Ethanolic extract of mango (Mangifera indica L.) malformed inflorescence as effective antifungal agent

Mango (Mangifera indica L.) suffers from floral and vegetative malformation and crop production is seriously affected. The anti-fungal activity of ethanolic extract of malformed mango inflorescence was observed at different concentrations (1000, 2000, 3000, 4000 and 5000 μl/ml) against 10 fungi, viz., Ustilago cynodontis, Cercospora cajani, Sphaerotheca sp., Cercospora sp., Alternaria solani, Bipolaris sp., Helminthosporium sp., Curvularia sp., Fusarium udum and Alternaria cajani. Spore germination of most of the fungi was inhibited at 5000 μg/ml. Some of them were also susceptible at 3000 or 4000 μg/ml concentration. Analysis of phenolic acids by high performance liquid chromatography (HPLC) showed 18 peaks in the extract, but only four could be identified, viz., capachin, gallic, benzoic and cinnamic acids. Because of the high efficacy of ethanolic extract of malformed mango inflorescence, its use under field conditions to control some plant diseases has been suggested.     

Antifungal and high-performance liquid chromatography analysis of the crude extracts of Acorus calamus,Tinospora cordifolia and Celestrus paniculatus

The antifungal activity of methanolic crude extracts of Acorus calamus, Tinospora cordifolia and Celestrus paniculatus was investigated against Alternaria solani, Curvularia lunata, Fusarium sp., Bipolaris sp. and Helminthosporium sp. at different concentrations (1000, 2000, 3000, 4000 and 5000 μg/ml). At 5000 μg/ml, crude extract of T. cordifolia is found to be highly effective against Helminthosporium sp. followed by A. calamus against A. solani. On the other hand, at 5000 μg/ml, C. paniculatus showed better activity against A. solani and Helminthosporium followed by A. calamus against A. solani at 4000 μg/ml. At 5000 μg/ml, all the three crude extracts showed least activity against fungus C. lunata and Fusarium sp. except A. calamus that showed better activity against C. lunata. The increase in the production of phenolic acid in the extract can be correlated with the induction of resistance in treated plants against phytopathogenic fungi. High-performance liquid chromatography analysis of the crude extract of medicinal plants showed six different phenolic acids (benzoic acid, cinnamic acid, caffeic acid, ferulic acid, gallic acid and tannic acid) present in varying amounts. The results of the study provide scientific basis for the use of the plant extract in the future development as antioxidant, antibacterial, antifungal and anti-inflammatory agent.     

Jatrorrhizine and columbamine alkaloids isolated from Argemone mexicana are inhibitory to spore germination of some plant pathogenic fungi

The isomeric alkaloids jatrorrhizine and columbamine isolated from the whole plant of Argemone mexicana were assessed against spore germination of some fungi, e.g. Alternaria cajani, Bipolaris sp., Helminthosporium sp., Fusarium udum and Curvularia sp. Both the alkaloids were effective against most of the fungi tested. Jatrorrhizine was highly effective against spore germination of Bipolaris sp., Fusarium udum and Curvularia sp. at 5000 ppm concentration, whereas columbamine was highly effective against Alternaria cajani, Helminthosporium sp. and Fusarium udum at 5000 ppm.     

Antifungal activity of ethanolic extract of Archu (Rheum emodi) on powdery mildew (Erysiphe cichoracearum) and its role in the induction of resistance in balsam (Impatiens balsamania)

Rheum emodi, vernacularly known as Archu, is one of the important high altitude medicinal plants widely distributed in Himalayan regions. Though widely used in Ayurveda for curing various human diseases, its use in plant diseases is limited. Ethanolic extract of Rheum rhizome was assayed against spore germination of Alternaria solani, Heliminthosporium penniseti and Curvularia palliscens. The inhibition of spore germination was concentration dependent. Maximum inhibition was obtained at 4000 and 5000 ppm followed by 3000, 2000 and 1000 ppm. However, the extract was highly effective in the pre-inoculation treatment against powdery mildew (Erysiphe cichoracearum) of balsam (Impatiens balsamania) under field conditions. High performance liquid chromatographic (HPLC) analysis of balsam leaves showed increased synthesis of phenolic acids, which has been correlated with induced resistance in inhibiting the disease intensity of balsam powdery mildew.     

Antifungal activity of two alkaloids of Zephyranthes citrina and their field efficacy against powdery mildew (Erysiphe cichoracearum) of balsam

The anti-fungal activity of two alkaloids isolated from bulbs of Zephyranthes citrina was observed against 10 fungi, viz, Aternaria solani, A. triticina, Curvularia lunata, C. maculuns, Cercospora malvacearum, Erysiphe sp., Fusarium udum, Helminthosporium pisi, H. speciferum and Ustilago cynodontis. Different concentrations (200, 400, 600, 800, 1000 µg/ml of alkaloids (A and B) were used. Spore germination was inhibited at 600, 800, 1000 µg/ml. B alkaloid was used against Erysiphe cichoracearum causing powdery mildew in balsam (Impatiens balsamina) in the field as pre- and post-inoculation treatments at 1000, 1500, 2000 µg/ml doses. The extract was effective in both pre- and post-inoculation treatments. Foliar application of this alkaloid resulted in inducing synthesis of phenolic acid in the leaves of balsam. Maximum phenolics were detected in the leaves treated with 1500 µg/ml in both pre- and post-inoculation treatments. The increase in the production of phenolics in treated leaves of balsam can be correlated with the induction of resistance in treated plants against powdery mildew. The significant efficacy of the alkaloid under field conditions opens the possibility of its use by farmers for also controlling other diseases.     

The mixture of tertiary and quaternary alkaloids isolated from Argemone ochroleuca inhibits spore germination of some fungi

The mixture of tertiary and quaternary alkaloids isolated from Argemone ochroleuca was separately assessed against spore germination of some plant pathogenic fungi, e.g. Alternaria alternata, Alternaria brassicae, Alternaria cajani, Bipolaris sp., Curvularia lunata, Curvularia sp., Colletotrichum musae, Fusarium udum, Helminthosporium sp., Helminthosporium pennisetti and Helminthosporium speciferum. Spore germination of Fusarium udum and Helminthosporium sp. was completely inhibited at very low concentration (200 ppm). A similar effect was observed on A. alternata, C. musae and H. pennisetti at 600, 800 and 1000 ppm. With quaternary alkaloids, Curvularia sp. and Colletotrichum musae were most sensitive as complete inhibition of spore germination was observed at 400, 600, 800 and 1000 ppm and a similar effect was observed with A. brassicae and A. cajani at 600, 800 and 1000 ppm. The remaining fungi were also highly sensitive to the mixture at different concentrations.     

Antifungal efficacy of some ethyl acetate extract fractions of Cyperus rotundus rhizomes against spore germination of some fungi

Plant products play an important role as safe and ecofriendly method in controlling various plant diseases. Ethyl acetate fractions of Cyperus rotundus rhizomes have been found highly effective against some species of Alternaria (A. alternata, A. brassicola, A. solani, Alternaria chearanthi), Colletotrichum (C. musae, Colletotrichum sp.), Curvularia (C. lunata, C. maculans, C. pallescens, C. pennisetti), Helminthosporium (H. pennissetti, H. spiciferum, H. echinoclova and Heterosporium colocasiae) in vitro. A. brassisicola was highly sensitive to all the fractions at all the concentrations. Fractions 4–7 inhibited complete spore germination at 2000 and 3000 μg/ml as compared to control. However, 100% spore germination inhibition was found in Colletotrichum species in all the fractions at 2000 and 3000 μg/ml but at lower concentrations 50–60% spore germination and 90% reduction of germ tube elongation were observed. Curvularia species was highly sensitive to all concentrations of all the fractions as 50–100% spore germination inhibition was recorded at 500 μg/ml dose. Some species of Helminthosporium were sensitive at 3000 μg/ml in some fractions but other test fractions showed least efficacy. Fractions 4–7 were 100% inhibitory for H. colocasiae. Germ tube elongation was also affected by 60–90%. The germ tube branching and their elongation were affected in almost all species at 30 to 95%. The high efficacy of ethyl acetate fractions of rhizomes of C. rotundus against some fungi indicates that they can be very well tried under field conditions against some important plant diseases as an ecofriendly method of plant disease control.     

The effect of decreasing alkalinity on microbial community dynamics in a sulfate-reducing bioreactor as analyzed by PCR-SSCP

Alkalinity refers to the substance that can react with acid in an aqueous system, which measures its capac-ity to neutralize hydrogen iron (H ). Alkalinity influ-ences greatly the stability and treatment ability of an-aerobic bioreactor. It has been well …     

The effect of decreasing alkalinity on microbial community dynamics in a sulfate-reducing bioreactor as analyzed by PCR-SSCP

PCR-single-strand conformation polymorphism (SSCP) and Southern blotting techniques were adopted to investigate microbial community dynamics in a sulfate-reducing bioreactor caused by decreasing influent alkalinity. Experimental results indicated that the sulfate-removal rate approached 87% in 25 d under the conditions of influent alkalinity of 4000 mg/L (as CaCO₃) and sulfate-loading rate of 4.8 g/(L·d), which indicated that the bioreactor started up successfully. The analysis of microbial community structure in this stage showed that Lactococcus sp., Anaerofilum sp. and Kluyvera sp. were dominant populations. It was found that when influent alkalinity reduced to 1000 mg/L, sulfate-removal rate decreased rapidly to 35% in 3 d. Then influent alkalinity was increased to 3000 mg/L, the sulfate-removal rate rose to 55%. Under these conditions, the populations of Dysgonomonas sp., Sporobacte sp., Obesumbacterium sp. and Clostridium sp. got to rich, which predominated in the community together with Lactococcus sp., Anaerofilum sp. and Kluyvera sp. However, when the alkalinity was decreased to 1500 mg/L, the sulfate-removal rate rose to and kept stable at 70% and populations of Dysgonomonas sp., Sporobacter sp. and Obesumbacterium sp. died out, while some strains of Desulfovibrio sp. and Clostridium sp. increased in concentration. In order to determine the minimum alkalinity value that the system could tolerate, the influent alkalinity was decreased from 1500 to 400 mg/L secondly. This resulted in the sulfate-removal rate, pH value and effluent alkalinity dropping quickly. The amount of Petrotoga sp., Prevotella sp., Kluyvera sp. and Neisseria sp. reduced obviously. The result data from Southern blotting indicated that the amount of sulfate-reducing bacteria (SRBs) decreased with influent alkalinity dropping. Analysis of the microbial community structure and diversity showed that the SRBs populations were very abundant in the inoculated activated sludge and the alkalinity decrease caused the reduction of the populations noted. Most of resident populations in the bioreactor were fermentative acidogenic bacteria (FABs), among which the phylum Firmicute was in the majority, but SRBs were very few. This community structure demonstrates the cooperation between SRBs and FABs, which sustains the system’s high sulfate-removal and operation stability.     

Two plant alkaloids isolated from<Emphasis Type="Italic">Corydalis longipes</Emphasis> as potential antifungal agents

The alkaloids N-methylhydrasteine hydroxylactam and 1-methoxyberberine chloride were isolated from Corydalis longipes. Both alkaloids showed high efficacy individually (in concentration of 50-150 ppm) and also in a 1:1 mixture against spore germination of some fungi, viz. Alternaria alternata, A. brassicae, Curvularia maculans, Curvularia sp., Colletotrichum gloeosporioides, Colletotrichum sp., Helminthosporium speciferum, H. pennisetti, Helminthosporium sp., and Ustilago cynodontis. The antifungal effect of single compounds was dose-dependent. If the mutual ratio of the two components in the mixture was changed from 1:1 to a major content of any of the two compounds, the inhibitory effect on spore germination decreased.     

Acquired resistance of Alternaria solani and Sclerotium rolfsii to Polyoxin-D

Acquired resistance to the antibiotic polyoxin-D was studied in two phytopathogenic fungi, Alternaria solani Sorauer, and Sclerotium rolfsii Sacc. The ED₅₀ value of the antibiotic for A. solani was 100 μg/ml and S., rolfsii 200 μg/ml. A. solani and S. rolfsii could be trained to tolerate concentrations upto 1.000 μg/ml and 2.200 μg/ml respectively. The acquired resistance in both cases was not lost on continued subculturing in fungicide-free, media. On transfer to fungicide-free media, Polyoxinresistant strains of both the fungi showed faster growth rate and appreciable reduction in sporulation compared to the original strains. The adapted strain of A. solani showed cross-resistance to Cycloheximide and Difolatan but not to Hinosan and Bayleton.     

Production of N-Succinyl-l-diaminopimelic Acid by Auxotrophic Mutants of Aerobacter aerogenes and Serratia marcescens

α,ε-Diaminopimelic acid (DAP)-requiring mutants isolated from Aerobacter aerogenes ATCC 8308 and Serratia marcescens ATCC 19180 were found to accumulate N-succinyl-l-diaminopimelic acid (SDAP) which was an intermediate in the biosynthesis of lysine in Escherichia coli. SDAP was isolated from the culture broth and identified by the behavior in paper chromatography, melting point, elementary analysis, infrared spectrum, and optical rotation.

The culture conditions for SDAP production by A. aerogenes KY 7049 (DAP^?) and S. marcescens KY 8921 (DAP^?/Lys^?) were investigated. A. aerogenes KY 7049 has an absolute requirement for DAP together with a relative requirement for l-lysine. High levels of DAP (2000~4000 μg/ml) were proved to be favorable for SDAP accumulation, while if lysine along with DAP was added to the fermentation medium, optimal level of DAP for SDAP production was relatively low (about 200 μg/ml at 200 μg/ml of lysine). A variety of compounds which may conceivably affect the course of a fermentation process, i.e., carbon source, inorganic nitrogen source, amino acids, vitamines, precursors, were screened at optimal levels of lysine and DAP. Thus, the amount of SDAP accumulation reached a level of 19.9 mg/ml with the medium containing 10% glucose and 2000 μg/ml of DAP. S. marcescens KY 8921 requires either DAP or lysine for growth. Optimal level of DAP and lysine for SDAP accumulation was 50~100μg/ml.     

Evaluation of lead and chromium tolerance and accumulation level in Gomphrena celosoides: a novel metal accumulator from lead acid battery waste contaminated site in Nigeria

Abstract

Biology, tolerance, and metal (Pb and Cr) accumulating ability of Gomphrena celosoides were studied under hydroponic conditions. The seedlings were raised in Hoagland’s solution containing different concentrations of Pb (0, 500, 1000, 1500, 2000, 3000, 4000, and 5000?mg l^?1) and Cr (0, 50, 100, 150, 200, 300, and 400?mg l^?1). Biomass and metal accumulation in different plant parts were determined at seven (7) and fourteen (14) days after stress. Antioxidant enzyme activities, protein, and proline contents were estimated in stressed and unstressed plants. Gomphrena celosoides was able to tolerate Pb and Cr concentrations up to 4000 and 100?mg l^?1, respectively in hydroponic solution. Metal accumulation was concentration and duration dependent with the highest Pb (21,127.90 and 117,985.29?mg kg^?1) and Cr (3130.85 and 2428.90?mg kg^?1) in shoot and root, respectively found in the plants exposed to 5000?mg l^?1 Pb and 400?mg l^?1 Cr for 14?days. Proline, antioxidant enzyme activities, and protein contents were the highest in plant exposed to higher Pb and Cr concentrations for 7 and 14?days. Gomphrena celosoides could be considered as Pb and Cr accumulator with proline and increase in antioxidant enzyme activities being the tolerance mechanisms.     

Effect of Initial Density of Meloidogyne hapla on its Pathogenic Potential and Reproduction in Three Species of Medicinal Plants

The pathogenic potential and reproduction fitness of Meloidogyne hapla on three species of medicinal plants, Angelica koreana, Peucedanum japonicum and Astragalus membranaceus was determined in potted soil under greenhouse conditions. Three weeks old seedlings were inoculated with population density (Pi) of 1000; 2000; 3000; 4000; 5000 and 10000 juveniles (J₂)/kg soil. A significant damage was observed in shoot and root length, weight and root‐diameter of these plants by all Pi levels at 90‐day postinoculation. Damage increased with increase in Pi up to 5000 J₂/kg soil. At 5000 Pi caused 34.8, 34.1 and 33.3% reduction in root weight of Ang. koreana, P. japonicum and Ast. membranaceus, respectively. Greater root gall severity was observed on Ang. koreana and P. japonicum than on Ast. membranaceus at all Pi levels. At 5000 Pi, root gall severity was 5.0, 5.0, and 3.0 on Ang. koreana, P. japonicum and Ast. membranaceus, respectively. Increasing rate of Pi exponentially reduced reproductive factor (Rf) of M. hapla on all of these medicinal plants. However, Rf was higher on Ang. koreana and P. japonicum than on Ast. membranaceus at all Pi levels. The host status of these medicinal plants renders them unsuitable for their use in crop rotation system in M. hapla‐infested fields.     

Comparison of concomitant and sequential inoculation of Steinernema sp. in the management of root-knot (Meloidogyne incognita) nematode infecting eggplant (Solanum melongena)

Culture of Steinernema sp. was maintained on Corcyra cephalonica larvae. Steinernema sp. (at 50, 500, 1000, 2500, 5000, 10,000 and 20,000 ij’s /500?g soil) was concomitantly inoculated with 500 J₂ of Meloidogyne incognita/500?g soil to the eggplant seedlings in the pots filled with 4?kg sterilised soil. The simultaneous inoculation of M. incognita with either of the inoculum levels (1000, 2500, 5000 and 10,000 J₃/500?g soil) of Steinernema sp. significantly reduced the damage caused by M. incognita in terms of plant growth parameters, viz. plant length, dry weight, number of flowers and weight of fruits. Moreover, the highest improvement in plant growth parameters, viz. plant length, dry weight, number of flowers and weight of fruits, was recorded in plants inoculated with 5000 J₃ of Steinernema sp./500?g soil followed by 2500, 1000 and 10,000 J₃/500?g soil. The highest reduction in the reproduction factor and number of galls/root system was recorded in the plants treated with 5000 J₃ Steinernema sp./500?g soil followed by 2500, 1000 and 10,000 J₃/500?g soil. Comparison of concomitant and sequential inoculations showed that the sequential inoculation (both prior and after) of Steinernema sp. at different inoculum levels (1000, 2500, 5000, 10,000 and 20,000 ij’s/500?g soil) was more effective in the management of root-knot nematode than the concomitant inoculation. Therefore, the application of Steinernema sp. might be useful for suppression of nematode pest on eggplant and may be used as an alternative for chemicals.     

Differences in the microflora of scarified and unscarified seeds ofKarwinskia humboldtiana (Rhamnaceae)

Seeds ofKarwinskia humboldtiana obtained from a 1997 collection in the locality of Villa de García Nuevo (León, Mexico) were contaminated with spores of filamentous fungi, bacteria and yeasts. The concentration of microorganisms in unscarified seeds ranged from 3.0×10³ to 7.5×10³ CFU/g. Predominant were bacterial isolates of the generaAeromonas sp.,Bacillus, andPseudomonas; from filamentous fungi were identifiedAlternaria, Aspergillus niger, Cladosporium sp.,Fusarium sp.,Mucor sp.,Penicillium commune, Trichothecium sp.; from yeastsRhodotorula sp. andSaccharomyces cerevisiae. Seed scarification significantly reduced the microbial contamination. Of the original fungal isolates, only two were identified on scarified seeds,viz. Cladosporium sp. andSaccharomyces cerevisiae; although a relatively high incidence of a unidentifiable ofPenicillium sp. was found, the bacterial spectrum was not altered. Treatment of scarified seeds with Vitavax 200 WP and Pomarsol Forte 80 WP (3 mg/g seeds) augmented germination by 10–19% compared to treated unscarified seeds, and by 16–31% compared to untreated unscarified seeds.     

Production of extracellular polysaccharides by a Rhizobium species from the root nodules of Melilotus alba

The Rhizobium sp., isolated from the root nodules of the leguminous fodder herb Melilotus alba, produced large amounts of extracellular polysaccharides (EPS) (963.5 μg/ml) in a yeast extract mannitol medium. Growth and EPS production started simultaneously, but EPS production reached its maximum during the stationary phase of growth of the bacteria, at 20 hours. EPS production was increased with all of the thirteen sugars tested. Different nitrogen sources, such as nitrates, glutamic acid, casamino acid and L-asparagine, increased the EPS production although it was inhibited by glycine, nitrite and ammonium salts. Among the vitamins and metal ions, only pyridoxal phosphate and ZnSO₄ promoted EPS production. Attempts were made to optimize the cultural requirements for growth and maximum EPS production. Maximum EPS production (1457.0 μg/ml) was obtained when the medium was supplemented with glucose (1%), pyridoxal phosphate (2 μ g/ml), ZnSO₄ × 7 H₂O (10 μg/ml) and glutamic acid (0.1%). Under these conditions, the production was increased by 254.3% compared to the control. The EPS contained arabinose, xylose and rhamnose monomers. The presence of arabinose and xylose in the EPS produced by a Rhizobium sp. was uncommon.     

Studies on root nodules of leguminous plants bioproduction of indole acetic acid by a Rhizobium sp. from a twiner Clitoria ternatea L.

The Rhizobium sp. isolated from the root nodules of Clitoria ternatea L., a leguminous twiner, produced a high amount of IAA (16.4 μg/ml) from tryptophan in an unsupplemented basal medium. The production of IAA started simultaneously with the growth and had no different growth and production phase. The growth and production were parallel and increased up to 45–50 h. The IAA production by the Rhizobium sp. was increased by 520% when the medium was supplemented with fructose (1.5%), MnSO₄ (1.0 μg/ml), riboflavin (0.10 μg/ml) and Triton X-100 (0.01%). The possible role of the rhizobial production of IAA on the rhizobia-legume symbiosis is discussed.     

Elicitation of alkaloids in in vitro PLB (protocorm-like body) cultures of Pinellia ternata

The objective of this study was to determine the effect of two endophytic bacterial elicitors (Pseudomonas sp. and Enterobacter sp.) on the production of alkaloids in protocorm-like bodies (PLBs) of Pinellia ternata Breit. Both bacterial strains increased the growth rate of P. ternata PLBs. Pseudomonas sp. promoted the differentiation of the PLBs, whereas Enterobacter sp. inhibited PLB differentiation. The bacterial strains increased guanosine production in PLBs by 9–166%, inosine production by 2–33%, and trigonelline production by 114–1140% compared to the control. For Pseudomonas sp., guanosine and trigonelline production was greater when bacterial extracts were added to the PLB suspension cultures rather than living cells (co-culture treatment). Inosine production was similar in both the bacterial extract and co-culture treatments. For the Enterobacter sp., guanosine, inosine, and trigonelline production tended to be greatest when living cells were added to the PLB suspension cultures rather than bacterial extracts. These results suggest that Pseudomonas sp. and Enterobacter sp. could increase alkaloid yield from P. ternata under field or tissue culture conditions. We also observed that Pseudomonas sp. and Enterobacter sp. produced some of the same alkaloids as their host plants. Additional study needs to be done to determine if these endophytic bacteria could be used to produce alkaloids in the fermentation industry.     

The use of hydrogen peroxide to control postharvest decay on 'Galia' melons

Sanosil-25, a disinfectant containing 48% hydrogen peroxide and silver salts as stabilising agents, inhibited the mycelial growth of the two main decays causing fungi of melons, Alternaria alternata and Fusarium solani in vitro at concentrations between 5000 and 10 000 μl litre^-1. However, in in vivo experiments, Sanosil-25 markedly decreased decay at a concentration of 5000 μl litre^-1 when incorporated into a wax treatment and produced no phytotoxic effect. This treatment may provide an alternative to imazalil which, although more effective, gives problems with residue levels. A concentration of 10 000 μl litre^-1 proved phytotoxic.