棒状腐败乳杆菌Lc7的生物学特性及益生作用

【目的】对分离自健康成人粪便样本的棒状腐败乳杆菌(Loigolactobacillus coryniformis)Lc7进行分类学鉴定和益生潜力评估。【方法】基于16SrRNA基因和基因组核心基因构建系统发育树,对Lc7进行分类学鉴定;通过耐酸和胆汁酸盐、粘附、抗氧化和抑菌实验,以及溶血、明胶酶活性和抗菌药物敏感性实验,评估Lc7的益生特性。同时,构建小鼠溃疡性结肠炎模型,评估Lc7的体内抗炎潜力。【结果】Lc7鉴定为L. coryniformis,在酸和胆汁酸盐的连续作用下,Lc7的存活率为70.17%。Lc7对HT-29细胞的粘附指数为56.33 CFU/cell,其自聚集和疏水性分别为80%和40%;Lc7对福氏志贺菌和鼠伤寒沙门菌等7个常见致病菌均有较强的抑制能力;对1,1-二苯基-2-苦基肼(1,1-diphenyl-2-picryl-hydrazyl,DPPH)和羟自由基(hydroxyl radicals,·OH)的清除率分别为91.70%和48.53%;Lc7无溶血现象和明胶酶活性,对选取的大多数抗生素均敏感。在小鼠结肠炎实验中,Lc7干预组小鼠结肠长度明显长于模型组(...     

黔西水城晚二叠世长兴期钙藻和有孔虫及其古环境意义

在黔西水城地区的K576井长兴组共鉴定钙藻3属3种,包括Gymnocodium bellerophontis、Permocalculus sp.和Tauridiumkurdistanensis;有孔虫8属10种,其中(虫筳)类2属2种,有孔虫动物群主要由Reichelinasp.indet.、Nankinella sp.、Pachyphloia schwageri、Pachyphloia sp.、Geinitzina sp.、Nestellorella sp. indet.、Howchinella sp.、Hemigordius aff. saranensis、Hemigordius sp.和Midiella sp. indet.组成。将本井按照生物特征分为有孔虫-钙藻-介形虫组合、有孔虫-腕足类-介形虫组合、介形虫-双壳类-腹足类组合、有孔虫-钙藻-双壳类组合、有孔虫-腕足类-介形虫组合、有孔虫-钙藻-双壳类组合和介形虫组合等7个组合。按照层序地层划分、垂向沉积序列特征和测井资料的分析,有孔虫-钙藻-介形虫组合(SQ3-3)和有孔虫-腕足类-介形虫组合(SQ3-4)时期地层为三角洲前...     

The potato StMKK5-StSIPK module enhances resistance to Phytophthora pathogens through activating the salicylic acid and ethylene signalling pathways

Mitogen-activated protein kinase (MAPK) cascades play pivotal roles in plant responses to both biotic and abiotic stress. A screen of a Nicotiana benthamiana cDNA virus-induced gene silencing (VIGS) library for altered plant responses to inoculation with Phytophthora infestans previously identified an NbMKK gene, encoding a clade D MAPKK that we renamed as NbMKK5, which is involved in immunity to P. infestans. To study the role of the potato orthologous gene, referred to as StMKK5, in the response to P. infestans, we transiently overexpressed StMKK5 in N. benthamiana and observed that cell death occurred at 2 days postinfiltration. Silencing of the highly conserved eukaryotic protein SGT1 delayed the StMKK5-induced cell death, whereas silencing of the MAPK-encoding gene NbSIPK completely abolished the cell death response. Further investigations showed that StMKK5 interacts with, and directly phosphorylates, StSIPK. Furthermore, both StMKK5 and StSIPK trigger salicylic acid (SA)- and ethylene (Eth)-related gene expression, and co-expression of the salicylate hydroxylase NahG with the negative regulator of Eth signalling CTR1 hampers StSIPK-triggered cell death. This observation indicates that the cell death triggered by StMKK5-StSIPK is dependent on the combination of SA- and Eth-signalling. By introducing point mutations, we showed that the kinase activity of both StMKK5 and StSIPK is required for triggering cell death. Genetic analysis showed that StMKK5 depends on StSIPK to trigger plant resistance. Thus, our results define a potato StMKK5-SIPK module that positively regulates immunity to P. infestans via activation of both the SA and Eth signalling pathways.     

融入生态安全的兰西城市群土地利用功能布局优化

区域生态安全对协调土地利用和生态环境的关系、促进资源的合理配置以及实现区域协调发展具有重要的意义。以兰西城市群作为典型地区,将形态空间格局分析(MPSA)应用到土地利用功能优化配置研究中,通过构建MSPA-GMOP-PLUS耦合模型,将生态安全纳入城市群土地利用功能和空间结构优化中,为区域布局优化提供了新思路。结果表明：(1)生态源地对区域生态安全具有重要意义,2020年兰西城市群共有20个生态源地,生态核心总面积为43355.5 km²。此外,在土地利用功能分区中,生态保护区面积占比仍为最大(30.7%),生态边缘区面积也占19.2%,为生态保护提供了缓冲地带。(2)基于MSPA的仿真模拟可以限制城市群不平衡的发展趋势,优于传统土地利用规划模式。新模式下的生产、生活、生态用地占比分别由传统模式的30.17%、11.25%、55.58%优化至31.92%、10.70%和57.38%。优化结果不仅保证了优质生产空间的稳定性,而且有效确保了生态空间的可持续性。此外,不同开发情景的下土地利用布局结果,可以作为提高土地资源利用率和制定土地利用空间规划的参考。     

滇南地区桃树种植模式对土壤有机碳组分及碳库管理指数的影响

经果林种植可改变土壤质量、改善生态环境,同时具有较高的经济效益。合理的种植模式可通过物种间的互补性提高资源利用效率,改善土壤碳库质量并提高综合效益。为探讨桃树种植模式对土壤有机碳组分及碳库管理指数的影响,以云南省开远市不同桃树种植模式(桃树单种-SP和桃树南瓜套种-PP)为研究对象,以毗邻的天然林地(CK)为对照,分析不同种植模式下活性碳库,即高锰酸钾氧化有机碳(POXC)、颗粒有机碳(POC),惰性有机碳库即矿物结合态有机碳(MAOC)在0—40 cm土层的分布情况,明确土壤有机碳组分与土壤理化性质的关系;计算碳库活度指数(CPAI)、碳库指数(CPI)以及碳库管理指数(CPMI),明确不同桃树种植模式的碳库变化情况。结果表明：桃树种植模式和对照的土壤有机碳组分的含量均随着土层深度的增加而减少,平均土壤有机碳(SOC)含量为：14.68 g/kg(CK)>9.57 g/kg(PP)>8.58 g/kg(SP)。平均活性有机碳组分所占比例与POC/MAOC均表现为：SP>CK>PP,PP的活性有机碳比例较低,具有较高的有机碳稳定性。两种桃树种植模式的CPMI在1...     

植物乳杆菌谷氨酸脱羧酶催化pH范围的理性改造及高效转化生产g-氨基丁酸

γ-氨基丁酸可由谷氨酸脱羧酶(glutamate decarboxylase, GAD)催化谷氨酸一步合成,反应体系成分简单、环境友好。然而,绝大多数GAD酶催化pH偏酸性且反应范围狭小,需要加入无机盐维持最适催化环境,增加了生产附加成分。此外,随着产物γ-氨基丁酸的生成,溶液pH会逐渐上升,不利于GAD酶的持续转化。本研究首先从实验室保藏的一株高产γ-氨基丁酸的植物乳杆菌(Lactobacillus plantarum)中克隆得到谷氨酸脱羧酶LpGAD,基于酶蛋白表面电荷修饰,选择9个位点进行定点突变及组合突变,酶学性质表征结果显示三突变体LpGAD^{S24R/D88R/Y309K}在催化pH区间内酶活力整体提高,尤其拓宽了在偏中性pH 6.0下的酶活,为野生酶的1.68倍。接下来,通过分子动力学模拟解析了酶活提高的机理。此外,将Lpgad与Lpgad^{S24R/D88R/Y309K}突变基因分别在谷氨酸棒杆菌(Corynebacterium glutamicum) E01中过表达,通过优化确定了摇瓶最适转化条件为反应温度40 ℃,菌体量OD₆₀₀=20,底物L-谷氨酸100.0 g/L,5-磷酸吡哆醛添加量为100 μmol/L。5 L发酵罐中,不调节pH,通过分批投料底物L-谷氨酸,γ-氨基丁酸产量高达402.8 g/L,较对照菌株提高了1.63倍。本研究成功拓宽了LpGAD的pH催化范围及酶活,提高了γ氨基丁酸的转化效率,为实现其规模化工业生产奠定了基础。     

-cyano-substituted analogoues of decarboxylated S-adenosylmethionine as enzyme activated, irreversible inhibitors of S-adenosylmethionine decarboxylase

A pair of -cyano analogues of decarboxylated S-adenosylmethionine (2a and 2b) were synthesized as potential enzyme activated, irreversible inhibitors of the[pyruvoyl enzyme S-adenosylmethionine decarboxylase (AdoMet-DC). Each of these analogues acts as an irreversible inactivator for ADoMet-DC from Escherichia coli (IC₅₀ values of 9 and 50 μM, respectively). These analogues also inactivate human AdoMet-DC, with K_I values of 246.6 and 7.2 μM, and k_inact values of 0.29 and 0.03 min⁻¹, respectively.     

Preparation of the pure diastereomeric forms of S- (5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2- cyclopentene and their evaluation as irreversible inhibitors of S-adenosylmethionine decarboxylase from Escherichia coli

The conformationally restricted S-adenosylmethionine analogue AdoMac (S-(5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2-cyclopentene has been shown to act as an enzyme activated, irreversible inhibitor of theEscherichia coli form of the enzyme S-adenosylmethionine decarboxylase. Inactivation of the enzyme is presumably initiated by formation of an imine linkage between the inhibitor and the terminal pyruvate of the enzyme, followed by base-catalyzed elimination of methylthioadenosine and generation of a latent electrophile. Removal of the driving force for the elimination of methylthioadenosine resulted in a reversibly binding inhibitor. Thus, the thioether analogue corresponding to AdoMac, and the corresponding dihydro derivative (H₂-AdoMac), reversibly inhibit the enzyme. AdoMac was resolved into its four pure diastereomeric forms, and each diastereomer was evaluated as an irreversible inhibitor of the enzyme. The K_I values for the individual diastereomers range between 3.83 and 39.6 μM, with the cis-1S,4R diastereomer being the most potent inhibitor. However, the k_inact values for the four diastereomers are not significantly different, suggesting that the binding of each diastereomer to the enzyme is configuration-dependent, while the subsequent inactivation likely proceeds through a single intermediate which is formed from each of the four diastereomers. Since each pure diastereomer represents a distinct conformational mimic exhibiting restricted sidechain rotation, the data suggests that these and related analogues may be useful as conformational probes for the catalytic site of AdoMet-DC.     

洞庭平原褐家鼠年龄分组及种群年龄动态分析

本文采用1986年10月—1989年12月在洞庭平原收集的褐家鼠(Rattus norvegicus)标本,以胴体重作指标,参考繁殖特征,将褐家鼠划分5个年龄组:Ⅰ.幼体组,雌鼠<60克,雄鼠<70克;Ⅱ.亚成体组,雌鼠60—99克,雄鼠70—119克;Ⅲ.成体Ⅰ组,雌鼠100—139克,雄鼠120—169克;Ⅳ.成体Ⅱ组,雌鼠140—189克,雄鼠170—219克;Ⅴ.老体组,雌鼠>190克,雄鼠>220克。种群年龄结构的季节动态特征是:开春时以Ⅲ、Ⅳ组占优势,初夏时Ⅰ、Ⅱ组明显增加,7、8、9月各年龄组比例较均匀,冬季以Ⅱ、Ⅲ组为主。还探讨了年龄与体重、体长、尾长及繁殖率的相互关系。