New heptaene antibiotic, flavomycin, formed by Act. flavus var. geptinicus var. nov]

An actinomycetous culture LIA-0734 was isolated from a soil sample. By its morphologo-cultural properties it was close to Act. flavus and differed from the latter in the sporophores, colour of the substrate mycelium on synthetic media amd markedly pronounced antagonism with respect to yeasts and yeast-like fungi. The culture was classified as Act. flavus var. geptinicus var. nov. The actinomycete produced new aromatic heptaens: flavomycins A and B. Their physico-chemical and biological characteristics and singularity are presented.     

New antibiotic, parvulomycin, produced by a culture of Actinomyces parvullus var. chromogenes var. nov]

Actinomycete LIA-O784 was isolated from a soil sample. By its morphological and cultural properties the isolate was close to Act. parvullus but differed from it in synthesis of melanoid pygment, thyrosinase, hydrogen sulphide and pronounced antifungal activity. The actinomycete was classified as a new variant and designated as Actinomyces parvullus var. chromogenes var. nov. The culture produced a new polyglycoside antibiotic named parvulomycin. The physico-chemical characteristics of the antibiotic is presented.     

New tetraene antibiotic, abkhazomycin]

The cultures of Act. L10-0740 and L10-0772 were isolated from a soil sample. By their morphological and cultural features they were close to Act. badiocolor and differed from the latter by their antibiotic properties. Because of this they were classified as a new variant of Act. badiocolor var. abhasus var. nov. The cultures produced a new tetraen antibiotic, named abkhazomycin. Its physico-chemical properties are presented.     

Polymorphism of a culture of the chromomycin producer, Actinomyces aburaviensis var. verrucosus]

Polymorphism of the chromomycin-producing orgnaism Act. aburaviensis var. verrucosus 144--3 was studied. The stable variants differed in the morphologo-cultural properties, assimilation of the carbon sources, the component composition of the luminescence substances and the quantitative property of the antibiotic production. A substance with violet luminescence was found in the culture of varient I in addition to chromomycin. Similarity of the phenotypes observed in variants 1 and 2 when grown on complex organic media is explained by intensive chromomycin production by variant 1 on such media. Active colonies may be selected according to the colour due to chromomycin. Forms with an activity almost 4 times higher than that of the initial culture were found among the colonies of variant 2.     

Adriamycin, 14-hydroxydaunomycin, a new antitumor antibiotic from S. peucetius var. caesius. Reprinted from Biotechnology and Bioengineering, Vol. XI, Issue 6, Pages 1101-1110 (1969)

Streptomyces peucetius var. caesius, obtained from S. peucetius, the daunomycin producing microorganism, by mutagenic treatment, differs from the parent culture by the color of the vegetative and aerial mycelia and by its antibiotic producing ability. S. peucetius var. caesius accumulates adriamycin in submerged and aerated culture on a medium containing glucose, brewer's yeast, and inorganic salts both in shake flasks and in stirred fermenters. Isolation of the product is performed by solvent extraction, chromatography on buffered cellulose columns, and crystallization as the hydrochloride. The new antitumor agent, adriamycin, is the 14-hydroxy derivative of daunomycin.     

A comparative study of Antithamnion tenuissimum and three varieties of A. cruciatum, including var. scandinavicum var. nov. (Rhodophyceae)

Antithamnion cruciatum (C. Agardh) Nägeli var. scandinavicum var. nov. is described from material collected on the Danish and Swedish coasts. The variety was found growing on small stones, shells and on Zostera at depths between 0.5 to ca. 10m. A comparative study was made with var. cruciatum , var. radicans (J. Agardh) Collins et Hervey and Antithamnion tenuissimum (Hauck) Schiffner, including examination of type material and laboratory cultures of isolates from the Mediterranean and the Swedish west coast. Variety scandinavicum differs from the other varieties mainly by absence or rare development of gland cells, and sparsely branched whorl-branches. A. tenuissimum is distinguished from A. cruciatum by its shorter thallus, a sympodial type of branching, apparent absence of gland cells and adaxial-monostichous branchlets. Asexual reproduction was found to occur commonly in culture in the two species and their varieties, the morphology of which is discussed.     

复轮生暗孢小克银汉霉新变种及其与原变种之间的配合

多年前分离自湖北神农架林中采回的一份土样的一株小克银汉霉Cu137最近经我们研究定名为复轮生暗孢小克银汉霉新变种(Cunninghamella phaeospora Boedijn var.multiverticillata var. nov.)。它与暗孢小克银汉霉原变种(Cunninghamella phaeospora Boedijnvar. phaeospora)的异同在于:(1)两者均具有单轴分枝、分枝单生或成对的孢子枝,不同的是新变种的孢子枝除单轴分枝外还可轮状分枝,或单轴与轮状混合分枝,轮状分枝可多达3轮,每轮由3-8分枝组成,原变种则完全不形成轮状分枝;(2)两者的孢子枝主轴上的顶生泡囊均具各种形状并略呈棱角状,但新变种的侧枝上的饱囊常常是棒形的,原变种的侧枝上的饱囊则很少棒形;(3)新变种的假根很发达,可自菌丝、匍匐菌丝、孢子枝、泡囊等部位发生,而原变种的假根一般不很发达,而且只从菌丝或甸旬菌丝发生;(4)两者的小型孢子囊在形状、大小、刺等方面一致,但新变种的小型孢子囊无色至淡褐色,原变种的小型孢子囊则为淡褐色至深褐色;(5)新变种的菌落白色,至多在老后污白色至极浅的淡褐色,原变种的菌落自始即为深灰色。除暗孢小克银汉霉原变种外,同属中还有另外三个分类群与复轮生暗孢小克银汉霉较为接近:印度小克银汉霉(Cunninghamella indica Baijal & B. S. Mehrotra)与印度刺孢小克银汉霉[Cunninghamella echinulata(Thaxt.) Thaxt. ex Blakeslee var. indica Baijal & B. S. Mehrotra]因菌落白色及孢子枝分枝棒形,轮生小克银汉霉(Cunninghamella verticillaata Paine)因孢子枝具数轮轮生分枝而与复轮生暗孢小克银汉霉这个新变种有一定的相似。但是,新变种无论是顶生泡囊还是侧生泡囊的直径都较上述三个分类群小得多;新变种的顶生饱囊往往不规则形状且略带棱角,而上述三个分类群则不是这样。还有,新变种的孢子枝分枝方式与印度小克银汉霉的颇为不同,新变种的小型孢子囊比印度刺孢小克银汉霉的小得多,其小型孢子囊主要球形至近球形,与轮生小克银汉霉的椭圆形至卵形也不一样。此外,新变种的假根系统极其发达,与同属中任何一种均不相同。将复轮生暗孢小克银汉霉新变种Cu137(+)与暗孢小克银汉霉原变种的一株分离自香港山边土壤的Cu 136(一)配合时只形成配子囊,但与原变种的模式菌株CBS 692.68(-)配合时则可形成少量的接合孢子囊。为了获得更多的接合孢子囊以观察它们是否可育的,我们将Cu 137和CBS 692.68同时接种到许多培养皿上,然后制片观察。用姆指轻轻、重复压盖玻片,可使接合孢子囊破裂以观察其内含物。结果,从100个以上的接合孢子囊中,仅在其中4个发现有接合孢子的形成,其余的都是空的和不育的。由于原变种尚未发现(+)配合型菌株,新变种则未发现(-)配合型菌株,原变种与原变种之间及新变种与新变种之间配合形成的有性型尚未获得,而且全世界均未见有报道,无法和新变种与原变种之间配合形成的有性型相比较,因此目前尚不能肯定新变种与原变种之间形成的有性型在形态上是否能代表这个种的有性型。但是,尽管如此,暗孢小克银汉霉的这两个变种之间配合形成的有性型的形态仍然是很有特征性的,足以把它与其它种的已知有性型相区别。     

中国水仙(Narcissus tazetta var.chinensis)的组织培养

以栽培的中国水仙Narcissus tazetta var.Chinensis鳞茎为实验材料,采取“无液处理”的简捷方法获取外植体,并筛选出中国水仙离体鳞茎培养的最佳培养基。     

褐脉少花龙葵毛状根的诱导、培养及其澳洲茄胺的产生

利用发根农杆茵的遗传转化和液体培养技术,研究了褐脉少花龙葵(solanum nigrum L.Var.Dauciforum)毛状根的诱导和离体培养及其澳洲茄胺的产生以及液体培养过程中培养基中N源和钙的消耗变化.结果表明.发根农杆菌ATCC15834感染褐脉少花龙葵叶片外植体5 d后产生毛状根.感染25d后,约90%的叶片外植体产生毛状根.毛状根能在无外源生长调节剂的MS固体和体培养基上自主生长.PCR扩增结果显示发根农杆菌Ri质粒的rilB和rolC基因已在少花龙葵毛状根基因组中整合并得到表达.所产生的毛状根能产生药用次生物质澳洲茄胺,其含量约为非转化植株根的1.3倍,达到582.05μg/g干重.少花龙葵毛状根液体培养0-5 d内处于生长迟滞期、5-15 d为快速生长期、15d后进入生长平台期.培养基的硝态氮和铵态氮在毛状根液体培养过程中被逐渐吸收和消耗,至培养15 d时铵态氮被消耗殆尽.而硝态氮仍剩余44.7%;培养基中钙的浓度在培养过程中虽逐渐降低,但在培养25d时仍未被完全消耗,其浓度约为起始浓度的43.5%.该结果为今后设计合适的培养基来规模培养褐脉少花龙葵毛状根生产药用次生物质澳洲茄胺提供了可能性.     

Adriamycin, 14-hydroxydaunomycin, a new antitumor antibiotic from S. peucetius var. caesius

Streptomyces peucetius var. caesius, obtained from S. peucetius, the daunomycin producing microorganism, by mutagenic treatment, differs from the parent culture by the color of the vegetative and aerial mycelia and by its antibiotic, producing ability. S. peucetius var. caesius accumulates adriamycin in submerged and aerated culture on a medium containing glucose, brewer's yeast, and inorganic, salts both in shake flasks and in stirred fementers. Isolation of the product is performed by solvent extraction, chromatography on buffered cellulose columns, and crystallization as the hydrochloride. The new antitumor agent, adriamycin, is the 14-hydroxv derivative of daunomyein.     

中国水仙遗传转化研究进展(综述)

中国水仙是我国传统名花,其栽培种一直存在三个问题：品种单一,繁殖速度有限,病毒积累严重。生物技术的发展为解决中国水仙目前存在的问题提供了新的途径。本文从外植体的预处理、愈伤组织和小鳞茎的诱导与分化、生根及移栽等方面概述中国水仙的组织培养进展,较详尽地阐述中国水仙基因工程的研究进展,并对中国水仙未来研究方向进行展望。     

Isolation and identification of Bacillus subtilis strain YB-05 and its antifungal substances showing antagonism against Gaeumannomyces graminis var. tritici

In this study, 98 putative Bacillus strains were isolated from wheat rhizospheric soil. Among the isolated strains, six showed strong inhibitory effects against the wheat take-all pathogen, Gaeumannomyces graminis var. tritici. One of the strains that showed significant inhibitory activity, YB-05, was identified as Bacillus subtilis based on a phylogenetic analysis of its 16S rDNA gene sequence, the results of the PCR analysis and cloning of its antifungal genes, its morphological characteristics and its physiological and biochemical properties. When tested with a dual-culture, cup–disc method and laboratory greenhouse studies, strain YB-05 was found to be superior to chemical treatment for control of the plant pathogen G. graminis var. tritici. After liquid culture, various antimicrobial substances in the culture medium were detected by high-performance liquid chromatography and high-resolution mass spectrometry, and the existence of their corresponding genes was verified by PCR analysis.     

Larvicidal activity of extracts from Artemisia species against Culex pipiens L. mosquito: comparing endemic versus ubiquist species for effectiveness

The larvicidal activity of ethanolic leaf extracts from two Artemisia species, Artemisia campestris var. glutinosa and A. molinieri, on mosquito Culex pipiens Linnaeus (Diptera, Culicidae) larvae was investigated. Since A. molinieri is a rare and protected species confined to temporary ponds of Southern France, its toxic activity may help to value this species and to finance its conservation. A. molinieri extracts showed a higher larvicidal activity (from 50 ppm (K=9.488, DDL=4, P<0.001)) than those from A. campestris var glutinosa (from 500 ppm (K=9.488, DDL=4, P<0.01)) after 48 h of exposure. Calculated lethal concentrations, after 48 h of exposure, (LC(50)) were low, 9091 and 9898 ppm for A. molinieri and A. campestris var. glutinosa extracts, respectively, but using a non-pollutant solvent (ethanol). However, A. molinieri may be valued as an environmentally friendly biocide and developing its culture may be of interest for both pesticide activity and conservation purpose.     

Competitive interactions between Microdochium nivale var. majus, M. nivale var. nivale and Fusarium culmorum in planta and in vitro

Microdochium nivale var. majus and var. nivale are economically important fungal pathogens of cereal seedlings, stem bases and ears, as is the toxigenic species Fusarium culmorum. Competition experiments on seedlings support an earlier report of differential host preference between the varieties of M. nivale on wheat and rye seedlings at 15 degrees C, but showed that it does not extend across a broad range of temperatures. The studies showed that, although interaction is disadvantageous to the less virulent pathogen, it does not confer an advantage to the more virulent pathogen. In mixed inoculum experiments on wheat seedlings at 15 degrees C and 20 degrees C, F. culmorum suppressed the growth of both varieties of M. nivale. However, if M. nivale var. majus became established on the seedlings, it was able to co-suppress colonization of wheat seedlings by F. culmorum. In contrast M. nivale var. nivale did not suppress F. culmorum significantly. The growth of M. nivale var. majus and F. culmorum was also co-suppressed in liquid culture. Significantly, the accumulation of deoxynivalenol mycotoxin was also reduced in the mixed in vitro culture compared with axenic culture of F. culmorum. However, in vitro interaction studies on solidified media were of only limited use in predicting the outcome of competitions in planta.     

Effect of storage conditions on population variability and activity of Streptomyces recifensis var. lyticus

The results of the study on survival and variation of Streptomyces recifensis var. lyticus 2435 producing lytic enzymes are presented. The culture was maintained for 2.5 years under a layer of vaseline oil, at a temperature of -20 degrees C and in lyophilized state. It was shown that irrespective of the storage method strain 2435 preserved its viability. However, the most intensive growth was observed in the lyophilized cultures. During the storage the content of the productive colonies characteristic of the morphological type culture in the population decreased while the number of the low active variants increased. Lyophilization of the strain spores in the sucrose-gelatine medium provided insignificant morphological variation of the culture and preservation of the initial level of its lytic activity against a number of test-microbes except S. aureus and M. lysodeikticus. Storage of the culture under vaseline oil and at a temperature of -20 degrees C resulted in lowering of its lytic activity against all the test-microbes used. For long-term maintenance of Streptomyces recifensis var. lyticus 2435 the method of lyophilization in the sucrose-gelatine medium is recommended.     

Several conditions of formation and isolation of violet pigment from culture fluid of Nocardia fructiferi var. ristomycin

A new water-soluble photochromogenic fluorescent violet pigment with the indicator properties was isolated from the culture fluid of Nocardia fructiferi var. ristomycini, strain 76. By its solubility and mobility in various solvent systems and by the absorption spectrum the new pigment is close but not identical to such a pigment as rubrocyanin. The presence of microelements, i.e. iron and magnesium in the medium was shown to be necessary for production of violet pigment.     

绿僵菌大孢变种的生物学特征及其对蛴螬的毒力研究

以报道较多的蛴螬病原真菌 布氏白僵菌、绿僵菌小孢变种作参照菌,系统研究了新分离鉴定的绿僵菌大孢变种的生物学特性,测定了其在偏低的自然气温下对蛴螬的致病能力.结果表明,绿僵菌大孢变种与参照菌的培养要求接近,最适宜的温度为25 ℃.3种供试的培养基中,PPDA为最适宜的培养基,其次为SDAY.在偏低的温度环境下绿僵菌大孢变种对蛴螬的毒杀能力强于布氏白僵菌,土壤和喷雾处理后累计僵虫率分别达88.23%和76.47%,有很好的田间应用前景.     

Colonization of barley roots by endophytic fungi and their reduction of take-all caused by Gaeumannomyces graminis var. tritici

Fungal root endophytes obtained from natural vegetation were tested for antifungal activity in dual culture tests against the root pathogen Gaeumannomyces graminis var. tritici. Fifteen isolates, including Acremonium blochii, Acremonium furcatum, Aspergillus fumigatus, Cylindrocarpon sp., Cylindrocarpon destructans, Dactylaria sp., Fusarium equiseti, Phoma herbarum, Phoma leveillei, and a sterile mycelium, selected based on the dual culture test, were inoculated on barley roots in growth tubes under axenic conditions, both in the absence and presence of G. graminis var. tritici. All isolates colonized the rhizosphere and very often the root cortex without causing disease symptoms and without affecting plant growth. Eight isolates significantly reduced the symptoms caused by G. graminis var. tritici, and 6 of them reduced its presence in the roots.     

UV-B辐射增强对海洋大型藻与微型藻种群生长关系的影响

选用孔石莼和青岛大扁藻为海洋大型藻和微型藻的代表,通过室内添加模拟试验研究了UV-B辐射增强对孔石莼(重量固定)与青岛大扁藻(密度不同)种群生长关系的影响。结果表明:(1)在单养情况下,4个UV-B辐射剂量都对孔石莼的生长产生抑制作用;对青岛大扁藻生长的影响却不同,低剂量(U-1)的UV-B辐射对青岛大扁藻的生长有促进作用,而高剂量的UV-B辐射则有显著的抑制作用;且因初始接种密度不同而各异。(2)在共养情况下,微藻对孔石莼的生长表现出一定的抑制作用,随着微藻初始接种密度的增加,其抑制作用亦增加;反之,在共培养的初始阶段(6 d内)孔石莼对微藻的生长也有抑制作用,但后期阶段(9 d后)表现出促进作用。(3)在共培养的同时附加UV-B辐射处理,随着初始接种密度的增加,青岛大扁藻对孔石莼生长的抑制作用更加明显;同时,与共养相比较,孔石莼对微藻生长的抑制作用亦趋于明显。