Effect of mineral fertilizer, pig manure, and Azospirillum rugosum on growth and nutrient contents of Lactuca sativa L

Benefits from the application of plant growth-promoting bacteria in agriculture largely depend on the complex interactions between several factors including the nature of fertilizers selected. This study was designed to determine the fine tuning between the inoculated bacteria and different fertilizers and their effect on the growth of lettuce plants (Lactuca sativa L.). Plant growth promotion by a novel species of the genus Azospirillum, namely A. rugosum IMMIB AFH-6, was tested by biochemical, bioassay, and greenhouse studies. The treatments used in the greenhouse study were; unfertilized control (Blank), half recommended dose of chemical fertilizer (1/2CF), full recommended dose of chemical fertilizer (1CF), pig manure fertilizer (PMF), pig manure fertilizer + half recommended dose of chemical fertilizer (PMF + 1/2CF), and pig manure fertilizer + full recommended dose of chemical fertilizer (PMF + 1CF). All these treatments when inoculated with A. rugosum IMMIB AFH-6 inoculation were, respectively, In-Blank, In-1/2CF, In-1CF, In-PMF, In-PMF + 1/2CF, and In-PMF + 1CF. Significant increase in plant biomass and shoot N, P, Ca, and Fe was shown in the In-Blank treatment. Plant growth in soil amended with PMF and A. rugosum IMMIB AFH-6 was significantly lower than in soil treated with the chemical fertilizer, but inoculation combined with chemical fertilizer significantly elevated the plant biomass. The In-PMF + 1/2CF treatment showed the highest yield. A. rugosum IMMIB AFH-6 facilitated the accumulation of trace minerals in higher concentrations when PMF was combined with 1CF. To examine the benefits of inoculation by A. rugosum IMMIB AFH-6, we have proposed a new type of data analysis which considers both biomass and nutrient content of plants. This new type of analysis has shown the importance of the mineral content of plant

Deinococcus ficus sp nov., isolated from the rhizosphere of Ficus religiosa L

A pale-pink strain (CC-FR2-10(T)) from the rhizosphere of the sacred tree Ficus religiosa L. in Taiwan was investigated by using a polyphasic taxonomic approach. The cells were Gram-positive, rod-shaped and non-spore-forming. Phylogenetic analyses using the 16S rRNA gene sequence of the isolate indicated that the organism belongs to the genus Deinococcus, the highest sequence similarities being found with Deinococcus grandis (96.1%), Deinococcus radiodurans (94.3%),Deinococcus radiopugnans (93.2%), Deinococcus indicus (93.0%), Deinococcus proteolyticus (92.5%), Deinococcus murrayi (92.4%) and Deinococcus geothermalis (90.7%). The DNA-DNA relatedness with respect to D. grandis DSM 3963(T) was 17.9%. Chemotaxonomic data revealed that strain CC-FR2-10(T) contains only menaquinone MK-8 as the respiratory quinone, unknown phosphoglycolipids as the predominant polar lipids and 16:1 omega 7c, 17:1 omega 8c and 17:1 omega 9c iso as the predominant fatty acids. The biochemical and chemotaxonomic properties demonstrate that strain CC-FR2-10(T) represents a novel species, for which the name Deinococcus ficus sp. nov. is proposed. The type strain is CC-FR2-10(T) (= CCUG 53391(T) = CIP 108832(T))

Transfer of Flexibacter sancti, Flexibacter filiformis, Flexibacter japonensis and Cytophaga arvensicola to the genus Chitinophaga and description of Chitinophaga skermanii sp nov

Analysis of the 16S rRNA gene sequences of species currently assigned to the genus Flexibacter has shown extensive intrageneric phylogenetic heterogeneity. It has been shown in previous studies that the species [Flexibacter] sancti, [Flexibacter] filiformis and [Flexibacter] japonensis were most closely related to Chitinophaga pinensis. In addition, [Cytophaga] arvensicola and species of the genus Terrimonas also clustered into this phylogenetic group. Although the similarities of 16S rRNA gene sequences were low (88.5-96.4%), there is no evidence for clear phenotypic differences between these organisms that justify assignment to different genera. A proposal is made to transfer these species to the genus Chitinophaga as Chitinophaga sancti comb. nov., Chitinophaga filiformis comb. nov., Chitinophaga japonenis comb. nov. and Chitinophaga arvensicola comb. nov. on the basis of phylogenetic and phenotypic data. Furthermore, a novel species is described within this genus, Chitinophaga skermanii sp. nov., with strain CC-SG1B(T) (=CCUG 52510(T)=CIP 109140(T)) as the type strain

Effect of free and encapsulated Pseudomonas putida CC-FR2-4 and Bacillus subtilis CC-pg104 on plant growth under gnotoblotic conditions

A study was performed to investigate the efificiency of microbial inoculants after encapsulating in alginate supplemented with humic acid on plant growth. Two promising plant growth promoting rhizobacteria were identified by 16S rDNA sequencing as Pseudomonas putida CC-FR2-4 and Bacillus subtilis CC-pg104, which were further characterized by biochemical analyses and inoculated to Lectuca sativa L. seedlings as free cells and entrapped in beads. Significant increase in shoot height after 21 days of growth was observed with encapsulated CC-pg104 inoculated plants. Highest increase in root length was observed with CC-pg104 free-cell inoculated plants, followed by plants inoculated with encapsulated CC-pg104. Results clearly demonstrated that inoculation of the encapsulated bacterial isolates promoted plant growth similar to their respective free cells and could be a novel and feasible technique for application in agricultural industry. (c) 2006 Elsevier Ltd. All rights reserved

Application of wastewater from paper and food seasoning industries with green manure to increase soil organic carbon: A laboratory study

This laboratory scale experiment was designed to study the suitability of organic wastes from paper and food seasoning industries to improve the soil organic carbon for rice cultivation. Lignin-rich wastewater from paper industry and nitrogen-rich effluent from a food industry at suitably lower concentrations were used at two levels of green manure to enhance the soil organic carbon fraction over time. Both the groups of soils with or without Sesbania were incubated under submerged condition at 25 degrees C for 15 days. Wastewaters from paper industry (WP), food industry (WS), and a combination of WP + WS were added separately to both the treatment groups in flasks. After 103 days of incubation, from all the three treatments and control, total organic carbon and alkali-soluble organic carbon fractions were analyzed. Results indicated that in all the three treatments containing green manure amended with industrial wastewaters, the organic carbon content increased significantly. The alkali-soluble organic carbon fraction was increased by 59% in the soil amended with green manure containing WS and by 31% in the treatment without green manure compared to control. The paper mill waste water namely, WP, increased the organic carbon only in the soil containing green manure by 63%. The combined treatment of WP + WS with green manure increased alkali-soluble organic carbon fraction by 90% compared to control, while in the treatment without green manure, the organic carbon increase was 71%. Overall, the combined treatment WP + WS with green manure could increase the alkali-soluble organic carbon fraction more than all other treatments. Hence, wastewater rich in organics from paper and food industries can be efficiently used to temporarily increase the soil organic carbon content. (C) 2007 Elsevier Ltd. All rights reserved

Impacts of monosodium glutamate industrial wastewater on plant growth and soil characteristics

Research into utilization of monosodium glutamate industrial wastewater (MSGW) as a plant nutrient source was undertaken. The physico-chemical and microbiological characteristics of MSGW were analyzed in detail. Effect of MSGW on early growth of Chinese cabbage (Brassica rapa L cv. Pekinensis) and maize (Zea mays L. cv. Bright Jean) was tested by the seed germination bioassay. Subsequently, in a greenhouse pot experiment using the same plant species, effects of MSGW application rates on the plant biomass yield, nitrogen content and soil properties were analyzed. The MSGW was characterized by high levels of N (56.7 g l(-1)), organic C (344.6 g l(-1)), total solids (600 g l(-1)) and other minerals. At MSGW concentrations below 1%, germination indices for both the plant species were significantly (p<0.01) higher than the control. Further, the greenhouse study results indicated significant increase in the plant biomass yield at MSGW application rates of 5000 and 7500Iha(-1). As the MSGW dose increased, the biomass yield decreased, decreasing the N-use efficiency. Maize showed significantly higher wastewater N-use efficiency compared to the Chinese cabbage. Although the total culturable bacterial and fungal counts in the raw MSGW were low, addition of MSGW to the soil increased the soil microbial activities and soil respiration. Soil organic C was also increased by the addition of MSGW, due to the presence of significant amounts of organic C in the wastewater. This preliminary study demonstrates that by proper management of the pH and optimization of application rate, MSGW can be utilized as a nutrient source for plant growth. Further long-term field studies to evaluate the environmental impact of MSGW usage in agriculture are being designed to reduce the environmental risks associated with the reuse of this underutilized wastewater in the agriculture. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved

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Study on the morphological and biocompatible properties of chitosan grafted silk fibre reinforced PVA films for tissue engineering applications

The current study delineates the preparation of novel chitosan grafted silk fibre reinforced Poly (vinyl alcohol) (PVA) composite films with desirable properties. Although silk fibroin has been extensively used for various biomedical applications, its properties could be further re-tailored for its suitability in the field of regenerative medicine. Chitosan was successfully grafted over silk, via acylation with succinic anhydride and thereby the fibres were incised and used for the preparation of the films. The grafted silk fibre reinforced PVA films were subjected to FTIR studies, microscopic analysis by atomic force microscopy (AFM) and optical microscopy techniques, X-ray diffraction (XRD) analysis and further evaluated for in vitro biocompatibility studies. The composite films demonstrated improved surface roughness with increasing concentration of the fibre and its dispersion in the polymer matrix was observed. Furthermore, in vitro biocompatibility and cellular behaviour such as adhesion and proliferation of mouse fibroblasts as well as astrocyte cells was studied and the results showed improved proliferative activity, when compared to the pristine PVA films. These results were further supported by the results confirmed by MTT assay demonstrating the films to be non-toxic. The efficiency and feasibility of the films to be used for tissue engineering, was further evaluated by haemocompatibility studies using human erythrocytes, thus making them a potential material to be used for biomedical applications. © 201

包含菌體及腐植酸的海藻酸鈣膠囊

本發明為提供一種用於將菌體固定化於微膠囊材料之方法，特別是關於將菌體與腐植酸共同固定化於天然微膠囊材料，例如海藻酸鈣膠囊中之方法。本發明亦關於根據該方法所製得之包含菌體及腐植酸的海藻酸鈣膠囊，以其所製成之微生物肥料或土壤改良劑，及其用以促進植物生長與改善菌體根部拓殖化作用之用途。 【創作特點】 本發明目的及優點將部分描述於下，或可由描述中顯而易見。 於一方面，本發明係關於一種製備菌體微膠囊之方法，該方法包含：將具有植物生長促進功能之菌體先行培養於液體培養基中以增殖菌體；及將該經增殖之菌體與腐植酸萃取物(HA)共同固定化於海藻酸鈣膠囊內，而製得「包含菌體及腐植酸之海藻酸鈣膠囊」。於一項具體態樣中，可將經過或未經乾燥之菌體微膠囊直接施用於作物根圈或土壤中，做為作物之微生物肥料、土壤改良劑或植物生長促進製劑。 本發明所稱之「包含菌體及腐植酸之海藻酸鈣膠囊」，其所包含之「菌體」為具有促進植物生長的活性的特徵者，其中包括如：(1)固氮酶、(2)礦物磷溶解作用、(3)產生吲哚－3－乙酸(indole－3－acetic acid；IAA；auxin)、(4)產生載鐵物質(siderophore)及(5)有機質分解活性。習知這些活性與PGPR的功能有關，包括：生物肥料作用(biofertilization)、植生激素作用(phytostimuiation)、生物防治(biocontrol)等，另外習知有機質的分解與土壤改良相關。將上述功能發揮作用的前提為菌體可於植物根圈(rhizosphere)內進行根拓殖化作用(root colonization)。 於另一方面，依據本發明可製得一種有序性的微生態結構(micro－ecologic structure)，亦即本發明所稱之「包含菌體及腐植酸之海藻酸鈣膠囊」之組成物，其包括菌體、HA萃取物及海藻酸鈣。本發明之「包含菌體及腐植酸之海藻酸鈣膠囊」可另包含其他具有功能之微生物、酶、發酵物或化學物質，以提高此膠囊之促進植物生長作用之功能。另一方面，「包含菌體及腐植酸之海藻酸鈣膠囊」視需要可另包含供培養微生物的培養物或穩定劑，以利微生物之生長繁殖或生理維持。 於一項具體態樣中，本發明提出一種固定化菌體於微膠囊的方法，其包含在海藻酸鹽聚合物之硬化作用尚未啟動前，先將腐植酸萃取液與菌體混合為「HA/菌體懸浮液」；將此「HA/菌體懸浮液」與海藻酸鈉混合，成為一「HA/菌體/海藻酸鈉」混合液，此時之「HA/菌體/海藻酸鈉」混合液尚未行硬化或交聯作用；以及將此一「HA/菌體/海藻酸鈉」混合液逐滴滴入氯化鈣水溶液以進行硬化作用而形成「包含微生物及腐植酸之海藻酸鈣膠囊」。 於另一項具體態樣中，本發明提出一種菌體微膠囊之保存及控制釋放方法，其中係將「包含微生物及腐植酸之海藻酸鈣膠囊」添加少量抗凍劑，經過冷凍乾燥成為乾燥膠囊。此一型態之膠囊施用植物根圈內可因水澎潤作用，使膠囊回復為近未乾燥前的膠囊狀態，以土壤中水份含量之乾燥與潮濕控制膠囊的復性速度，復性後之膠囊可以進行正常之釋放作用。 於另一方面，本發明提出一種促進植物生長作用之方法，其係將本發明「包含菌體及腐植酸的海藻酸鈣膠囊」施用於植物根圈土壤。於一項具體態樣中，將本發明「包含菌體及腐植酸之海藻酸鈣膠囊」施用於土壤、堆肥或含有機質之廢棄物等，可發揮其提高有機質分解、促進堆肥腐熟及土壤改良的目的。 本發明之「含有微生物菌體及腐植酸的海藻酸鈣微膠囊」可用以提供保護微域環境，保護菌體免於受到存於土壤的原生動物與嗜菌體等生物的侵襲、土壤中有害物質的毒害之菌體大量流失。並利用該「含有微生物菌體及腐植酸的海藻酸鈣微膠囊」製成控制釋放的生物製劑，可避免游離菌體施用後根圈內之菌體數量的減少，因此可以提高微生物之根拓殖化(root colonization)效應。 本發明之再一方面在於，係利用「含有菌體及腐植酸的海藻酸鈣微膠囊」以提供菌體掩護作用，可避免游離菌體施用根圈後植物防衛系統之對於菌體之根拓殖化的抑制，並利用防護免疫作用(immuno－protection)的機制，以改善微生物之根拓殖化作用、提高促進植物生長的效益。 依照本發明之具體實施態樣，用以固定化之菌體菌種為；(1)不屬於已知之人體病原菌、(2)非植物病原菌體、(3)未經任何人工基因改造、(4)游離細胞狀態具有植物生長促進功能、(5)菌體被固定化於完整之海藻酸鈣膠囊內可存活的功效者。 本發明的其他特性將在下面詳細揭示具體實施例觀察後變得顯而易見