苏云金芽胞杆菌clpP基因缺失对碱刺激敏感性的影响

[目的]比较苏云金芽胞杆菌与枯草芽胞杆菌在碱性培养条件下生长情况,明确clpp基因在碱刺激条件下的作用.[方法]采用同源重组技术敲除苏云金芽胞杆菌HD73菌株clpP基因,通过在不同pH下生长曲线的测定明确了clpP基因缺失突变体对碱性环境的敏感性,测定clpp基因的缺失对芽胞形成率、芽胞萌发效率和盐胁迫的影响.[结果]苏云金芽胞杆菌在碱刺激后,当培养基pH值为8.9-9.1时可以恢复生长,而枯草芽胞杆菌在pH值为8.2-8.4时可以恢复生长,说明苏云金芽胞杆菌对碱性环境适应能力更强,这有助于作为病原菌的Bt适应昆虫中肠的碱性环境.clpp基因缺失对芽胞形成率和萌发效率没有明显的影响.在将培养基中NaOH终浓度调节至30 mmol/L NaOH时,clpp基因缺失突变体的生长较出发菌株慢.说明ClpP在苏云金芽胞杆菌对碱性环境的适应过程中具有重要作用.     

球孢白僵菌(Beauveria bassiana)培养特征多样性与寄主及地理来源的相关性分析

球孢白僵菌不同菌株微观萌发特征及宏观培养特性表现出高度的多样性,2个性状集团经主成分分析后,得到微观萌发特征的主因子为单芽管萌发孢子比例、芽管总平均长度及芽管整齐度;宏观培养性状的主因子分别为察氏培养基上的菌落颜色、菌落基质颜色、菌落生长速率及液体培养液的pH值。由主因子指标进行典型相关分析,两性状集团总体相关不显著(p=0.1378),但菌落生长速率与单芽管萌发孢子比例及芽管总平均长度表现出显著的相关性。由主因子指标进行的聚类及Shannon-Wiener多样性指数统计表明,相同地理来源菌株间的相似性程度高于相同分离寄主间的菌株相似性。     

球孢白僵菌（Beauveria bassiana）培养特征多样性与寄主及处理来源的相关性分析

球孢白僵菌不同菌株微观萌发特征及宏观培养特性表现出高度的多样性。2个性状集团经主成分分析后,得到微观萌发特征的主因子为单芽管萌发孢子比例、芽管总平均长度及芽管整齐度;宏观培养性状的主因子分别为察氏培养基上的菌落颜色、菌落基质颜色、菌落生长速率及液体培养液的pH值,由主因子指标进行典型相关分析,两性状集团总体相关不显著（p=0.1378）,但菌落生长速率与单芽管萌发孢子比例及芽管总平均长度表现出显著的相关性,由主因子指标进行的聚类及Shannon-Wiener多样性指数统计表明,相同地理来源菌株间的相似性程度高于相同分离寄主间的菌株相似性。     

长柄石杉芽胞及芽胞植株形态研究

为研究石杉属植物芽胞形态特征以及芽胞在石杉属植物无性繁殖中的作用,对野外采集和移栽后新生的长柄石杉（Huperzia javanica）芽胞体、芽胞的形态特征、芽胞萌发形成芽胞植株的过程进行了描述,并对芽胞和芽胞残体发生数量进行计数。结果表明,芽胞体由芽胞和芽托部组成,前者是6个小叶围住的小芽,后者为具有6个鳞状叶的柄状体;芽胞脱落后一周开始萌发并进一步发育为芽胞植株;芽胞体发生枝条上平均有5个芽胞发生;芽胞发生具有一定程度的周期性,可以据此估计石杉植株株龄和生长速度。     

云南省4种蕨类植物提取液的抑菌活性

为了合理开发利用蕨类植物资源,本研究利用圆纸片法分析了云南省4种蕨类植物单芽狗脊蕨(Woodwardia unigemmata)、蜈蚣蕨(Pteris vittata)、鸡足山耳蕨(Polystichum jizhushanense)和灰绿耳蕨(Polystic-hum eximium)的提取液对金黄色葡萄球菌(Staphylococus aureus)、大肠杆菌(Escherichia coli)和枯草芽孢杆菌(Bacillus subtilis)的抑菌活性。抑菌试验结果表明,当4种蕨类植物提取液浓度分别为10%、8%、6%和4%时,对照组无抑菌圈出现,单芽狗脊蕨提取液对大肠杆菌和金黄色葡萄球菌无抑菌活性,灰绿耳蕨提取液对金黄色葡萄球菌无抑菌活性,其它的蕨类植物提取液对供试微生物都表现出不同程度的抑菌活性;并且这4种蕨类植物提取液至少对一种供试细菌具有抑菌活性,抑菌活性范围为8～14mm,平均抑菌圈直径为11.2mm,4种提取液对枯草芽孢杆菌抗菌活性效果最明显。由此,我们推测本研究中的4种蕨类植物均有抗菌物质存在,并将有望成为筛选抗菌新型药物的种质资源。     

下延叉蕨和芽胞叉蕨配子体发育的研究

利用光学显微镜详细观察了叉蕨属(Tectaria)下延叉蕨(Tectaria decurrens)和芽胞叉蕨(T.fauriei)的配子体发育过程,记录了配子体各发育阶段的特征。结果表明:(1)下延叉蕨和芽胞叉蕨的孢子均为单裂缝,具周壁,由周壁形成纹饰,孢子极面观椭圆形,赤道面观豆形或肾形。(2)孢子萌发方式为向心型。(3)原叶体发育方式为三叉蕨型。(4)成熟原叶体心脏形,两翼向斜上方扩展。(5)均具单细胞和多细胞毛状体,在丝状体或片状体阶段出现。研究认为,从配子体发育角度看,叉蕨属是较进化的陆生真蕨类;毛状体的类型、位置和出现时间等特征在叉蕨属种间存在差异,可作为该属种间分类的特征。     

四川海螺沟冰川土样芽胞杆菌资源分析

【背景】芽胞杆菌(Bacillus-like)是一类能形成具有强抗性芽胞且可在多种极端环境下存活的细菌,其产生的多种功能代谢产物在多种领域具有重要研究价值。由于冰川低温、寡营养的独特生态环境,其存在的芽胞杆菌可能具有特殊性,因此研究冰川芽胞杆菌有利于发掘新基因、丰富芽胞杆菌多样性。【目的】了解四川海螺沟冰川土壤芽胞杆菌资源,为挖掘芽胞杆菌新资源提供基础。【方法】采用纯培养法分离获得冰川土壤芽胞杆菌资源,利用16S r RNA基因进行系统发育分析,测定代表性菌株的生理生化特性并采用类平均法和欧氏距离模型进行聚类分析。【结果】共筛选到可培养细菌44株,经16S r RNA基因鉴定确定其中36株为芽胞杆菌,隶属于4个属的19个种,分别为芽胞杆菌属(Bacillus)11个种24株、类芽胞杆菌属(Paenibacillus)2个种3株、短芽胞杆菌属(Brevibacillus)4个种5株和赖氨酸芽胞杆菌属(Lysinibacillus)2个种4株,其中以芽胞杆菌属(Bacillus)为优势属。分离菌中仅3株芽胞杆菌可在4°C生存,7株能在50°C生长,大部分菌株在30°C下生长良好;有74%菌株能耐碱,有37%菌株能在无盐条件下生长。根据生理生化结果,采用类平均法和欧氏距离模型进行聚类分析,可分为3组,分别包含7种、4种和6种芽胞杆菌。第1组均可以水解七叶灵和利用葡萄糖,第2组均不能水解七叶灵和利用葡萄糖,第3组仅能共同利用葡萄糖。【结论】四川海螺沟冰川土壤蕴藏着较为丰富的芽胞杆菌资源,为芽胞杆菌新资源挖掘提供了资源保障。     

西双版纳热带山地常绿阔叶林蕨类植物的组成及生态特征

根据4个50m×50m样地(400个5m×5m小样方,共计1hm2取样面积)的详细调查及对各样地外蕨类植物区系的采集调查,研究了西双版纳热带山地常绿阔叶林蕨类组成特点及其数量特征等。结果表明:在所调查的5hm2滇南热带山地常绿阔叶林里记录有蕨类植物64种;在生活型组成上,以地面芽和地下芽为主;在叶特征上,以革质和纸质叶居多,叶形以一回羽状和二回羽状叶占优势;在森林群落中蕨类植物有明显的季相变化;在重要值上,狗脊、疏叶蹄盖蕨、苏铁蕨、光叶鳞盖蕨、清秀复叶耳蕨和假稀羽鳞毛蕨的重要值之和占重要值总和的84.02%,其中,狗脊的重要值占重要值总和的1/3以上,它们是该森林群落中占优势的蕨类种类,在生态分布上为该森林群落的"适宜种";研究还发现,勐腊凤尾蕨为该森林群落的确限种。     

基于可培养法分析亚热带植物内生与根际芽胞杆菌种类分布异质性

【背景】芽胞杆菌是农业上重要的微生物菌剂,对植物具有促生、防病、防虫等作用。【目的】了解亚热带植物内生和根际芽胞杆菌的种群分布,为其功能挖掘提供科学依据。【方法】采用可培养手段对甘蔗(Saccharum officinarum)、红麻(Hibiscus cannabinus L.)、黄麻(Corchorus capsularis L.)、黄秋葵(Abelmoschus esculentus)和玫瑰茄(Hibiscus sabdariffa)根际土壤和根内生芽胞杆菌进行分离,利用16S rRNA基因测序对分离菌株进行分类鉴定,并分析系统发育地位。【结果】共获得了可培养芽胞杆菌菌株144株,其中根内生芽胞杆菌82株,根际土壤62株。经16S rRNA基因测序鉴定为芽胞杆菌4个属37个种,分别为芽胞杆菌属、短芽胞杆菌属、类芽胞杆菌属和赖氨酸芽胞杆菌属。芽胞杆菌菌落数量和种类在根部及其根际土壤中差异较大,根际土壤中芽胞杆菌菌落数量远远大于根部,根际土壤中芽胞杆菌菌落含量范围为(0.2-370.0)×10~5CFU/g,而根部为(0.1-81.0)×10~3 CFU/g。根部分离获得的芽胞杆菌种类远大于根际土壤中,从作物根际土壤中共获得了芽胞杆菌19个种,从根部获得内生芽胞杆菌共32个种。阿氏芽胞杆菌(Bacillus aryabhattai)、仙草芽胞杆菌(Bacillus mesonae)和假蕈状芽胞杆菌(Bacillus pseudomycoides)同时存在于4种作物的根际土壤和根部,其他芽胞杆菌种类仅存在于1种作物的根际或者根部。【结论】亚热带作物根际土壤和根部芽胞杆菌种类和数量极为丰富,而且还存在可分离培养的芽胞杆菌潜在新物种,这为了解植物与根际微生物相互作用、根际环境生态平衡提供了理论基础和科学依据。     

枯草杆菌的芽胞在肉鸡肠道中的生活状态和分布

目的探讨枯草芽胞杆菌的芽胞在肉鸡肠道中的生活状态和分布。方法以20日龄AA肉鸡为研究对象,饲喂枯草芽胞菌剂,分别测定鸡粪中芽胞数量和鸡肠道不同部位的活菌数量。结果饲喂3h后,鸡粪中开始检测到芽胞的存在,24h达到最高值,直至饲喂120h后,肠道内的芽胞基本排除。排出芽胞总量为饲喂芽胞总数的3．0倍左右,同时研究还表明：芽胞在实验肉鸡的十二指肠2内开始萌发,并进行了繁殖,在小肠的后端,即小肠3和小肠4,活菌数量达到高峰。结论部分芽胞进入小肠后即可开始萌发,并进行生长繁殖,而且在肠道内有短暂滞留。     

Development and germination of rhizoidal gemmae of Bryum violaceum

Approximately four to six weeks after transferring gametophores of Bryum violaceum Crundwell & Nyholm to fresh soil, abundant new rhizoids with stalked gemmae were present at the bases of the gametophores. The development of gemmae on rhizoids was followed from single cell initials to multicellular, three-dimensional forms. Mature gemmae, the vegetative diaspores, were pale orange, purple or reddish brown and each had a uniseriate stalk of two to four cells. While remaining on soil, the rhizoidal gemmae showed no in situ germination. However, following the removal of gametophores with rhizoidal gemmae, or rhizoids alone with gemmae, and their placement upon wet filter paper in Petri dishes in light, the gemmae germinated. During germination, the protonemata emerged consistently from mature gemmae at their distal ends, revealing the existence of gemma polarity. In the case of immature gemmae, on the other hand, the protonemata emerged from any surface cell indicating that gemma polarity had not yet been established.     

Asexual reproduction in a population of a leafy hepatic species Lophozia silvicola Buch in central Norway

The seasonal pattern and degree of asexual reproduction of a boreal hepatic species, Lophozia silvicola Buch, were studied in a lowland population of the species in central Norway. Asexual reproduction was quantified as numbers of 1–2 -celled gemmae produced per individual shoot, with average production ranging from 1000 to 7000 gemmae per shoot. Despite a slightly increasing trend in early spring, average numbers of gemmae present on shoots were fairly constant during the growing season. Germinability of gemmae was high in early spring, but declined rapidly towards the end of the season, suggesting an increasing proportion of gemmae in the dormant stage in the diaspore bank. Despite repeated sampling, average shoot density remained fairly constant in L. silvicola colonies during the growth season. Gemma production did not correlate with shoot density. Local colonization efficiency was estimated as frequency of occupied substrate patches and proportion of substrate area occupied by L. silvicola in a 10×20 m plot. Colonizable substrates included decaying wood, bare soil and rock surfaces. Colonies of L. silvicola were small, and differences in colony sizes between the substrate types were not significant. Colonies were found in 38% of the substrate patches, but total cover was low, less than 0.1% of all colonizable substrate.     

Three modes of asexual reproduction in the moss Octoblepharum albidum

Abstract

Populations of Octoblepharum albidum growing in very moist and shaded habitats were found to have three kinds of asexual reproduction. First, the production of foliar gemmae along the upper half of leaf margins. Second, the production of protonema gemmae on protonemata originated from foliar gemmae. Third, the production of buds and new shoots at leaf tips. The formation of new shoots is further repeated at the leaf tips of the new shoots, giving the plant the character of a 'walking moss'.     

GROWTH,DEVELOPMENT AND GEMMA FORMATION IN THE LIVERWORT SCAPANIA NEMOROSA AS INFLUENCED BY L-ARGININE,L-HISTIDINE AND L-GLUTAMIC ACID

New procedures of aseptic bryophyte culture which permitted the alteration of the culture medium during the life cycle were used to investigate development and gemma differentiation in Scapania nemorosa from the aspect of organic nutrition. Cultures started from gemmae (asexual reproductive structures) were treated with l-arginine, l-histidine and l-glutamic acid upon reaching each of 3 developmental stages: Cell Mass stage, Juvenile stage, and Early Adult stage. Ten weeks after each treatment, quantitative determinations of growth and gemma formation, as well as a qualitative appraisal of the effect of treatment on development, were made on representative cultures. In addition, an amino acid analysis was made of young plants which had not produced gemmae, of older plants which had produced gemmae, and of gemmae. The results of the culture experiments indicate that all 3 amino acids are stimulatory to production of gemmae and that the degree to which this is so is contingent on the ontogenetic stage treated. Moreover, the qualitative data, in conjunction with the amino acid analysis, indicate that each compound influences gemma formation in a somewhat different manner.     

Growth and Dormancy in Lunularia cruciata (L.) Dum.: IV. LIGHT AND TEMPERATURE CONTROL OF RHIZOID FORMATION IN GEMMAE

The first sign of initiation of growth in dormant gemmae ofL. cruciata is the formation of rhizoids. Gemmae in the cupcannot ‘germinate‘ until exposed to substrate conditionsallowing the outward diffusion of a growth inhibitor. Rhizoidproduction depends on temperature and light. With long lightperiods rhizoids are formed over a wide range of temperatures.Transference to darkness after 2 h white light causes about50 per cent of gemmae to produce rhizoids, and these are formedonly between 20 and 25 °C. Outside these temperature limitsthe percentage of gemmae with rhizoids soon drops to zero. Althoughrhizoid production is prevented in total darkness, gemmae remainalive for well over 6 months. Red light for as little as 5 spromoted, and far-red light inhibited, rhizoid formation inthe dark. Coumarin and indol-3yl-acetic acid can substitutefor light and partly reverse the effect of far-red irradiation.     

中国藓类植物无性繁殖体的初步观察

无性生殖在苔藓植物的生活史中起着重要的作用,并且常通过各种无性繁殖体来完成。无性繁殖体的形态常被用来辅助鉴定一些不育的藓类植物。本文通过对38种藓类植物的无性繁殖体进行显微观察,结果显示：无性繁殖体在不同的藓类植物之间已经过多次演化;无性繁殖体的形态在种内是相对稳定的,且与其着生位置、配子体的分枝方式密切相关,而与植物的系统位置以及生境并无直接的关系;无性繁殖体的颜色与其胞壁的厚度以及表面纹饰密切相关。此外,无性繁殖体的产生常常与假根、原丝体有共存关系。在研究中发现,藓类植物的无性繁殖体主要包括原丝体芽胞、无性芽胞（叶生芽胞、中肋芽胞、枝生芽胞）、芽体、假根芽胞和假根状块茎;其中原丝体芽胞和无性芽胞最为常见。     

ROLE OF ASYMMETRIC CELL DIVISION IN PTERIDOPHYTE DIFFERENTIATION. II. EFFECT OF CA2+ ON ASYMMETRIC CELL DIVISION,RHIZOID ELONGATION,AND ANTHERIDIUM DIFFERENTIATION IN VITTARIA GEMMAE

Gametophytes of Vittaria graminifolia reproduce vegetatively by means of gemmae. Each gemma consists of a linear array of six cells: four body cells and a knob-shaped terminal cell at each end. When gemmae are shed from the gametophyte onto Knop's mineral medium, the two terminal cells do not divide, but elongate to form primary rhizoids. The body cells undergo asymmetric cell division, and the smaller daughter cells differentiate into either secondary rhizoids or prothalli. When gibberellic acid is included in the medium, antheridia are formed as a result of asymmetric cell division instead of vegetative structures. We studied the effect of Ca²⁺ on asymmetric cell division, rhizoid elongation, and antheridium formation in gemmae cultured on Knop's mineral medium and variations of Knop's medium. Ca²⁺ inhibited the onset of cell division and rhizoid elongation, but was required for differentiation of antheridia. Treatments which lowered the Ca²⁺ content of gemmae (EGTA and dilute HCl extraction, culture on verapamil-containing and Ca²⁺-deficient medium) caused an early onset of cell division and rhizoid elongation. The stimulation of growth was most pronounced when gemmae were deprived of Ca²⁺ during the first 24 hr of culture. The proportion of cell divisions which differentiated into antheridia in response to GA was greatly reduced when the Ca²⁺ status of gemmae was lowered with verapamil and Ca²⁺-EGTA buffers.     

GEMMIFEROUS FERN GAMETOPHYTES—VITTARIACEAE

Approximately 10 % of all fern species reproduce vegetatively in the gametophytic stage by means of gemmae. Gametophyte morphologies in these species depart radically from the commonly figured heart-shaped type and expand considerably the opportunities for physiological and morphological studies utilizing fern gametophytes. Original observations on four species of vittarioid ferns are presented and compared with earlier observations on gametophytes of this family. Vittarioid gametophytes grow from a discontinuous marginal meristem which results in a much branched thallus of indeterminant growth. Aerial branches of the gametophytes terminate in gemma production, which proceeds by a regular and predictable sequence of events. The sequence may differ considerably among species but is remarkably constant within species. Archegonia are produced on short ventral branches, and antheridia are produced primarily on germinating gemmae. Ananthacorus angustifolius is the only known member of the Vittariaceae which does not produce gemmae and is considered to represent the primitive condition. In this species antheridia are scattered over the thallus, suggesting that a change in the mode of control of antheridium production may have evolved in the family along with gemma production.     

Growth and Dormancy in Lunularia cruciata (L.) Dum: V. THE CONTROL OF GROWTH BY A NATCRAL, ENDOGENOUS INHIBITOR

The production of an endogenous growth inhibitor by Lunulariahas been demonstrated in tests with diffusates from gemmae andthalli. Increased growth made by bisected gemmae compared withintact controls suggests that self-inhibition also occurs andpoints to the growing tip as the locus of inhibitor production.More inhibitor is produced in short-day (optimum growth) thanin long-day (dormancy inducing) conditions, but even in shortdays the growth of thalli depends on conditions allowing theinhibitor to diffuse away. Thalli prevented from doing thisby resting the apical 2 mm on non-wettable cover slips stoppedgrowth and showed morphological changes of incipient dormancy.Some of the ecological implications of these tests are discussed.     

Growth and Dormancy in Lunularia cruciata (L.) Dum.: I. RESUMPTION OF GROWTH AND ITS CONTINUATION

Lunularia cruciata may become dormant at three stages in itslife history: mature thallus, gemma, and spore. The resumptionof growth and its continuation in various conditions have beenstudied in thalli and gemmae. Air-dry, mature thalli of theIsrael strain, planted on a suitable medium, produce adventitiousbranches ventrally from the region immediately posterior tothe existing meristem, which itself fails to resume growth.When dormant gemmae are taken from the gemma-cup, however, theexisting apical cells and meristems simply recommence growing,so that the new growth is continuous with the old. Except inthe case of mature thalli aroused from dormancy, apical dominanceis pronounced, and branching occurs only by bifurcation. Thisdominance can be broken by applying sucrose solution to thegrowing tips—possibly a plasmolytic effect. The growth in area of freshly planted gemmae accelerates forabout 25 days before its relative rate slows to any great extent.During the first half of this period, growth is due exclusivelyto the expansion of existing cells, but subsequently cell numbersincrease rapidly from the 8–10,000 present in the dormantgemma. Cell numbers were estimated by using a modified macerationtechnique, in which chelation followed prolonged fixation. Lunularia grows successfully at quite low light intensities.Of the mineral nutrient deficienccs investigated, lack of Plimits growth most severely, although N-deficiency also restrictsit to a very low level. Gemma-cup production appears to be unaffected by light intensity,at least within levels permitting growth. There is, however,a large temperature effect, cup production decreasing markedlyabove 12° C.