Promotion of plant growth and in situ degradation of phenol by an engineered Pseudomonas fluorescens strain in different contaminated environments

We show that Pseudomonas fluorescens strain P13, a plant growth-promoting bacterium, enhanced the growth of corn in uncontaminated soil but not in contaminated soil, perhaps because of its inability to reduce phytotoxicity. Another bacterial strain, Pseudomonas aeruginosa strain SZH16, showed in situ phenol-degrading activity and contained a plasmid loaded with a gene encoding for catechol 2, 3-dioxygenase, an important enzyme in the degradation pathway of aromatic compounds. We implanted this biodegradation ability into strain P13, using horizontal gene transfer techniques using strain SZH16 as the donor and P13 as the recipient, to generate a phenol-degrading transconjugant which obtained the effective plasmid from strain SZH16. Introduction of the transconjugant P13 strain into an artificially phenol-spiked soil promoted the growth of corn and in situ phenol degradation, and the increase in plant biomass correlated with the decrease in soil phenol content. Furthermore, the transconjugant P13 strain was also found to stimulate corn growth and reduce phenol concentration in water containing phenol and in historically contaminated field soils, indicating that the transconjugant strain could promote plant growth in both contaminated and uncontaminated environments. The transconjugant P13 strain was more efficient than either strain P13 or SZH16, and shows how plant growth-promoting bacteria which show no, or only limited, ability to degrade organic pollutants may be modified. This technique is attractive for many environmental remediation and agronomic applications.     

In‐situ remediation of contaminated farmland by horizontal transfer of degradative plasmids among rhizosphere bacteria

Horizontal transfer of catabolic genes for pollutant degradation among rhizobacteria plays an important role in environmental bioremediation, but lacks support from field trial data. To address this problem, Pseudomonas fluorescens strain TP13 was inoculated into the soils of a phenol‐contaminated farmland on which tomato seedlings were growing in April 2009, 2010, 2011 and 2012. Results consistently showed that introduction of TP13 strain significantly reduced phenol content and increased plant biomass after 20 days, compared with controls. Strain TP13 was able to colonize the plant rhizosphere and the number of rhizosphere bacteria which were grown on phenol and contained the plasmids containing the gene encoding for catechol 2, 3‐dioxygenase (C23O) increased gradually in the later stages of the experiment. The increase in magnitude of the plasmid‐containing rhizosphere bacteria correlated well with plant biomass, while the number of plasmid‐containing rhizosphere bacteria and phenol content was strongly negatively correlated. Furthermore, six strains (T1‐T6) of rhizosphere bacteria were isolated and found to possess large plasmids containing identical C23O genes and similar HindIII restriction patterns. Sequence alignment showed that the C23O genes from strains T1‐T6 contained almost identical sequences and the sequence of the C23O of strain T1 was the same as that of strain TP13. These data indicated that the plasmids were transferred from strain TP13 to these rhizosphere bacteria and that horizontal gene transfer stimulated phenol degradation and plant growth in the contaminated farmland. This is an important finding for in situ remediation of contaminated farmland.     

Potential gene exchange between Bacillus thuringiensis subsp. kurstaki and Bacillus spp. in soil in situ

The possible transfer of genes from Bacillus thuringiensis subsp. kurstaki (Btk) to indigenous Bacillus spp. was investigated in soil samples from stands of cork oak in Orotelli (Sardinia, Italy) collected 5 years after spraying of the stands with a commercial insecticidal preparation (FORAY 48B) of Btk. Two colonies with a morphology different from that of Btk were isolated and identified as Bacillus mycoides by morphological and physiological characteristics and by 16S rDNA analysis. Amplification by the polymerase chain reaction (PCR) of the DNA of the two isolated B. mycoides colonies with primers used for the identification of the Btk cry genes showed the presence of a fragment of 238 bp of the cry1Ab9 gene that had a similarity of 100% with the sequence of the cry1Ab9 gene present in GenBank, indicating that the isolates of B. mycoides acquired part of the sequence of this gene from Btk. No cells of Btk or B. mycoides carrying the 238-bp fragment of the cry1Ab9 gene were isolated from samples of unsprayed control soil. However, the isolates of B. mycoides were not able to express the partial Cry1Ab protein. Hybridization with probes for IS231 and the cry1Ab9 gene suggested that the inverted repeated sequence, IS231, was probably involved in the transfer of the 238-bp fragment from Btk to B. mycoides. These results indicate that transfer of genes between introduced Btk and indigenous Bacillus spp. can occur in soil under field conditions.     

Growth promotion of plants by plant growth-promoting rhizobacteria under greenhouse and two different field soil conditions

This study was conducted with sugar beet in greenhouse and field at two soil type with different organic matter (containing 2.4 and 15.9% OM, referred as the low- and high-OM soil) conditions in order to investigate seed inoculation of sugar beet, with five N₂-fixing and two phosphate solubilizing bacteria in comparison to control and mineral fertilizers (N and P) application. Three bacterial strains dissolved P; all bacterial strains fixed N₂ and significantly increased growth of sugar beet. In the greenhouse, inoculations with PGPR increased sugar beet root weight by 2.8-46.7% depending on the species. Leaf, root and sugar yield were increased by the bacterial inoculation by 15.5-20.8, 12.3-16.1, and 9.8-14.7%, respectively, in the experiment of low- and high-OM soil. Plant growth responses were variable and dependent on the inoculants strain, soil organic matter content, growing stage, harvest date and growth parameter evaluated. The effect of PGPR was greater at early growth stages than at the later. Effective Bacillus species, such as OSU-142, RC07 and M-13, Paenibacillus polymyxa RC05, Pseudomonas putida RC06 and Rhodobacter capsulatus RC04 may be used in organic and sustainable agriculture.     

Increased degradation of phenanthrene in soil by Pseudomonas sp. GF3 in the presence of wheat

A phenanthrene-degrading bacterial strain Pseudomonas sp. GF3 was examined for plant-growth promoting effects and phenanthrene removal in soil artificially contaminated with low and high levels of phenanthrene (0, 100 and 200 mg kg⁻¹) in pot experiments. Low and high phenanthrene treatments significantly decreased the growth of wheat. Inoculation with bacterial strain Pseudomonas sp. GF3 was found to increase root and shoot growth of wheat. Strain GF3 was able to degrade phenanthrene effectively in the unplanted and planted soils. Over a period of 80 days the concentration of phenanthrene in soil in which wheat was grown was significantly lower than in unplanted soil (p<0.05). At the end of the 80-d experiments, 62.2% and 42.3% of phenanthrene had disappeared from planted soils without Pseudomonas sp. GF3 when the phenanthrene was added at 100 and 200 mg kg⁻¹ soil, respectively, but 84.8% and 70.2% of phenanthrene had disappeared from planted soils with the bacterial inoculation. The presence of vegetation significantly enhances the dissipation of phenanthrene in the soil. There was no significant difference in soil polyphenol oxidase activities among the applications of 0, 100 and 200 mg kg⁻¹ of phenanthrene. However, the enzyme activities in planted and unplanted soils inoculated with the strain Pseudomonas sp. GF3 were significantly higher than those of non-inoculation controls. The bacterial isolate was also able to colonize and develop in the rhizosphere soil of wheat after inoculation.     

Lignin and cellulose degradation and nitrogen dynamics during decomposition of three leaf litter species in a Mediterranean ecosystem

Cellulose and lignin degradation dynamics was monitored during the leaf litter decomposition of three typical species of the Mediterranean area, Cistus incanus L., Myrtus communis L. and Quercus ilex L., using the litter bag method. Total N and its distribution among lignin, cellulose and acid-detergent-soluble fractions were measured and related to the overall decay process. The litter organic substance of Cistus and Myrtus decomposed more rapidly than that of Quercus. The decay constants were 0.47 year⁻¹, 0.75 year⁻¹ and 0.30 year⁻¹ for Cistus, Myrtus and Quercus, respectively. Lignin and cellulose contents were different as were their relative amounts (34 and 18%, 15 and 37%, 37 and 39% of the overall litter organic matter before exposure, for Cistus, Myrtus and Quercus, respectively). Lignin began to decrease after 6 and 8 months of exposure in Cistus and Myrtus, respectively, while it did not change significantly during the entire study period in Quercus. The holocellulose, in contrast, began to decompose in Cistus after 1 year, while in Quercus and Myrtus immediately. Nitrogen was strongly immobilized in all the litters in the early period of decay. Its release began after the first year in Cistus and Myrtus and after 2 years of decomposition in Quercus. These litters still contained about 60, 20 and 90% of the initial nitrogen at the end of the experiment (3 years). Prior to litter exposure nitrogen associated with the lignin fraction was 65, 54 and 37% in Cistus, Myrtus and Quercus, while that associated with the cellulose fraction was 30, 24 and 28%. Although most of the nitrogen was not lost from litters, its distribution among the litter components changed significantly during decomposition. In Cistus and Myrtus the nitrogen associated with lignin began to decrease just 4 months after exposure. In Quercus this process was slowed and after 3 years of decomposition 8% of the nitrogen remained associated with lignin or lignin-like substances. The nitrogen associated with cellulose or cellulose-like substances, in contrast, began to decrease from the beginning of cellulose decomposition in all three species. At the end of the study period most of the nitrogen was not associated to the lignocellulose fraction but to the acid-detergent-soluble substance (87, 88 and 84% of the remaining litter nitrogen).     

Rhizosphere and endophytic bacteria for induction of systemic resistance of banana plantlets against bunchy top virus

Bunchy top is one of the most important viral diseases affecting banana production worldwide. Several approaches have been made to control or combat the bunchy top virus disease in banana, transmitted by the banana aphid, Pentalonia nigronervosa Coq. No tactic is able to completely protect the plants against this viral infection. To ameliorate this problem, an attempt was made to use different bacterial strains, which could be inoculated to the roots of micropropagated plantlets to avoid the post transplanting problems. Virus indexed micropropagated plantlets of banana cv. Virupakshi (AAB) were subjected to root colonization followed by foliar spraying with three bacterial strains viz., Pseudomonas fluorescens strains Pf1, CHA0 and Bacillus subtilis strain EPB22 along with an un-inoculated control during primary and secondary hardening stage in the nursery and at the time of transplanting, 3rd, 5th and 7th month after planting in the field. Microbe inoculated plantlets showed improved vegetative growth, physiological attributes, PR—proteins and phenol contents besides reducing banana bunchy top disease (BBTD) incidence in the field. These results indicate that, transplant mixes amended with beneficial bacterial strains can enhance growth of banana plants, besides reducing the damage caused by banana bunchy top virus.     

Co-inoculation of selected nodule endophytic rhizobacterial strains with Rhizobium tropici promotes plant growth and controls damping off in common bean

Production of common bean(Phaseolus vulgaris)is limited by the occurrence of damping off(rhizoctoniosis),which is caused by the fungus Rhizoctonia solani.However,the co-inoculation of plant growth-promoting rhizobacteria(PGPR)involved in biological control along with diatomic nitrogen(N2)-fixing rhizobia can enhance N nutrition and increase production.In this context,finding microorganisms with synergistic effects that perform these two roles is of fundamental importance to ensure adequate yield levels.The aim of this study was to evaluate the effects of co-inoculation of nodule endophytic strains of the genera Bacillus,Paenibacillus,Burkholderia,and Pseudomonas with Rhizobium tropici CIAT 899,an N2-fixing rhizobial strain,on the biocontrol of damping off and growth promotion in common bean plants.Greenhouse experiments were conducted under axenic conditions using the common bean cultivar Pérola.The first experiment evaluated the potential of the 14 rhizobacterial strains,which were inoculated alone or in combination with CIAT 899,for the control of R.solani.The second experiment evaluated the ability of these 14 rhizobacterial strains to promote plant growth with three manners of N supply:co-inoculation with CIAT 899 at low mineral N supply(5.25 mg N mL^-1),low mineral N supply(5.25 mg N mL^-1),and high mineral N supply(52.5 mg N mL^-1).The use of rhizobacteria combined with rhizobia contributed in a synergistic manner to the promotion of growth and the control of damping off in the common bean.Co-inoculation of the strains UFLA 02-281/03-18(Pseudomonas sp.),UFLA 02-286(Bacillus sp.),and UFLA 04-227(Burkholderia fungorum)together with CIAT 899 effectively controlled damping off.For the common bean,mineral N supply can be replaced by the co-inoculation of CIAT 899 with plant growth-promoting strains UFLA 02-281/02-286/02-290/02-293.Nodule endophytes UFLA02-281/02-286 are promising for co-inoculation with CIAT 899 in the common bean,promoting synergy with rhizobial inoculation and protection against disease.     

Autumn growth and cold hardening of winter wheat under simulated climate change

Abstract

Plant responses to elevated CO₂ are governed by temperature, and at low temperatures the beneficial effects of CO₂ may be lost. To document the responses of winter cereals grown under cold conditions at northern latitudes, autumn growth of winter wheat exposed to ambient and elevated levels of temperature (+2.5°C), CO₂ (+150 µmol mol^?1), and shade (?30%) was studied in open-top chambers under low light and at low temperatures. Throughout the experiment, temperature dominated plant responses, while the effects of CO₂ were marginal, except for a positive effect on root biomass. Increased temperature resulted in increased leaf area, total biomass, total root biomass, total stem biomass, and number of tillers, but also a lower content of total sugars and a weaker tolerance to frost. The loss of frost tolerance was related to the larger size of plants grown at elevated temperature. The 30% light reduction under shading did not affect the growth, sugar content, or frost tolerance of winter wheat. At the low temperatures found at high latitudes during autumn, the atmospheric CO₂ increase is unlikely to enhance autumn growth of winter wheat to any significant extent, while a temperature increase may have important and major effects on its development and growth.     

Colonisation of seminal roots of wheat and barley by egg-parasitic nematophagous fungi and their effects on Gaeumannomyces graminis var. tritici and development of root-rot

This study provides evidence that egg-parasitic nematophagous fungi, Pochonia chlamydosporia, Pochonia rubescens and Lecanicillium lecanii, can also reduce root colonisation and root damage by a fungal pathogen. Interactions of nematophagous fungi with the take-all fungus, Gaeumannomyces graminis var. tritici (Ggt), and their influence on severity of the root disease it causes were studied in laboratory and pot experiments. In Petri dish experiments the three nematophagous fungi reduced colonisation of barley roots by Ggt and also reduced necrotic symptoms. On the contrary, root colonisation by nematophagous fungi was unaffected by Ggt. In growth tube experiments, the three nematophagous fungi again reduced Ggt root colonisation and increased effective root length of barley seedlings. This was true for both simultaneous and sequential inoculation of nematophagous fungi versus Ggt. In the pot experiments the inoculum of the tested fungi in soil was applied in the same pot, as a mixture or in layers, or in coupled pots used for wheat grown with a split-root system. The nematophagous fungi P. chlamydosporia (isolate 4624) and L. lecanii (isolate 4629), mixed with Ggt or in split root systems with the pathogen, promoted growth of wheat (i.e. increased shoot weight), although no disease reduction was found. In split root systems, lower levels of peroxidase activity were found in seedlings inoculated with Ggt in combination with the nematophagous isolates 4624 and 4629 than when the take-all fungus was applied alone.Our results show that nematophagous fungi reduce root colonisation by Ggt, root damage and stress induced senescence in Ggt-inoculated plants.     

Growth promotion and increased potassium uptake of cotton and rape by a potassium releasing strain of Bacillus edaphicus

A potassium-releasing bacterial strain Bacillus edaphicus NBT was examined for plant-growth-promoting effects and nutrient uptake on cotton and rape in K-deficient soil in pot experiments. Inoculation with bacterial strain B. edaphicus NBT was found to increase root and shoot growth of cotton and rape. Strain NBT was able to mobilize potassium efficiently in both plants when illite was added to the soil. In cotton and rape growing in soils treated with insoluble potassium and inoculated with strain NBT, the potassium content was increased by 30 and 26%, respectively. Bacterial inoculation also resulted in higher N and P contents of above ground plant components. The bacterial isolate was also able to colonize and develop in the rhizosphere soil of cotton and rape after root inoculation.     

Root colonization and growth promotion of wheat and maize by Pseudomonas aurantiaca SR1

Wheat (Triticum aestivum L.), rice (Oryza sativa) and maize (Zea mays L.) are the most important cereals worldwide. However, in the last few years, soil has been submitted to both use and handling pressures due to the increase in agricultural practices, which are leading to its degradation. The use of plant growth-promoting rhizobacteria (PGPR) as inoculants constitutes a biological alternative for sustainable production. Pseudomonas aurantiaca SR1 was formulated as an inoculant in order to evaluate its growth promotion effect in the field when applied on maize and wheat seeds at the sowing time. P and N fertilization treatments were also included in the assays. P. aurantiaca SR1 colonized the root system of both crops and it persisted at appropriate population densities. It also showed a significant plant growth-promoting effect that was reflected in the yield. Another relevant finding was that both crops, when inoculated with P. aurantiaca SR1, presented higher yields with fertilization doses lower than those conventionally applied. This indicated its potential use as a reasonable alternative for crop production, with a minimization of the ecological impact.     

水体氮浓度对狐尾藻和金鱼藻片段萌发及生长的影响

为了厘清水体氮浓度对沉水植物片段萌发及生长的影响,通过模拟控制试验,设计了水体总氮浓度分别为0 (CK)、 0.5 mg/L (N1)、 2 mg/L (N2)、 8 mg/L (N3)和12 mg/L (N4）共5个处理,研究了水体不同氮浓度条件下狐尾藻 (Myriophyllum spicatum) 和金鱼藻 (Ceratophyllum demersum)片段(3节)萌芽数、 萌发位置、 芽长和生物量的差异。结果表明, 水体氮浓度对不同沉水植物片段萌发的影响存在差异,较高的水体氮浓度不利于狐尾藻片断萌发,而对金鱼藻片段却有一定程度的促进;氮浓度处理促进了狐尾藻顶端优势,却抑制了金鱼藻的顶端优势。狐尾藻以0.5 mg/L处理的芽长较长,生物量以2 mg/L处理最大;而金鱼藻以8 mg/L处理芽长最长,生物量以2 mg/L处理最大。由此可见,不同沉水植物对水体氮适应性存在差异,水体氮浓度较低时,沉水植物断枝可以进行萌发和生长, 而当氮浓度超过2 mg/L时,对沉水植物断枝萌发及生长反而有抑制作用;金鱼藻片段比狐尾藻对水体氮浓度的耐受性强。     

田菁茎瘤固氮根瘤菌在小麦体内的定殖及营养元素相关miRNA的表达

【目的】随着化肥过度使用引起的环境问题的出现,无公害农业的推广,以及新型肥料研究领域不断拓宽,微生物肥料,尤其是植物根际促生菌的研究成为近年来的热点。然而微生物肥料的增产机理还基本停留在作物农学性状的表观调查上,没有从分子水平进行研究。因此,本文用田菁茎瘤固氮根瘤菌（Azorhizobium caulinodans ORS571）侵染小麦,探索GFP-A.caulinodans在小麦幼苗组织中的分布与定殖规律以及营养元素相关miRNAs在小麦与A.caulinodans互作中的作用机制。【方法】使用A.caulinodans侵染小麦种子（品种为小偃22）,将接菌6 d后的小麦幼苗的根和接菌12 d后的叶制作玻片,利用激光共聚焦显微镜对样品进行逐层扫描,检测GFP-A.caulinodans在幼苗不同组织中的分布与定殖情况。同时,采集接菌后0 h、 6 h、 12 h、 24 h、 48 h、 72 h、 96 h的小麦幼根取样,用Trizol法提取总RNA。利用试剂盒进行加尾和反转录反应,将样品总RNA中的miRNA合成为cDNA,使用-tubulin作为内参基因,进行实时定量PCR反应,使用2-CT方法计算相对表达量。利用psRNATarget 在线软件,采用默认参数,对miRNA的靶基因进行预测。【结果】 1）激光共聚焦结果显示,GFP-A.caulinodans可定殖于根的表皮细胞、 细胞间隙、 根尖破损处和根毛,在根维管组织和叶片气孔部位,也发现有GFP-A.caulinodans存在。2）实时定量PCR分析结果表明,6条与营养元素代谢有关的miRNA表达发生变化,其中miR164、 miR167和miR827相对表达量呈现出先上调后下调的趋势,miR169 和miR398相对表达量也基本呈现出这一趋势。miR164、 miR167、 miR169和miR398的相对表达量在接菌12 h时上调至最高点,分别为对照的4.13、 2.84、 2.46和3.99倍; miR827相对表达量在接菌24 h时达到最高点,为对照的2.17倍。miR399相对表达量呈现出先下调后上调的趋势,在接菌24 h时降至最低点,为对照的0~21倍。3）通过靶基因预测,6条miRNA的靶基因分别编码了NAC1转录因子、 生长素响应因子、 HAP转录因子、 Cu/Zn 超氧化物歧化酶、 蛋白缀合酶PHO2和SPX-MFS亚家族蛋白。【结论】GFP-A.caulinodans能够从根毛和根尖破损处等部位进入小麦幼苗根部定殖,并可通过向上迁移到达叶片,在气孔处定殖。接种A.caulinodans可不同程度增加小麦根中响应氮素、 磷素、 微量元素的miRNAs相对表达量,增强小麦幼苗对营养元素的吸收和利用,促进小麦根的形态建成。     

Reduction of fungal growth and lignin decomposition in needle litter by avian excreta

The effects of excessive addition of excreta from the Great Cormorant Phalacrocorax carbo, a colonial piscivorous bird, on the growth and the ability of fungi to decompose needle litter of Chamaecyparis obtusa were examined by a pure-culture test. Colony growth rate, mass loss of needle litter, and utilization patterns of lignin and carbohydrates were investigated and compared for 22 species in basidiomycetes, ascomycetes, and zygomycetes. Colony growth rate of basidiomycetes decreased on medium supplemented with excreta (excreta medium) as compared to control medium without excreta, whereas such a difference was not found for ascomycetes. Mass loss of needle litter caused by basidiomycetes was generally higher than those caused by ascomycetes and zygomycetes. Basidiomycetes decomposed both lignin and carbohydrates in various proportions, whereas ascomycetes and zygomycetes decomposed carbohydrates selectively. Mass loss of litter caused by basidiomycetes and ascomycetes was lower when incubated on excreta medium than on control medium. Mass loss of lignin and nitrogen caused by basidiomycetes was lower on excreta medium than on control medium, whereas such differences were not found for ascomycetes. Mass loss of carbohydrate was not different between the media for basidiomycetes or ascomycetes.     

Selection of Variovorax paradoxus-like bacteria in the mycosphere and the role of fungal-released compounds

At two occasions (2004 and 2006), a similar cluster of culturable bacteria was found to be selected in the mycosphere of the basidiomycetous fungus Laccaria proxima (Agaricales, Tricholomataceae) in the field. The bacteria, identified as related to Variovorax paradoxus, comprised 7.3-9% of the total culturable bacterial community in the L. proxima mycosphere, but were not found in corresponding bulk soil (<0.3%). One strain, denoted HB44, was selected in order to unravel the basis of the V. paradoxus mycosphere competence in in vitro experiments with the former Laccaria laccata, which was recently reclassified as Lyophyllum sp. strain Karsten (Agaricales, Tricholomataceae). In liquid culture experiments, L. strain Karsten was shown to be an avid producer of glycerol, next to acetate and formate, which constituted the most abundant carbonaceous compounds released. Strain HB44 was able to grow avidly at the expense of the glycerol liberated by the fungus, as evidenced by proton NMR analysis of the fungal exudates in the medium before and after bacterial growth. In sterilized field soil, strain HB44 survived significantly better in the presence than in the absence of L. strain Karsten. Addition of a glycerol series to the sterilized soil (without the fungus) resulted in the persistence or growth of strain HB44, but only if the pH of this soil was previously set at 5.5. Thus, we provide evidence for the contention that tricholomataceous fungi can create specific niches in soil for the V. paradoxus-related strain HB44 and that glycerol may be a main carbon source that drives the selection of this organism.     

Fate, tree growth effect and potential impact on soil microbial communities of mycorrhizal and bacterial inoculation in a forest plantation

The knowledge of the survival of inoculated beneficial fungal and bacterial strains in the field and the effects of their release on the indigenous microbial communities has been of great interest since the practical use of selected natural or genetically modified microorganisms has been developing. The aim of this study was to monitor, 4 years after plantation into the field site, the effects of Douglas fir (Pseudotsuga menziesii) co-inoculation with the mycorrhiza helper bacterial strain Pseudomonas fluorescens BBc6R8 and/or the fungal strain Laccaria bicolor S238N on seedling growth and on the indigenous bacterial and ectomycorrhizal communities using quantitative and qualitative approaches. The field persistence of the inoculated strains was also monitored. The seedling shoot volume estimate was statistically significantly higher in the fungal inoculated plots in comparison to the non-inoculated plots but no treatment-related changes in the quantitave or qualitative microbial measurements were observed and the inoculated strains could not be detected after 4 years.     

Production of lignocellulose-degrading enzymes and degradation of leaf litter by saprotrophic basidiomycetes isolated from a Quercus petraea forest

Due to the production of lignocellulose-degrading enzymes, saprotrophic basidiomycetes can significantly contribute to the turnover of soil organic matter. The production of lignin- and polysaccharide-degrading enzymes and changes of the chemical composition of litter were studied with three isolates from a Quercus petraea forest. These isolates were capable of fresh litter degradation and were identified as Gymnopus sp., Hypholoma fasciculare and Rhodocollybia butyracea. Within 12 weeks of incubation, H. fasciculare decomposed 23%, R. butyracea 32% and Gymnopus sp. 38% of the substrate dry mass. All fungi produced laccase and Mn-peroxidase (MnP) and none of them produced lignin peroxidase or other Mn-independent peroxidases. There was a clear distinction in the enzyme production pattern between R. butyracea or H. fasciculare compared to Gymnopus sp. The two former species caused the fastest mass loss during the initial phase of litter degradation, accompanied by the temporary production of laccase (and MnP in H. fasciculare) and also high production of hydrolytic enzymes that later decreased. In contrast, Gymnopus sp. showed a stable rate of litter mass loss over the whole incubation period with a later onset of ligninolytic enzyme production and a longer lasting production of both lignin and cellulose-degrading enzymes. The activity of endo-cleaving polysaccharide hydrolases in this fungus was relatively low but it produced the most cellobiose hydrolase. All fungi decreased the C/N ratio of the litter from 24 to 15-19 and Gymnopus sp. also caused a substantial decrease in the lignin content. Analytical pyrolysis mass spectrometry of litter decomposed by this fungus showed changes in the litter composition similar to those caused by white-rot fungi during wood decay. These changes were less pronounced in the case of H. fasciculare and R. butyracea. All fungi also changed the mean masses of humic acid and fulvic acid fractions isolated from degraded litter. The humic acid fraction after degradation by all three fungi contained more lignin and less carbohydrates. Compared to the decomposition by saprotrophic basidiomycetes, litter degradation in situ on the site of fungal isolation resulted in the relative enrichment of lignin and differences in lignin composition revealed by analytical pyrolysis. It can most probably be explained by the participation of non-basidiomycetous fungi and bacteria during natural litter decomposition.     

Upper and lower levels of boron for cotton and wheat as influenced by calcium carbonate

Abstract

Pot studies were conducted to study effect of the boron (B) levels (0, 0.12, 0.25, 0.50, 0.75, 1.0, 2.0 3.0, 4.0, 5.0 and 10?mg?B?kg^?1) and calcium carbonate (CaCO₃) (0, 1.25, 2.5 and 5.0%) on cotton and wheat crops in cropping system. In absence of CaCO_3, reduced growth of cotton and wheat were observed when B was applied @ 2.0?mg?kg^?1 soil. Necrosis of leaves in cotton and purple coloration of plumule in wheat was observed @ 10.0?mg?B?kg^?1. Irrespective of CaCO₃, 0.57 and 7.67?mg?kg^?1 Hot Water Soluble B (HWS-B), 62 and 940?mg?kg^?1 B dry matter (DM) of leaves, 45 and 210?mg?B?kg^?1 DM of petioles and 20 and 51?mg?B?kg^?1 DM of sticks, produced 90 and 50%of the maximum dry matter yield (DMY) of cotton, respectively. The values for wheat were 0.66 and 6.71?mg HWS-B kg^?1, 7.94 and 27.0?mg?B?kg^?1 grain and 15.3 and 170?mg?B?kg^?1 straw, respectively.