Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid-state fermentation system

Two methods were compared for determining the concentration of penetrative biomass during growth of Rhizopus oligosporus on an artificial solid substrate consisting of an inert gel and starch as the sole source of carbon and energy. The first method was based on the use of a hand microtome to make sections of approximately 0.2- to 0.4-mm thickness parallel to the substrate surface and the determination of the glucosamine content in each slice. Use of glucosamine measurements to estimate biomass concentrations was shown to be problematic due to the large variations in glucosamine content with mycelial age. The second method was a novel method based on the use of confocal scanning laser microscopy to estimate the fractional volume occupied by the biomass. Although it is not simple to translate fractional volumes into dry weights of hyphae due to the lack of experimentally determined conversion factors, measurement of the fractional volumes in themselves is useful for characterizing fungal penetration into the substrate. Growth of penetrative biomass in the artificial model substrate showed two forms of growth with an indistinct mass in the region close to the substrate surface and a few hyphae penetrating perpendicularly to the surface in regions further away from the substrate surface. The biomass profiles against depth obtained from the confocal microscopy showed two linear regions on log-linear plots, which are possibly related to different oxygen availability at different depths within the substrate. Confocal microscopy has the potential to be a powerful tool in the investigation of fungal growth mechanisms in solid-state fermentation.     

英国 Hampsfell 蕨菜草地生态系统的第一性生产量的研究

 4月中、下旬出现在英国的Hampsfell(英格兰的西北部)地面的蕨菜,以前两个月的生长速度最快,9月叶片达最大高度,但其重量高峰则出现在8月。地上部分生物量也以8月为最高,平均792克·米-2,其后逐渐下降,于11月地面部分枯死。地下部分的生物量季节变化不很明显,但随着地上部分的生长,地下部分的生物量也有所增长。土层深度是影响蕨菜生物量的最重要因子。蕨菜草地生态系统的第一性生产量平均为778克·米-2·年-1,在土深地段平均为918克·米-2·年-l,在浅土地段为525克·米-2·年-l。     

沙埋对花棒种子萌发和幼苗生长的影响

为探讨花棒(Hedysarum scopariium)种子萌发和幼苗生长对沙埋的响应,并为固沙造林、水土保持提供理论基础,研究了6种沙埋深度(0、1、2、3、4和5cm)对花棒种子萌发和幼苗生长的影响.结果表明,埋深对花棒幼苗出土率、首次出苗时间、单株叶片数、幼苗生长高度以及生物量的分配均有极显著影响(P<0.001).种子出苗率在2 cm沙埋下达到最高(95.6%),在5cm沙埋下最低(43.4%);幼苗最大高度(11.6cm)、绝对株高(13.9cm)和最大地上生物量(26.7mg)均出现在2cm的埋深,幼苗最小高度(3.3cm)、最小根长(4.3cm)和最小地上生物量(5.3mg)出现在5cm的埋深;生物量分配随沙埋深度增大而更多地分配给地下部分.2cm的沙埋是花棒种子萌发和幼苗生长的最佳深度.     

Differentiation and anaerobiosis in standing liquid cultures of Streptomyces coelicolor

Streptomyces coelicolor differentiates on solid agar media by forming aerial hyphae that septate into spores. We here show that differentiation also occurs in standing liquid minimal media. After a period of submerged growth, hyphae migrate to the air interface, where they become fixed by a rigid reflecting film. Colonies that result from these hyphae form sporulating aerial hyphae. In addition, submerged hyphae in the liquid minimal medium may attach to the surface. Liquid standing cultures easily become anoxic only 1 to 2 mm below the surface. Yet, biomass increases, implying the existence of metabolic pathways supporting anaerobic growth.     

不同水位梯度下克隆植物大米草的生长繁殖特性和生物量分配格局

通过温室模拟控制实验,研究了不同水位梯度处理对外来克隆植物大米草(Spartina anglica) 衰退种群的形态性状、克隆生长性状、生物量积累及生物量分配格局的影响.实验共设置5种水位处理,分别为淹没至大米草株高的1/2, 淹没至株高的1/3,表面水淹0 cm,低于土表5 cm (-5 cm),以及低于土表10 cm (-10 cm).结果表明:水位梯度对大米草的形态性状、克隆生长性状、生物量的积累与分配格局等部分指标均有显著影响(p < 0.05);株高、克隆分株数、根状茎节数、根状茎总长、间隔子长度及生物量在0 cm至1/3株高水位梯度处理下均显著高于-10 cm和1/2株高水位处理(p < 0.05),但与-5 cm处理无显著差异(p > 0.05);-10 cm水位处理的地上生物量分配和根状茎生物量分配显著低于其它处理(p < 0.05).综合大米草衰退种群的形态特征、克隆生长特性、生物量积累及分配格局对水位处理的响应格局,认为大米草种群较适宜的水位梯度为表面积水至淹没株高的1/3处,而水位的变化可能是大米草种群自然衰退的原因之一.     

岩生植物金发草生长特征研究

研究了重庆地区金发草在3种基质种生境中的生长特征.结果表明,种生境下金发草基径、冠幅、高度及根系面积、最长根和根深差异极显著.3种基质中,紫色土中金发草的地上部分基径(15.18cm)、冠幅(3086.77cm²)和高度(66.8cm)均为最大值,而紫色砂岩金发草的基径(10.89cm)、冠幅(1868.79cm²)和高度(60.7cm)均为最小值,但差异不显著.砂岩中金发草的根系比紫色土中分布广泛,差异显著,说明岩石生境中金发草将较多生物量投入到根系,通过增加根系的生长,提高其在岩石上的固着能力,扩大根系吸收面积,忍耐岩石基质的干燥和贫瘠.     

Improvement of dry matter digestibility of water hyacinth by solid state fermentation using white rot fungi

Feeding value of water hyacinth biomass colonized by three species of white rot fungi during solid-state fermentation was investigated. All three organisms proved to be efficient degraders and enhanced dry matter digestibility. Loss of organic matter was maximum (23.6+/-0.1% dry wt) after 48 days by P. ostreatus. C. indica showed maximum cellulose degradation (18.5+/-0.1% dry wt) than other two fungi after 48 days of incubation. In all cases, an extensive removal of hemicellulose at the initial growth period and a delayed degradation of lignin were observed. Hemicellulolysis was maximum (46.3+/-0.1% dry wt) by C. indica, but delignification (14.2+/-0.2% dry wt) by P. sajor-caju after 48 days. The amount of reducing sugar in the degraded biomass decreased at early stages, but increased as degradation progressed in all three cases (maximum 1.1+/-0.05% dry wt after 48 days by C. indica). Soluble nitrogen content increased only during 16-32 days of incubation (highest 1.1+/-0.1% dry wt after 32 days by P. sajor-caju). Crude protein of the bioconverted biomass increased gradually up to 32 days but decreased thereafter (maximum 10.3+/-0.1% dry wt after 32 days by P. sajor - caju). Per cent change in in vitro dry matter digestibility of degraded substrates enhanced gradually after 8 days and reached maximum after 32 days but thereafter decreased (highest + 20.4+/-0.3% dry wt by P. sajor-caju). The results demonstrated the efficient degrading capacity of the test fungi and their potential use in conversion of water hyacinth biomass into mycoprotein-rich ruminant feed, more so by P. sajor-caju.     

Biomass of extramatrical ectomycorrhizal mycelium and fine roots in a young Norway spruce stand — a study using ingrowth bags with different substrates

Background and aims

The partitioning of below ground carbon inputs into roots and extramatrical ectomycorrhizal mycelium (ECM) is crucial for the C cycle in forest soils. Here we studied simultaneously the newly grown biomass of ECM and fine roots in a young Norway spruce stand.

Methods

Ingrowth mesh bags of 16 cm diameter and 12 cm height were placed in the upper soil and left for 12 to 16 months. The 2 mm mesh size allowed the ingrowth of fungal hyphae and roots whereas a 45 μm mesh size allowed only the ingrowth of hyphae. The mesh bags were filled with either EA horizon soil, pure quartz sand or crushed granite. Controls without any ingrowth were established for each substrate by solid tubes (2010) and by 1 μm mesh bags (2011). The fungal biomass in the substrates was estimated by the PLFA 18:2ω6,9 and ECM biomass was calculated as difference between fungal biomass in mesh bags and controls.

Results

The maximum ECM biomass was 438 kg ha^?1 in October 2010 in 2 mm mesh bags with EA substrate, and the minimum was close to zero in 2011 in 45 μm mesh bags with quartz sand. The high P content of the crushed granite did not influence the ECM biomass. Fine root biomass reached a maximum of 2,343 kg ha^?1 in October 2010 in mesh bags with quartz sand after 16 months exposure. In quartz sand and crushed granite, ECM biomass correlated positively with fine root biomass and the number of root tips, and negatively with specific root length.

Conclusion

The ratio of ECM biomass/fine root biomass in October ranged from 0.1 to 0.3 in quartz sand and crushed granite, but from 0.7 to 1.8 in the EA substrate. The results for the EA substrate suggest a large C flux to ECM under field conditions.     

Life cycle and growth ofPotamogeton crispus L. in a shallow pond,ojaga-ike

The life cycle and growth ofPotamogeton crispus L. were studied in a shallow pond, Ojaga-ike. With respect to the shoot elongation and seed and turion formations, the life cycle of this plant in the pond could be divided into following five stages: germination, inactive growth, active growth, reproductive and dormant stages. It was suggested that the plant showed these successive stages depending mainly upon water temperature. The turions germinated on the bottom in autumn when the water temperature fell below ca. 20 C. The plant showed hardly any growth during winter (December—early March) when the temperature was below 10 C. In the spring when the bottom water temperature rose to above 10 C (mid-March), the plant started to grow again and the shoot elongated rapidly at the rate of 4.2 cm day⁻¹ until the shoot apex reached the pond surface in late April. Both the increment of node number and the internodal elongation were associated with this rapid shoot growth. On 10 May (last sampling date), the mean values of shoot length, internodal length and the number of nodes estimated for 10 predominant plants were 238.2±5.6 cm, 7.1±0.8 cm and 34.9±4.0 cm, respectively. The turion formation and flowering occurred during the period from mid-April to mid-May when the surface water temperature ranged 19 and 22 C. The dry weight of a plant reached the maximum mean value of 1180 mg on 10 May. At its peak biomass, an individual plant produced 1–10 turions (5.5 on average) of which the mean individual turion dry weight was 53.2 mg. The turion dry weight accounted for ca. 42% of the total plant biomass m⁻² at that time.     

几种狼尾草属观赏植物在北京地区的生长特性

在北京地区气候条件下评价了狼尾草‘紫光’（Pennisetum alopecuroides（L．）Spreng‘Ziguang’）、绒毛狼尾草（Pennisetum setaceum（Forssk．）Chiov．）、羽绒狼尾草（Pennisetum villosum R．Br．ex Fresen）、狼尾草‘小兔子’（Pennisetum alopecuroides Desv．ex Ham‘Little Bunny’）和东方狼尾草（Pennisetum orientale Willd．ex Rich．）的田间生长状况、越冬存活率、根系生物量分布和繁殖特性。结果表明,这5种植物在北京地区气候条件下,能正常发芽、生长、抽穗、开花和结实。其中狼尾草‘紫光’、绒毛狼尾草植株高大,分蘖旺盛,盛花期花序高度达135CH和156cm,植株茎数达156和217个／株,地上生物量达608．7和535．3g/株,根系密集分布空间在距地表10～40cm,距茎秆0～30cm。狼尾草‘小兔子’和东方狼尾草株型矮小,株高为40cm,植株茎数分别为162和108个／株,地上生物量分别为124．2和39．8g／株,根系密集分布空间为距地表10～30cm,距茎秆15～30cm。羽绒狼尾草的茎秆半匍匐生长,分蘖能力强,盛花期花序高为79cm,植株茎数为389个／株,地上生物量高达619．9g／株,根系密集区为距地表0～30am,距茎秆0～30cm。狼尾草‘紫光’、狼尾草‘小兔子’和东方狼尾草在北京地区能安全越冬,成活率为100％,羽绒狼尾草和绒毛狼尾草在自然条件下不能越冬,成活率为0。5种狼尾草都可以播种和分株的方式扩繁。     

Conidiation of Aspergillus niger in continuous culture

Summary Conidiation of Aspergillus niger was studied in carbon-limited and nitrogen-limited chemostat culture. Under citrate-limitation conidiation intensity varied inversely with dilution rate. Conidiophores were less complex than in aerial conidiation and at high dilution rates conidia occasionally developed from modified hyphal tips. Conidiation was difficult to achieve under glucose-limitation. At the low dilution rates that allowed limited conidiation steady state could not be maintained due to onset of autolysis. At higher dilution rates when steady state was readily obtained conidiation did not occur. The maximum yield constants under citrate-limitation and glucose-limitation were respectively 0.145 and 0.4 mg dry weight/mg substrate, while the relative specific maintenance values were 0.045 and 0.018 mg substrate/mg dry weight/h. Under ammonium-limitation with citrate as the carbon source there was no conidiation. When nitrate became the limiting nitrogen source conidiophore initiation occurred but biomass production was low and wash-out occurred at D=0.034 h^-1.     

Factors affecting the bioconversion of Philippine tung seed by black soldier fly larvae for the production of protein and oil-rich biomass

A systematic study on the use of Philippine tung (Reutealis trisperma) seed as a substrate for the cultivation of black soldier fly larvae (Hermetia illucens) was performed. The characteristics of Reutealis trisperma seed from two different locations: West Java and Papua, were determined. The seed has a relatively high oil (37.6–39.2%, dry weight) and protein content (14.9–28.2%, dry weight). The effect of cake content in the substrate (0–20%, wet weight), moisture content in the substrate (50–70%, wet weight), feeding rate (50–100?mg/larva/d), lighting condition (dark-light) and substrate depth in a rearing container (4–10?cm) was performed. An optimum prepupal biomass productivity of 123.4?g/m²/d was obtained (20%, wet weight of cake content in the substrate, 60%, wet weight of moisture content in the substrate, 100?mg/larva/d, dark, 6?cm substrate depth). The protein and oil content of the biomass were also determined to evaluate the effect of Reutealis trisperma seed as a substrate for the cultivation of black soldier fly larvae to produce protein and oil-rich biomass. The oil content in the biomass was also extracted and the fatty acid composition was identified. The prepupal biomass has a relatively high amount of protein (45%, dry weight) and oil content (26.6%, dry weight) and is suitable for cattle feed application.     

Yield and species density of grasslands during restoration management

Abstract. The relation between the dry matter production and species density of 27 grasslands with different fertilization histories in the Netherlands was studied. The range in species density was 12–28 species on 150 m², the average annual dry matter production in 1983–1985 varied between 3.3 and 12.8 ton /ha. The results show an optimum curve and confirm the hump-backed model proposed by other researchers for a great variety of vegetation types. It is concluded that a high species density can be expected when the annual dry matter production above a stubble height of 5 cm, is between 4 and 6 ton/ha (ca. 4.6 - 6.0 ton seasonal maximum standing biomass). Species density can be expected to decrease when the dry matter production exceeds values of 6 - 7 ton ha^-1yr^-1 (ca. 6.0 - 6.7 ton seasonal maximum standing biomass). The implications of this relation for grassland restoration, aimed at an increase in species density, are discussed. The results agree with other grassland data based on the seasonal maximum standing biomass. The difference between annual production estimated by harvesting the biomass after mowing at 5 cm above the soil surface (mostly in two cuts), and the maximum seasonal standing biomass must be taken into account. It is proposed to compare data on the basis of the estimation that 62 ± 5% of the annual yield is harvested at the first cut, and that there is a linear relation between the total standing biomass, y (g/m²), and the biomass harvested above a stubble height of 5 cm, x (g/m²) that could be described by the regression line y = 1.10 x + 189.6 (r = 0.89, p < 0.01, n = 34).     

中国东北羊草草原生长季内产量生态模拟及信息参数应用

通过对中国北方羊草草原生物量动态、生物量垂直空间格局及其与环境因子相互关系等主要产量生态数量特征的模拟与内在相关性的研究 ,结果表明 ,草地地上生物量的生长规律呈“单峰”型 ,最大地上生物量出现在 8月 5日 ,其值为1 97.3g· m- 2 干物质 ,而后下降 ;在达到峰值前 ,符合 logistic模型 ,进一步分析模型有关特征值获得了草地有效管理期为返青后的第 73天到第 1 1 9天等十分重要的产量生态信息参数。生长季内地上生物量动态与前一个月的平均气温 ( R=0 .82 87)和积累降雨量 ( R=0 .8932 )均呈极显著正相关 ,这是实施科学水肥管理的重要参数 ;而地上部生物量最大绝对增长速率 ( AGR)出现在 6月 2 0日至 7月 5日 ,平均为 3.35 33g· m- 2 · d- 1干物质 ;而地上部生物量最大相对增长速率( RGR)出现在 5月 2 0日至 6月 5日 ,平均为 0 .0 6 6 2 g· g- 1· d- 1干物质 ;在生长后期绝对增长速率和相对增长速率均出现负值 ,这表明地上部生物量的生长效率在生长初期最高。地上生物量垂直空间格局由下向上呈幂函数变化 ,其模型为 :Bn=a Xb,其中 93%的产量集中在 4 0 cm以下 ,这对不同的家畜的选择利用与刈割利用提供了依据 ;不同种群对草原牧草产量形成的作用是不同的 ,羊草种群对草原牧草产量形成的正向     

Glycogen and trehalose accumulation during colony development in Streptomyces antibioticus

Streptomyces antibioticus accumulated glycogen and trehalose in a characteristic way during growth on solid medium. Glycogen storage in the substrate mycelium took place during development of the aerial mycelium. The concentration of nitrogen source in the culture medium influenced the time at which accumulation started as well as the maximum levels of polysaccharide stored. Degradation of these glycogen reserves was observed near the beginning of sporulation. The onset of sporogenesis was always accompanied by a new accumulation of glycogen in sporulating hyphae. During spore maturation the accumulated polysaccharide was degraded. No glycogen was observed in aerial non-sporulating hyphae or in mature spores. Trehalose was detected during all phases of colony development. A preferential accumulation was found in aerial hyphae and spores, where it reached levels up to 12% of the cell dry weight. The possible roles of both carbohydrates in the developmental cycle of Streptomyces are discussed.     

The effect of electric field stimulation on the biosorption of uranium by non-living biomass derived from Kluyveromyces marxianus IMB3

Summary Non-living biomass from the thermotolerant, ethanol-producing yeast strain Kluyveromyces marxianus IMB3 is capable of uranium biosorption. The biomass has an observed biosorption capacity of 115mg uranium/g dry weight of biomass with a calculated value of 127mg uranium/g dry weight. Following exposure of the biomass to electric fields of 2,500 V/cm for 20msec. the maximum biosorption capacity (observed or calculated) for uranium did not differ significantly for the untreated biomass. However, at lower residual concentrations of uranium (<10mg/L) the capacity of the treated biomass for uranium was significantly increased above values obtained with untreated material.     

Vertical distribution and abundance of invertebrates within the sandy substrate of a low-gradient headwater stream

SUMMARY. 1 The vertical distribution of invertebrates (>0.053 mm) was studied in a sandy-bottomed, first-order stream on the Coastal Plain of Virginia, U.S.A. Invertebrate species composition, abundance and bio-mass were determined monthly over one year at sediment depth intervals of 0–1, 1–5, 5–15,15–30 and 30–40 cm.
2. The subsurface community was numerically dominated by species of Chironomidae, Nematoda and Crustacea, while much of the biomass was due to early instars of several species of Trichoptera.
3. Invertebrate density and biomass decreased significantly with depth in the substrate (ANOVA; P <0.05). Annual mean density decreased from 1,346,844 individuals m⁻³ at the surface to 13,578 individuals m⁻³ at 15–30 cm. Annual mean biomass decreased from 66.30 g m⁻³ at the surface to 0.44 g m⁻³ at 15–30 cm.
4. Dissolved oxygen decreased markedly from the surface to the 5 cm depth in the substrate, anaerobic conditions often occurring below 10 cm. Density and biomass both showed a significant positive relationship with dissolved oxygen concentration (Linear regression; P <0.05).
5. Physical forces were important in structuring the subsurface invertebrate community. Besides low dissolved oxygen concentration, sediment scouring resulting from storm discharge dramatically reduced density and biomass     

淹水深度对互叶白千层幼苗气体交换、叶绿素荧光和生长的影响

实验设置对照、浅淹(水位高出土壤表面5cm)和深淹(水位高出土壤表面20cm)3种处理,研究了淹水深度对互叶白千层幼苗的气体交换、叶绿素荧光和生长状况的影响。研究结果表明,随着淹水深度的增加,互叶白千层受到的胁迫程度有所增强。经过270d的淹水处理,浅淹组和深淹组的株高和生物量有所下降,分别为对照的90.86%、64.58%和74.52%、36.46%。浅淹组植株叶绿素含量、净光合速率、气孔传导率和蒸腾速率略有下降,分别为对照的95.39%、94.26%、90.02%和88.94%。深淹组植株在淹水后180d内上述参数显著下降,分别为对照的79.44%、73.54%、61.79%和71.46%,随后逐渐接近对照组。浅淹组PSⅡ的最大光化学量子效率(Fv/Fm)比较稳定,与对照组基本相同。深淹组在淹水后150d内Fv/Fm稍有下降,随后恢复到对照水平。浅淹组植株光化学淬灭系数(qP)稍有下降,非光化学淬灭(NPQ)略有上升,分别为对照的96.63%和105.66%。深淹组植株在淹水后120d内qP显著下降,NPQ明显上升,分别为对照的94.51%和126.66%,随后逐渐接近对照组。另外,淹水过程中,互叶白千层形成不定根和产生发达的通气组织,淹水植株的根孔隙度显著高于对照。     

Buoyant Densities and Dry-Matter Contents of Microorganisms: Conversion of a Measured Biovolume into Biomass

Several isolates of bacteria and fungi from soil, together with cells released directly from soil, were studied with respect to buoyant density and dry weight. The specific volume (cubic centimeters per gram) of wet cells as measured in density gradients of colloidal silica was correlated with the percent dry weight of the cells and found to be in general agreement with calculations based on the partial specific volume of major cell components. The buoyant density of pure bacterial cultures ranged from 1.035 to 1.093 g/cm³, and their dry-matter content ranged from 12 to 33% (wt/wt). Average values proposed for the conversion of bacterial biovolume into biomass dry weight are 1.09 g/cm³ and 30% dry matter. Fungal hyphae had buoyant densities ranging from 1.08 to 1.11 g/cm³, and their dry-matter content ranged from 18 to 25% (wt/wt). Average values proposed for the conversion of hyphal biovolume into biomass dry weight are 1.09 g/cm³ and 21% dry matter. Three of the bacterial isolates were found to have cell capsules. The calculated buoyant density and percent dry weight of these capsules varied from 1.029 g/cm³ and 7% dry weight to 1.084 g/cm³ and 44% dry weight. The majority of the fungi were found to produce large amounts of extracellular material when grown in liquid cultures. This material was not produced when the fungi were grown on either sterile spruce needles or membrane filters on an agar surface. Fungal hyphae in litter were shown to be free from extracellular materials.     

Growth and fatty acid composition of Nannochloropsis sp. grown mixotrophically in fed-batch culture

The cell growth and eicosapentaenoic acid (EPA) yields of Nannochloropsis sp. were enhanced in the fed-batch cultures. With feeding glucose solution, the biomass reached 1.1 g dry wt l(-1) after 10 days' culture, which was 40% higher than that obtained in the batch culture (0.8 g dry wt l(-1)). With supplement of nitrate solution, the biomass reached 1 g dry wt l(-1), and reached the stationary phase 2 days earlier than the others. The maximum of biomass (1.2 g dry wt l(-1)) was obtained with the supplement of the mixture of glucose and nitrate solution. The EPA yields of Nannochloropsis sp. after 10 days' growth in the fed-batch cultures were 52 mg l(-1), 43 mg l(-1) and 56 mg l(-1) with, respectively, addition of nitrate, glucose and both together. In batch culture only 35 mg EPA l(-1) was obtained.