Discovery of novel intermediate forms redefines the fungal tree of life

Fungi are the principal degraders of biomass in terrestrial ecosystems and establish important interactions with plants and animals. However, our current understanding of fungal evolutionary diversity is incomplete and is based upon species amenable to growth in culture. These culturable fungi are typically yeast or filamentous forms, bound by a rigid cell wall rich in chitin. Evolution of this body plan was thought critical for the success of the Fungi, enabling them to adapt to heterogeneous habitats and live by osmotrophy: extracellular digestion followed by nutrient uptake. Here we investigate the ecology and cell biology of a previously undescribed and highly diverse form of eukaryotic life that branches with the Fungi, using environmental DNA analyses combined with fluorescent detection via DNA probes. This clade is present in numerous ecosystems including soil, freshwater and aquatic sediments. Phylogenetic analyses using multiple ribosomal RNA genes place this clade with Rozella, the putative primary branch of the fungal kingdom. Tyramide signal amplification coupled with group-specific fluorescence in situ hybridization reveals that the target cells are small eukaryotes of 3-5?μm in length, capable of forming a microtubule-based flagellum. Co-staining with cell wall markers demonstrates that representatives from the clade do not produce a chitin-rich cell wall during any of the life cycle stages observed and therefore do not conform to the standard fungal body plan. We name this highly diverse clade the cryptomycota in anticipation of formal classification.     

东寨港红树林丛枝菌根真菌侵染和孢子密度及其影响因子研究

丛枝菌根(AM)结构存在于多种红树植物根内,但目前对于红树林中AM真菌的研究还未能完全揭示其存在的原因和状况。研究通过对海南东寨港红树林中不同潮位16种红树植物根内AM侵染情况、AM真菌孢子密度以及土壤理化因子开展调查,以了解东寨港红树林中AM真菌的存在状况及其影响因素。结果表明12种红树植物与AM真菌存在共生关系,其中海漆(Excoecaria agallocha)、许树(Clerodendrum inerme)、水黄皮(Pongamia pinnata)和黄槿(Hibiscus tiliaceus)侵染率较高,均高于55%;16种植物根际土壤中均检测到AM真菌孢子,每25 g土壤的平均孢子密度为(25.7±2.7)个。方差分析表明,不同宿主植物根内AM侵染情况差异显著;潮位对AM侵染率和孢子密度均具有显著影响,表现为高潮位显著高于中、低潮位,而中低潮位间无显著差异。线性回归分析表明土壤含水率、电导率和有效磷含量可能是影响东寨港红树林生境中AM侵染率和孢子密度的主要土壤因子。     

FAST台址环境对GPS-RTK测量精度的影响及应对措施

FAST（Five-hundred-meter Aperture Spherical radio Telescope,500 m口径球反射面射电望远镜）是我国正在贵州建设的重大科学项目,建成后将成为世界上最大的单口径射电望远镜.其中FAST馈源一次支撑测量精度要求17 mm,将采用GPS-RTK与全站仪两种测量方式,本文主要通过实测数据分析FAST台址洼地环境对GPS-RTK测量精度造成的影响,并根据FAST台址地形图和馈源舱的运行轨迹制定相应的解决方案.     

影响AM真菌孢子萌发因素的研究

在离体条件下培养AM真菌孢子,测定不同培养基pH值、成分以及培养温度条件下的孢子萌发率,优化AM真菌孢子的培养环境.结果表明：培养基pH6.0时,孢子萌发率最高,过高或过低都会抑制孢子的萌发.AM真菌在土壤中储存两个月最有利于孢子萌发.培养基中添加人工合成植物激素GR24对孢子萌发率有极显著影响,浓度450 pg/L时,孢子萌发率最高.低浓度的精胺能够增加孢子菌丝的生长点个数,提高孢子的萌发率,含量过高时（＞40 mg/L）抑制孢子的萌发.培养温度30℃较适合AM真菌孢子的萌发,低于或高于30℃孢子萌发率有所下降.本试验结果将为构建AM真菌-毛状根双重培养体系提供适宜的培养条件.     

Genome sequence and gene compaction of the eukaryote parasite Encephalitozoon cuniculi.

Microsporidia are obligate intracellular parasites infesting many animal groups. Lacking mitochondria and peroxysomes, these unicellular eukaryotes were first considered a deeply branching protist lineage that diverged before the endosymbiotic event that led to mitochondria. The discovery of a gene for a mitochondrial-type chaperone combined with molecular phylogenetic data later implied that microsporidia are atypical fungi that lost mitochondria during evolution. Here we report the DNA sequences of the 11 chromosomes of the approximately 2.9-megabase (Mb) genome of Encephalitozoon cuniculi (1,997 potential protein-coding genes). Genome compaction is reflected by reduced intergenic spacers and by the shortness of most putative proteins relative to their eukaryote orthologues. The strong host dependence is illustrated by the lack of genes for some biosynthetic pathways and for the tricarboxylic acid cycle. Phylogenetic analysis lends substantial credit to the fungal affiliation of microsporidia. Because the E. cuniculi genome contains genes related to some mitochondrial functions (for example, Fe-S cluster assembly), we hypothesize that microsporidia have retained a mitochondrion-derived organelle.     

地道药材凤丹根际AM真菌与丹皮酚含量的相关性分析

凤丹(Paeonia ostii)含有重要药用价值的次生代谢产物丹皮酚.丛枝菌根真菌(AM真菌)与凤丹存在密切的共生关系,菌根真菌能够直接或间接影响植物次级代谢过程.采用HPLC法测定凤丹中丹皮酚的含量、碱解离-酸性品红染色法测定侵染率、曲利苯兰染色法测定菌丝量、湿筛沉淀法测定孢子密度.结果发现,凤丹根际丛枝菌根侵染率、孢子密度和菌丝量与丹皮酚含量呈显著的正相关.本文通过AM真菌的孢子密度、菌丝量、侵染率与凤丹丹皮酚含量的相关性分析,表明AM真菌是凤丹中丹皮酚含量的促进因素,凤丹体内AM真菌侵染越强,根际土壤中AM孢子产生越多,土壤菌丝量越高,则丹皮酚含量积累越高.研究AM真菌对丹皮酚次级代谢物生成的影响,对进一步研究AM真菌对丹皮酚在药用价值方面提供重要依据.     

网状球囊霉孢子萌发和菌丝生长条件的优化

为了确定AM真菌孢子萌发和菌丝生长的最佳水平条件,采用L16(45)多因子正交实验对影响网状球囊霉孢子萌发和菌丝生长的因素进行了优化选择.结果表明:优组合为A4B2C2D3E3,即pH 8,水琼脂质量浓度10 g/L,培养周期10 d,后熟条件20 d,培养温度28℃为最佳组合.同时发现在优组合下孢子不仅萌发得早,萌发率高,菌丝生长也很旺盛.     

加压介质对蓝色犁头霉孢子存活率及甾体转化的影响

分别以N2、高纯空气[V（O2）：V（N2）=21：79]和CO2为加压介质，研究高压条件下蓝色犁头霉孢子存活率的变化情况。在3种不同加压介质下，蓝色犁头霉孢子存活率显示出相同的变化规律，随压力升高，孢子存活率逐步降低；压力一定时，蓝色犁头霉孢子存活率随保压时间的延长而降低；在相同压力和保压时间下，蓝色犁头霉孢子在高纯空气中的存活率明显高于在N2和CO2中的孢子存活率，其中蓝色犁头霉孢子在CO2中的存活率始终低于其他两种加压介质；在适宜的压力和加压介质条件下，压力可显著提高蓝色犁头霉氢化可的松产量。     

The control of moldy risk during rice storage based on multivariate linear regression analysis and random forest algorithm

Clarifying the mechanism of fungi growth is of great significance for maintaining the quality during grain storage. Among the factors that affect the growth of fungi spores, the most important factors are temperature, moisture content and storage time. Therefore, through this study, a multivariate linear regression model among several important factors, such as the spore number and ambient temperature, rice moisture content and storage days, were developed based on the experimental data. In order to build a more accurate model, we introduce a random forest algorithm into the fungal spore prediction during grain storage. The established regression models can be used to predict the spore number under different ambient temperature, rice moisture content and storage days during the storage process. For the random forest model, it could control the predicted value to be of the same order of magnitude as the actual value for 99% of the original data, which have a high accuracy to predict the spore number during the storage process. Furthermore, we plot the prediction surface graph to help practitioners to control the storage environment within the conditions in the low risk region.     

Ribosomal RNA sequence suggests microsporidia are extremely ancient eukaryotes

The microsporidia are a group of unusual, obligately parasitic protists that infect a great variety of other eukaryotes, including vertebrates, arthropods, molluscs, annelids, nematodes, cnidaria and even various ciliates, myxosporidia and gregarines. They possess a number of unusual cytological and molecular characteristics. Their nuclear division is considered to be primitive, they have no mitochondria, their ribosomes and ribosomal RNAs are reported to be of prokaryotic size and their large ribosomal subunit contains no 5.8S rRNA. The uniqueness of the microsporidia may reflect their phylogenetic position, because comparative sequence analysis shows that the small subunit rRNA of the microsporidium Vairimorpha necatrix is more unlike those of other eukaryotes than any known eukaryote 18S rRNA sequence. We conclude that the lineage leading to microsporidia branched very early from that leading to other eukaryotes.     

组培条件对金发藓Polytrichum commune (Bryopsida:Musci)孢子萌发的影响(英文)

在组培条件下探讨了培养基类型,蔗糖浓度,光照强度,温度和铅离子对金发藓孢子萌发的影响。结果表明:含有丰富营养成分的培养基有利于孢子的两极萌发,而含有低浓度营养成分的培养基则有利于孢子的萌发。在含有矿质离子的培养基中,光对孢子萌发没有影响。然而,在没有矿质离子的SM培养基中,光能显著地抑制孢子的萌发。在w=0.5%的蔗糖浓度和20～30℃的温度范围内,孢子萌发率较高。随着铅浓度的升高,孢子萌发率显著降低。组培条件下金发藓孢子萌发所需的上述条件也符合其野外生长的环境特征。研究结果揭示了对金发藓孢子萌发起促进或抑制的环境因子,有助于在野外或组培条件下人为地培养金发藓。     

几种丝状真菌原生质体的形成与再生

本文用商品混合酶酶解的方法,研究了黑曲霉、绿色木霉、桔青霉头孢霉、粗糙脉孢霉、紫色红曲霉、白地霉、凤尾菇、冬菇、平菇等10种13株丝状真菌原生质体的制备及再生,对这些真菌原生质体形成和再生过程中的形态学变化进行了观察,并通过透射电子显微镜的观察证明所得到的制备物确实为没有细胞壁的原生质体.实验表明,用商品纤维素酶、蜗牛酶、溶菌酶等配制成的混合酶液,可成功地从上述分属于子囊菌纲、担子菌纲、半知菌纲的丝状真菌菌丝体制备出原生质体.实验还说明,巯基乙醇对多数丝状真菌原生质体的形成并不是必需的.在所有供试菌株中均观察到由原生质体直接长出菌丝及由原生质体先形成酵母芽链状再长出菌丝这两类再生方式.     

三种微生物实验方法的改进

对微生物实验教学过程中的菌种分离纯化、菌种保藏、微生物显微摄影等3种常规实验方法进行了改进。菌种分离纯化改进方法是结合“10倍稀释法”和“挑单孢法”设计的,是一种适用于单细胞微生物及产孢子的真菌和放线菌的新方法,该法简单,可以实时观察孢子萌发、菌丝生长。细菌分裂等现象;茸种保藏方法是在4℃冰箱保藏方法基础上的一种简易菌种保藏方法,操作简单,易掌握。无需特别设备,尤其适宜于大、中专院校及一些基层单位作为保存菌种用;显微摄影改进方法就是数码相机直接通过目镜照相,该法操作简单易学,能够及时记录实验结果,在数码相机比较普及的时代,更能体现实验改革与时俱进的特点。     

A mitochondrial remnant in the microsporidian Trachipleistophora hominis

Microsporidia are obligate intracellular parasites of several eukaryotes. They have a highly complex and unique infection apparatus but otherwise appear structurally simple. Microsporidia are thought to lack typical eukaryotic organelles, such as mitochondria and peroxisomes. This has been interpreted as support for the hypothesis that these peculiar eukaryotes diverged before the mitochondrial endosymbiosis, which would make them one of the earliest offshoots in eukaryotic evolution. But microsporidial nuclear genes that encode orthologues of typical mitochondrial heatshock Hsp70 proteins have been detected, which provides evidence for secondary loss of the organelle or endosymbiont. In addition, gene trees and more sophisticated phylogenetic analyses have recovered microsporidia as the relatives of fungi, rather than as basal eukaryotes. Here we show that a highly specific antibody raised against a Trachipleistophora hominis Hsp70 protein detects the presence, under light and electron microscopy, of numerous tiny ( approximately 50 x 90 nm) organelles with double membranes in this human microsporidial parasite. The finding of relictual mitochondria in microsporidia provides further evidence of the reluctance of eukaryotes to lose the mitochondrial organelle, even when its canonical function of aerobic respiration has been apparently lost.     

丛枝菌根真菌的增殖技术比较分析

以三叶草作宿主植物,采用单孢接种和多孢接种．接种摩西球囊霉、地表球囊霉、地球囊霉、缩球囊霉和网状球囊霉,在土培、沙培和液培3种条件下进行增殖．收获后分别测定丛枝着生率、泡囊的数量和产孢量等重要的生产指标．研究结果表明,土培法、沙培法、液培法都可用来生产接种剂．但生产效果存在显著差异．所测定的各个指标在液培条件下都较高．沙培法次之,土培法最低．参试的5个菌种都对三叶草表现出亲和性．其中摩西球囊霉和地表球囊霉各项指标都较高．适于生产菌剂;多孢接种较单孢接种具有显著的数量优势．单孢接种可用于菌种纯化．     

油菜内生真菌的分离鉴定

本试验通过对健康油菜组织表面消毒、内生真菌的分离培养和鉴定,获得以下结果共获得内生真菌45种, 其中子囊菌1种,半知菌43种(丝孢纲40种,腔孢纲3种),担子菌1种,分离物中37个种分属于20个不同属,8个种归属待定.从分离几率来看,没有明显优势的种类存在,在叶部分出率最高的是Alternaria alternata, 分离几率仅为7.04%.不同油菜器官的内生真菌种类有一定的差异,从叶部分离出25种,根部分离出12种、花器11种、茎12种、幼苗2种.以上结果显示油菜内生真菌复杂、具有丰富的种类多样性.     

氢氧化钠对苏云金芽孢杆菌芽孢及其萌发的影响

【目的】了解碱对细菌芽孢及萌发的影响及机制,进一步认识芽孢的抗性,为寻找更好的杀灭芽孢的方法提供理论参考。【方法】应用单细胞拉曼光谱采集经NaOH处理后的单个苏云金芽孢杆菌(Bt)芽孢的拉曼光谱,并应用微分干涉差显微镜成像术分析其萌发动态。【结果】1.0mol/L NaOH处理30min、45min和60min后,Bt芽孢的存活率分别为25.0%、3.6%和1.8%左右;芽孢吡啶二羧酸(DPA)和蛋白质的特征峰有微弱的降低,蛋白质二级结构α螺旋特征峰(1 280/1 652cm~(–1))的位置和信号强度与对照没有明显的差异。在10mmol/L丙氨酸下,单个芽胞开始大量释放CaDPA的时间(Tlag)、快速释放CaDPA所需的时间(ΔT_(release))和芽胞皮层水解所需的时间(ΔT_(lys))分别是对照的3.5~5倍、12~13倍和1.5~2倍;在月桂胺触发下,T_(lag)值和ΔT_(release)值分别是对照的4倍和11倍左右。外源CaDPA无法触发处理30 min后的芽孢的萌发,而对照则萌发极快。【结论】NaOH处理没有破坏Bt芽孢的内膜,也没有引起成孢内CaDPA泄漏,但令与芽孢萌发相关的蛋白质受损,受损最严重的是皮层水解酶,对芽胞萌发产生严重的影响。     

平行气流真菌孢子释放强度试验研究

真菌是室内环境中普遍存在的空气微生物污染物,可对人体产生许多危害.采用自行开发的平行气流真菌孢子释放强度测定装置(PAFST),测量了5种典型风速下,4种常见真菌孢子柑桔青霉(Penicillium citrinum)、扩张青霉(Penicillium expansum)、黑曲霉(Aspergillus niger)和枝孢属(Cladosporium Spp.)在3种材料表面的释放强度.结果表明气流的作用形式是影响孢子释放强度的主要因素;在琼脂表面,风速0.9m/s时,柑桔青霉的释放强度为(7±9)个/cm2;枝孢属在风速4.7m/s时,释放强度为(50±35)个/cm2;在无纺布表面柑桔青霉和扩张青霉的最小释放气流速度是0.3m/s,黑曲霉是2.4m/s,枝孢属为9.4m/s;当达到孢子释放最大强度时,风速增加释放强度将不再增大.     

不同培养方式对白腐菌降解染料体系抑杂菌效果的影响

研究了非灭菌环境白腐菌降解染料体系的抑菌方法。采用3种不同碳氮摩尔比液体培养基。试验结果表明,当采用白腐菌孢子作为种子在非灭菌环境进行培养时,3种碳氮摩尔比液体培养基在培养2 d后全部感染酵母菌,使体系失去降解染料能力;而采用先灭菌环境培养后非灭菌环境脱色的方法却获得了较好的脱色效果,3种液体培养基中的白腐菌对染料的脱色率均在70%以上,与灭菌环境得到的结果基本相当。因此,当采用白腐菌孢子作为种子进行培养时,只有采用灭菌环境培养的方法,才能使白腐菌在非灭菌环境具有较高的降解染料的能力。     

吉林省西部湿地天然水中的铁细菌

从吉林省西部天然水中分离出Fb₁和Fb₂两种铁细菌 . Fb₁细胞为椭圆形, 细胞成对排列, 有单生、 双生鞭毛和芽孢, 细胞大小为0 .7~ 1.0 μm; Fb₂为杆状, 呈多个细胞相互连结, 无鞭毛和芽孢, 细胞大小为(0.4~ 0.7) μm×(1.5~2.1) μm. 两种菌均为革兰氏阴性, 且都具有荚膜, 菌浓度约为 10 000 000个/mL, 在牛肉膏蛋白胨培养基中生长20 h可达到稳定期, 24 h后进入衰退期. 实验结果表明, 两种菌的基本生理生化性质相似.