Morphology and ultrastructure of Interfilum and Klebsormidium (Klebsormidiales,Streptophyta) with special reference to cell division and thallus formation

Representatives of the closely related genera, Interfilum and Klebsormidium, are characterized by unicells, dyads or packets in Interfilum and contrasting uniseriate filaments in Klebsormidium. According to the literature, these distinct thallus forms originate by different types of cell division, sporulation (cytogony) versus vegetative cell division (cytotomy), but investigations of their morphology and ultrastructure show a high degree of similarity. Cell walls of both genera are characterized by triangular spaces between cell walls of neighbouring cells and the parental wall or central space among the walls of a cell packet, exfoliations and projections of the parental wall and cap-like and H-like fragments of the cell wall. In both genera, each cell has its individual cell wall and it also has part of the common parental wall or its remnants. Therefore, vegetative cells of Interfilum and Klebsormidium probably divide by the same type of cell division (sporulation-like). Various strains representing different species of the two genera are characterized by differences in cell wall ultrastructure, particularly the level of preservation, rupture or gelatinization of the parental wall surrounding the daughter cells. The differing morphologies of representatives of various lineages result from features of the parental wall during cell separation and detachment. Cell division in three planes (usual in Interfilum and a rare event in Klebsormidium) takes place in spherical or short cylindrical cells, with the chloroplast positioned perpendicularly or obliquely to the filament (dyad) axis. The morphological differences are mainly a consequence of differing fates of the parental wall after cell division and detachment. The development of different morphologies within the two genera mostly depends on characters such as the shape of cells, texture of cell walls, mechanical interactions between cells and the influence of environmental conditions.     

<Emphasis Type="Italic">Entransia</Emphasis> and <Emphasis Type="Italic">Hormidiella</Emphasis>, sister lineages of <Emphasis Type="Italic">Klebsormidium</Emphasis> (Streptophyta), respond differently to light,temperature, and desiccation stress

The green-algal class Klebsormidiophyceae (Streptophyta), which occurs worldwide, comprises the genera Klebsormidium, Interfilum, Entransia, and Hormidiella. Ecophysiological research has so far focused on the first two genera because they are abundant in biological soil crust communities. The present study investigated the photosynthetic performances of Hormidiella attenuata and two strains of Entransia fimbriata under light, temperature, and desiccation stress. Their ultrastructure was compared using transmission electron microscopy. The two Entransia strains showed similar physiological responses. They used light more efficiently than Hormidiella, as indicated by higher oxygen production and relative electron transport rate under low light conditions, lower light saturation and compensation points, and higher maximum oxygen production during light saturation. Their requirement for low light levels explains the restriction of Entransia to dim limnetic habitats. In contrast, Hormidiella, which prefers drier soil habitats, responded to light gradients similarly to other aero-terrestrial green algae. Compared to Entransia, Hormidiella was less affected by short-term desiccation, and rehydration allowed full recovery of the photosynthetic performance. Nevertheless, both strains of Entransia coped with low water availability better than other freshwater algae. Photosynthetic oxygen production in relation to respiratory consumption was higher in low temperatures (Entransia: 5 °C, Hormidiella: 10 °C) and the ratio decreased with increasing temperatures. Hormidiella exhibited conspicuous triangular spaces in the cell wall corners, which were filled either with undulating cell wall material or with various inclusions. These structures are commonly seen in various members of Klebsormidiophyceae. The data revealed significant differences between Hormidiella and Entransia, but appropriate adaptations to their respective habitats.     

Isolation,identification and growth conditions of photosynthetic bacteria found in seafood processing wastewater

Four photosynthetic bacteria, isolated from 14 samples taken from seafood processing plants, were identified as species of Rhodocyclus gelatinosus, belonging to the purple, non-sulphur bacteria of the family Rhodospirillaceae. Cultivation in synthetic medium under four different conditions indicated that all four strains gave maximum carotenoid and bacteriochlorophyll synthesis under anaerobic conditions in the light, with values of 11 to 12.6 and 102 to 108 mg/g dry cell wt, respectively. These values are 87% higher than the pigment content obtained from aerobic cultivation, although the cell biomass of all strains (1.7 to 2.3 g/l) was 22 to 38% higher under aerobic conditions. Protein content was always between 32 and 43%. The specific growth rates of all isolates in aerobic cultivation (0.04 to 0.06 h^-1) were twice those in anaerobic conditions in the light. No growth occurred in anaerobic conditions in the dark.     

Solar UV-B radiation, associated with ozone depletion, inhibits the Antarctic terrestrial microalga, Stichococcus bacillaris

This study shows that increased UV-B arising from stratospheric ozone depletion over Antarctica reduced cell viability and the maximum quantum yield of photochemistry (F _v/F _m) in a unicellular terrestrial microalga, Stichococcus bacillaris. In the UV waveband, rates of F _v/F _m decline increased with decreasing wavelength. Photosynthetically active radiation (PAR) also reduced F _v/F _m, though less than UV radiation. Further experiments under different ozone column thicknesses showed a significantly greater decline in cell viability and F _v/F _m under ozone depletion compared with non-depleted conditions. The inhibitory effects of ambient solar radiation suggest that S. bacillaris is unlikely to inhabit soil surfaces, but colonises shaded areas beneath soil surface particles. During periods of ozone depletion, increases in the ratio of UV-B:PAR may reduce the thickness of the sub-surface zone where light conditions are suitable for colonisation by this alga.     

Biological control of Phytophthora blight in red pepper (Capsicum annuum L.) using Bacillus subtilis

Phytophthora blight is one of the most important devastating diseases of red pepper plants. Forty-one bacterial isolates were obtained from rhizosphere soil and subsequently tested for antagonistic activity under in vitro and in vivo conditions. Among the 41 isolates tested, 12 exhibited a maximum antagonistic activity in dual culture assay. These 12 isolates were further screened for disease suppression on red pepper plants in both natural and greenhouse conditions. All the antagonists showed varying levels of antagonism, whereas the isolates R33 and R13 exhibited the maximum (86.8 and 71%) ability to reduce the disease severity in in vivo conditions. Based on the 16S rDNA sequencing, the most effective isolates were identified as Bacillus subtilis. In addition, the isolates were also screened for siderophores, hydrogen cyanide and hydrolytic enzymes. Further, the isolates increased the root and shoot length of the red pepper, which is an added advantage of the isolates while performing the desired function.     

Isolation of thermotolerant,halotolerant, facultative biosurfactant-producing bacteria

Several facultative bacterial strains tolerant to high temperature and salinity were isolated from the oil reservoir brines of an Iranian oil field (Masjed-I Soleyman). Some of these isolates were able to grow up to 60°C and at high concentration of NaCl (15% w/v). One of the isolates grew at 40°C, while it was able to grow at 15% w/v NaCl. Tolerances to NaCl levels decreased as the growth temperatures were increased. Surfactant production ability was detected in some of these isolates. The use of biosurfactant is considered as an effective mechanism in microbial-enhanced oil recovery processes detected in some of these isolates. The surfactant producers were able to grow at high temperatures and salinities to about 55°C and 10% w/v, respectively. These isolates exhibited morphological and physiological characteristics of the Bacillus genus. The partial sequencing of the 16S ribosomal deoxyribonucleic acid gene of the selected isolates was assigned them to Bacillus subtilis group. The biosurfactant produced by these isolates caused a substantial decrease in the surface tension of the culture media to 26.7 mN/m. By the use of thin-layer chromatography technique, the presence of the three compounds was detected in the tested biosurfactant. Infrared spectroscopy and ¹H nuclear magnetic resonance analysis were used, and the partial structural characterization of the biosurfactant mixture of the three compounds was found to be lipopeptidic in nature. The possibility of use of the selected bacterial strains reported, in the present study, in different sectors of the petroleum industry has been addressed.     

Growth and photosynthetic characteristics of several <Emphasis Type="Italic">Cosmarium</Emphasis> strains (Zygnematophyceae,Streptophyta) isolated from various geographic regions under a constant light-temperature regime

Numerous detailed studies have been made of climatically and environmentally influenced macroalgal geographic distribution patterns. However, so far, there have been only a few intrinsic investigations of the geographic distributions of microalgae. In order to investigate the physiological differences among geographically different microalgal strains, six Cosmarium strains were collected from various climate areas and studied. They were grown under a constant light-temperature regime (16°C and 30 μmol photons m⁻² s⁻¹) and nutrient supply. The arctic representative, C. crenatum var. boldtianum, and the typical tropical desmid, C. beatum, behaved like algae adapted to high light intensities, as judged from the distinctly high values of photosynthetic capacity and saturating irradiance measured, in accordance with the high solar radiation prevailing in their sampling areas. The arctic taxon appeared more optimally suited to the low cultivation temperature, as evidenced by the relatively high values of growth rates, maximum quantum yield and photosynthetic efficiency measured. The cosmopolitan taxa, C. meneghinii and C. punctulatum var. subpunctulatum, exhibited a high maximum quantum yield and photosynthetic efficiency concomitantly during growth, which explained their ubiquitous distribution. Nevertheless, two clones belonging to C. punctulatum var. subpunctulatum, collected from polar and mountainous tropical regions, differed significantly with regard to cell volume, growth rates, surface area to volume ratio and photosynthetic parameters. The physiological differences between the Cosmarium strains were in accordance with their geographic origin; they are discussed in detail in this study. Moreover, these differences were maintained despite the long-term cultivation under identical and constant laboratory conditions.     

Mycolytic effect of fluorescent Pseudomonas in biocontrolling of fungal phytopathogenic Curvularia lunata,Fusarium oxysporum,Alternaria padwickii and Rhizoctonia solani

About 50 bacterial strains, each of Pseudomonas fluorescens, from different rhizospheric soil of different plants were screened for antagonistic activity against Curvularia lunata, Fusarium oxysporum, Alternaria padwickii, Rhizoctonia solani causing black kernel, kernel spotting, root rots, stackburn and sheath blight diseases of rice (Oryza sativa L.). Out of the 50 isolates, 15 isolates were found to be effective in lysing the cell wall of the above-mentioned putative pathogens tested in vitro. These Pseudomonas isolates produced mycolytic enzymes, viz. β-1,3-glucanases, β-1,4-glucanases and lipases. P. fluorescens PAK1 and PAK12 among the strains were more effective for the production of these enzymes while PAK12 produce good level of β-1,3-glucanases, β-1,4-glucanases and lipases against tested fungal pathogens. These findings demonstrate a mechanism of antagonism by P. fluorescens against different fungal plant pathogens.     

Physiological characterisation of Colletotrichum gloeosporioides,the incitant of anthracnose disease of noni in India

Ten isolates of Colletotrichum gloeosporioides were collected from different noni growing areas of Tamil Nadu, Karnataka and Kerala in India and their pathogenicity was proved under glass house conditions. Effect of different pH levels, temperature, light intensity and media were tested against the growth of C. gloeosporioides under in vitro. The results indicated that the growth of C. gloeosporioides was maximum in pH range of 6.50–7.00 and temperature range of 25–30°C. Exposure of the fungus to alternate cycles of 12 h light and 12 h darkness resulted in the maximum mycelial growth of C. gloeosporioides compared to the 24 h exposure to continuous light and 24 h exposure to continuous dark. Among the different media tested, host leaf extract medium supported significantly the maximum growth of all the 10 isolates of C. gloeosporioides followed by potato dextrose agar. Further, the strains were found to vary morphologically between the isolates under the study.     

Investigation of the cyt gene in Bacillus thuringiensis and the biological activities of Bt isolates from the soil of China

The presence of cyt genes was investigated in 80 type strains of Bacillus thuringiensis and 143 isolates obtained from soil samples of China by PCR amplification using two pairs of primers for the cyt1 and cyt2 genes. Three type strains of serotypes H11ac, H14 and H36, eight isolates belonging to H3, H14, H18 and H21, and one isolate of unknown serotype harbored cyt genes. We also tested the cytolytic activity for mammal cells, the hemolytic activity for sheep erythrocytes and insecticidal activity against mosquitoes of five isolates that contained cyt genes but did not belong to B. thuringiensis serovar israelensis. The protein profiles of the five isolates were different from those of the type strains of B. thuringiensis serovar israelensis, and among the five isolates, only Y-5 showed mosquitocidal activity against larvae of Culex quinquefasciatus. All five of the isolates exhibited hemolytic activity, but only three could cause the cell death of A549 cells. The cytopathological changes induced by NX-4 in some A549 cells were characterized with cell-ballooning.     

A new microbial consortia containing entomopathogenic fungus,Beauveria bassiana and plant growth promoting rhizobacteria,Pseudomonas fluorescens for simultaneous management of leafminers and collar rot disease in groundnut

The virulence of 20 isolates of Beauveria bassiana (Balsamo) Vuillemin to larvae of the leafminer, Aproaerema modicella, was tested in the laboratory. Leafminer larvae were sprayed with a standard concentration of 1×10⁸ condia/mL of each B. bassiana isolate. All the B. bassiana isolates tested were pathogenic to A. modicella and the mortality varied between 16.7 and 68.9%. Beauveria bassiana isolate B2 was found to be the most virulent followed by isolate B4 which resulted in 59% mortality. Beauveria isolate B2 was selected for dose–response mortality studies with four different doses (1×10², 1×10⁴, 1×10⁶ and 1×10⁸ conidia/mL). Among the various doses tested, 1×10⁸ conidia/mL produced the highest mortality (70.7%). In addition, morphogenesis of the insect pest in all stages, larval, pupal and adult was greatly affected due to fungal infection. Further, B. bassiana isolate B2 and two Pseudomonas fluorescens strains, TDK1 and Pf1 were tested alone and in combination for suppression of groundnut leafminer and collar rot disease and promotion of plant growth and yield both under glasshouse and field conditions. The mixture of B. bassiana and P. fluorescens strains significantly reduced the leafminer and collar rot disease incidences when applied as talc-based formulation through seed, soil and foliar application under glasshouse and field conditions.     

Isolation of hydrogen-producing bacteria from granular sludge of an upflow anaerobic sludge blanket reactor

H₂-producing bacteria were isolated from anaerobic granular sludge. Out of 72 colonies (36 grown under aerobic conditions and 36 under anaerobic conditions) arbitrarily chosen from the agar plate cultures of a suspended sludge, 34 colonies (15 under aerobic conditions and 19 under anaerobic conditions) produced H₂ under anaerobic conditions. Based on various biochemical tests and microscopic observations, they were classified into 13 groups and tentatively identified as follows: From aerobic isolates,Aeromonas spp. (7 strains),Pseudomonas spp. (3 strains), andVibrio spp. (5 strains); from anaerobic isolates,Actinomyces spp. (11 strains),Clostridium spp. (7 strains), andPorphyromonas sp. When glucose was used as the carbon substrate, all isolates showed a similar cell density and a H₂ production yield in the batch cultivations after 12h (2.24–2.74 OD at 600 nm and 1.02–1.22 mol H₂/mol glucose, respectively). The major fermentation by-products were ethanol and acetate for the aerobic isolates, and ethanol, acetate and propionate for the anaerobic isolates. This study demonstrated that several H₂ producers in an anaerobic granular sludge exist in large proportions and their performance in terms of H₂ production is quite similar.     

产毒与不产毒铜绿微囊藻对模拟酸雨及紫外辐射的生理响应

为了比较研究酸雨与紫外辐射对淡水水体常见藻华蓝藻的生理学影响,选取铜绿微囊藻(Microcystis aeruginosa)产毒(FACHB-905)与不产毒(FACHB-469)株系作为实验材料,通过人工模拟酸雨,研究了不同p H处理后2藻株的光合生理变化以及对紫外辐射的敏感性的异同。实验设置3个p H梯度,p H7.10为对照组(正常培养基培养的藻体),两模拟酸雨处理组(p H5.65和p H4.50);两种辐射处理,可见光处理(PAR)以及全波长辐射处理(PAB)。研究结果表明,905藻株细胞粒径在各p H处理下都要显著高于469藻株,模拟酸雨处理显著降低了两藻株细胞的平均粒径及体积,但叶绿素含量显著提高;酸雨处理同时也引起细胞死亡率的增加,表现为藻体有效光化学效率显著降低,生长速率显著受到抑制,低p H下呈负增长,且这种抑制程度在469下更为显著。高的可见光以及紫外辐射处理,使两株系有效光化学效率随p H的降低而呈降低趋势,其中469藻株降低至更低的水平,且高光辐射以及紫外诱导的抑制率要显著高于905藻体,这可能与469藻株较低的光保护色素有关(较低的类胡萝卜素以及紫外吸收物质)。在未来全球变化背景下,不同种类的浮游植物对环境变化的响应及适应能力不同,可改变水体的群落结构和种群丰度,铜绿微囊藻905较469较强的耐受酸雨以及紫外辐射的能力,可能会使该株系在竞争力上占据优势。     

Characterisation,screening and selection of Bacillus subtilis isolates for its biocontrol efficiency against major rice diseases

A Soil bacterium, Bacillus subtilis, isolated from the rhizosphere of rice, showed high biocontrol activity against blast, sheath blight and bacterial blight. In the present study, four B. subtilis strains isolated from paddy soil were studied under laboratory, greenhouse and field conditions. Among the four strains assayed, UASP17 gave maximum inhibition of paddy pathogens and was validated under field trials. B. subtilis (UASP17) under different doses and methods of applications was evaluated for two seasons at Agriculture Research Station (UAS, Raichur). UASP17 was effective in reducing the severity of blast (9.00% and 15.57%), bacterial leaf blight (BLB) (5.00% and 6.11%) and sheath blight (11.93% and 4.17%) diseases for two seasons. The application of bioagent also increased the paddy grain yield (61.00 and 64.30?Q/ha) in the two seasons, respectively. Taken together, these results indicate that B. subtilis UASP17 as seedling dip for 30?min (10?mL/L of water) prior to transplanting and 2.50?L/ha foliar application was effective in managing the diseases of paddy.     

Early seedling and seasonal N2-fixing symbiotic activity of two soybean [Glycine max (L.) Merr.] cultivars inoculated with Bradyrhizobium strains of diverse origin

In areas with a short growing season the poor adaptability of soybean [Glycine max Meer. (L.)] to cool soil conditions is considered the primary yield limiting factor. Soybean requires temperatures in the 25 to 30°C range for optimum N₂-fixation and yield. Field studies were conducted in 1990 and 1991 at Montreal, Quebec to determine whether adaptability to cool soil conditions, with respect to earlier symbiosis establishment and function, existed among either Bradyrhizobium strains or soybean genotypes. An early maturing isoline of the soybean cultivar Evans and the cultivar Maple Arrow were inoculated with one of four strains isolated from the cold soils of Hakkaido, northern Japan, or the commercially used strains 532C or USDA110, at two planting dates. Plot biomass and nodulation were assessed at seedling (V2), and flowering(R2) growth stages and harvest maturity. Soybean genotypes did not differ for pre-flowering nodulation or N₂-fixation in the cool spring conditions of the first year. Seasonal N₂-fixation rates were also determined at the final harvest by the N-balance and ¹⁵N-isotope dilution methods. Significantly higher symbiotic activity was found for two of the four Hakkaido strains and was reflected in higher final soybean seed yield and total N₂-fixation for the growing season, as compared to the two commercial strains. Planting 14 days earlier resulted in greater early vegetative and total seasonal N₂ fixation and yield in the second year when soil temperatures were warmer, emphasizing the need for the development of soybean-Bradyrhizobium combinations superior in nodule development and function under cool soil conditions.     

The effects of ultraviolet radiation on photosynthetic performance,growth and sunscreen compounds in aeroterrestrial biofilm algae isolated from building facades

The effects of artificial ultraviolet radiation [UVR; 8 W m⁻² ultraviolet-A (UVA), 0.4 W m⁻² ultraviolet-B (UVB)] on photosynthetic performance, growth and the capability to synthesise mycosporine-like amino acids (MAAs) was investigated in the aeroterrestrial green algae Stichococcus sp. and Chlorella luteoviridis forming biofilms on building facades, and compared with the responses of two green algae, from soil (Myrmecia incisa) and brackish water (Desmodesmus subspicatus). All species exhibited decreasing quantum efficiency (F _v/F _m) after 1–3 days exposure to UVR. After 8–12 days treatment, however, all aeroterrestrial isolates exhibited full recovery under UVA and UVA/B. In contrast, D. subspicatus showed only 80% recovery after treatment with UVB. While Stichococcus sp. and C. luteoviridis exhibited a broad tolerance in growth under all radiation conditions tested, M. incisa showed a significant decrease in growth rate after exposure to UVA and UVA/B. Similarly D. subspicatus grew with a reduced rate under UVA, but UVA/B led to full inhibition. Using HPLC, an UV-absorbing MAA (324 nm-MAA) was identified in Stichococcus sp. and C. luteoviridis. While M. incisa contained a specific 322 nm-MAA, D. subspicatus lacked any trace of such compounds. UV-exposure experiments indicated that all MAA-containing species are capable of synthesizing and accumulating these compounds, thus supporting their function as an UV-sunscreen. All data well explain the conspicuous ecological success of aeroterrestrial green algae in biofilms on facades. Biosynthesis and accumulation of MAAs under UVR seem to result in a reduced UV-sensitivity of growth and photosynthesis, which consequently may enhance survival in the environmentally harsh habitat.     

Mycoparasitism studies of Trichoderma harzianum strains against Rhizoctonia solani: evaluation of coiling and hydrolytic enzyme production

The genus Trichoderma is a potential biocontrol agent against several phytopathogenic fungi. One parameter for its successful use is an efficient coiling process followed by a substantial production of hydrolytic enzymes. The interaction between fifteen isolates of Trichoderma harzianum and the soil-borne plant pathogen, Rhizoctonia solani, was studied by light microscopy and transmission electron microscopy (TEM). Macroscopic observations of fungal growth in dual cultures revealed that growth inhibition of the pathogen occurred soon after contact with the antagonist. All T. harzianum isolates tested exhibited coiling around the hyphae of R. solani. The strains ALL23, ALL40, ALL41, ALL43 and ALL49 did not differ in coiling frequency and gave equal coiling performances. No correlation between coiling frequency and the production of cell wall-degrading chitinases, N-acetyl-β-d-glucosaminidase and β-1,3-glucanases, was found.     

Biological spectrum of Trichoderma harzianum Rifai isolates to control fungal diseases of tomato (Solanum lycopersicon L.)

Three Trichoderma harzianum isolates viz., Th-Sks, Th-Ke and Th-Ar collected from respective states of India viz., Rajasthan, Kerala and Andhra Pradesh were evaluated for the management of six fungal diseases namely damping off, Fusarium wilt, Rhizoctonia wilt, early leaf spot, late blight and Septoria leaf spot in tomato. During in vitro analysis, T. harzianum isolates inhibited the pathogens’ growth. Isolate Th-Sks was the most virulent antagonist against all the test pathogens and exhibited maximum of 79.47% growth inhibition of Phytophthora infestans. Isolate Th-Sks proved most effective at suppression efficacy in the range of 95–100% and 91–100% against all diseases under glasshouse and in the field conditions, respectively. Tomato seeds treatment with isolate Th-Sks also promoted plant height (78.23 cm) and fruits yield (290 g/plant) during field trial and data were found to be not-significantly different from other isolates. Thus, it is concluded that isolate Th-Sks can be utilised as a biocontrol agent for management of fungal diseases in tomato.     

Survival of rhizobia on arrowleaf clover seeds under stresses of seed-coat toxins,heat and desiccation

Eighteen strains of Rhizobium leguminosarum bv. trifolii that had been found to be heat- and desiccation-stress tolerant in soil were tested for their resistance to Trifolium vesiculosum, Savi (arrowleaf clover) coat toxins using an agar plate technique and for survival on seed. They were further tested for their tolerance to seed coat toxins in combination with heat and desiccation stresses. The zone of inhibition in agar ranged from no inhibition to 21 mm diameter of growth inhibition around seed. Inoculation on seed under conditions of 28 °C and 100% relative humidity for 7 days resulted in a range of survival from nearly 100% to less than 1%. The correlation between zone of growth inhibition and survival on seed was not statistically significant. Strains that were desiccation or heat tolerant in soil were not necessarily desiccation or heat tolerant on seed but strains that were heat tolerant in soil were better able to survive on glass beads at 37 °C than strains that were not heat tolerant in soil. The zone of rhizobial inhibition on an agar medium was not a valid method to screen for isolates better able to survive on seed and the ability of rhizobia to withstand heat or drought stress on seed was not related to their ability to survive in soil. This revised version was published online in August 2006 with corrections to the Cover Date.