Shuanghuanglian, a traditional Chinese medicine, is well-known for its bioactive compounds, such as flavonoids, which offer significant health benefits. However, the production of Shuanghuanlian generates substantial pharmaceutical residues, which are often discarded as waste, posing significant environmental and economic challenges. To date, research on repurposing these medicine residues has been limited. This study utilized beneficial microbes to efficiently extract and utilize the residual bioactive compounds. Notably, a newly isolated Lactiplantibacillus plantarum strain LLB exhibited remarkable efficiency in converting flavonoid glycosides (e.g., phillyrin and luteoloside) into their corresponding aglycones. When combined synergistically with Bacillus subtilis and Saccharomyces cerevisiae, strain LLB maintained robust flavonoid glycoside conversion while enhancing lactobacilli viability in the fermented medicine residues. As a feed additive for broiler chickens, the fermented residue not only boosted antioxidant (superoxide dismutase) and anti-inflammatory (IL-10) markers, but also preserved growth performance and meat quality. Furthermore, the fermented residue modulated the gut microbiome, increasing Rikenella while reducing Elusimicrobiota and Parabacteroides abundances. Our findings demonstrate that microbial transformation of Shuanghuanglian residues offers a sustainable strategy for waste valorization and a novel feed additive for enhancing animal health.

Fungi produce various bioactive secondary metabolites (SMs) as protective and weaponized tools to enhance survival in shared ecological niches. By mimicking a competitive ecosystem, cocultivation has been proven to be particularly successful in stimulating SM discovery. Here, we reported the identification of four novel metabolites, epiclactones A and B, epioxochromane and aoergostane, from the coculture of two biotechnologically important strains, Aspergillus oryzae and Epicoccum dendrobii. Transcriptome and metabolome analyses revealed widespread silent gene activation during fungal-fungal interaction. The majority of differentially expressed gene clusters were summarized for both strains. Based on these highly activated biosynthetic pathways, we suggested that a bidirectional chemical defense occurred under cocultivation. E. dendrobii enhanced the production of the spore inhibitor, fumigermin. Moreover, A. oryzae highly accumulated the antifungal agent kojic acid with a yield of up to 1.10 g/L. This study provides an excellent example for the discovery of hidden natural products by cocultivation.

为明确我国黄淮产区地黄种质间遗传多样性,解析种质间的亲缘关系,为该区地黄品种选育和种质鉴定提供依据,利用毛细管电泳技术和108对SSR引物对地黄种质进行分析,以获取种质间遗传相似系数、多态性引物、种质分辨率,利用软件TASSEL 3.0和Fig Tree(V 1.4.3)进行聚类分析和作图,选取多态性好、稳定性强的引物构建地黄种质的SSR指纹图谱.结果表明,共获得多态性引物50对,其片段长度在107～365 bp,种质间遗传相似系数在0.552～0.984,平均为0.729,野生种质的平均遗传相似系数明显低于地方品种和当前主栽品种.聚类结果显示,17个地黄种质分为3个亚群.选取并利用多态性强和分辨率高、扩增稳定的15对引物构建了地黄种质的SSR指纹图谱,实现了对每个地黄种质的快速鉴定.综上,SSR分析是实现地黄鉴真、聚类、DNA指纹图谱构建的一种有效方法.

[背景]粪臭素是畜牧堆肥中有机污染物的主要成分,造成养殖场及周边环境恶化,粪臭素污染问题亟待解决,利用微生物降解粪臭素是一种环保节能的有效方法.[目的]分离鉴定粪臭素高效降解菌株,研究其降解特性,为粪臭素降解提供高效的菌种资源,为该菌株应用于臭味污染环境的净化提供基础.[方法]以粪臭素为唯一碳源的无机盐培养基作为培养基质,从猪粪堆肥样品中分离筛选粪臭素高效降解菌株,通过形态特征和16SrRNA基因序列分析进行分离菌株的初步鉴定,分析其生长规律及粪臭素降解特性,并利用气相色谱质谱联用(GC-MS)对菌株代谢粪臭素的产物进行分析.[结果]从样品中分离获得一株能以粪臭素为唯一碳源的细菌YKSW-6菌株,形态学和16S rRNA基因序列分析初步鉴定该菌株为戈登氏红球菌(Rhodococcus gordoniae).接种量为10％时,该菌培养14 h对100 mg/L的粪臭素降解率达到100％.其能够利用D-山梨醇、溴-丁二酸等18种碳源,对亚碲酸钾、溴酸钾等13种化学敏感物具有抗性.菌株YKSW-6在5％接种量、温度30-42℃和pH值为6.0-9.0时对100 mg/L的粪臭素降解效率均能达到100％,菌株生长和降解粪臭素的最佳条件为:pH 7.2,温度37℃,转速180r/min.GC-MS结果表明,粪臭素在菌株的作用下C2先被氧化,转变为3-甲基羟基吲哚,随后进一步被氧化为N-(2-乙酰基苯基)甲酰胺.同时中间产物还有苯乙醛和苯乙酸.[结论]红球菌YKSW-6为目前已报道的降解粪臭素能力较强的菌株,丰富了粪臭素降解菌种的资源库,为实际环境微生物修复应用提供了理论参考.

为了探究生物除臭剂在生猪养殖场的应用效果并对其进行安全性评价,该试验使用复合微生物除臭剂,在生猪养殖场以NH3和H2S释放量为指标,进行大气环境和密闭环境试验,并对该除臭剂进行小鼠和家兔急性毒性检测.结果表明:生物除臭剂在大气环境下对猪粪中NH3和H2S的去除率可达76.13％和82.61％;在密闭环境试验中去除率可达80.48％和68.42％,均比对照组有显著提高(P＞0.05).其对昆明种雄、雌性小鼠急性经口毒性LD50＞40 g/kg体重、对家兔皮肤无刺激.说明该生物除臭剂在实际应用中具有高效的除臭能力和安全性.

RNAi technology, known as a revolutionary technology in the history of pesticides, has been identified as a very promising novel approach for crop protection, which is of great significance for achieving the sustainable agricultural development of the United Nations Food and Agriculture Organization. Although many studies have shown that RNA biopesticides have strong application prospects, its stability seriously restricts the commercial use. As the core component of RNAi, double-stranded RNA (dsRNA) is unstable in its natural form. Therefore, how to ensure the stability of dsRNA is one of the most significant challenges in realizing the commercial use of RNA biopesticides. Nanomaterials such as cationic polymers and lipofectamine can improve the stability of dsRNA in the environment, which has been proved. This paper reviews the recent research progress of nanomaterials that can be used to improve the environmental stability of dsRNA, and discusses the advantages and limitations of different nanomaterials combined with dsRNA, which provides reference for the selection of dsRNA nanoformulations.

Some microbial strains are ideal producers of extracellular enzymes that can be used in various industries. However, in many fields, especially in the pharmaceutical field, these enzymes need to be recovered and purified through multistep processes and tedious procedures before they can be used. The recovery process is difficult and increases the cost of enzyme production. Therefore, reducing purification steps will greatly benefit the utilization of microbial enzymes. The 35 M strain of Bacillus amyloliquefaciens, which has high extracellular protease production, was isolated from a phosphate mine. When cultured in a medium with soybean meal as the main component, the maximum activity of extracellular protease reached 16,992 U/mL. SDS-PAGE showed that there were two main proteins in the fermentation supernatant, with a paucity of other defined protein bands. Mass spectrometry and zymogram analysis showed that the two main bands were two proteases, corresponding to alkaline protease (AprM) and neutral protease (NprM), respectively. Gene cloning, sequencing, and further comparisons were used to confirm AprM and NprM correspond to these proteases from B. amyloliquefaciens. Notably, SDS-PAGE and zymogram analysis showed that NprM had obviously higher catalytic efficiency toward casein than did AprM. Strain 35 M is a promising protease producer with great potential for applications in industrial protease production. Additionally, this study demonstrates strain 35 M may be particularly well suited to use in degrading anti-nutritional factors in soybean meal, so as to improve the nutritional value of soybean meal.

采用盆栽试验探讨了不同剂量棘孢木霉SFC-3菌剂对小麦生理生化指标及生防效果的影响.结果 表明:施用SFC-3菌剂对小麦的株高和主根长度没有明显影响(P＞0.05),但可提高小麦的出芽率,当施菌剂量为1.0～2.0 g/kg时小麦的出芽率显著高于对照(P＜0.05);施用SFC-3菌剂小麦叶片中叶绿素含量和可溶性蛋白含量与对照相比均显著增加(P＜0.05),其中施菌剂量为1.0 g/kg时最高.而小麦叶片中的MDA含量在施菌剂量为1.0 g/kg时显著低于对照和其他施菌处理(P＜0.05);当施菌剂量为0.5～1.0 g/kg时,小麦叶片中SOD和POD活性显著高于对照,进一步增加施菌剂量,小麦叶片中SOD和POD活性均随之显著降低(P＜0.05).当施菌剂量为1.5 g/kg和2.0 g/kg时每克根中小麦孢囊线虫2龄幼虫数量最少,发病级别最低,其相对防效分别为88.68％和87.04％,其次为施菌剂量1.0 g/kg的处理,其相对防效为80.50％.施用不同剂量的棘孢木霉SFC-3菌剂对小麦生理生化指标均有不同程度的影响,并且当施菌剂量为1.0 g/kg时对小麦生长和孢囊线虫的防治效果最好.

Fungi from unique environments exhibit special physiological characters and plenty of bioactive natural products. However, the recalcitrant genetics or poor transformation efficiencies prevent scientists from systematically studying molecular biological mechanisms and exploiting their metabolites. In this study, we targeted a guanophilic fungus Amphichorda guana LC5815 and developed a genetic transformation system. We firstly established an efficient protoplast preparing method by conditional optimization of sporulation and protoplast regeneration. The regeneration rate of the protoplast is up to about 34.6% with 0.8 M sucrose as the osmotic pressure stabilizer. To develop the genetic transformation, we used the polyethylene glycol-mediated protoplast transformation, and the testing gene AG04914 encoding a major facilitator superfamily transporter was deleted in strain LC5815, which proves the feasibility of this genetic manipulation system. Furthermore, a uridine/uracil auxotrophic strain was created by using a positive screening protocol with 5-fluoroorotic acid as a selective reagent. Finally, the genetic transformation system was successfully established in the guanophilic fungus strain LC5815, which lays the foundation for the molecular genetics research and will facilitate the exploitation of bioactive secondary metabolites in fungi.

霉酚酸是世界上应用最广泛的免疫抑制剂之一,市场需求巨大.目前为止,主要是通过真菌发酵的方式进行霉酚酸的工业生产,而用于生产的菌株多是经过诱变的高产短密青霉菌.本文从霉酚酸的研究应用现状、化学合成以及生物合成途径、遗传调控、发酵生产以及市场分析等方面对霉酚酸的研究及产业化进展进行了系统综述.为该药物的新颖衍生物开发、提高产率以及应用先进生物技术智能化创制提供重要参考和依据.

Siderophores are small molecular iron chelators and participate in the multiple cellular processes in fungi. In this study, biosynthesis gene clusters of coprogens and dimerumic acids were identified by transcriptional level differences of genes related to iron deficiency conditions in Metarhizium robertsii . This leads to the characterization of new coprogen metachelin C ( 1 ) and five known siderophores metachelin A ( 2 ), metachelin A-CE ( 3 ), metachelin B ( 4 ), dimerumic acid 11-mannoside ( 5 ), and dimerumic acid ( 6 ). The structure of metachelin C ( 1 ) was elucidated by NMR spectroscopy and HR-ESI-MS analysis. Genetic deletions of mrsidA , and mrsidD abolished the production of compounds 1 – 6 that implied their involvement in the biosynthesis of coprogen and dimerumic acid. Interestingly, NRPS gene mrsidD is responsible for biosynthesis of both coprogen and dimerumic acid, thus we proposed plausible biosynthetic pathways for the synthesis of coprogen and dimerumic acid siderophores. Therefore, our study provides the genetic basis for understanding the biosynthetic pathway of coprogen and dimerumic acid in Metarhizium robertsii .

As an indispensable essential amino acid in the human body, lysine is extremely rich in edible mushrooms. The α-aminoadipic acid (AAA) pathway is regarded as the biosynthetic pathway of lysine in higher fungal species in Agaricomycetes. However, there is no deep understanding about the molecular evolutionary relationship between lysine biosynthesis and species in Agaricomycetes. Herein, we analyzed the molecular evolution of lysine biosynthesis in Agaricomycetes. The phylogenetic relationships of 93 species in 34 families and nine orders in Agaricomycetes were constructed with six sequences of LSU, SSU, ITS (5.8 S), RPB1, RPB2, and EF1-α datasets, and then the phylogeny of enzymes involved in the AAA pathway were analyzed, especially homocitrate synthase (HCS), α-aminoadipate reductase (AAR), and saccharopine dehydrogenase (SDH). We found that the evolution of the AAA pathway of lysine biosynthesis is consistent with the evolution of species at the order level in Agaricomycetes. The conservation of primary, secondary, predicted tertiary structures, and substrate-binding sites of the enzymes of HCS, AAR, and SDH further exhibited the evolutionary conservation of lysine biosynthesis in Agaricomycetes. Our results provide a better understanding of the evolutionary conservation of the AAA pathway of lysine biosynthesis in Agaricomycetes.

Edible mushrooms are important nutraceutical sources of foods and drugs, which can produce various nutritional ingredients including all essential amino acids. The method of rapid screening for the strains producing specific functional components is very indispensable. Homocitrate synthase is one of the key enzymes in the α-aminoadipate pathway for lysine biosynthesis and has preferable sequence conservation in Agaricales. Based on the blast of homocitrate synthase homologous genes of strains of Agaricales, we achieved combinations of degenerate primers as molecular markers to rapidly screen the lysine-producing edible mushrooms. The experimental results revealed that the consistency between PCR amplification and HPLC analysis attained 82 and 75% in strains of Agaricales and Polyporales, respectively. The finding showed that the molecular marker has higher universality for screening edible mushroom resources of Agaricales. This PCR-based approach shows excellent potential in evaluating and discriminating edible wild-grown mushrooms with high lysine content in Agaricales.

为考察磷灰石、石灰和黑麦草对Cd污染土壤的修复效果,通过盆栽试验研究了不同剂量磷灰石(0、6、12、24 g·kg-1风干土,编号为CK、L1、L2、L3)和石灰(1、2、4 g·kg-1风干土,编号为S1、S2、S3)对Cd胁迫下黑麦草生物量、根形态、黑麦草Cd含量、Cd富集量、富集系数、土壤pH、土壤有效态Cd含量等的影响.结果表明,磷灰石、石灰处理(L1,S1除外)显著增加了黑麦草生物量、根长、根表面积、根体积、根尖数、Cd富集量和土壤pH,显著降低了黑麦草Cd含量、Cd富集系数、根平均直径和土壤有效态Cd含量.相关性分析结果表明:除根平均直径为正相关关系外,黑麦草根长、根表面积、根体积、根尖数与土壤有效态Cd含量、黑麦草地上部分及根系Cd含量、Cd富集系数呈显著或极显著负相关关系;除根平均直径为负相关关系外,其他根指标与黑麦草地上部分及根系生物量、Cd富集量呈极显著正相关关系.其中,L3处理黑麦草地上部分及根系生物量达到CK处理(不添加改良剂)的176.0倍和174.4倍;根平均直径相比CK降低了29.4％,根长、根表面积、根体积和根尖数分别为CK处理的20.2、21.0、21.3倍和24.5倍;黑麦草地上部分及根系Cd富集量达到CK处理的89.6倍和100.9倍.研究表明,根形态受土壤有效态Cd含量的影响显著,并可通过影响黑麦草生物量及Cd吸收决定其Cd富集量,施用24 g·kg-1的磷灰石并种植黑麦草能很好地修复Cd污染土壤.

Siderophores are small molecular iron chelators and participate in the multiple cellular processes in fungi. In this study, we discovered and identified five amphiphilic coprogen siderophores including three new natural products according to LC-MS-guided separation strategy from Trichoderm hypoxylon. The structures of three new coprogens were elucidated by NMR spectroscopy, and high-resolution (HR)-ESI-MS analysis. Genetic deletions of dfcA and dfcB abolished the production of compounds 1–5 that implied their involvement in the biosynthesis of coprogens. Interestingly, cultivations of ΔdfcA and ΔdfcB mutants with pathogenic fungi Fusarium oxysporum and Mucor corcinelloides showed the weaker inhibitions in comparison to wild type that demonstrated coprogen’s role in combating the pathogenic fungi. Our study not only enriched the diversities of siderophores but also provided an approach for finding the rare amphiphilic coprogen siderophores in fungi. Furthermore, this work provided a basis for investigation on the biosynthesis of fungal amphiphilic siderophores and their ecological roles in nature. • A series of amphiphilic coprogens were found. • The gene cluster of amphiphilic coprogens and ecological roles were elucidated.

为了获得具有防治根结线虫作用的菌株,采用几丁质平板透明圈初筛和离体拮抗试验复筛的方法,得到一株具有生防潜力的真菌菌株SFC-3.通过形态学特征观察和真菌内转录间隔区序列(Internal transcribed spacer,ITS)分析鉴定,确定菌株SFC-3为棘孢木霉(Trichoderma asperellum),其几丁质酶活为2.07 U/mL.根结线虫虫卵寄生实验和二龄幼虫致死率实验表明:菌株SFC-3对虫卵的寄生率最高达到78.86％,高于SFC-1、SFC-22等其他9株菌,在第7天时对根结线虫虫卵孵化的相对抑制率最高达到96.51％,对二龄幼虫的校正死亡率达到93.37％.通过筛选抗根结线虫菌株并研究了其生防效果,证实了菌株SFC-3有比较优秀的抗根结线虫性能.

葡萄糖氧化酶作为一种重要的氧化还原酶,在饲料、畜牧等行业应用广泛,可用于改善动物饲料的消化利用率.本文对葡萄糖氧化酶的发酵生产工艺、提取纯化工艺、酶学特性改进工艺及质量控制工艺作了简单介绍,以期为葡萄糖氧化酶的工业化生产和应用提供帮助.

为了探讨新型快速秸秆还田促腐菌剂对小麦秸秆还田在水稻田的施用剂量和促生效果,进行了相关的大田试验.本试验在常规施肥和小麦秸秆粉碎还田的基础上设置了M1(CK),M2(30 kg/hm2),M3(75 kg/hm2)和M4(120 kg/hm2)4个梯度的促腐菌剂施用剂量,研究了不同施用剂量对水稻成产因素以及水稻产量的影响.研究表明,施用新型快速秸秆还田促腐菌剂不仅可以促进秸秆降解,且对每公顷水稻的穗数、穗粒数及产量影响显著.与对照组M1相比,M3(75 kg/hm2)组促产效果最好,其中每公顷穗数为466.35万穗,比对照提高了16.09％;穗粒数为86.03比对照提高了12.12％.在实际产量方面M3组为7510.35 kg/hm2,比对照提高了12.94％.土壤理化性质表明,促腐菌剂显著提高了土壤中碱解氮、有效磷、速效钾的含量,与对照组相比分别提高了93.33％、18.13％和46.93％.该试验评价了施用不同剂量新型快速秸秆还田促腐菌剂对小麦秸秆在水稻田降解及水稻产量的影响,为新型快速秸秆还田促腐菌剂的研发奠定了理论基础.

糖尿病作为一种复杂的代谢疾病,严重威胁着人类的健康,而2型糖尿病因其发病原因复杂,一直以来备受关注.现代研究发现肉桂提取物及其活性成分具有抗菌、抗肿瘤等多种药理作用,药用价值极高,对2型糖尿病可以产生很好的改善作用,能够从多方面改善2型糖尿病的症状,临床研究表明肉桂提取物及其活性成分对于改善胰岛素抵抗和脂质代谢紊乱效果良好.本文将结合肉桂提取物及其活性成分的临床研究和基础研究从降血糖、抑制糖基化终产物形成、阻止糖尿病并发症等方面对其改善作用进行论述,以期为肉桂提取物及其活性成分在临床上对2型糖尿病的治疗提供帮助.

烟梗是烟草工业的重要副产物,也是宝贵的自然资源.本研究首先利用白腐菌漆酶对烟梗丝进行预处理,提升了添加烟梗丝的卷烟品质;然后分别以木质素、纤维素、半纤维素和果胶的降解率为响应值,采用Box-Behnken设计建立方程模型,对漆酶、纤维素酶、半纤维素酶和果胶酶组成的复合酶预处理烟梗丝条件进行了优化.结果 表明:每100g烟梗丝加入30U漆酶,在料液比为35％、温度为30℃、酶解pH为5处理48h的条件下预处理的烟梗丝对提升卷烟品吸效果最佳,烟梗丝中木质素、纤维素、半纤维素和果胶的降解率分别为20.16％、15.10％、7.20％和12.40％;为获得与之相同的各组分降解率,响应面法优化漆酶复合酶最佳处理条件为:每100g烟梗丝加入漆酶14.72U、纤维素酶1.00U、半纤维素酶1.00U、果胶酶8.45U.验证发现烟梗丝各组分降解率实测值与理论值无显著性差异,且显微结构观察显示复合酶处理后的烟梗丝表面致密结构被破坏,孔洞数量明显增加.本研究获得的白腐菌漆酶预处理后的烟梗丝在卷烟中的添加能有效改善卷烟品质,且漆酶复合酶的使用大幅减少了漆酶的用量,降低了漆酶预处理烟梗丝的成本,为废弃烟梗生物质的资源化利用提供了重要依据.