拮抗烟草疫霉的枝穗霉菌株筛选

【背景】烟草疫霉(Phytophthora nicotianae)引起的烟草黑胫病(tobacco black shank)是烤烟生产上重要的土传根茎病害之一,生产上防治困难。【目的】筛选对病原菌具有强拮抗能力的有益微生物菌株是开展生物防治的基础。【方法】采用平板对峙法筛选对烟草疫霉具有拮抗作用的枝穗霉菌株。根据枝穗霉在烟草疫霉菌落上的覆盖程度和产孢量,以及对烟草疫霉菌丝、孢囊梗和孢子囊的缠绕情况,将枝穗霉的拮抗能力划分为强、中等、弱和无4个等级。【结果】供试8种65株枝穗霉中,6株具有强拮抗能力、27株具有中等拮抗能力、22株具有弱拮抗能力、10株无拮抗能力;不同枝穗霉菌株对烟草疫霉的抑制率大小为20.0%-86.7%。【结论】粉红枝穗霉(Clonostachys rosea)菌株7901、11361和亚麻生枝穗霉(C. byssicola)5072、6729、7507及条孢枝穗霉(C. grammicospora)6730对烟草疫霉具有强拮抗能力,这为后续盆栽试验及作用机理研究等提供了种质资源。     

谷氨酸脱氢酶与醇脱氢酶的共表达及其在制备(R)-2-氯-1-苯乙醇中的应用

【背景】醇脱氢酶AdhS能催化不对称还原反应制备(R)-2-氯-1-苯乙醇,但由于自身再生辅酶NADH的能力不足,需要辅酶再生酶协助其再生NADH。谷氨酸脱氢酶能以谷氨酸为底物,再生辅酶NAD(P)H,具有辅酶再生酶的潜力。【目的】克隆表达谷氨酸脱氢酶基因gdhA,构建谷氨酸脱氢酶GdhA与醇脱氢酶AdhS的大肠杆菌共表达体系,提高AdhS制备(R)-2-氯-1-苯乙醇的转化效率。【方法】从枯草芽孢杆菌(Bacillus subtilis) 168中克隆基因gdhA,并在大肠杆菌(Escherichia coli) BL21(DE3)中表达,分析辅酶再生活力;再与醇脱氢酶AdhS共表达,优化表达条件;分析不同辅酶再生方案对制备(R)-2-氯-1-苯乙醇的转化效率的影响。【结果】谷氨酸脱氢酶GdhA再生NADH的比活力为694 U/g。经GdhA与AdhS的共表达及表达条件优化后,制备(R)-2-氯-1-苯乙醇的转化效率达465 U/L。经比较,GdhA协助再生辅酶NADH,可使AdhS制备(R)-2-氯-1-苯乙醇的转化效率提高到约3倍。【结论】谷氨酸脱氢酶GdhA为NADH高效再生酶,与醇脱氢酶AdhS共表达可显著提高AdhS制备(R)-2-氯-1-苯乙醇的转化效率。     

连作花魔芋软腐病株与健株根域丛枝菌根真菌群落多样性

【目的】解析不同连作年限花魔芋软腐病株、健株根域的丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)群落多样性。【方法】使用AMF 18S SSU rRNA基因特异引物AMV4.5NF/AMDGR对正茬及连作2年和3年的软腐病株、健株魔芋根系和根际土壤DNA扩增建库,通过高通量测序和生物信息学分析探究魔芋软腐病与其根域AMF群落多样性的关系。【结果】魔芋根系具有明显的AMF菌丝、泡囊和丛枝等结构。在相同连作年限条件下,健株根系AMF总侵染率、侵染强度和孢子密度均显著高于病株(P<0.05);在不同连作年限条件下,病株根系AMF总侵染率和侵染强度随连作年限延长而降低。从所有样品中共鉴定到9属53种AMF,其中有49个已知种和4个新种。球囊霉属(Glomus)和类球囊霉属(Claroideoglomus)是AMF群落的优势属,其AMF种分别占总AMF种数的41.5%和26.4%;丰度最高的Paraglomus sp.VTX00308是所有样品的共有种。连作、软腐病及二者的交互作用显著影响根系AMF群落的Shannon指数和Simpson指数及根际土壤AMF的Chao1指数(P<0.05)。通过丰度差异分析发现6个在连作软腐病发生后丰度差异显著的AMF种(P<0.05);NMDS分析表明,不同连作年限的魔芋软腐病株与健株之间的根域AMF菌种组成、相对丰度和群落结构存在差异。相关性分析表明,软腐病发病率和病情指数与魔芋根系和根际土壤AMF的Shannon指数、根系AMF的Chao1和Simpson指数以及AMF总侵染率、侵染强度和孢子密度极显著负相关(P<0.01)。【结论】比对健株,连作魔芋软腐病株根际土壤AMF孢子密度以及根系AMF侵染率、种数和多样性均降低,其群落结构显著改变。     

代谢工程改造热带假丝酵母生产1,2,4-丁三醇

【背景】 1,2,4-丁三醇属于手性多羟基醇,是一种重要的有机合成的化学中间体,以木糖为原料经四步酶反应是目前研究最多的生物合成路线。然而大肠杆菌的鲁棒性较弱,对发酵液中一些抑制剂的耐受性不是很好,同时存在严重的碳代谢抑制。近年来,鲁棒性较好的酵母菌成为更有吸引力的宿主,其中热带假丝酵母具有天然的木糖代谢途径,可以更好地利用木糖。【目的】在热带假丝酵母中构建从木糖到1,2,4-丁三醇的代谢途径。【方法】在热带假丝酵母中敲除木糖还原酶基因GRE3,从而阻断自身的木糖代谢途径。将来源于Caulobacter crescentus的木糖脱氢酶基因(xylB)和木糖酸脱水酶基因(xylD)及来源于Lactococcus lactis的酮酸脱羧酶基因(kdcA)克隆至C. tropicalis 207中,得到重组菌C. tropicalis BT,在此基础上考察重组菌代谢木糖合成1,2,4-丁三醇的能力,确定限速步骤,并通过增加关键基因xylD与kdcA的拷贝数提高1,2,4-丁三醇产量。【结果】在30℃、200 r/min、接种量1%、以30 g/L木糖为底物的情况下,重组菌的1,2,4-丁三醇的产量达到了1.2 g/L,在5 L发酵罐中的产量达到了3.7 g/L。【结论】在热带假丝酵母中实现以木糖为底物的1,2,4-丁三醇代谢途径,并通过在基因组上增加关键基因xylD与kdcA的拷贝数,获得了一株高产1,2,4-丁三醇的重组酵母菌株,这为后续在热带假丝酵母中进一步提高1,2,4-丁三醇产量奠定了基础。     

高磷土壤中丛枝菌根真菌的分离鉴定

【背景】丛枝菌根(arbuscular mycorrhiza,AM)真菌具有广泛的寄主范围、环境适应性和优良的植物促生能力。然而,土壤的高磷水平严重抑制了AM真菌生长及AM形成。【目的】分离鉴定出耐较高有效磷含量的华南土著AM真菌菌株,为菌根学研究工作提供新颖材料。【方法】采用经典形态学和分子系统学方法鉴定高磷土壤中AM真菌。【结果】从有效磷含量为53-131 (平均值±标准差为88.2±17.6) mg/kg的根区土壤中鉴定出7属25种AM真菌,包括无梗囊霉属(Acaulospora) 12种、球囊霉属(Glomus) 7种、隔球囊霉属(Septoglomus) 2种、近明球囊霉属(Claroideoglomus) 1种、根孢囊霉属(Rhizophagus) 1种、硬囊霉属(Sclerocystis) 1种和类球囊霉属(Paraglomus) 1种,其中幼套近明球囊霉(Claroideoglomus etunicatum)和蜜色无梗囊霉(Acaulospora mellea)是优势种。在(87.7±8.0) mg/kg的高磷水平下,AM真菌仍能形成丛枝和泡囊。但当有效磷含量达到(99.7±1.2) mg/kg时,菌根侵染率和丛枝丰度显著下降,但仍能够形成泡囊。【结论】从广州市南沙区有效磷含量为(88.2±17.6) mg/kg的耕地植物根区土壤中,鉴定出具有耐高磷潜力的7属25种AM真菌,幼套近明球囊霉和蜜色无梗囊霉等分离株可作为后续高磷抑制机制解析及耐高磷AM真菌菌剂研发工作的试验菌株。     

米修链霉菌TF78对香蕉枯萎病的田间防效及根际土壤微生物的影响

【背景】香蕉枯萎病是香蕉生产上的毁灭性病害,生物防治是遏制该病害发生的有效手段。在前期的研究中,从健康香蕉根际土壤中分离获得一株对香蕉枯萎病具有良好盆栽防治效果的生防菌——米修链霉菌(Streptomyces misionensis) TF78,但其对香蕉枯萎病的田间生防潜力和对土壤微生物环境的影响尚不清楚。【目的】评价米修链霉菌TF78对香蕉枯萎病的田间防治效果,明确其对香蕉根际土壤微生物群落的影响。【方法】选取两块发病香蕉园,测定该生防菌株对香蕉枯萎病的防治效果,并利用扩增子测序技术分析施用菌剂组和空白对照组共12份香蕉根际土壤的微生物多样性和丰度。【结果】米修链霉菌TF78对两块香蕉园的田间防效分别达55.30%和45.32%。该生防菌株处理组的物种稀释曲线坡度大于空白对照组,并显著富集了优势种群梳霉门(Kickxellomycota),消减了绿弯菌门(Chloroflexi)、酸杆菌门(Acidobacteria)和苔藓杆菌(Bryobacter)的丰度,对土壤中优势种群变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)、芽单胞菌门(Gemmatimonadetes)及木霉属(Trichoderma)、鞘氨醇单胞菌属(Sphingomonas)、寡养单胞菌属(Stenotrophomonas)和芽孢杆菌属(Bacillus)的相对丰度影响不显著。【结论】米修链霉菌TF78塑造了不利于香蕉枯萎病菌Fusarium oxysporum f.sp.cubense存活的土壤环境,有效降低了田间香蕉枯萎病的发生,同时对土壤中大部分具有重要生态功能和抑菌功能的优势微生物种群影响不显著。该研究结果为米修链霉菌TF78的进一步开发应用奠定了基础。     

拮抗匍枝根霉的生防菌R1B的筛选鉴定和抑菌活性分析

为防治梨软腐病,从不同植物内生菌中筛选匍枝根霉拮抗菌,同时研究其生防活性。用对峙培养法筛选拮抗匍枝根霉的内生菌;采用果实打孔接种法检验R1B对梨软腐病的防治效果,初步探讨其抗菌活性物质。结果显示来自霍山石斛的菌株R1B对匍枝根霉具有较好的拮抗活性,生理生化和分子鉴定表明R1B与贝莱斯芽孢杆菌具有最近的亲缘关系。菌株R1B几乎完全抑制匍枝根霉菌丝的生长;接种匍枝根霉4 d后,R1B完全抑制梨软腐病的发生,而对照全部腐烂。R1B中含有抗菌肽合成基因bacA(溶杆菌素)、ituC(伊枯草菌素)、bmyB(杆菌抗霉素)及fenD(丰原素)。抗菌二肽溶杆菌素以及伊枯草菌素和丰原素可能是菌株R1B合成的抗菌物质。R1B具有防治梨软腐病的良好潜力。     

夏季故宫养心殿空气中真菌的调查与分析

【目的】在无法实现洁净环境的古建筑内,文物易遭受霉菌的破坏,尤其是在闷热的夏季。探明空气中真菌的种类对文物、游客的安全具有重要意义。【方法】采用自然沉降法与撞击法对夏季养心殿正殿内代表性的6个取样位置的气生真菌进行培养并进行ITS1 rDNA序列分析。【结果】利用自然沉降法测得气生真菌22种,以枝孢属(Cladosporium)、曲霉属(Aspergillus)和青霉属(Penicillium)为优势类群,在2个位置(佛堂二层与西暖阁)空气真菌污染超标;而撞击法测得100余种,腐生营养型真菌比例较高,优势类群为链格孢属(Alternaria)、Cladosporium、木霉属 (Trichoderma)、根霉属(Rhizopus)、Aspergillus和Penicillium,所有6个位置均超标。通过对环境因子与真菌多样性的相关性分析发现,养心殿内真菌丰度与温度、湿度及悬浮颗粒物有着密切关系。在相对湿度较低的6月,温度对丰度影响较大;高湿度时,悬浮颗粒物与湿度对真菌丰度影响更大。丝状真菌的丰度与小粒径悬浮颗粒物、相对湿度存在显著正相关,而空气中的酵母菌与温度相关性更高。【结论】本研究对养心殿正殿空气中真菌的种属进行了鉴定,并分析了与环境因子的相关性,为预防、开放展览以及修缮提供了科学依据。     

不同浓度壳聚糖对炭黑曲霉和硫色镰刀菌生长和发育的影响

【背景】壳聚糖是广泛存在于甲壳动物的一种多糖，具有广谱的抗真菌活性，但壳聚糖是否影响炭黑曲霉(Aspergillus carbonarius)和硫色镰刀菌(Fusarium sulphureum)生长和发育尚未见报道。【目的】明确不同浓度壳聚糖对A. carbonarius和F. sulphureum生长和发育的影响。【方法】通过在PDA培养基中添加不同浓度壳聚糖，测定两种真菌的菌落直径、生物量和菌丝干重，观察产孢量、孢子萌发和芽管长度，比较抑菌的差异。【结果】壳聚糖处理可显著改变两种真菌的菌落形态，处理浓度越高菌落皱缩和变形越明显；壳聚糖还可以有效抑制两种真菌的菌落生长、菌丝干重和菌丝生物量，抑制效果呈明显的浓度依赖，对F. sulphureum的抑制效果更好。壳聚糖可抑制两种真菌的产孢量、孢子萌发和芽管伸长，处理浓度越高抑制效果越好，对F. sulphureum的抑制效果更为明显。壳聚糖对A. carbonarius和F. sulphureum的EC₅₀值分别为0.12 mg/mL和0.075 mg/mL。【结论】壳聚糖可有效抑制A. carbonarius和F. sulphureum的生长发育，抑制效果呈浓度依赖，F. sulphureum对壳聚糖更为敏感。     

中国毛霉目的一个新纪录属—单囊霉属

单囊霉属(Haplosporangium)是被孢霉科(Mortierellaceae)内的一个小属,由Thaxter在1914年建立的。它的主要特征为孢囊梗从高度分化、通常分隔的菌丝上长出。本文首次在中国报道了两个种(极尖单囊霉Haplosporangium attenuatissima sp. nov.和单孢单囊霉H.decipens Thaxter),其中极尖单囊霉是新种,其特征为孢囊梗基部强烈膨大、顶端极细,孢子囊内含1-9个孢子。     

Systematic significance of mature embryo of bamboos

The mature embryo of seven species belonging to five genera of Indian bamboos is described. In all these the basic pattern of embryo organisation is same: the scutellar and coleoptilar bundles are not separated by an internode, the epiblast is absent, the lower portion of the scutellum and the coleorhiza are separated by a cleft and the margins of embryonic leaves overlap. The features unique to fleshy fruited bamboos are: presence of a massive scutellum, the juxtaposition of plumule and radicle and the occurrence of a bud in the axil of the coleoptile. The fleshy fruit bearing bamboos should be classified into one group, the tribeMelocanneae. Evidence is provided to recognise additional groups in the subfamilyBambusoideae.     

Taxonomic status of the species of the green algal genusNeochloris

Komárek has recently reviewed the various species assigned to the green algal genusNeochloris Starr (Chlorococcales, Chlorococcaceae) and removed those with uninucleate vegetative cells to a new genus,Ettlia. Watanabe & Floyd, unaware ofKomárek's work, also reviewed the species ofNeochloris and distributed them among three genera—Neochloris, Chlorococcopsis gen. nov., andParietochloris gen. nov.—on the basis of details of the covering of the zoospore and the arrangement of the basal bodies of the flagellar apparatus. This paper reconciles these two treatments and makes additional recommendations at the ranks of genus, family, order, and class.     

Analysis of twin of eyeless regulation during early embryogenesis in Drosophila melanogaster

The Drosophila Pax6 homolog twin of eyeless (toy) is so far the first zygotically expressed gene involved in eye morphogenesis in Drosophila. The study of its expression during embryogenesis is therefore informative of the initial events of eye development in Drosophila. We have analyzed how the initial expression domain of toy at cellular blastoderm is regulated. We show that the three maternal patterning systems active in the cephalic region (the anterior, terminal and dorsal-ventral systems) cooperate with zygotically activated gap genes to shape the initial expression domain of toy. Whereas Bicoid, Dorsal and Torso signaling synergistically act as activators, Hunchback, Knirps and Decapentaplegic act as repressors.     

广西青藓科分类学修订

通过对采自广西24个县(市)的1 147份青藓科植物标本的逐一鉴定及相关文献的查阅,确认有广西青藓科植物11属、44种,其中包括广西青藓科植物新记录属1属,即拟异叶藓属(Pseudokindbergia),新记录种7种,分别为匐枝青藓(Brachythecium procumbens)、阔叶尖喙藓(Oxyrrhynchium latifolium)、泛生尖喙藓(O. vagans)、拟异叶藓(Pseudokindbergia dumosa)、华东细喙藓(Rhynchostegiella sinensis)、长肋拟青藓(Sciurohypnum populeum)和弯叶拟青藓(S. reflexum)。该文提供了修订后的广西青藓科植物名录,并对其中的新记录属、种的主要形态学识别特征、生境和地理分布等进行了详细描述。     

The effect of cyanobacteria and azolla on the performance of rice under different levels of fertilizer nitrogen

Cyanobacteria and/or azolla were inoculated, with urea at 0, 72 or 144 kg N/ha, in plots in which azolla-free Indica rice var. IR 28 was grown. Productive tillers, yield and nitrogen contents of grain and straw positively responded to inoculation with cyanobacteria or azolla, even with fertilizer-N up to 144 kg N/ha. Inoculation improved colonization by cyanobacteria. Azolla were superior to the asymbiotic cyanobacteria in enhancing rice performance. Urea at a rate of 72 kg N/ha was found to support the best colonizations when applied with cyanobacteria or azolla or, to give maximum rice yields, both inoculants.     

Evaluation of antiprotozoal and antimycobacterial activities of the resin glycosides and the other metabolites of Scrophularia cryptophila

Resin glycosides are secondary metabolites exclusive to the convolvulaceous plants. In this study, crypthophilic acids A–C (1–3), the first resin glycosides occurring in another family (Scrophulariaceae), and the other constituents of Scrophularia cryptophila were examined for in vitro antiprotozoal and antimycobacterial potentials. Except for crypthophilic acid B (2), all tested compounds exhibited growth-inhibitory effect against Trypanosoma brucei rhodesiense, with l-tryptophan (6) and buddlejasaponin III (7) being the most potent ones (IC₅₀'s 4.1 and 9.7 μg/ml). In contrast, the activity towards Trypanosoma cruzi was poor, and only crypthophilic acid C (3), 6 and 7 were trypanocidal at concentrations above 40 μg/ml. With the exception of 2 and 6, all compounds were active against Leishmania donovani. Harpagide (4) and 3 emerged as the best leishmanicidal agents (IC₅₀'s 2.0 and 5.8 μg/ml). Only compounds 3, 6 and 7 showed antimalarial activity against Plasmodium falciparum with IC₅₀ values of 4.2, 16.6 and 22.4 μg/ml. Overall the best and broadest spectrum activity was presented by compounds 3 and 7, as they inhibited all four parasitic protozoa. None of the isolates had significant activity against Mycobacterium tuberculosis (MICs >100 μg/ml) or were toxic towards mammalian (L6) cells. This is the first report of antiprotozoal activity for natural resin glycosides, as well as for harpagide (4), acetylharpagide (5), tryptophan (6) and buddlejasaponin III (7).     

Apoptotic Regulation of Caspase Activity

Intracellular cysteine aspartate-specific proteases (caspases) play both signaling and effector roles in realizing the program of cell death. Caspases function as proteolytic cascades unique for each cell type and signal triggering apoptosis. All parts of the proteolytic cascades are duplicated and controlled by feedback signals. Amplification cycles between pairs of caspases (the third and the sixth, the ninth and the third, the twelfth and the sixth, and others) help multiply the initial apoptotic signal. The presence of physiological inhibitors of apoptosis that directly interact with caspases creates a multilevel regulatory network of apoptosis in cell. The caspase proteolytic cascades are also regulated by sphingolipid secondary messengers, among them ceramide, sphingosine, and their phosphates. Moreover, an association of the caspase signaling with ubiquitin-dependent proteolysis is shown in cells. In particular, the use of extracellular activators and inhibitors of caspases allows irreversible activation of apoptosis in tumor cells or the prevention of apoptosis in cortical neurons under neurodegenerative diseases.     

Phylogeny of cardinalfishes (Teleostei: Gobiiformes: Apogonidae) and the evolution of visceral bioluminescence

The cardinalfishes (Apogonidae) are a diverse clade of small, mostly reef-dwelling fishes, for which a variety of morphological data have not yielded a consistent phylogeny. We use DNA sequence to hypothesize phylogenetic relationships within Apogonidae and among apogonids and other acanthomorph families, to examine patterns of evolution including the distribution of a visceral bioluminescence system. In conformance with previous studies, Apogonidae is placed in a clade with Pempheridae, Kurtidae, Leiognathidae, and Gobioidei. The apogonid genus Pseudamia is recovered outside the remainder of the family, not as sister to the superficially similar genus Gymnapogon. Species sampled from the Caribbean and Western Atlantic (Phaeoptyx, Astrapogon, and some Apogon species) form a clade, as do the larger-bodied Glossamia and Cheilodipterus. Incidence of visceral bioluminescence is found scattered throughout the phylogeny, independently for each group in which it is present. Examination of the fine structure of the visceral bioluminescence system through histology shows that light organs exhibit a range of morphologies, with some composed of complex masses of tubules (Siphamia, Pempheris, Parapriacanthus) and others lacking tubules but containing chambers formed by folds of the visceral epithelium (Acropoma, Archamia, Jaydia, and Rhabdamia). Light organs in Siphamia, Acropoma, Pempheris and Parapriacanthus are distinct from but connected to the gut; those in Archamia, Jaydia, and Rhabdamia are simply portions of the intestinal tract, and are little differentiated from the surrounding tissues. The presence or absence of symbiotic luminescent bacteria does not correlate with light organ structure; the tubular light organs of Siphamia and chambered tubes of Acropoma house bacteria, those in Pempheridae and the other Apogonidae do not.     

Anti-fungal effects of phenolic amides isolated from the root bark of Lycium chinense

Four phenolic amides, dihydro-N-caffeoyltyramine (1), trans-N-feruloyloctopamine (2), trans-N -caffeoyltyramine (3), and cis-N-caffeoyltyramine (4), were isolated from an ethyl acetate extract of the root bark of Lycium chinense Miller. All had an anti-fungal effect; compounds 1-3 were potent at 5-10 microg ml(-1) and were without hemolytic activity against human erythrocyte cells. Compound 4 was active at 40 microg ml(-1). All four compounds impeded the dimorphic transition of pathogen, Candida albicans.