Analysis of Chuanxiong Rhizoma substrate on production of ligustrazine in endophytic Bacillus subtilis by ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry

Ligustrazine was the active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma. However, the content of ligustrazine is very low. We proposed a hypothesis that ligustrazine was produced by the mutual effects between endophytic Bacillus subtilis and the Ligusticum chuanxiong Hort. This study aimed to explore whether the endophytic B. subtilis LB5 could make use of Chuanxiong Rhizoma fermentation matrix to produce ligustrazine and clarify the mechanisms of action preliminarily. Ultra high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry analysis showed the content of ligustrazine in Chuanxiong Rhizoma was below the detection limit (0.1 ng/mL), while B. subtilis LB5 produced ligustrazine at the yield of 1.0268 mg/mL in the Chuanxiong Rhizoma‐ammonium sulfate fermentation medium. In the fermented matrix, the reducing sugar had a significant reduction from 12.034 to 2.424 mg/mL, and rough protein content increased from 2.239 to 4.361 mg/mL. Acetoin, the biosynthetic precursor of ligustrazine, was generated in the Chuanxiong Rhizoma‐Ammonium sulfate (151.2 mg/mL) fermentation medium. This result showed that the endophytic bacteria B. subtilis LB5 metabolized Chuanxiong Rhizoma via secreted protein to consume the sugar in Chuanxiong Rhizoma to produce a considerable amount of ligustrazine. Collectively, our preliminary research suggested that ligustrazine was the interaction product of endophyte, but not the secondary metabolite of Chuanxiong Rhizoma itself.     

一株西沙群岛野生诺尼种子的内生细菌CICC 10594的分离与鉴定

以中国西沙群岛野生诺尼种子的内生细菌菌株CICC 10594为研究对象,利用基于形态学观察、生理生化特征鉴定及系统发育学分析的微生物多相分类鉴定技术,对CICC 10594的分类地位进行了鉴定与分析,结果显示,CICC10594为阴沟肠杆菌.     

植物深色有隔内生真菌（DSE）的研究进展

对公开发表的有关植物深色有隔内生真菌（Dark Separate Endophytes,DSE）研究资料比较分析显示,DSE在世界各种生态类型的植物根内广泛存在,但目前对此类真菌生态功能的研究还处于起步阶段。为了给DSE的更深入的研究提供有启发意义的思路和框架,文章回顾和讨论了DSE的研究历史、类群组成、生态分布和可能的生物学功能。认为DSE-植物共生应当具有多方面的功能而非仅限于促进植物对营养的吸收。强调这一类群鲜为人知的根定植真菌的广布性和潜在的生态学意义,以引发研究者对此类真菌的广泛兴趣。     

Biodiversity and application prospects of fungal endophytes in the agarwood-producing genera,Aquilaria and Gyrinops (Thymelaeaceae): A review

Agarwood is originated from the resinous part of Aquilaria and Gyrinops plants and has been a precious biomaterial for applications in traditional medicine, perfumery, cosmetics, and religious purposes all over the world. In the wild, the formation of agarwood is related to the defense mechanism of the tree in response to physical damage that allows further microbial infestation into its wood, while having the whole tree covered with agarwood would take up a long time, and it rarely happens. For Aquilaria and Gyrinops, the presence of endophytes is mainly found derived from the tree. The isolated endophytes could be important sources of natural products, while some could contribute to the formation of agarwood in the tree, which is safe for the environment and human health. This review summarized the biodiversity of fungal endophytes recorded in Aquilaria and Gyrinops and their potential effects on host trees. Till now, 67 endophytic genera have been isolated from Aquilaria and Gyrinops, and 18 ones were found responsible for the promotion of agarwood formation. Additionally, 92 compounds have been reported to be produced by the agarwood endophytes, and 52 ones displayed biological activities, most of which have anti-inflammatory, anti-bacterial, and anti-cancer activities. Nevertheless, fungal endophytes are promising agents that deserved to be further studied and scaled up to a commercial level for the production of agarwood oil, but the role of endophytes in the agarwood host trees needs to be furtherly investigated in future studies.     

The Role of Endophytes in Combating Fungal- and Bacterial-Induced Stress in Plants

Plants are subjected to multifaceted stresses that significantly jeopardize crop production. Pathogenic microbes influence biotic stress in plants, which ultimately causes annual crop loss worldwide. Although the use of pesticides and fungicides can curb the proliferation of pathogens in plants and enhance crop production, they pollute the environment and cause several health issues in humans and animals. Hence, there is a need for alternative biocontrol agents that offer an eco-friendly mode of controlling plant diseases. This review discusses fungal- and bacterial-induced stress in plants, which causes various plant diseases, and the role of biocontrol defense mechanisms, for example, the production of hydrolytic enzymes, secondary metabolites, and siderophores by stress-tolerant fungi and bacteria to combat plant pathogens. It is observed that beneficial endophytes could sustain crop production and resolve the issues regarding crop yield caused by bacterial and fungal pathogens. The collated literature review indicates that future research is necessary to identify potential biocontrol agents that can minimize the utility of synthetic pesticides and increase the tenable agricultural production.     

OSMAC Strategy Integrated with Molecular Networking for Accessing Griseofulvin Derivatives from Endophytic Fungi of Moquiniastrum polymorphum (Asteraceae)

Three endophytic fungi isolated from Moquiniastrum polymorphum (Less.) G. Sancho (Asteraceae) were cultivated using the one strain many compounds (OSMAC) strategy to evaluate the production of griseofulvin derivatives. Extracts obtained were analyzed by HPLC–MS/MS and the chromatographic and spectrometric data used to elaborate a feature-based molecular network (FBMN) through the GNPS platform. This approach allowed the observation of differences such as medium-specific and strain-specific production of griseofulvin derivatives and variations of cytotoxic activity in most extracts. To evaluate the efficiency of the OSMAC approach allied with FBMN analysis in the prospection of compounds of biotechnological interest, griseofulvin and 7-dechlorogriseofulvin were isolated, and the relative concentrations were estimated in all culture media using HPLC–UV, allowing for the inference of the best strain–medium combinations to maximize its production. Malt extract-peptone broth and Wickerham broth media produced the highest concentrations of both secondary metabolites.     

莲瓣兰内生真菌利用碳源的生理学特性研究

对分离自莲瓣兰根内的5株内生真菌利用碳源的生理学特性进行研究，方法是三点种植实验，培养5d，测量碳源培养基上内生真菌菌落直径，同时，进行液体培养5d，将菌丝过滤，烘干至恒重，计算菌丝干重。结果：这5株内生真菌利用碳源能力各不同；其中，属于木霉属的LP161菌株对各种碳源均有很好的利用能力，尤其在纤维素培养基上生长最快；并且，LP161菌株在葡萄糖液体培养基内生长最快，培养5d后菌丝干重达到1．18g/100ml。     

Rare C25 Steroids Produced by Penicillium chrysogenum P1X,a Fungal Endophyte of Huperzia serrata

Three new metabolites, norcyclocitrinol A ( 1 ), erythro‐11α‐hydroxyneocyclocitrinol ( 2 ), and pesudocyclocitrinol A ( 3 ), along with six known analogs, i.e., neocyclocitrinols A–D ( 4 – 7 , resp.), cyclocitrinol ( 8 ), and 24‐epicyclocitrinol ( 9 ), were isolated and identified from the culture broth of Penicillium chrysogenum P1X, a fungal endophyte of Huperzia serrata. Compounds 1 – 9 were identified by spectroscopic methods to share the same C₂₅‐steroid skeleton featuring an unusual bicyclo[4.4.1] A/B ring system. In particular, 1 represents the first example of a C₂₅ steroid with a bisnor C‐atom side chain. All compounds were evaluated for their cytotoxic activities against HeLa and HepG2 cell lines. However, none of them exhibited a significant cytotoxicity at a concentration of 20 μM .     

石斛共生真菌木霉菌拮抗作用的初步研究

为探讨拮抗真菌及其次生代谢产物对石斛病原菌生长的影响,从药用植物石斛中分离、鉴定和筛选出对常见病原真菌镰刀菌(Fusarium)有拮抗作用的木霉菌(Trichoderma).通过平板对峙培养,观察其对镰刀菌生长的影响;通过几丁质水解圈实验,检测木霉几丁质酶的活力.该研究所筛选获得的木霉生防菌株,可应用于石斛的栽培管理,...     

蛇足石杉外植体表面消毒及内生菌消除方法

为了获得蛇足石杉的无菌材料,使用双氧水、酒精、升汞等消毒剂对外植体进行表面消毒,使用混合抗菌溶液及不同温度的热水浴处理外植体以消除内生菌.结果表明:茎段经预处理后,用47 ℃热水浴处理1 h,再经70 %酒精处理30 s, 0.1 %升汞处理7 min后,能达到较好的表面消毒及内生菌消除的效果;孢子囊和茎尖经预处理后,再经过70%酒精处理30 s,0.1 %升汞处理4 min即可达到较好的表面消毒效果;消毒剂结合温度处理能降低内生菌的污染率;孢子囊和茎尖几乎不含内生菌,是可用于组织培养的较好的外植体.