Commenced in January 2007
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Edition: International
Paper Count: 55

Search results for: rhizosphere

55 Isolation and Characterization of Indigenous Rhizosphere Bacteria Producing Gibberellin Acid from Local Soybeans in Three Different Areas of South Sulawesi

Authors: Asmiaty Sahur, Ambo Ala, Baharuddin Patanjengi, Elkawakib Syam'un


This study aimed to isolate and characterize the indigenous Rhizosphere bacteria producing Gibberellin Acid as plant growth isolated from local soybean of three different areas in South Sulawesi, Indonesia. Several soil samples of soybean plants were collected from the Rhizosphere of local soybeans in three different areas of South Sulawesi such as Soppeng, Bone and Takalar. There were 56 isolates of bacteria were isolated and grouped into gram-positive bacteria and gram negative bacteria .There are 35 isolates produce a thick slime or slimy when cultured on media Natrium Broth and the remaining of those produced spores. The results showed that of potential bacterial isolated produced Gibberellin Acid in high concentration. The best isolate of Rhizosphere bacteria for the production of Gibberellin Acid is with concentration 2%. There are 4 isolates that had higher concentration are AKB 19 (4.67 mg/ml) followed by RKS 17 (3.80 mg/ml), RKS 25 (3.70 mg / ml) and RKS 24 (3.29 mg/ml) respectively.

Keywords: rhizosphere, bacteria, gibberellin acid, soybeans

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54 Biodiversity of Plants Rhizosphere and Rhizoplane Bacteria in the Presence of Petroleum Hydrocarbons

Authors: Togzhan D. Mukasheva, Anel A. Omirbekova, Raikhan S. Sydykbekova, Ramza Zh. Berzhanova, Lyudmila V. Ignatova


Following plants-barley (Hordeum sativum), alfalfa (Medicago sativa), grass mixture (red fescue-75%, long-term ryegrass - 20% Kentucky bluegrass - 10%), oilseed rape (Brassica napus biennis), resistant to growth in the contaminated soil with oil content of 15.8 g / kg 25.9 g / kg soil were used. Analysis of the population showed that the oil pollution reduces the number of bacteria in the rhizosphere and rhizoplane of plants and enhances the amount of spore-forming bacteria and saprotrophic micromycetes. It was shown that regardless of the plant, dominance of Pseudomonas and Bacillus genera bacteria was typical for the rhizosphere and rhizoplane of plants. The frequency of bacteria of these genera was more than 60%. Oil pollution changes the ratio of occurrence of various types of bacteria in the rhizosphere and rhizoplane of plants. Besides the Pseudomonas and Bacillus genera, in the presence of hydrocarbons in the root zone of plants dominant and most typical were the representatives of the Mycobacterium and Rhodococcus genera. Together the number was between 62% to 72%.

Keywords: pollution, root system, micromycetes, identification

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53 Rhizosphere Microbiome Involvement in the Natural Suppression of Soybean Cyst Nematode in Disease Suppressive Soil

Authors: M. Imran Hamid, Muzammil Hussain, Yunpeng Wu, Meichun Xiang, Xingzhong Liu


The rhizosphere microbiome elucidate multiple functioning in the soil suppressiveness against plant pathogens. Soybean rhizosphere microbial communities may involve in the natural suppression of soybean cyst nematode (SCN) populations in disease suppressive soils. To explore these ecological mechanisms of microbes, a long term monoculture suppressive soil were taken into account for further investigation to test the disease suppressive ability by using different treatments. The designed treatments are as, i) suppressive soil (S), ii) conducive soil (C), iii) conducive soil mixed with 10% (w/w) suppressive soil (CS), iv) suppressive soil treated at 80°C for 1 hr (S80), and v) suppressive soil treated with formalin (SF). By using an ultra-high-throughput sequencing approach, we identified the key bacterial and fungal taxa involved in SCN suppression. The Phylum-level investigation of bacteria revealed that Actinobacteria, Bacteroidetes, and Proteobacteria in the rhizosphere soil of soybean seedlings were more abundant in the suppressive soil than in the conducive soil. The phylum-level analysis of fungi in rhizosphere soil indicated that relative abundance of Ascomycota was higher in suppressive soil than in the conducive soil, where Basidiomycota was more abundant. Transferring suppressive soil to conducive soil increased the population of Ascomycota in the conducive soil by lowering the populations of Basidiomycota. The genera, such as, Pochonia, Purpureocillium, Fusarium, Stachybotrys that have been well documented as bio-control agents of plant nematodes were far more in the disease suppressive soils. Our results suggested that the plants engage a subset of functional microbial groups in the rhizosphere for initial defense upon nematode attack and protect the plant roots later on by nematodes to response for suppression of SCN in disease-suppressive soils.

Keywords: disease suppressive soil, high-throughput sequencing, rhizosphere microbiome, soybean cyst nematode

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52 The Response of Soil Biodiversity to Agriculture Practice in Rhizosphere

Authors: Yan Wang, Guowei Chen, Gang Wang


Soil microbial diversity is one of the important parameters to assess the soil fertility and soil health, even stability of the ecosystem. In this paper, we aim to reveal the soil microbial difference in rhizosphere and root zone, even to pick the special biomarkers influenced by the long term tillage practices, which included four treatments of no-tillage, ridge tillage, continuous cropping with corn and crop rotation with corn and soybean. Here, high-throughput sequencing was performed to investigate the difference of bacteria in rhizosphere and root zone. The results showed a very significant difference of species richness between rhizosphere and root zone soil at the same crop rotation system (p < 0.01), and also significant difference of species richness was found between continuous cropping with corn and corn-soybean rotation treatment in the rhizosphere statement, no-tillage and ridge tillage in root zone soils. Implied by further beta diversity analysis, both tillage methods and crop rotation systems influence the soil microbial diversity and community structure in varying degree. The composition and community structure of microbes in rhizosphere and root zone soils were clustered distinctly by the beta diversity (p < 0.05). Linear discriminant analysis coupled with effect size (LEfSe) analysis of total taxa in rhizosphere picked more than 100 bacterial taxa, which were significantly more abundant than that in root zone soils, whereas the number of biomarkers was lower between the continuous cropping with corn and crop rotation treatment, the same pattern was found at no-tillage and ridge tillage treatment. Bacterial communities were greatly influenced by main environmental factors in large scale, which is the result of biological adaptation and acclimation, hence it is beneficial for optimizing agricultural practices.

Keywords: tillage methods, biomarker, biodiversity, rhizosphere

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51 Rhizosphere Microbial Communities in Fynbos Endemic Legumes during Wet and Dry Seasons

Authors: Tiisetso Mpai, Sanjay K. Jaiswal, Felix D. Dakora


The South African Cape fynbos biome is a global biodiversity hotspot. This biome contains a diversity of endemic shrub legumes, including Polhillia, Wiborgia, and Wiborgiella species, which are important for ecotourism as well as for improving soil fertility status. This is due to their proven N₂-fixing abilities when in association with compatible soil bacteria. In fact, Polhillia, Wiborgia, and Wiborgiella species have been reported to derive over 61% of their needed nitrogen through biological nitrogen fixation and to exhibit acid and alkaline phosphatase activity in their rhizospheres. Thus, their interactions with soil microbes may explain their survival mechanisms under the continued summer droughts and acidic, nutrient-poor soils in this region. However, information regarding their rhizosphere microbiome is still unavailable, yet it is important for Fynbos biodiversity management. Therefore, the aim of this study was to assess the microbial community structures associated with rhizosphere soils of Polhillia pallens, Polhillia brevicalyx, Wiborgia obcordata, Wiborgia sericea, and Wiborgiella sessilifolia growing at different locations of the South African Cape fynbos, during the wet and dry seasons. The hypothesis is that the microbial communities in these legume rhizospheres are the same type and are not affected by the growing season due to the restricted habitat of these wild fynbos legumes. To obtain the results, DNA was extracted from 0.5 g of each rhizosphere soil using PowerSoil™ DNA Isolation Kit, and sequences were obtained using the 16S rDNA Miseq Illumina technology. The results showed that in both seasons, bacteria were the most abundant microbial taxa in the rhizosphere soils of all five legume species, with Actinobacteria showing the highest number of sequences (about 30%). However, over 19.91% of the inhabitants in all five legume rhizospheres were unclassified. In terms of genera, Mycobacterium and Conexibacter were common in rhizosphere soils of all legumes in both seasons except for W. obcordata soils sampled during the dry season, which had Dehalogenimonas as the major inhabitant (6.08%). In conclusion, plant species and season were found to be the main drivers of microbial community structure in Cape fynbos, with the wet season being more dominant in shaping microbial diversity relative to the dry season. Wiborgia obcordata had a greater influence on microbial community structure than the other four legume species.

Keywords: 16S rDNA, Cape fynbos, endemic legumes, microbiome, rhizosphere

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50 Effect of Fertilization and Combined Inoculation with Azospirillum brasilense and Pseudomonas fluorescens on Rhizosphere Microbial Communities of Avena sativa (Oats) and Secale Cereale (Rye) Grown as Cover Crops

Authors: Jhovana Silvia Escobar Ortega, Ines Eugenia Garcia De Salamone


Cover crops are an agri-technological alternative to improve all properties of soils. Cover crops such as oats and rye could be used to reduce erosion and favor system sustainability when they are grown in the same agricultural cycle of the soybean crop. This crop is very profitable but its low contribution of easily decomposable residues, due to its low C/N ratio, leaves the soil exposed to erosive action and raises the need to reduce its monoculture. Furthermore, inoculation with the plant growth promoting rhizobacteria contributes to the implementation, development and production of several cereal crops. However, there is little information on its effects on forage crops which are often used as cover crops to improve soil quality. In order to evaluate the effect of combined inoculation with Azospirillum brasilense and Pseudomonas fluorescens on rhizosphere microbial communities, field experiments were conducted in the west of Buenos Aires province, Argentina, with a split-split plot randomized complete block factorial design with three replicates. The factors were: type of cover crop, inoculation and fertilization. In the main plot two levels of fertilization 0 and 7 40-0-5 (NPKS) were established at sowing. Rye (Secale cereale cultivar Quehué) and oats (Avena sativa var Aurora.) were sown in the subplots. In the sub-subplots two inoculation treatments are applied without and with application of a combined inoculant with A. brasilense and P. fluorescens. Due to the growth of cover crops has to be stopped usually with the herbicide glyphosate, rhizosphere soil of 0-20 and 20-40 cm layers was sampled at three sampling times which were: before glyphosate application (BG), a month after glyphosate application (AG) and at soybean harvest (SH). Community level of physiological profiles (CLPP) and Shannon index of microbial diversity (H) were obtained by multivariate analysis of Principal Components. Also, the most probable number (MPN) of nitrifiers and cellulolytics were determined using selective liquid media for each functional group. The CLPP of rhizosphere microbial communities showed significant differences between sampling times. There was not interaction between sampling times and both, types of cover crops and inoculation. Rhizosphere microbial communities of samples obtained BG had different CLPP with respect to the samples obtained in the sampling times AG and SH. Fertilizer and depth of sampling also caused changes in the CLPP. The H diversity index of rhizosphere microbial communities of rye in the sampling time BG were higher than those associated with oats. The MPN of both microbial functional types was lower in the deeper layer since these microorganisms are mostly aerobic. The MPN of nitrifiers decreased in rhizosphere of both cover crops only AG. At the sampling time BG, the NMP of both microbial types were larger than those obtained for AG and SH. This may mean that the glyphosate application could cause fairly permanent changes in these microbial communities which can be considered bio-indicators of soil quality. Inoculation and fertilizer inputs could be included to improve management of these cover crops because they can have a significant positive effect on the sustainability of the agro-ecosystem.

Keywords: community level of physiological profiles, microbial diversity, plant growth promoting rhizobacteria, rhizosphere microbial communities, soil quality, system sustainability

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49 Phytoremediation Potenciality of ‘Polypogon monspeliensis L. in Detoxification of Petroleum-Contaminated Soils

Authors: Mozhgan Farzami Sepehr, Farhad Nourozi


In a greenhouse study, decontamination capacity of the species Polypogon monspoliensis, for detoxification of petroleum-polluted soils caused by sewage and waste materials of Tehran Petroleum Refinery. For this purpose, the amount of total oil and grease before and 45 days after transplanting one-month-old seedlings in the soils of five different treatments in which pollution-free agricultural soil and contaminated soil were mixed together with the weight ratio of respectively 1 to 9 (% 10), 2 to 8 (%20), 3 to 7 (%30) , 4 to 6 (%40), and 5 to 5 (%50) were evaluated and compared with the amounts obtained from control treatment without vegetation, but with the same concentration of pollution. Findings demonstrated that the maximum reduction in the petroleum rate ,as much as 84.85 percent, is related to the treatment 10% containing the plant. Increasing the shoot height in treatments 10% and 20% as well as the root dry and fresh weight in treatments 10% , 20% , and 30% shows that probably activity of more rhizosphere microorganisms of the plant in these treatments has led to the improvement in growth of plant organs comparing to the treatments without pollution.

Keywords: phytoremediation, total oil and grease, rhizosphere, microorganisms, petroleum-contaminated soil

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48 Study on the Enhancement of Soil Fertility and Tomato Quality by Applying Concentrated Biogas Slurry

Authors: Fang Bo Yu, Li Bo Guan


Biogas slurry is a low-cost source of crop nutrients and can offer extra benefits to soil fertility and fruit quality. However, its current utilization mode and low content of active ingredients limit its application scale. In this report, one growing season field research was conducted to assess the effects of concentrated biogas slurry on soil property, tomato fruit quality, and composition of the microflora in both non-rhizosphere and rhizosphere soils. The results showed that application of concentrated slurry could cause significant changes to tomato cultivation, including increases in organic matter, available N, P, and K, total N, and P, electrical conductivity, and fruit contents of amino acids, protein, soluble sugar, β-carotene, tannins, and vitamin C, together with the R/S ratios and the culturable counts of bacteria, actinomycetes, and fungi in soils. It could be concluded as the application is a practicable means in tomato production and might better service the sustainable agriculture in the near future.

Keywords: concentrated slurry, fruit quality, soil fertility, sustainable agriculture

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47 Effect of Chemical Fertilizer on Plant Growth-Promoting Rhizobacteria in Wheat

Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline


The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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46 Combined Aplication of Indigenous Pseudomonas fluorescens and the AM Fungi as the Potential Biocontrol Agents of Banana Fusarium wilt

Authors: Eri Sulyanti, Trimurti Habazar, Eti Farda Husen, Abdi Dharma, Nasril Nasir


In this study, combination of some biocontrol agents with different mechanisms was an alternative to improve the effectiveness of the biological control agents. Single and combined applications of indigenous Pseudomonas fluorescens and Arbuscular Mychorrhizae Fungi (AM Fungi) isolates were tested to induce the resistance on susceptible Cavendish banana against F.oxysporum f. sp. cubense race 4 under greenhouse conditions. These isolates originally isolated from healthy banana rhizosphere at endemic Fusarium wilt areas in the centre of production banana in West Sumatra. These researches were conducted with Randomized Block Design with 16 treatments and 10 replications. The treatments were three indigenous isolates of Pseudomonas fluorescens (Par1-Cv, Par4-Rj1, Par2-Jt1) and 3 isolates of AM Fungi (Gl1BuA4, Gl2BuA6, and Gl1KeP3. The biocontrol agents were applied as single agents and combination two of them. This study demonstrated that the application of combination biocontrol organisms Pseudomonas fluorescens and AM Fungi provided were more effective than single application. The combination of Par1-Cv and Gl1BuA4 isolates was the most effective to control Fusarium wilt and followed by the combination of Par1-Cv and Gl2BuA6 and Par2-Jt1 and Gl1P3.

Keywords: pseudomonad fluorescens (Pf), arbuscular mychorrhizae fungi (AM Fungi) indigenous isolates, fusarium oxysporum f. sp. cubense, soil rhizosphere

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45 Plant Microbiota of Coastal Halophyte Salicornia Ramossisima

Authors: Isabel N. Sierra-Garcia, Maria J. Ferreira, Sandro Figuereido, Newton Gomes, Helena Silva, Angela Cunha


Plant-associated microbial communities are considered crucial in the adaptation of halophytes to coastal environments. The plant microbiota can be horizontally acquired from the environment or vertically transmitted from generation to generation via seeds. Recruiting of the microbial communities by the plant is affected by geographical location, soil source, host genotype, and cultivation practice. There is limited knowledge reported on the microbial communities in halophytes the influence of biotic and abiotic factors. In this work, the microbiota associated with the halophyte Salicornia ramosissima was investigated to determine whether the structure of bacterial communities is influenced by host genotype or soil source. For this purpose, two contrasting sites where S. ramosissima is established in the estuarine system of the Ria de Aveiro were investigated. One site corresponds to a natural salt marsh where S. ramosissima plants are present (wild plants), and the other site is a former salt pan that nowadays are subjected to intensive crop production of S. ramosissima (crop plants). Bacterial communities from the rhizosphere, seeds and root endosphere of S. ramossisima from both sites were investigated by sequencing bacterial 16S rRNA gene using the Illumina MiSeq platform. The analysis of the sequences showed that the three plant-associated compartments, rhizosphere, root endosphere, and seed endosphere, harbor distinct microbiomes. However, bacterial richness and diversity were higher in seeds of wild plants, followed by rhizosphere in both sites, while seeds in the crop site had the lowest diversity. Beta diversity measures indicated that bacterial communities in root endosphere and seeds were more similar in both wild and crop plants in contrast to rhizospheres that differed by local, indicating that the recruitment of the similar bacterial communities by the plant genotype is active in regard to the site. Moreover, bacterial communities from the root endosphere and rhizosphere were phylogenetically more similar in both sites, but the phylogenetic composition of seeds in wild and crop sites was distinct. These results indicate that cultivation practices affect the seed microbiome. However, minimal vertical transmission of bacteria from seeds to adult plants is expected. Seeds from the crop site showed higher abundances of Kushneria and Zunongwangia genera. Bacterial members of the classes Alphaprotebacteria and Bacteroidia were the most ubiquitous across sites and compartments and might encompass members of the core microbiome. These findings indicate that bacterial communities associated with S. ramosissima are more influenced by host genotype rather than local abiotic factors or cultivation practices. This study provides a better understanding of the composition of the plant microbiota in S. ramosissima , which is essential to predict the interactions between plant and associated microbial communities and their effects on plant health. This knowledge is useful to the manipulations of these microbial communities to enhance the health and productivity of this commercially important plant.

Keywords: halophytes, plant microbiome, Salicornia ramosissima, agriculture

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44 Rhizoremediation of Contaminated Soils in Sub-Saharan Africa: Experimental Insights of Microbe Growth and Effects of Paspalum Spp. for Degrading Hydrocarbons in Soils

Authors: David Adade-Boateng, Benard Fei Baffoe, Colin A. Booth, Michael A. Fullen


Remediation of diesel fuel, oil and grease in contaminated soils obtained from a mine site in Ghana are explored using rhizoremediation technology with different levels of nutrient amendments (i.e. N (nitrogen) in Compost (0.2, 0.5 and 0.8%), Urea (0.2, 0.5 and 0.8%) and Topsoil (0.2, 0.5 and 0.8%)) for a native species. A Ghanaian native grass species, Paspalum spp. from the Poaceae family, indicative across Sub-Saharan Africa, was selected following the development of essential and desirable growth criteria. Vegetative parts of the species were subjected to ten treatments in a Randomized Complete Block Design (RCBD) in three replicates. The plant-associated microbial community was examined in Paspalum spp. An assessment of the influence of Paspalum spp on the abundance and activity of micro-organisms in the rhizosphere revealed a build-up of microbial communities over a three month period. This was assessed using the MPN method, which showed rhizospheric samples from the treatments were significantly different (P <0.05). Multiple comparisons showed how microbial populations built-up in the rhizosphere for the different treatments. Treatments G (0.2% compost), H (0.5% compost) and I (0.8% compost) performed significantly better done other treatments, while treatments D (0.2% topsoil) and F (0.8% topsoil) were insignificant. Furthermore, treatment A (0.2% urea), B (0.5% urea), C (0.8% urea) and E (0.5% topsoil) also performed the same. Residual diesel and oil concentrations (as total petroleum hydrocarbons, TPH and oil and grease) were measured using infra-red spectroscopy and gravimetric methods, respectively. The presence of single species successfully enhanced the removal of hydrocarbons from soil. Paspalum spp. subjected to compost levels (0.5% and 0.8%) and topsoil levels (0.5% and 0.8%) showed significantly lower residual hydrocarbon concentrations compared to those treated with Urea. A strong relationship (p<0.001) between the abundance of hydrocarbon degrading micro-organisms in the rhizosphere and hydrocarbon biodegradation was demonstrated for rhizospheric samples with treatment G (0.2% compost), H (0.5% compost) and I (0.8% compost) (P <0.001). The same level of amendment with 0.8% compost (N-level) can improve the application effectiveness. These findings have wide-reaching implications for the environmental management of soils contaminated by hydrocarbons in Sub-Saharan Africa. However, it is necessary to further investigate the in situ rhizoremediation potential of Paspalum spp. at the field scale.

Keywords: rhizoremediation, microbial population, rhizospheric sample, treatments

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43 Iron Influx, Its Root-Shoot Relations and Utilization Efficiency in Wheat

Authors: Abdul Malik Dawlatzai, Shafiqullah Rahmani


Plant cultivars of the same species differ in their Fe efficiency. This paper studied the Fe influx and root-shoot relations of Fe at different growth stages in wheat. The four wheat cultivars (HD 2967, PDW 233, PBW 550 and PDW 291) were grown in pots in Badam Bagh agricultural researching farm, Kabul under two Fe treatments: (i) 0 mg Fe kg⁻¹ soil (soil with 2.7 mg kg⁻¹ of DTPA-extractable Fe) and (ii) 50 mg Fe kg⁻¹ soil. Root length (RL), shoot dry matter (SDM), Fe uptake, and soil parameters were measured at tillering and anthesis. Application of Fe significantly increased RL, root surface area, SDM, and Fe uptake in all wheat cultivars. Under Fe deficiency, wheat cv. HD 2967 produced 90% of its maximum RL and 75% of its maximum SDM. However, PDW 233 produced only 69% and 60%, respectively. Wheat cultivars HD 2967, and PDW 233 exhibited the highest and lowest value of root surface area and Fe uptake, respectively. The concentration difference in soil solution Fe between bulk soil and root surface (ΔCL) was maximum in wheat cultivar HD 2967, followed by PBW 550, PDW 291, and PDW 233. More depletion at the root surface causes steeper concentration gradients, which result in a high influx and transport of Fe towards root. Fe influx in all the wheat cultivars increased with the Fe application, but the increase was maximum, i.e., 4 times in HD 2967 and minimum, i.e., 2.8 times in PDW 233. It can be concluded that wheat cultivars HD 2967 and PBW 550 efficiently utilized Fe as compared to other cultivars. Additionally, iron efficiency of wheat cultivars depends upon uptake of each root segment, i.e., the influx, which in turn depends on depletion of Fe in the rhizosphere during vegetative phase and higher utilization efficiency of acquired Fe during reproductive phase that governs the ultimate grain yield.

Keywords: Fe efficiency, Fe influx, Fe uptake, Rhizosphere

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42 Fungicidal Action of the Mycogenic Silver Nanoparticles Against Aspergillus niger Inciting Collar Rot Disease in Groundnut (Arachis hypogaea L.)

Authors: R. Sarada Jayalakshmi Devi B. Bhaskar, S. Khayum Ahammed, T. N. V. K. V. Prasad


Use of bioagents and biofungicides is safe to manage the plant diseases and to avoid human health hazards which improves food security. Myconanotechnology is the study of nanoparticles synthesis using fungi and their applications. The present work reports on preparation, characterization and antifungal activity of biogenic silver nanoparticles produced by the fungus Trichoderma sp. which was collected from groundnut rhizosphere. The culture filtrate of Trichoderma sp. was used for the reduction of silver ions (Ag+) in AgNO3 solution to the silver (Ag0) nanoparticles. The different ages (4 days, 6 days, 8 days, 12 days, and 15 days) of culture filtrates were screened for the synthesis of silver nanoparticles. Synthesized silver nanoparticles were characterized using UV-Vis spectrophotometer, particle size and zeta potential analyzer, Fourier Transform Infrared Spectrophotometer (FTIR) and Transmission Electron Microscopy. Among all the treatments the silver nitrate solution treated with six days aged culture filtrate of Trichoderma sp. showed the UV absorption peak at 440 nm with maximum intensity (0.59) after 24 hrs incubation. The TEM micrographs showed the spherical shaped silver nanoparticles with an average size of 30 nm. The antifungal activity of silver nanoparticles against Aspergillus niger causing collar rot disease in groundnut and aspergillosis in humans showed the highest per cent inhibition at 100 ppm concentration (74.8%). The results points to the usage of these mycogenic AgNPs in agriculture to control plant diseases.

Keywords: groundnut rhizosphere, Trichoderma sp., silver nanoparticles synthesis, antifungal activity

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41 Transcriptomic Response of Calmodulin Encoding Gene (CaM) in Pesticide Utilizing Talaromyces Fungal Strains

Authors: M. D. Asemoloye, S. G. Jonathan, A. Rafiq, O. J. Olawuyi, D. O. Adejoye


Calmodulin is one of the intracellular calcium proteins that regulates large spectrum of enzymes and cellular functions including metabolism of cyclic nucleotides and glycogen. The potentials of calmodulin gene in fungi necessitates their genetic response and their strong cassette of enzyme secretions for pesticide degradation. Therefore, this study was carried out to investigate the ‘Transcriptomic’ response of calmodulin encoding genes in Talaromyces fungi in response to 2, 2-dichlorovinyl dimethyl phosphate (DDVP or Dichlorvos) an organophosphate pesticide and γ-Hexachlorocyclohexane (Lindane) an organochlorine pesticide. Fungi strains isolated from rhizosphere from grasses rhizosphere in pesticide polluted sites were subjected to percentage incidence test. Two most frequent fungi were further characterized using ITS gene amplification (ITS1 and ITS4 combinations), they were thereafter subjected to In-vitro DDVP and lindane tolerance tests at different concentrations. They were also screened for presence and expression of calmodulin gene (caM) using RT-PCR technique. The two Talaromyces strains had the highest incidence of 50-72% in pesticide polluted site, they were both identified as Talaromyces astroroseus asemoG and Talaromyces purpurogenum asemoN submitted in NCBI gene-bank with accession numbers KY488464 and KY488468 respectively. T. astroroseus KY488464 tolerated DDVP (1.23±0.023 cm) and lindane (1.11±0.018 cm) at 25 % concentration while T. purpurogenum KY488468 tolerated DDVP (1.33±0.061 cm) and lindane (1.54±0.077 cm) at this concentration. Calmodulin gene was detected in both strains, but RT-PCR expression of caM gene revealed at 900-1000 bp showed an under-expression of caM in T. astrorosues KY488464 but overexpressed in T. purpurogenum KY488464. Thus, the calmodulin gene response of these fungal strains to both pesticides could be considered in monitoring the potentials of fungal strains to pesticide tolerance and bioremediation of pesticide in polluted soil.

Keywords: Calmodulin gene, pesticide, RT-PCR, talaromyces, tolerance

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40 AMF activates PDH 45 and G-proteins Genes to Alleviate Abiotic Stress in Tomato Plants

Authors: Deepak Bhardwaj, Narendra Tuteja


Global climate change is impacting large agrarian societies, especially those in countries located near the equator. Agriculture, and consequently, plant-based food, is the hardest hit in tropical and sub-tropical countries such as India due to an increased incidence of drought as well as an increase in soil salinity. One method that holds promise is AMF-rich biofertilizers which assist in activating proteins which in turn help alleviate abiotic stress in plants. In the present study, we identified two important species of (arbuscular mycorrhizal fungus) AMF belonging to Glomus and Gigaspora from the rhizosphere of the important medicinal plant Justicia adathoda. These two species have been found to be responsible for the abundance of Justicia adathoda in the semi-arid areas of the Jammu valley located in northern India, namely, the Union Territory of Jammu and Kashmir. We isolated the species of Glomus and Gigaspora from the rhizosphere of Justicia adathoda and used them as biofertilizers for the tomato plant. Significant improvements in the growth parameters were observed in the tomato plants inoculated with Glomus sp. and Gigaspora sp. in comparison with the tomato plants that were grown without AMF treatments. Tomato plants grown along with Glomus sp. and Gigaspora sp. have been observed to withstand 200 mM of salinity and 25% PEG stress. AMF also resulted in an increased concentration of proline and antioxidant enzymes in tomato plants. We also examined the expression levels of salinity and drought stress-inducible genes such as pea DNA helicase 45 (PDH 45) and genes of G-protein subunits of the tomato plants inoculated with and without AMF under stress and normal conditions. All the stress-inducible genes showed a significant increase in their gene expression under stress and AMF inoculation, while their levels were found to be normal under AMF inoculation without stress. We propose a model of abiotic stress alleviation in tomato plants with the help of external factors such as AMF and internally with the help of proteins like PDH 45 and G-proteins.

Keywords: AMF, abiotic stress, g-proteins, PDH-45

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39 Which Mechanisms are Involved by Legume-Rhizobia Symbiosis to Increase Its Phosphorus Use Efficiency under Low Phosphorus Level?

Authors: B. Makoudi, R. Ghanimi, A. Bargaz, M. Mouradi, M. Farissi, A. Kabbaj, J. J. Drevon, C. Ghoulam


Legume species are able to establish a nitrogen fixing symbiosis with soil rhizobia that allows them, when it operates normally, to ensure their necessary nitrogen nutrition. This biological process needs high phosphorus (P) supply and consequently it is limited under low phosphorus availability. To overcome this constraint, legume-rhizobia symbiosis develops many mechanisms to increase P availability in the rhizosphere and also the efficiency of P fertilizers. The objectives of our research works are to understand the physiological and biochemical mechanisms implemented by legume-rhizobia symbiosis to increase its P use efficiency (PUE) in order to select legume genotypes-rhizobia strains combination more performing for BNF under P deficiency. Our studies were carried out on two grain legume species, common bean (Phaseolus vulgaris) and faba bean (Vicia faba) tested in farmers’ fields and in experimental station fewer than two soil phosphorus levels. Under field conditions, the P deficiency caused a significant decrease of Plant and nodule biomasses in all of the tested varieties with a difference between them. This P limitation increased the contents of available P in the rhizospheric soils that was positively correlated with the increase of phosphatases activities in the nodules and the rhizospheric soil. Some legume genotypes showed a significant increase of their P use efficiency under P deficiency. The P solubilization test showed that some rhizobia strains isolated from Haouz region presented an important capacity to grow on solid and liquid media with tricalcium phosphate as the only P source and their P solubilizing activity was confirmed by the assay of the released P in the liquid medium. Also, this P solubilizing activity was correlated with medium acidification and the excretion of acid phosphatases and phytases in the medium. Thus, we concluded that medium acidification and excretion of phosphatases in the rhizosphere are the prominent reactions for legume-rhizobia symbiosis to improve its P nutrition.

Keywords: legume, phosphorus deficiency, rhizobia, rhizospheric soil

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38 The Prevalence of Citrus Specific Nematode Tylenchulus semipenetrans Cobb 1913 on the Coast of the Black Sea in Georgia

Authors: E.Tskitisvili, L. Jgenti, I. Eliava, T. Tskitishvili, N. Bagathuria, M. Gigolashvili


The fight against dangerous nematode diseases that have world economic importance requires accurate data about the prevalence of these pests. In the point of view of the International Convention on Biological Diversity, the identification of the plant invasion causing dangerous pathogen in the early stages of invasion on new territory is the most important part of the program, which aims to monitor the Bio-Agro Coenosis and Bio-Control. Citrus nematode-specific belongs to the pathogen species, which can cause epiphytotics particularly for large areas and cause irreparable damage to citrus plantations. This paper provides a brief tour of the spread of citrus nematodes on the Black Sea coast (Adjara and Abkhazia). Also the bio-ecological monitoring data to detect the potential sources of invasion for evaluating the current conditions of the citrus nematodes prevalence. Through 2006-2010, the material was gained by structural monitoring system during the citrus vegetation period on tangerines, lemon and oranges from nine points of the study area. Mature forms of Tylenchulus semipenetrans Cobb, 1913 were observed in almost all of the samples of the root system, the peak of larvae was observed in late spring and outumn. 92 forms of nematode has been detected in the rhizosphere belonging to 8 Orders: Areolaimida, Dorylaimida, Enoplida, Mononchida, Tylenshida, Monshysterida, Rhabditida, Aphelenchida, 23 families and 40 genera. 75 forms are identified as species. It is estimated the number of nematodes fauna and ecological groups. To detect possible sources of invasion we obtained additional materials in 2013-2014 from citrus plantations planted in 2011, where is planted tangerine trees introduced from Spain and Japan. The fauna of rhizosphere is identified and Tylenchulus semipenetrans Cobb, 1913 is not detected.

Keywords: Citrus nematodes, infection, bioecological monitoring, epiphytotics

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37 Microbial Inoculants to Increase the Biomass and Nutrient Uptake of Tithonia Cultivated as Hedgerow Plants to Control Erosion in Ultisols

Authors: Nurhajati Hakim, Kiki Amalia, A. Agustian, H. Hermansah, Y. Yulnafatmawita


Ultisols require greater amounts of fertilizer application compared to other soils and susceptible to erosion. Unfortunately, the price of synthetic fertilizers has increased over time during the years, making them unaffordable for most Indonesian farmers. While terrace technique to control erosion very costly.Over the last century, efforts to reduce reliance on synthetic agro-chemicals fertilizers and erosion control have recently focused on Tithonia diversifolia as a fertilizer alternative, and as hedgerow plant to control erosion. Generally known by its common name of tree marigold or Mexican sunflower, this plant has attracted considerable attention for its prolific production of green biomass, rich in nitrogen, phosphorous and potassium (NPK). In pot experiments has founded some microbial such as Mycorrhizal, Azotobacter, Azospirillum, phosphate solubilizing bacterial (PSB) and fungi (PSF) are expected to play an important role in biomass production and high nutrient uptake of this plant. This issue of importance was pursued further in the following investigation in field condition. The aim of this study was to determine the type of microbial combination suitable for Tithonia cultivation as hedgerow plants in Ultisols which have higher biomass production and nutrient content, and decline soil erosion. The field experiment was conducted with 6 treatments in a randomized block design (RBD) using 3 replications. The treatments were: Tithonia rhizosphere without microbial inoculated (A); Inokulanted by Mycorrhizal + Azotobacter + Azospirillium (B); Mycorrhizal + PSF (C); Mycorrhizal + PSB(D); Mycorrhizal + PSB + PSF(E);and without hedgerow Tithonia (F).The microbial substrates were inoculated into the Tithonia rhizosphere in the nursery. The young Tithonia plants were then planted as hedgerow on Ultisols in the experimental field for 8 months, and pruned once every 2 months. Soil erosion were collected every rainy time. The differences between treatments were statistically significant by HSD test at the 95% level of probability. The result showed that treatment C (mycorrhizal + PSB) was the most effective, and followed by treatment D (mycorrhizal + PSF) in producing higher Tithonia biomass about 8 t dry matter 2000 m-2 ha-1 y-1 and declined soil erosion 71-75%.

Keywords: hedgerow tithonia, microbial inoculants, organic fertilizer, soil erosion control

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36 Biochar-induced Metals Immobilization in the Soil as Affected by Citric Acid

Authors: Md. Shoffikul Islam, Hongqing Hu


Reducing trace elements' mobility and bioavailability through amendment addition, especially biochar (BC), is a cost-effective and efficient method to address their toxicity in the soil environment. However, the low molecular weight organic acids (LMWOAs) in the rhizosphere could affect BC's efficiency to stabilize trace metals as the LMWOAs could either mobilize or fix metals in the soils. Therefore, understanding the BC's and LMWOAs' interaction mechanisms on metals stabilization in the rhizosphere is crucial. The present study explored the impact of BC derived from rice husk and citric acid (CA) and the combination of BC and CA on the redistribution of cadmium (Cd), lead (Pb), and zinc (Zn) among their geochemical forms through incubation experiment. The changes of zeta potential and X-ray diffraction (XRD) pattern of BC and BC-amended soils to investigate the probable mechanisms of trace elements' immobilization by BC under the CA attack were also examined. The rice husk BC at 5% (w/w) was mixed with the air-dry soil (an Anthrosols) contaminated with Cd, Pb, and Zn in the plastic pot. The 2, 5, 10, and 20 mM kg-1 (w/v) of CA were added separately into the pot. All the ingredients were mixed thoroughly with the soil. A control (CK) treatment was also prepared without BC and CA addition. After 7, 15, and 60 days of incubation with 60% (w/v) moisture level at 25 °C, the incubated soils were determined for pH and EC and were sequentially extracted to assess the metals' transformation in soil. The electronegative charges and XRD peaks of BC and BC-amended soils were also measured. Compared to CK, the application of BC, low level of CA (2 mM kg-1 soil) (CA2), and BC plus the low concentration of CA (BC-CA2) considerably declined the acid-soluble Cd, Pb, and Zn in which BC-CA2 was found to be the most effective treatment. The reversed trends were observed concerning the high levels of CA (>5-20 mM kg-1 soil) and the BC plus high concentrations of CA treatments. BC-CA2 changed the highest amounts of acid-soluble and reducible metals to the oxidizable and residual forms with time. The most increased electronegative charges of BC-CA2 indicate its (BC-CA2) highest Cd, Pb, and Zn immobilizing efficiency, probably through metals adsorption and fixation with the negative charge sites. The XRD study revealed the presence of P, O, CO32-, and Cl1- in BC, which might be responsible for the precipitation of CdCO3, pyromorphite, and hopeite in the case of Cd, Pb, and Zn immobilization, respectively. The findings depicted that the low concentration of CA increased metals' stabilization, whereas the high levels of CA enhanced their mobilization. The BC-CA2 emerged as the best amendment among treatments for metals stabilization in contaminated soils.

Keywords: Biochar, citric acid, immobilization, trace elements contaminated soil

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35 Mechanisms of Metals Stabilization in the Soil by Biochar Material as Affected by the Low Molecular Weight Organic Acids

Authors: Md. Shoffikul Islam, Hongqing Hu


Immobilizing trace elements by reducing their mobility and bioavailability through amendment application, especially biochar (BC), is a cost-effective and efficient method to address their toxicity in the soil environment. However, the low molecular weight organic acids (LMWOAs) in the rhizosphere could affect BC's efficiency to immobilize trace metals as the LMWOAs could either mobilize or fix metals in the soils. Therefore, understanding the BC's and LMWOAs' interaction mechanisms on metals stabilization in the rhizosphere is crucial. The present study examined the impact of BC derived from rice husk, tartaric acid (TA), and oxalic acid (OA), and the combination of BC and TA/OA on the changes of cadmium (Cd), lead (Pb), and zinc (Zn) among their geochemical forms through incubation experiment. The changes of zeta potential and X-ray diffraction (XRD) pattern of BC and BC-amended soils to investigate the probable mechanisms of trace elements' immobilization by BC under the attacks of TA and OA were also examined. The rice husk BC at 5% (w/w) was mixed with the air-dry soil (an Anthrosols) contaminated with Cd, Pb, and Zn in the plastic pot. The TA and OA each at 2, 5, 10, and 20 mM kg-1 (w/v) were added separately into the pot. All the ingredients were mixed thoroughly with the soil. A control (CK) treatment was also prepared without BC, TA, and OA addition. After 7, 15, and 60 days of incubation with 60% (w/v) moisture level at 25 °C, the incubated soils were determined for pH and EC and were sequentially extracted to assess the metals' transformation in soil. The electronegative charges and XRD peaks of BC and BC-amended soils were also measured. The BC, low level of TA (2 mM kg-1 soil), and BC plus the low concentration of TA (BC-TA2) addition considerably declined the acid-soluble Cd, Pb, and Zn in which BC-TA2 was found to be the most effective treatment. The trends were reversed concerning the high levels of TA (>5-20 mM kg-1 soil), all levels of OA (2-20 mM kg-1 soil), and the BC plus high levels of TA/OA treatments. BC-TA2 changed the highest amounts of acid-soluble and reducible metals to the oxidizable and residual fractions with time. The most increased electronegative charges of BC-TA2 indicate its (BC-TA2) highest metals' immobilizing efficiency, probably through metals adsorption and fixation with the negative charge sites. The XRD study revealed the presence of P, O, CO32-, and Cl1- in BC, which might be responsible for the precipitation of CdCO3, pyromorphite, and hopeite concerning Cd, Pb, and Zn immobilization, respectively. The findings demonstrated that the low level of TA increased metals immobilization, while the high levels of TA and all levels of OA enhanced their mobilization. The BC-TA2 was the best treatment in stabilizing metals in soil.

Keywords: biochar, immobilization, low molecular weight organic acids, trace elements contaminated soil

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34 Crop Genotype and Inoculum Density Influences Plant Growth and Endophytic Colonization Potential of Plant Growth-Promoting Bacterium Burkholderia phytofirmans PsJN

Authors: Muhammad Naveed, Sohail Yousaf, Zahir Ahmad Zahir, Birgit Mitter, Angela Sessitsch


Most bacterial endophytes originate from the soil and enter plants via the roots followed by further spread through the inner tissues. The mechanisms allowing bacteria to colonize plants endophytically are still poorly understood for most bacterial and plant species. Specific bacterial functions are required for plant colonization, but also the plant itself is a determining factor as bacterial ability to establish endophytic populations is very often dependent on the plant genotype (cultivar) and inoculums density. The effect of inoculum density (107, 108, 109 CFU mL-1) of Burkholderia phytofirmans strain PsJN was evaluated on growth and endophytic colonization of different maize and potato cultivars under axenic and natural soil conditions. PsJN inoculation significantly increased maize seedling growth and tuber yield of potato at all inoculum density compared to uninoculated control. Under axenic condition, PsJN inoculation (108 CFU mL-1) significantly improved the germination, root/shoot length and biomass up to 62, 115, 98 and 135% of maize seedling compared to uninoculated control. In case of potato, PsJN inoculation (109 CFU mL-1) showed maximum response and significantly increased root/shoot biomass and tuber yield under natural soil condition. We confirmed that PsJN is able to colonize the rhizosphere, roots and shoots of maize and potato cultivars. The endophytic colonization increased linearly with increasing inoculum density (within a range of 8 x 104 – 3 x 107 CFU mL-1) and were highest for maize (Morignon) and potato (Romina) as compared to other cultivars. Efficient colonization of cv. Morignon and Romina by strain PsJN indicates the specific cultivar colonizing capacity of the bacteria. The findings of the study indicate the non-significant relationship between colonization and plant growth promotion in maize under axenic conditions. However, the inoculum level (109 CFU mL-1) that promoted colonization of rhizosphere and plant interior (endophytic) also best promoted growth and tuber yield of potato under natural soil conditions.

Keywords: crop genotype, inoculum density, Burkholderia phytofirmans PsJN, colonization, growth, potato

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33 Broad Survey of Fine Root Traits to Investigate the Root Economic Spectrum Hypothesis and Plant-Fire Dynamics Worldwide

Authors: Jacob Lewis Watts, Adam F. A. Pellegrini


Prairies, grasslands, and forests cover an expansive portion of the world’s surface and contribute significantly to Earth’s carbon cycle. The largest driver of carbon dynamics in some of these ecosystems is fire. As the global climate changes, most fire-dominated ecosystems will experience increased fire frequency and intensity, leading to increased carbon flux into the atmosphere and soil nutrient depletion. The plant communities associated with different fire regimes are important for reassimilation of carbon lost during fire and soil recovery. More frequent fires promote conservative plant functional traits aboveground; however, belowground fine root traits are poorly explored and arguably more important drivers of ecosystem function as the primary interface between the soil and plant. The root economic spectrum (RES) hypothesis describes single-dimensional covariation between important fine-root traits along a range of plant strategies from acquisitive to conservative – parallel to the well-established leaf economic spectrum (LES). However, because of the paucity of root trait data, the complex nature of the rhizosphere, and the phylogenetic conservatism of root traits, it is unknown whether the RES hypothesis accurately describes plant nutrient and water acquisition strategies. This project utilizesplants grown in common garden conditions in the Cambridge University Botanic Garden and a meta-analysis of long-term fire manipulation experiments to examine the belowground physiological traits of fire-adapted and non-fire-adapted herbaceous species to 1) test the RES hypothesis and 2) describe the effect of fire regimes on fine root functional traits – which in turn affect carbon and nutrient cycling. A suite of morphological, chemical, and biological root traits (e.g. root diameter, specific root length, percent N, percent mycorrhizal colonization, etc.) of 50 herbaceous species were measuredand tested for phylogenetic conservatism and RES dimensionality. Fire-adapted and non-fire-adapted plants traits were compared using phylogenetic PCA techniques. Preliminary evidence suggests that phylogenetic conservatism may weaken the single-dimensionality of the RES, suggesting that there may not be a single way that plants optimize nutrient and water acquisition and storage in the complex rhizosphere; additionally, fire-adapted species are expected to be more conservative than non-fire-adapted species, which may be indicative of slower carbon cycling with increasing fire frequency and intensity.

Keywords: climate change, fire regimes, root economic spectrum, fine roots

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32 Monitoring of Potato Rot Nematode (Ditylenchus destructor Thorne, 1945) in Southern Georgia Nematode Fauna Diversity of Rhizosphere

Authors: E. Tskitishvili, L. Jgenti, I. Eliava, T. Tskitishvili, N. Bagathuria, M. Gigolashvili


The nematode fauna of 20 agrocenosis (soil, tuber of potato, green parts of plant, roots) was studied in four regions in South Georgia (Akhaltsikhe, Aspindza, Akhalkalaki, Ninotsminda). In all, there were registered 173 forms of free-living and Phyto-parasitic nematodes, including 132 forms which were specified according to their species. A few exemplars of potato root nematode (Ditylenchus destructor) were identified in soil samples taken in Ninotsminda, Akhalkalaki and Aspinda stations, i.e. invasion is weak. Based on our data, potato Ditylenchus was not found in any of the researched tubers, while based on the data of previous years the most of tubers were infested. The cysts of 'golden nematodes' were not found during inspection of material for detection of Globoderosis

Keywords: ditylenchus, monitoring, nematoda, potato

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31 The Potential of Rhizospheric Bacteria for Mycotoxigenic Fungi Suppression

Authors: Vanja Vlajkov, Ivana PajčIn, Mila Grahovac, Marta Loc, Dragana Budakov, Jovana Grahovac


The rhizosphere soil refers to the plant roots' dynamic environment characterized by their inhabitants' high biological activity. Rhizospheric bacteria are recognized as effective biocontrol agents and considered cardinal in alternative strategies for securing ecological plant diseases management. The need to suppress fungal pathogens is an urgent task, not only because of the direct economic losses caused by infection but also due to their ability to produce mycotoxins with harmful effects on human health. Aspergillus and Fusarium species are well-known producers of toxigenic metabolites with a high capacity to colonize crops and enter the food chain. The bacteria belonging to the Bacillus genus has been conceded as a plant beneficial species in agricultural practice and identified as plant growth-promoting rhizobacteria (PGPR). Besides incontestable potential, the full commercialization of microbial biopesticides is in the preliminary phase. Thus, there is a constant need for estimating the suitability of novel strains to be used as a central point of viable bioprocess leading to market-ready product development. In the present study, 76 potential producing strains were isolated from the rhizosphere soil, sampled from different localities in the Autonomous Province of Vojvodina, Republic of Serbia. The selective isolation process of strains started by resuspending 1 g of soil samples in 9 ml of saline and incubating at 28° C for 15 minutes at 150 rpm. After homogenization, thermal treatment at 100° C for 7 minutes was performed. Dilution series (10-1-10-3) were prepared, and 500 µl of each was inoculated on nutrient agar plates and incubated at 28° C for 48 h. The pure cultures of morphologically different strains indicating belonging to the Bacillus genus were obtained by the spread-plate technique. The cultivation of the isolated strains was carried out in an Erlenmeyer flask for 96 h, at 28 °C, 170 rpm. The antagonistic activity screening included two phytopathogenic fungi as test microorganisms: Aspergillus sp. and Fusarium sp. The mycelial growth inhibition was estimated based on the antimicrobial activity testing of cultivation broth by the diffusion method. For the Aspergillus sp., the highest antifungal activity was recorded for the isolates Kro-4a and Mah-1a. In contrast, for the Fusarium sp., following 15 isolates exhibited the highest antagonistic effect Par-1, Par-2, Par-3, Par-4, Kup-4, Paš-1b, Pap-3, Kro-2, Kro-3a, Kro-3b, Kra-1a, Kra-1b, Šar-1, Šar-2b and Šar-4. One-way ANOVA was performed to determine the antagonists' effect statistical significance on inhibition zone diameter. Duncan's multiple range test was conducted to define homogenous groups of antagonists with the same level of statistical significance regarding their effect on antimicrobial activity of the tested cultivation broth against tested pathogens. The study results have pointed out the significant in vitro potential of the isolated strains to be used as biocontrol agents for the suppression of the tested mycotoxigenic fungi. Further research should include the identification and detailed characterization of the most promising isolates and mode of action of the selected strains as biocontrol agents. The following research should also involve bioprocess optimization steps to fully reach the selected strains' potential as microbial biopesticides and design cost-effective biotechnological production.

Keywords: Bacillus, biocontrol, bioprocess, mycotoxigenic fungi

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30 Surface Active Phthalic Acid Ester Produced by a Rhizobacterial Strain

Authors: M. L. Ibrahim, A. Abdulhamid


A surface active molecule synthesized by a rhizobacterial strain Bacillus lentus isolated from Cajanus cajan was investigated. The bioemulsifier was extracted, purified and partially characterized using standard methods. Surface properties of the bioemulsifier were determined by studying the emulsification index, solubility test and stability studies. Partial purification of the bioemulsifier was carried out using FT-IR analysis, Silica-gel column chromatography and thin layer chromatography. GC-MS analysis was carried out to detect the composition and mass of the lipids and esters. The isolate showed an emulsifying activity of 57% and surface activity of 36mm. The stability studies revealed that the bioemulsifier had better stability at temperature of 70oC, 8% pH and 8% NaCl concentration. FT-IR indicated the bioemulsifier to contain peptide and aliphatic chain, TLC revealed the compound to be ninhydrin positive and Column chromatography showed the presence of three amino acids namely; glutamine, valine and cysteine. GC-MS indicated the lipid moiety to contain aliphatic chain ranging from C9-C16 and two major peaks of 1,2-benzenedicarboxylic acid diethyl octyl ester. Therefore, surface active agent from Bacillus lentus can be used effectively in a wide range of applications such as in MEOR and in the biosynthesis of plasticizers for industrial uses.

Keywords: Bacillus lentus, bioemulsifiers, phthalic acid ester, Rhizosphere

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29 Diversity, Phyto Beneficial Activities and Agrobiotechnolody of Plant Growth Promoting Bacillus and Paenibacillus

Authors: Cheba Ben Amar


Bacillus and Paenibacillus are Gram-positive aerobic endospore-forming bacteria (AEFB) and most abundant in the rhizosphere, they mediated plant growth promotion and disease protection by several complex and interrelated processes involving direct and indirect mechanisms that include nitrogen fixation, phosphate solubilization, siderophores production, phytohormones production and plant diseases control. In addition to their multiple PGPR properties, high secretory capacity, spore forming ability and spore resistance to unfavorable conditions enabling their extended commercial applications for long shelf-life. Due to these unique advantages, Bacillus species were the most an ideal candidate for developing efficient PGPR products such as biopesticides, fungicides and fertilizers. This review list all studied and reported plant growth promoting Bacillus species and strains, discuss their capacities to enhance plant growth and protection with special focusing on the most frequent species Bacillus subtilis, B. pumilus ,B. megaterium, B. amyloliquefaciens , B. licheniformis and B. sphaericus, furthermore we recapitulate the beneficial activities and mechanisms of several species and strains of the genus Paenibacillus involved in plant growth stimulation and plant disease control.

Keywords: bacillus, paenibacillus, PGPR, beneficial activities, mechanisms, growth promotion, disease control, agrobiotechnology

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28 Rejuvenation of Peanut Seedling from Collar Rot Disease by Azotobacter sp. RA2

Authors: Ravi R. Patel, Vasudev R. Thakkar


Use of plant growth-promoting rhizobacteria (PGPR) to increase the production and decrees disease occurrence is a recent method in agriculture. An RA2 rhizospheric culture was isolated from peanut rhizosphere from Junagadh region of Gujarat, India and showed different direct and indirect plant growth promoting activity like indole acetic acid, gibberellic acid, siderophore, hydrogen cyanide, Ammonia and (1-Aminocyclopropane-1-Carboxylate) deaminase production, N2 fixation, phosphate and potassium solubilization in vitro. RA2 was able to protect peanut germinating seedling from A. niger infection and reduce collar rot disease incidence 60-35% to 72-41% and increase germination percentage from 70-82% to 75-97% in two varieties GG20 and GG2 of peanut. RA2 was found to induce resistance in A. hypogaea L. seedlings via induction of different defense-related enzymes like phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, lipoxygenase and pathogenesis related protein like chitinase, ß – 1,3- glucanase. Jasmonic acid one of the major signaling molecules of inducing systemic resistance was also found to induced due to RA2 treatments. RA2 bacterium was also promoting peanut growth and reduce A. niger infection in pot studies. 16S rDNA sequence of RA2 showed 99 % homology to Azotobacter species.

Keywords: plant growth promoting rhizobacteria, peanut, aspergillus niger, induce systemic resistance

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27 Constructed Wetlands: A Sustainable Approach for Waste Water Treatment

Authors: S. Sehar, S. Khan, N. Ali, S. Ahmed


In the last decade, the hunt for cost-effective, eco-friendly and energy sustainable technologies for waste water treatment are gaining much attention due to emerging water crisis and rapidly depleting existing water reservoirs all over the world. In this scenario, constructed wetland being a “green technology” could be a reliable mean for waste water treatment especially in small communities due to cost-effectiveness, ease in management, less energy consumption and sludge production. Therefore, a low cost, lab-scale sub-surface flow hybrid constructed wetland (SS-HCW) was established for domestic waste water treatment.It was observed that not only the presence but also choice of suitable vegetation along with hydraulic retention time (HRT) are key intervening ingredients which directly influence pollutant removals in constructed wetlands. Another important aspect of vegetation is that it may facilitate microbial attachment in rhizosphere, thus promote biofilm formation via microbial interactions. The major factors that influence initial aggregation and subsequent biofilm formation i.e. divalent cations (Ca2+) and extra cellular DNA (eDNA) were also studied in detail. The presence of Ca2+ in constructed wetland demonstrate superior performances in terms of effluent quality, i.e BOD5, COD, TDS, TSS, and PO4- than in absence of Ca2+. Finally, light and scanning electron microscopies coupled with EDS were carried out to get more insights into the mechanics of biofilm formation with or without Ca addition. Therefore, the same strategy can be implemented in other waste water treatment technologies.

Keywords: hybrid constructed wetland, biofilm formation, waste water treatment, waste water

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26 Arsenic Speciation in Cicer arietinum: A Terrestrial Legume That Contains Organoarsenic Species

Authors: Anjana Sagar


Arsenic poisoned ground water is a major concern in South Asia. The arsenic enters the food chain not only through drinking but also by using arsenic polluted water for irrigation. Arsenic is highly toxic in its inorganic forms; however, organic forms of arsenic are comparatively less toxic. In terrestrial plants, inorganic form of arsenic is predominantly found; however, we found that significant proportion of organic arsenic was present in root and shoot of a staple legume, chickpea (Cicer arientinum L) plants. Chickpea plants were raised in pot culture on soils spiked with arsenic ranging from 0-70 mg arsenate per Kg soil. Total arsenic concentrations of chickpea shoots and roots were determined by inductively coupled plasma-mass-spectrometry (ICP-MS) ranging from 0.76 to 20.26, and 2.09 to 16.43 µg g⁻¹ dry weight, respectively. Information on arsenic species was acquired by methanol/water extraction method, with arsenic species being analyzed by high-performance liquid chromatography (HPLC) coupled with ICP-MS. Dimethylarsinic acid (DMA) was the only organic arsenic species found in amount from 0.02 to 3.16 % of total arsenic shoot concentration and 0 to 6.93 % of total arsenic root concentration, respectively. To investigate the source of the organic arsenic in chickpea plants, arsenic species in the rhizosphere of soils of plants were also examined. The absence of organic arsenic in soils would suggest the possibility of formation of DMA in plants. The present investigation provides useful information for better understanding of distribution of arsenic species in terrestrial legume plants.

Keywords: arsenic, arsenic speciation, dimethylarsinic acid, organoarsenic

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