双[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯(militarine)和秋水仙碱的含量为综合权重指标,对其微波提取工艺进行考察。同时,利用高效液相色谱技术建立13批不同产地山慈菇的指纹图谱,明确其主要成分含量分布情况。最后,运用滤纸片扩散法,对制备的山慈菇提取物进行抗菌活性评价。结果表明:山慈菇的最佳微波提取工艺条件:乙醇浓度55%、料液比1:30 g/mL、提取时间17 min。在此条件下,山慈菇的提取率为12.83%。指纹图谱结果表明,山慈菇中3种化学成分的含量分布趋势一致,其中militarine为13批山慈菇中的共有化学成分,而dactylorhin A和秋水仙碱分别在1个产地样品中未检出,表明不同产地山慈菇中主要化学成分分布具有差异性。抗菌实验结果显示,山慈菇提取物对胶孢炭疽菌、辣椒疫霉菌、灰葡萄孢菌、茄链格孢菌、荧光假单胞菌、水稻白叶枯菌和禾谷镰孢菌均没有显示抗菌活性(抑菌圈小于6 mm),对雷尔氏菌具有较好的抑制活性(抑菌圈8.5 mm),表现出一定的抗菌选择性。本研究确定了微波提取山慈菇的最佳工艺,建立的山慈菇中3种成分同步含量分析方法稳定可行,研究结果可为山慈菇药材的质量标准建立和综合开发利用提供参考。