枯草芽孢杆菌表达与调控工具相关研究进展

枯草芽孢杆菌作为革兰氏阳性模式微生物,由于其清晰的遗传背景、高效的分泌能力以及简单的培养条件等优势被广泛的应用于生物技术产业。近年来,随着代谢工程与合成生物学的发展,枯草芽孢杆菌相关表达系统与调控工具研究也取得了很大进展。围绕枯草芽孢杆菌动态调控工具的研究进展,分别从转录水平调控和转录后水平调控两个层面上进行综述,并对调控元件在生物技术中的应用进行了讨论。最后,对未来枯草芽孢杆菌表达与调控工具的发展进行了展望。     

解淀粉芽胞杆菌抗真菌活性研究进展

解淀粉芽胞杆菌(Bacillus amyloliquefaciens)具有很强的抑制植物病原真菌的能力。其菌体细胞能产生多种酶类、脂肽类抗生素、生物表面活性素、聚酮类化合物和抑菌蛋白,同时具有诱导植物产生系统抗性(ISR)的能力,因此在工农业、种植业、养殖业、食品加工业、果蔬的采后保鲜和饲料业等行业具有重要价值。本文对解淀粉芽胞杆菌抗真菌作用、抗真菌能力提高策略、抗菌化合物合成调节、抑制真菌机制及其引发的ISR等问题进行了深入探讨和综述。     

凝结芽胞杆菌生物学特性及其在家禽生产中的应用

作为一种新兴的微生物饲料添加剂,凝结芽胞杆菌既表现出类乳酸菌的益生特性,又具有芽胞杆菌抗逆性好、易储存的优点,有利于产业化及市场推广。凝结芽胞杆菌能够产生有机酸及多种酶类、修复受损的肠黏膜、抑制病原菌繁殖及促进营养物质的消化吸收,在家禽生产中受到广泛关注和应用。目前关于凝结芽胞杆菌的研究不够深入,其生物学特性及作用机制还在进一步研究中,还需加深其在畜牧业中的应用研究。本文主要对凝结芽胞杆菌近几年的研究进展及其在家禽生产中的应用进行综述,为今后凝结芽胞杆菌的研发及应用提供参考依据。     

Effect of a Bacillus subtilis strain on flax protection against Fusarium oxysporum and its impact on the root and stem cell walls

In Normandy, flax is a plant of important economic interest because of its fibres. Fusarium oxysporum, a telluric fungus, is responsible for the major losses in crop yield and fibre quality. Several methods are currently used to limit the use of phytochemicals on crops. One of them is the use of plant growth promoting rhizobacteria (PGPR) occurring naturally in the rhizosphere. PGPR are known to act as local antagonists to soil‐borne pathogens and to enhance plant resistance by eliciting the induced systemic resistance (ISR). In this study, we first investigated the cell wall modifications occurring in roots and stems after inoculation with the fungus in two flax varieties. First, we showed that both varieties displayed different cell wall organization and that rapid modifications occurred in roots and stems after inoculation. Then, we demonstrated the efficiency of a Bacillus subtilis strain to limit Fusarium wilt on both varieties with a better efficiency for one of them. Finally, thermo‐gravimetry was used to highlight that B. subtilis induced modifications of the stem properties, supporting a reinforcement of the cell walls. Our findings suggest that the efficiency and the mode of action of the PGPR B. subtilis is likely to be flax variety dependent.     

GeSUT4 mediates sucrose import at the symbiotic interface for carbon allocation of heterotrophic Gastrodia elata (Orchidaceae)

Gastrodia elata, a fully mycoheterotrophic orchid without photosynthetic ability, only grows symbiotically with the fungus Armillaria. The mechanism of carbon distribution in this mycoheterotrophy is unknown. We detected high sucrose concentrations in all stages of Gastrodia tubers, suggesting sucrose may be the major sugar transported between fungus and orchid. Thick symplasm‐isolated wall interfaces in colonized and adjacent large cells implied involvement of sucrose importers. Two sucrose transporter (SUT)‐like genes, GeSUT4 and GeSUT3, were identified that were highly expressed in young Armillaria‐colonized tubers. Yeast complementation and isotope tracer experiments confirmed that GeSUT4 functioned as a high‐affinity sucrose‐specific proton‐dependent importer. Plasma‐membrane/tonoplast localization of GeSUT4‐GFP fusions and high RNA expression of GeSUT4 in symbiotic and large cells indicated that GeSUT4 likely functions in active sucrose transport for intercellular allocation and intracellular homeostasis. Transgenic Arabidopsis overexpressing GeSUT4 had larger leaves but were sensitive to excess sucrose and roots were colonized with fewer mutualistic Bacillus, supporting the role of GeSUT4 in regulating sugar allocation. This is not only the first documented carbon import system in a mycoheterotrophic interaction but also highlights the evolutionary importance of sucrose transporters for regulation of carbon flow in all types of plant‐microbe interactions.     

外源微生物对木麻黄幼苗生长和土壤微生物群落的影响

将解淀粉芽孢杆菌(YB706)和伯克氏菌(BK8)外源添加至木麻黄盆栽苗中,运用Biolog生态板和磷脂脂肪酸(PLFA)技术,探究外施细菌能否改善连栽木麻黄土壤养分和微生物群落及植株生长。结果表明:与空白处理(CK)相比,YB706和BK8处理的木麻黄盆栽苗土壤碱解氮、速效磷显著增加,土壤全氮、全磷、全钾和速效钾无明显变化,幼苗株高分别增加59.1%和63.9%,BK8处理叶绿素含量提高81.9%。各处理平均颜色变化率(AWCD)呈现YB706>CK>BK8;除氨基酸类外,土壤微生物对不同碳源的利用率也表现为YB706>CK>BK8;经YB706和BK8处理的土壤微生物种类和数量均显著增加,除放线菌外,各类微生物PLFA总含量均为BK8>YB706>CK,与CK相比,土壤真菌/细菌有所提高。YB706和BK8处理的土壤微生物群落Simpson、Shannon、Brillouin和McIntosh指数均显著提高。表明外施YB706和BK8可促进木麻黄幼苗生长,且显著增加土壤速效养分含量,提高土壤微生物群落结构多样性,改善土壤微生物环境。     

芽胞杆菌治疗幽门螺杆菌感染疗效的Meta分析

目的评价芽胞杆菌(Bacillus)联合根除标准疗法治疗幽门螺杆菌(H.pylori)感染的有效性。方法收集关于芽胞杆菌联合治疗H.pylori感染的随机对照试验(RCT),检索时限为建库至2020年5月;对符合纳入标准的研究进行偏倚风险评价及Meta分析。结果最终纳入14个RCT。Meta分析结果显示芽胞杆菌联合治疗能提高H.pylori根除率(ITT分析:RR=1.13,95%CI:1.08~1.18,P0.001;PP分析:RR=1.13,95%CI:1.09~1.17,P0.001),降低不良反应发生率(RR=0.42,95%CI:0.35~0.50,P0.001)。根据亚组分析结果,芽胞杆菌联合H.pylori两种常规治疗方案[三联疗法(RR=1.22,95%CI:1.10~1.36,P=0.003),铋剂四联疗法(RR=1.11,95%CI:1.07~1.15,P0.001)]以及芽胞杆菌联合H.pylori常规治疗的两种疗程[10 d(RR=1.14,95%CI:1.04~1.24,P=0.006),14 d(RR=1.14,95%CI:1.07~1.21,P0.001)]均提高H.pylori根除率;而亚组分析芽胞杆菌种类中,地衣芽胞杆菌差异有统计学意义(RR=1.14,95%CI:1.10~1.19,P0.001),凝结芽胞杆菌与蜡样芽胞杆菌需扩大样本量进一步统计分析。结论芽胞杆菌联合疗法能有利于提高H.pylori根除率,并降低总不良反应的发生,相对于标准疗法有一定的意义。     

A statistical approach for the enhanced production of thermostable alkaline protease showing detergent compatibility activity from Bacillus circulans

Abstract

Response surface methodology (RSM) was employed to enhance the production of a thermostable alkaline protease from Bacillus circulans. Significant influences of peptone, yeast extract, and glucose on protease production were noted with a one-variable-at-a-time optimization strategy. Then, a full factorial central composite design (CCD) was applied to study the effects of glucose, peptone, and yeast extract to determine the optimal concentrations of these compounds for protease production by B. circulans under shake flask fermentation conditions. The statistical reliability and significance of the model was validated by an F-test for analysis of variance (ANOVA); enzyme production was improved significantly under optimized conditions. The enzyme was purified by ammonium sulphate fractionation, and gel filtration chromatography. Maximum enzyme activity was observed at 60°C temperature, and at pH 10. Alkaline protease from B. circulans showed excellent compatibility and stability in the presence of commercial detergents like Ariel, Surf Excel, Tide, Rin, Nirma, Wheel, and Doctor and showed excellent blood destaining effectiveness with commercial detergents.     

Molecular cloning and heterologous expression of an alkaline xylanase from Bacillus pumilus HBP8 in Pichia pastoris

The xynHB gene, encoding alkaline xylanase was cloned from Bacillus pumilus by a shot-gun method. The gene was cloned into vector pHBM905A, and expressed in Pichia pastoris GS115. Xylanase-secreting transformants were selected on plates containing RBB-xylan. Enzymatic activity in the culture supernatants was up to 644?U?mL^?1 and the optimal secretion time was 4 days at 25°C. SDS-PAGE showed two bands, of 32.2?kDa and 29.6?kDa, both larger than the predicted mass of 22.4?kDa based on its amino acid sequence. Zymogram analysis demonstrated that the enzyme in both bands could hydrolyze xylan. Deglycosylation by endoglycosidase H revealed that both were derived from the same protein but contain different extents of glycosylation (30 and 25%). The optimal pH and temperature of the enzyme was pH6–9 and 50°C, respectively.