Antimicrobial activity against select food-borne pathogens by phenolic antioxidants enriched in cranberry pomace by solid-state bioprocessing using the food grade fungus Rhizopus oligosporus

Listeria monocytogenes, Vibrio parahaemolyticus and Escherichia coli 0157:H7 are important food-borne pathogens. Control of these pathogens using synthetic antimicrobials such as currently approved antibiotics is challenging due to potential development of resistance. A profile of antimicrobials compared to a single compound could be potentially more effective. Cranberry pomace is a byproduct of the cranberry processing industry. Solid-state bioprocessing of cranberry pomace using the food grade fungus Rhizopus oligosporus improves phenolic and antioxidant profiles. We hypothesize that these phytochemicals mobilized during pomace bioprocessing could improve the antimicrobial functionality. The objective of this research was to use the food grade fungus R. oligosporus to release phenolic aglycones and relate its antioxidant functionality and diphenyl mobilization to antimicrobial activity against L. monocytogenes, V. parahaemolyticus and E. coli 0157:H7. Bioprocessing of pomace was done for 20 days with R. oligosporus. Total phenolics in water extracts were assayed using a Folin-Ciocalteu method. The antioxidant potential was measured using a β-carotene oxidation system (APF) and 1,1-diphenyl-2-picrylhydrazyl-radical (DPPH) system. Changes in phenolic profiles were analyzed using HPLC. Antimicrobial activities of the extracts during growth were tested using standard disk assays. Soluble phenolics, antioxidant activity and ellagic acid concentrations were enriched by bioprocessing and antimicrobial activity of the extracts against L. monocytogenes and V. parahaemolyticus correlated with highest soluble phenolics and APF in the same extracts. For E. coli 0157:H7 inhibition correlated with the extracts corresponding to highest DPPH and ellagic acid concentration. The bioprocessing-based antimicrobial activity depended on different phenolic functional properties of the extracts. Sensitivity towards soluble phenolics reflected in DPPH activity suggested inhibition by the disruption of the membrane by hyperacidification. Whereas, sensitivity to APF and ellagic acid suggests potential antimicrobial activity by membrane-transport disruption. The variation in sensitivity of pathogens has implications for designing new food grade antimicrobials. This bioprocessing strategy can be an innovative approach to produce broad spectrum antimicrobials against important food-borne pathogens.     

中国产石韦属植物孢子形态特征及分类学意义

利用光镜和扫描电镜,对石韦属(Pyrrosia)19种植物的孢子纹饰进行了研究。结果表明:19种石韦属孢子都为黄色,形态均为肾形,两侧对称,单裂缝,说明该属植物是一个自然类群。表面纹饰类型有3种,即瘤状、瘤状—疣状和瘤状—网脊状。孢子表面纹饰特征性状稳定,在种间存在较大差异。该研究结果可以为形态近似种的分类提供参考依据,同时也为石韦属属下分类系统的建立提供了重要证据。     

12种茶藨子属植物叶表皮微形态特征及其分类学意义

利用扫描电镜技术观察了茶藨子属7个亚属12种植物的叶表皮微形态。结果显示：叶表皮细胞分为多边形和不规则形两种类型;垂周壁式样为平直—弓形、浅波形、深波形;叶表皮角质层多为光滑或较光滑,少有脊状纹饰或乳突状凸起;表皮毛分为单毛和腺毛;黑果茶藨子、水葡萄、臭茶藨子的叶表面均分布不同密度的盾状腺毛,支持将臭茶藨子系划分为臭茶藨子亚属;气孔器均为无规则型,形状为圆形或椭圆形,多数种类的气孔仅分布在下表皮。茶藨子属植物叶表皮微形态特征可为属内亚属划分及种间分类提供一定的依据。     

23种顶蒴藓类孢子形态的观察

报道了中国２３种顶蒴藓类孢子的形态特征。观察结果显示：孢子形状有两类,球形和卵球形。孢子直径可分为４组：１０μm以下、１１－２０μm、２１－３０μm和３１μm以上。萌发孔有３种类型：近极薄壁区、单裂缝和无萌发孔。外壁纹饰有４种：棒状、瘤状、疣状和鼓槌状。孢子颜色有６种：黄褐色、黑褐色、金黄色、草绿色和紫红色。藓类孢子体积不同物种间有差异,但球形和卵球形是基本类型。苔藓孢子的萌发孔有３种类型：三裂缝为类     

Whey for mass production of Beauveria bassiana and Metarhizium anisopliae

Spore production of Beauveria bassiana and Metarhizium anisopliae was studied in a novel whey-based culture media. Spore yield and viability were determined for two B. bassiana (GHA-726 and CA-603) and two M. anisopliae (CA-1 and IMI 330189) isolates following production in three whey-based systems: solid, liquid, and a diphasic production system. Our study indicated that whey permeate can be used effectively for production of spores of entomopathogenic fungi. However, spore yield and viability were significantly influenced by fungal isolate, whey concentration, and the type of production process used. Under the conditions defined in the present study, spore yields ranging from 1.3 × 10⁹–10 × 10¹¹ spores l⁻¹ of whey medium could be obtained depending on the strain and production process used. Our study revealed that spores produced by all strains in whey-based solid and liquid media showed between 73–99 % viability; germination rates were comparable with those obtained using the standard SDA medium. In the two-stage production process, the viabilities of conidia produced by GHA-726, CA-603, and CA-1 were 35–86, 32–98, and 6–29 %, respectively; viability was correlated with whey concentration and isolates. Whey permeate can be used as a growth substrate for mass production of biocontrol fungi. We hypothesize that spore yield and viability could be improved by careful selection of whey content in the medium, incorporation of critical additives and optimization of culture conditions.     

Production of phenolic antioxidants by the solid-state bioconversion of pineapple waste mixed with soy flour using Rhizopus oligosporus

The ability of Rhizopus oligosporus to produce enhanced levels of free phenolics from pineapple residue mixed with soy flour as potential nitrogen source was investigated. Concurrently, phenolic-linked β-glucosidase and the antioxidant activity of the extracts were followed. Two treatments were studied: (A) 9 g of pineapple residue and 1 g of soy flour (P9); (B) 5 g of pineapple residue and 5 g of soy flour (P5). The increase of water extractable phenolics was 39.3% for P9 treatment and 79.4% for P5 treatment. During early stages of growth high antioxidant activity, low phenolic content and low β-glucosidase specific activity was observed. High antioxidant activity was likely due to the presence of insoluble polymeric phenolics, know for their high antioxidant activity. A marked decrease of the antioxidant activity of P5 treatment during late stages of growth was observed due to likely formation of free soluble phenolics. The moderate total phenolics content and high β-glucosidase specific activity of P9 treatment in late stages is likely the consequence of low nitrogen content in this treatment. The bioconversion of pineapple residue by R. oligosporus leads to enhanced levels of phenolic compounds, mainly for P5 treatment. This approach offers a novel strategy to enhance the value of pineapple wastes.     

Expansion of Rosa rugosa and Hippophaë rhamnoides in coastal grey dunes: Effects at different spatial scales

The study analysed the effects of shrub expansion on vegetation composition and plant species diversity in coastal grey dunes on the North Sea island Spiekeroog, comparing Rosa rugosa and Hippophaë rhamnoides. Species composition was recorded in plots of two spatial scales, 1 and 16 m², considering the full range of shrub cover from less than 10 to almost 100%. Although R. rugosa and H. rhamnoides established and spread in the same grey dune environment, the vegetation of the two shrubland types was much different. While the H. rhamnoides plots were relatively species-rich, characterised by remnant grey dune vegetation with many small, often annual, light-demanding species except in the densest shrubs, the R. rugosa plots were clearly species-poorer due to the loss of many typical grey dune species, including only few shade-tolerant taxa. The total number of species, the number of herbaceous species and of species typical for grasslands decreased with increasing cover of H. rhamnoides and R. rugosa at both spatial scales. For the number of shrubs and shrubland species, hardly any significant effects of shrub cover were observed in R. rugosa, while there were positive effects in H. rhamnoides. Both the Shannon index and evenness decreased with increasing cover of the two shrub species at both spatial scales. Here, the decline in species diversity was more improved in R. rugosa than in H. rhamnoides.     

Motility and chemotactic response of Pseudomonas fluorescens toward chemoattractants present in the exudate of Macrophomina phaseolina

Pseudomonas fluorescens strains (LAM1-hydrophilic) and (LAM2-hydrophobic) showed positive chemotaxis towards attractants (sugars, amino acids, polyols and organic acids) present in the exudate of Macrophomina phaseolina (a soilborne plant pathogenic fungus). The varied response of motility traits such as speed, rate of change in direction (RCDI) and net to gross displacement ratio (NGDR) was observed for different chemoattractants. Swimming speed of the strains was highest in 10-fold diluted exudate or 100–1000 μM strength of different attractants, but further dilutions significantly decreased the swimming speed (P = 0.05). Chemotactic response of P. fluorescens was positively correlated with swimming speed (P = 0.05; r = 0.76). Relative to control, the RCDI values decreased 1.5-fold in amino acids or sugars, and 1.2-fold in polyols or organic acids. With increase in swimming speed, the NGDR of both strains also increased, but the RCDI decreased. Both hydrophilic and hydrophobic strains did not show significant differences in their motility traits. The results demonstrate that M. phaseolina exudate contains chemical attractants that serve as signal for flagellar motility of P. fluorescens. Motile P. fluorescens strains thus may consume fungal exudate as nutrients, and thus spores could offer a niche for these bacteria in soil.     

Spore morphology as a taxonomic tool in the delimitation of three Asplenium L. species complexes (Aspleniaceae: Pteridophyta) in Cuba

As a contribution to the taxonomic study of Asplenium L. for the new "Flora de la República de Cuba", the spore morphology was studied into three complexes of hardly delimited species. Twenty-two collections have been studied by SEM and eighty-nine by LM. Shape, number of spores per sporangium and perispore ornamentation, specially fine details revealed by scanning electron microscopy, were the most valuable characters in the recognition of taxa. Irregularities in spore shape and size, unusual development of perispore wall surface elements, and collapsed protoplast characterize aborted spores and suggest the occurrence of a possible hybrid species.     

感染羽茅的香柱菌属内生真菌对丛枝菌根真菌孢子萌发的影响

内生真菌与丛枝菌根(AM)真菌是构成草原生态系统的重要组成部分.内生真菌会抑制其宿主植物的AM真菌侵染率.本研究以感染2种香柱菌属内生真菌[Epichloё gansuensis(Eg)和E. sibirica(Es)]的天然禾草羽茅为供试材料,进行体外纯培养的内生真菌培养滤液、感染内生真菌的羽茅叶片(包括鲜叶和枯叶)浸提液,以及根系分泌物对摩西球囊霉(Gm)和幼套球囊霉(Ge)2种AM真菌孢子萌发影响试验.结果表明: 香柱菌属内生真菌的培养滤液会显著抑制2种AM真菌孢子的萌发,而感染香柱菌属内生真菌的羽茅根系分泌物只对Ge孢子萌发有显著抑制作用,且上述抑制作用与内生真菌种类无关;鲜叶浸提液对Gm和Ge的孢子萌发率均无显著影响,而枯叶浸提液对Ge的孢子萌发有显著抑制作用.在自然生态系统中,香柱菌属内生真菌通常存在于宿主植物体内,可能通过影响宿主植物的根系分泌物来影响AM真菌孢子的萌发.     

Ambispora granatensis, a new arbuscular mycorrhizal fungus, associated with Asparagus officinalis in Andalucia (Spain)

A new dimorphic fungal species in the arbuscular mycorrhiza-forming Glomeromycota, Ambispora granatensis, was isolated from an agricultural site in the province of Granada (Andalucía, Spain) growing in the rhizosphere of Asparagus officinalis. It was propagated in pot cultures with Trifolium pratense and Sorghum vulgare. The fungus also colonized Ri T-DNA transformed Daucus carota roots but did not form spores in these root organ cultures. The spores of the acaulosporoid morph are 90-150 μm diam and hyaline to white to pale yellow. They have three walls and a papillae-like rough irregular surface on the outer surface of the outer wall. The irregular surface might become difficult to detect within a few hours in lactic acid-based mountings but are clearly visible in water. The structural central wall layer of the outer wall is only 0.8-1.5 μm thick. The glomoid spores are formed singly or in small, loose spore clusters of 2-10 spores. They are hyaline to pale yellow, (25)40-70 μm diam and have a bilayered spore wall without ornamentation. Nearly full length sequences of the 18S and the ITS regions of the ribosomal gene place the new fungus in a separate clade next to Ambispora fennica and Ambispora gerdemannii. The acaulosporoid spores of the new fungus can be distinguished easily from all other spores in genus Ambispora by the conspicuous thin outer wall.     

SPORE MORPHOLOGY IN THE GENUS BRUCHIA SCHWAEGR. (MUSCI)

Scanning electron microscopy observations of the spores of Bruchia have resulted in the recognition of four spore types based on the ornamentation of the distal spore surface: warty or verrucate, pitted, reticulate, and spinose. The proximal surface of the spores of all species, except B. brevipes, is characterized by a central aperture region surrounded by a triangular murus or rows of spinae forming a triangle and 1 or 2 smooth or verrucate collars. The ornamentation patterns observed are considered to be characteristic for the genus. Spore morphology alone can rarely be used to distinguish species but in conjunction with certain other characteristics, it is an important taxonomic feature. Spore morphology is a major characteristic used to define the limits of the highly variable species B. flexuosa (spinose spores) and to distinguish it from the closely related species B. texana (reticulate spores). Variations in ornamentation patterns within the genus support the recognition of two subgenera (subgenus Bruchia and subgenus Sporledera). Spore morphology also supports the close relationship between Bruchia and Trematodon and is sufficient to eliminate several questionable taxa from the genus.     

15个杜鹃花品种叶片解剖结构与植株耐热性的关系

以15个杜鹃花品种为试材,利用扫描电镜观察叶片17项解剖结构,应用变异系数、相关分析和聚类分析对叶片解剖结构进行筛选,采用隶属函数对各品种的耐热性进行综合评价,探讨杜鹃花叶片解剖结构与植株耐热性的关系,为杜鹃花耐热品种选育、引种栽培及栽培区划分提供细胞学理论依据.结果表明: 杜鹃花叶片为典型的异面叶,表皮细胞垂周壁略呈波浪状,气孔为无规则形,仅分布于下表皮,品种之间有16项解剖结构指标差异达到极显著水平,仅下表皮厚度为显著水平;叶片角质层厚度、气孔密度、气孔宽度、栅栏组织厚度和组织结构疏松度是影响耐热性的主要叶片解剖结构性状,而叶片厚度、气孔长度、孔径、气孔开度、海绵组织厚度、栅海比、上下表皮细胞长度和厚度、组织结构疏松度、中脉厚度与植株耐热性的关系不大.15个品种杜鹃花耐热性存在一定差异,其强弱顺序为:松江大桃红>状元红>绿色光辉>粉珍珠>外国红>蓝茵>笔止>大和之春>国旗红>玉玲珑>红珊瑚>宁波红>套瓣朱砂>爱丁堡>琉球红.15个品种杜鹃花抗热性可划分为4类:松江大桃红、状元红和绿色光辉为抗热类型;粉珍珠、外国红、蓝茵、笔止、大和之春、国旗红和玉玲珑为中度抗热类型;红珊瑚、宁波红、套瓣朱砂和爱丁堡为低抗热类型;琉球红为不抗热类型.杜鹃花耐热性的准确评价需参考形态结构、生理生化、遗传等因素,以及杜鹃花高温受害及受害后恢复状况.     

Streptomyces torulosus sp. n., an Unusual Knobby-Spored Taxon

Several strains of a streptomycete with unusual spore wall ornamentation were isolated from Iowa soils. They are further characterized by coiled chains of spores, gray aerial mycelium, melanin-like pigments, and the production of antibacterial and antifungal antibiotics. Spore wall ornamentations, as seen with the transmission electron microscope, do not correspond with any of the four generally recognized categories of spore wall ornamentation-smooth, warty, spiny, or hairy-but seem to represent an intermediate position between warty and spiny. This unusual ornamentation consists of blunt projections rising out of an otherwise smooth surface and is termed "knobby." To obtain further information on the nature of the knoblike projections and their possible status in representing a defined spore wall ornamentation category, scanning electron micrographs and transmission electron micrographs representing all sporewall ornamentation categories were compared. Differentiation between the several categories was more difficult with the scanning electron microscope than with the transmission electron microscope.     

Formation of spore ornamentation in two Sphaerophorus species

Two different ways of achieving a spore ornamentation have been demonstrated in Sphaereophorus , belonging to the Caliciales. In S. globosus the ornamentation is formed within the ascus by an external secondary spore wall in an ontogenetic process with several unique features. In S. murrayi the ornamentation is formed at a late stage, when the spores have been released from the asci. Carbonaceous material formed among the asci and paraphyses is added to the surface of the primary wall, and a very irregular ornamentation is formed. The name Sphaerophorus murrayi Ohlsson is validated.     

新疆曲尾藓属10种植物蒴齿和孢子的形态观察

该研究通过光学显微镜(LM)和扫描电镜(SEM)观察了曲尾藓属(Dicranum Hedw.)10种植物的蒴齿和孢子的形态特征。结果表明:(1)曲尾藓属10种植物的孢蒴均为圆柱形。(2)孢子球形、近球形、椭圆形或三角状卵形,颜色为黄色、棕色或黄褐色;近极面向内凹,纹饰为疣状、颗粒状或芽孢状。(3)曲尾藓属10种植物的蒴齿多为披针形,但也有阔披针形,颜色为杏黄色、深红色、褐黄色或红褐色;纹饰为条状或疣状,具穿孔成网状结构,一些种无穿孔成平滑结构,表面具细疣。该研究结果为曲尾藓属属下类群划分提供了分类依据,并为苔藓植物系统分类、演化研究提供基础资料。     

苔藓植物孢子形态的研究(一)——几种泥炭藓(Sphagnum)孢子的光学研究和电子显微镜观察

泥炭藓(Sphagnum)是水湿或沼泽地区丛生藓类。其植物体形态和构造分化简单,孢子体仅具由配子体组织发育的假蒴柄;孢蒴球形,盖裂,无蒴轴及蒴齿分化,因而为鲜纲中一个独立的亚纲,在苔藓系统排列中,列为原始的类型之一。     

烟气对泥炭藓孢子萌发影响的模拟研究

适量烟气能促进种子萌发,但对苔藓植物孢子的作用尚不清楚.选取采自长白山区泥炭地的粗叶泥炭藓和中位泥炭藓的孢蒴为试验材料,通过燃烧泥炭地植物产生烟气,制备烟溶液,分别与不同大小(大:直径为2.10～2.50 mm;小:直径为1.50～1.90 mm)以及不同保存时长(旧:4.3和6.3年;新:0.3年)的孢蒴进行两组双因素试验,经不同时长的烟溶液浸泡和萌发试验,模拟研究烟气、孢蒴大小和保存时长对苔藓植物孢子萌发的影响.结果表明: 烟溶液浸泡影响孢子萌发,培养10 d时,不同时长的烟溶液浸泡均可使孢子的萌发率提高5倍以上,小孢蒴孢子的萌发率高;培养21 d时,仅适度浸泡(3 d)表现出促进萌发的作用,孢蒴大小对孢子萌发率无影响;烟溶液浸泡对长时间保存(4.3和6.3年)的孢蒴孢子萌发无促进作用.研究表明,适量烟气可加速新泥炭藓孢子以及小孢蒴孢子的萌发.在存在不定期火烧干扰的泥炭地中,与对种子植物的作用类似,烟气可能在苔藓植物种群的有性更新和种群维持中发挥重要作用.     

祖先性状重建法揭示铁线蕨属植物孢子表面纹饰的形态多样性及其演化

蕨类植物孢子表面的纹饰形态复杂多样, 在分类学和古生物学研究中具有重要意义, 目前探索祖先性状演化常用形态学与系统发育学结合的方法。本文选取一回羽叶铁线蕨类植物为材料, 用扫描电子显微镜观察其孢子形态, 用5个叶绿体基因序列构建系统树, 用最大似然法和最大简约法来分析孢子性状的演化。结果表明: 一回羽叶铁线蕨类的孢子纹饰有5种类型, 即光滑、粗糙、颗粒、疣状及瘤状; 这一类群的祖先孢子纹饰有很大的可能是瘤状纹饰; 疣状纹饰可能是由瘤状纹饰演化而来; 孢子纹饰由简单向复杂演化, 越进化的物种其孢子纹饰越复杂。     

金针菇担孢子核相及遗传属性的研究

以3个不同的金针菇菌株为材料,研究了其担孢子的核相及遗传属性。荧光染色观察显示,担孢子核相以双核为主,双核孢子、单核孢子和无核孢子分别占80.2%、7.5%和12.3%。源于单孢分离物的菌丝为有隔膜、无锁状联合的多核菌丝。在交配试验中,源于不同菌株单孢分离物的菌丝原生质体的配对形成具锁状联合的菌落,而源于同一单孢分离物的菌丝原生质体的配对则形成无锁状联合的菌落,暗示担孢子中的两个核具有相同的交配型。RAPD分析显示,源于同一单孢分离物的菌丝原生质体为10个随机引物所扩增的图谱彼此完全相同,印证了担孢子中的双核是同质的。此外,观察表明,一个担子上着生有4个担孢子。因此,金针菇是一种具4个含同质双核担孢子的四极性蕈菌。