Crude extract of the cyanobacterium, Hapalosiphon sp., causes a cessation of root elongation and cell division in several plant species

The phytotoxic activities of the crude extract from the cyanobacterium, Hapalosiphon sp., on the initial growth and root cell division of several plant species were investigated. Although the germination percentages of the plants were not affected by the extract, their root and shoot growth were remarkably suppressed, depending on the extract concentrations. Roots were more sensitive to the extract than shoots and the sensitivity, determined by the I₅₀ concentrations between the susceptible and tolerant species, differed by 5 and 11 times in dicotyledonous and monocotyledonous plants, respectively. The mitotic index for wheat ( Triticum aestivum L. cv. Norin 61) root cells was not affected, but the mitotic index for onion ( Allium cepa L. cv. Raputa II) was reduced with the increasing concentration of the extract. In the onion root meristem, the number of cells in prophase, metaphase, anaphase, and telophase were decreased significantly with 6 g DW L⁻¹ (g dry weight of algae L⁻¹). This suggests that the extract might inhibit the processes regulating the entry of cells into mitosis.     

Effects of aqueous extracts and decomposition of Mikania micrantha H.B.K. debris on selected agronomic crops

Laboratory and greenhouse experiments were conducted to determine the effects of aqueous extract and the debris of Mile-a-Minute ( Mikania micrantha H.B.K.) on the germination and growth of four test species, viz. tomato ( Lycopersicon esculentum Mill.), Chinese cabbage ( Brassica chinensis L.), corn ( Zea mays L.) and long bean ( Vigna sesquipedalis L.). Germination percentage, radicle length and fresh weight of two crops, Chinese cabbage and tomato, decreased progressively when plants were exposed to increasing concentrations (12.5, 25, 37.5 and 50 g L⁻¹) of aqueous extract of Mikania , but did not affect those of corn and long bean seedlings. The emergence of tomato and Chinese cabbage was greatly reduced when Mikania debris was incorporated into the soil, irrespective of the amount of debris and the decomposition period. In contrast, the emergence of corn and long bean seedlings was not affected. The application of fertilizer (NPK 15 : 15 : 15) enhanced the fresh weight of all seedlings tested as compared with those not supplied with the fertilizer. Four compounds were identified in the leaf extract of Mikania , viz. caffeic acid, p -hydroxybenzaldehyde, resorcinol and vanillic acid.     

Allelopathic potential of Trifolium resupinatum L. (Persian clover) and Trifolium alexandrium L. (Berseem clover)

This experiment was conducted in order to determine if Persian clover ( Trifolium resupinatum ) and Berseem clover ( Trifolium alexandrium ) contained water-soluble and methanol-soluble constituents that could affect the seedling growth of four test species; namely, Amaranthus retroflexus , Convolvulus arvensis (field bindweed), Secale cereale , and Sinapis arvensis (wild mustard) . The above-ground tissues of the Persian and Berseem clover plants were collected during the vegetative growth stage and oven-dried. Three concentrations of aqueous and methanolic extracts were used at full-strength (33.3 g L⁻¹), half-strength (16.7 g L⁻¹), and quarter-strength (8.3 g L⁻¹). Distilled water was used as the control. The weed seeds were placed in Petri dishes containing the legume extract or distilled water (control). The seedling growth of the weed species declined with the increasing concentration of the clover extracts. Wild mustard exhibited the highest sensitivity to both the legume extracts. Compared to the aqueous extract, the methanolic extract caused a greater decline in the seedling growth of the weeds. Berseem clover was the stronger inhibitor of the weed seedling growth, as compared to Persian clover. Therefore, the amounts of allelochemicals present might differ in these two clover species. Field bindweed showed the least sensitivity to both the legume extracts. Field bindweed showed more tolerance to the allelochemicals produced by the clover species.     

Control of Penicillium expansum by plant volatile compounds

Nine plant volatiles were tested for their activity in vitro and in vivo against Penicillium expansum , the cause of blue mould of pear. In vitro spore germination and mycelial growth assay showed a consistent fungicidal activity by trans -2-hexenal, carvacrol, trans -cinnamaldehyde and citral, while hexanal (-)- carvone, p -anisaldehyde, eugenol and 2-nonanone exhibited a progressively lower inhibition. trans -2-Hexenal was the best inhibitor of conidial germination [MIC (minimum inhibitory concentration) = 24·6  µ L L⁻¹; ED₅₀ = 10·2  µ L L⁻¹], while carvacrol was the best inhibitor of mycelial growth (MIC = 24·6  µ L L⁻¹; ED₅₀ = 9  µ L L⁻¹). The four most active compounds in in vitro studies were tested in vivo as fumigants against blue mould on pear cv. Conference. Best control was achieved by trans -2-hexenal vapour treatments (12·5  µ L L⁻¹) when applied over a 24-h period, beginning 24 h after inoculation. In contrast, carvacrol (12·5–200  µ L L⁻¹), and trans -cinnamaldehyde (50–400  µ L L⁻¹) were ineffective and citral (200  µ L L⁻¹) showed only slight effect.     

Characterization of reactive oxygen species-involved oxidative damage in Hapalosiphon species crude extract-treated wheat and onion roots

Crude extract-induced oxidative damage using the cyanobacterium, Hapalosiphon sp., was investigated in wheat seedlings ( Triticum aestivum L. cv. Norin 61) and onion seedlings ( Allium cepa L. cv. Raputa II). The analysis of root cell viability or cell death using Evans blue uptake showed that the root-tip cells of wheat and onion lost viability after 24 h and 48 h treatment with 3 g dry weight (DW) L⁻¹ of the crude extract, respectively. Lipid peroxidation was induced in the roots of both species and the shoots of onion, whereas no increase in lipid peroxide formation was observed in the wheat shoots. In onion, the degree of random DNA fragmentation increased with the increasing concentration of the extract and laddering of the DNA was observed with 6 g DW L⁻¹ of the extract, but no apparent DNA ladder formation occurred in the wheat. Pretreatment for 1 h with the NADPH oxidase inhibitors, diphenyleneidonium or imidazole, reduced the crude extract-induced root cell death in both species. From the results, we suggest that the Hapalosiphon sp. crude extract might enhance reactive oxygen species (ROS) production, which causes membrane lipid damage and fragmentation of the DNA of plant cells, resulting in cell death and growth inhibition. The crude extract-mediated phytotoxic damage might be caused by ROS, triggered by NADPH oxidase.     

Inhibitory effects of aqueous extracts of black mustard on germination and growth of alfalfa

The present study investigated the allelopathic effects of various black mustard ( Brassica nigra L.) plant parts on alfalfa germination and seedling growth. Aqueous extracts of black mustard ( Brassica nigra L.) leaf, stem, flower and root plant parts were made to determine their effects on germination and dry weights of hypocotyl, and radicle length of 5-day-old alfalfa seedlings over a range of extract concentrations. All aqueous extracts significantly inhibited alfalfa seed germination and seedling growth when compared with the distilled water control. Increasing the aqueous extract concentrations of separated Brassica nigra plant parts significantly inhibited alfalfa germination, seedling length and weight. Based on significant radicle length reactions to aqueous extracts, the toxicity may be classified in the following order of decreasing inhibition: flower, leaf, mixture of all plant parts, root, and stem. Increasing the concentration of aqueous flower extracts significantly inhibited the water uptake by germinated alfalfa seeds. The greatest inhibition in water uptake occurred at a 12 g kg⁻¹ extract concentration for seeds soaked for 8 h.     

A rapid reliable test for screening aryloxyphenoxy- propionic acid resistance within Alopecurus myosuroides and Lolium spp. populations

A reliable seedling bioassay was developed and tested for the rapid screening for resistance to aryloxyphenoxypropionic (APP) herbicides in Alopecurus myosuroides and Lolium spp. populations. It is based upon the difference in coleoptile length of resistant and susceptible A. myosuroides and Lolium seedlings, respectively, exposed to fenoxaprop-P acid and diclofop acid solution for 6 days in a plastic box. A 6 mg L⁻¹ fenoxaprop-P acid solution was selected as the best concentration for a reliable screening of resistant biotypes within A. myosuroides populations. At this concentration, coleoptile lengths of susceptible and resistant seedlings were shorter and longer than 10 mm respectively. Similarly, resistant seedlings within Lolium populations were easily detected at 10 mg L⁻¹ diclofop acid. At this concentration, coleoptile lengths of susceptible and resistant seedlings were shorter and longer than 20 mm respectively. For both populations, the coleoptile length distributions appear to discriminate between two kinds of APP-resistant biotypes (highly and slightly resistant).     

Antifungal activity of bergenin, a constituent of Flueggea microcarpa

The antifungal activity of bergenin against some plant pathogenic fungi, namely, Alternaria alternata , A. brassicae , A. carthami , Fusarium udum , F. oxysporum f.sp. ciceri , Curvularia lunata and Erysiphe pisi , was studied. Bergenin as its monosodium salt was effective against all the fungi and the effective dose for complete inhibition of spore germination varied from 15 μg mL⁻¹ for F. udum to 125 μg mL⁻¹ for E. pisi . Experiments on the effect of bergenin on powdery mildew development under glasshouse conditions revealed that it can control powdery mildew of pea at 2000 μg mL⁻¹ by postinoculation treatment, the results being comparable with those of carbendazim (1000 μg mL⁻¹) and wettable sulfur (2000 μg mL⁻¹). It affected hyphal elongation and the number of primary and secondary branches.     

A semiselective medium for the isolation of Acidovorax valerianellae from soil and plant debris

Bacterial black spot of corn-salad ( Valerianella locusta ) caused by Acidovorax valerianellae appeared in France in the beginning of the 1990s. The disease is now widespread in the main area of corn-salad production and is responsible for high depreciation and significant economic losses every year. A semiselective medium is described for the detection of the bacterium in various environments and to identify primary sources of inoculum. The medium TSAV (tryptic soy broth, 3 g L⁻¹, agar 15 g L⁻¹, 5-fluorouracyl 5 mg L⁻¹, novobiocin 5 mg L⁻¹, propiconazole 5 mg L⁻¹) increased chances of obtaining cultures of A. valerianellae from naturally infested soil and from root-debris. Using TSAV, the pathogen was detected in root-debris up to 39 days after the harvest of a diseased crop. As only a few days usually separates harvest from the next sowing date, soil is likely to be an important source of inoculum for the next crop.     

Weed control in summer-sown soybeans with flumioxazin plus acetochlor and flumiclorac-pentyl plus clethodim

Field trials were conducted in Taigu, Shanxi province, China, to evaluate the efficacy of flumioxazin plus acetochlor and flumiclorac-pentyl plus clethodim applied to summer-sown soybeans at pre- and postemergence. It was demonstrated that tank-mixing flumioxazin at 50 g ai ha^-1 and acetochlor at 800 g ai ha^-1 created an effective soil-applied herbicide for weed control in soybean crops. The control efficacy was better than when the herbicides were applied individually, and no injury was caused to the soybeans. Flumiclorac-pentyl at 50 g ai ha^-1 plus clethodim at 70 g ai ha^-1 suppressed both broad-leaved weeds and grass weeds with an increased efficacy of more than 90%. Flumiclorac-pentyl applied alone or tank-mixed caused some injury to soybean seedlings, but the soybeans recovered 2–3 weeks after treatment and there was no reduction in the yield.     

Management of late leaf spot of groundnut (Arachis hypogaea) with chlorothalonil-tolerant isolates of Pseudomonas aeruginosa

Fifteen groundnut-associated bacterial isolates that inhibited by > 90% the in vitro conidial germination of Phaeoisariopsis personata , causal agent of late leaf spot disease of groundnut, were applied as a prophylactic spray (10⁸ cfu mL⁻¹) and tested for control of the disease in the glasshouse. Two groundnut seed-associated bacterial isolates, GSE 18 and GSE 19, identified as Pseudomonas aeruginosa , reduced the lesion frequency (LF) by up to 70%. A 90-day-old peat-based formulation of P. aeruginosa GSE 18 reduced LF measured 15 days postinoculation by up to 60%. Both P. aeruginosa GSE 18 and GSE 19 were tolerant to chlorothalonil (Kavach®) up to 2000  µ g mL⁻¹ in LB broth. In glasshouse trials, GSE 18 and GSE 19 tested in combination with reduced concentrations of chlorothalonil were highly efficient in management of the disease. The disease was completely controlled by chlorothalonil (> 250  µ g mL⁻¹), and in the presence of GSE 18 or GSE 19, 100  µ g mL⁻¹ chlorothalonil was equally effective. Application of rifamycin-resistant mutants of GSE 18 or GSE 19 together with chlorothalonil significantly increased the survival of these isolates in the groundnut phylloplane. In the field, a combination of GSE 18 and 500  µ g mL⁻¹ chlorothalonil reduced disease severity comparable to 2000  µ g mL⁻¹ chlorothalonil alone. Use of chlorothalonil-tolerant pseudomonads together with a quarter concentration of the recommended field dose of chlorothalonil doubled pod yield compared with the untreated unsprayed control.     

Synthesis and herbicidal activity of new 3-(3-bromothiophen-2-yl)-5-(2,6-difluorobenzyl)oxymethyl-5-methyl-4,5-dihydroisoxazole as a paddy field herbicide

A new compound 3-(3-bromothiophen-2-yl)-5-(2,6-difluorobenzyl)oxymethyl-5-methyl-4,5-dihydroisoxazole was synthesized and the herbicidal activity was assessed under glasshouse and flooded paddy field conditions. This compound demonstrated good rice selectivity and potent herbicidal activity against annual weeds at 125 g ai ha⁻¹ under greenhouse conditions. Soil application of this compound showed the complete control of barnyardgrass to the 4th leaf stage at 250 g ai ha⁻¹. Field trials indicated that this compound, combined with pyrazosulfuron-ethyl, controlled annual and perennial weeds rapidly with good tolerance by transplanted rice seedlings from postemergence and soil application. This compound showed a low mammalian and environmental toxicity in various toxicological tests.     

Factors affecting the control of Pythium ultimum damping-off of sugar beet by Pythium oligandrum

Application of Pythium oligandrum to a soil-based compost as a mycelial suspension (5 × 10² CFU g⁻¹ of dry compost) and oospore alginate pellets (10⁵ oospores/g of dry compost) controlled pre- and postemergence damping-off of sugar beet caused by Pythium ultimum to a level similar to metalaxyl seed treatment. Oospore seed treatments and aqueous suspensions of oospores applied to compost failed to control disease. Problems in the use of P. oligandrum oospore inocula for the control of damping-off were highlighted. It was shown that treatment of oospores with cellulase (20 g L⁻¹) increased germination approximately three-fold in comparison to untreated spores. Untreated and cellulase pretreated oospores were subsequently evaluated as seed treatments for their ability to control damping-off of sugar beet. The highest rate of pretreated oospores (10⁴ oospores/seed) gave levels of emergence and establishment in infested compost that were not significantly different from the uninfested controls, whereas seed treatment with untreated oospores gave no significant reduction in disease. In a trial carried out in a controlled environment to assess the effect of pH (4.5–8.0), P. oligandrum (10⁴ cellulase pretreated oospores/seed) was shown to control pre- and postemergence damping-off of sugar beet at pH 7.0 and 7.5 only.     

Resistance to Sphaerotheca pannosa in roses induced by 2,6-dichloroisonicotinic acid

The susceptible rose cv. Madelon and the partially resistant cv. Sonia both responded with reduced development of rose powdery mildew when they were treated with the synthetic inducer 2,6-dichloroisonicotinic acid (INA). The EC₅₀ for number of colonies cm⁻² was approximately 0.4 mg L⁻¹ in both cultivars when treated 4 days prior to inoculation. However, conspicuous differences were observed with respect to number of spores per cm². For sporulation, the EC₅₀ was 0.37 mg L⁻¹ in cv. Madelon and only 0.08 mg L⁻¹ in cv. Sonia. A comparison with the pathosystems cucumber/ Sphaerotheca fuliginea and red cabbage/ Peronospora parasitica is made and the importance of the observed phenomenon for the selection of parents in a breeding programme for (partial) resistance is discussed.     

Biological control of fusarium crown and root rot of tomato with antagonistic bacteria and integrated control when combined with the fungicide carbendazim

Two bacterial isolates, Bacillus megaterium (c96) and Burkholderia cepacia (c91), demonstrated to be antagonistic against Fusarium oxysporum f.sp. radicis-lycopersici , the causal organism of fusarium crown and root rot of tomato, were evaluated as biocontrol agents alone and when integrated with the fungicide carbendazim. In an initial screening, these isolates reduced disease incidence by 75 and 88%, respectively. In vitro , both biocontrol agents were highly tolerant to the fungicide carbendazim, commonly used to control fusarium diseases. Carbendazim reduced disease symptoms by over 50% when used at > 50  µ g mL⁻¹, but had little effect at lower concentrations. Combination of the bacterial isolates and carbendazim gave significant ( P  ≤ 0·05) control of the disease when plants were artificially inoculated with the pathogen. Application of carbendazim at a low concentration (1  µ g mL⁻¹) in combination with B. cepacia c91 reduced disease symptoms by 46%, compared with a reduction of 20% obtained with the bacterium alone and no control with the chemical treatment alone. A combination of B. megaterium c96 with an increased application rate of 10  µ g mL⁻¹ carbendazim significantly reduced disease symptoms by 84% compared with inoculated controls and by 77% compared with carbendazim treatment alone. In this experiment, the integrated treatment also slightly outperformed application of 100  µ g mL⁻¹ carbendazim, and bacteria applied without fungicide also provided good disease control.     

Sensitivity to cymoxanil in populations of Plasmopara viticola in northern Italy

In 1993, control failures were reported on grapevine in northern Italy under severe downy mildew pressure after postinfection application of cymoxanil in mixtures with copper or mancozeb. A monitoring survey was started immediately in Piedmont (north-western Italy) in order to determine the sensitivity of populations of Plasmopara viticola to cymoxanil from those vineyards where the fungicide was not controlling the disease satisfactorily. In 1994 and 1995, monitoring surveys were extended to north-eastern Italy, where cymoxanil mixtures were not performing as well as in the past. Sampled populations were tested on detached leaf discs and on whole potted plants under controlled conditions. In 1993, 12 populations, sampled in Piedmont, showed MIC values (minimum inhibitory concentration) varying from 10 to more than 100 mg L⁻¹ cymoxanil. With a baseline reference population having a MIC value of 3 mg L⁻¹, resistance factors ranged from 3 to more than 30. In 1994, 17 populations out of 27 sampled in Trentino (north-eastern Italy) showed MIC values of 100 mg L⁻¹ or higher and in 1995, 32 populations out of 38 showed the same behaviour. In similar experiments, the MIC values of populations from nontreated plots were between 3 and 10 mg L⁻¹. In whole potted plant tests, populations with MIC values higher than 200 mg L⁻¹ in a leaf disc test were not controlled by 500 mg L⁻¹ of cymoxanil. The results of our study suggest that resistance to cymoxanil in P. viticola may contribute to a lack of disease control in Italian vineyards.     

Reduced rates of tank mixtures for red sprangletop (Leptochloa chinensis [L.] Nees) and greater club-rush (Scirpus grossus [L.] f.) control in rice

A study on the efficacy of tank-mix combinations on the growth of Leptochloa chinensis (L.) Nees and Scirpus grossus (L.) f. at the 3–4-leaf stage was conducted under glasshouse conditions. Sethoxydim plus cyhalofop-butyl at 8 + 25, 16 + 25, 32 + 25, 32 + 12.5, and 32 + 6.25 g ai ha⁻¹ provided good control of L. chinensis , ranging from 89–95%. However, a comparison of the costs of herbicide mixtures revealed that sethoxydim at 32 g ai ha⁻¹ plus cyhalofop-butyl at 6.25 g ai ha⁻¹ were the most cost-effective among the combinations. For Scirpus grossus , bensulfuron plus 2,4-D dimethylamine at 2.5 + 18, 5.0 + 18, 1.25 + 36, 2.5 + 36, and 5.0 + 36 g ai ha⁻¹ gave good control that ranged from 88–93%. Tank-mixing bensulfuron with 2,4-D at 1.25 + 36 g ai ha⁻¹ was, however, the most cost-effective mixture compared to the other mixtures. None of the tank mixtures tested in this study caused injury to the rice seedlings. However, further field studies need to be undertaken to verify these findings.     

Effect of salinity on pythium damping-off of cucumber and on the tolerance of Pythium aphanidermatum

A study was conducted to investigate the effect of salinity on pythium damping-off and tolerance to salinity among Pythium aphanidermatum isolates from different geographical origins, genetic backgrounds and saline/non-saline soils. Increasing irrigation-water salinity from 0·01 to 5 dS m⁻¹ significantly increased mortality in cucumber seedlings inoculated with P. aphanidermatum and reduced dry weight of non-inoculated seedlings. In vitro tests in culture media amended with different concentrations of NaCl showed that growth of P. aphanidermatum, P. spinosum and P. splendens isolates was stimulated or unaffected at salinity levels stressful for cucumber (electrical conductivity = 5 dS m⁻¹). Significant differences were found in tolerance to salinity among 47 P. aphanidermatum isolates obtained from different greenhouses. The concentration of NaCl required to reduce growth of P. aphanidermatum isolates by 50% varied from 23 to 62 dS m⁻¹, with an average of 46 dS m⁻¹, however, oospore production was more sensitive to salinity and no oospores were produced above 20 dS m⁻¹. Analysis of tolerance of P. aphanidermatum isolates to salinity showed no relationship between tolerance to salinity and geographical origins or genetic data. Isolates of P. aphanidermatum from greenhouses with no salinity problems were as tolerant to salinity as isolates obtained from salinity-affected greenhouses, suggesting lack of evidence for ecological adaptation within P. aphanidermatum in Oman. Increased mortality in cucumber seedlings at higher salinity levels may imply a synergistic interaction between salinity stress and salinity-tolerant Pythium species on cucumber seedlings, resulting in greater seedling losses.     

Action of the fungicide phosphite on Eucalyptus marginata inoculated with Phytophthora cinnamomi

Although phosphite has been effective in the control of P. cinnamomi in E. marginata (jarrah) , the biochemical mechanisms behind phosphite protection are poorly understood. Using an aeroponics system, jarrah clones with moderate resistance to P . cinnamomi were treated with foliar applications of phosphite (0 and 5 g L⁻¹). The roots were inoculated with zoospores of P. cinnamomi at 4 days before and 0, 2, 5, 8 and 14 days after phosphite treatment. Root segments were then analysed for activity of selected host defence enzymes (4-coumarate coenzyme A ligase [4-CL], cinnamyl alcohol dehydrogenase [CAD]) and the concentration of soluble phenolics and phosphite. Lesion development was most effectively reduced when phosphite concentrations within the roots were highest (i.e. days 8–14). During this time, the levels of host defence enzymes remained relatively unchanged. Lesion development was also effectively restricted when phosphite concentrations within the roots were lowest (i.e. days 2 and 5); a significant increase in host defence enzymes was associated with this decrease in lesion development. It was concluded from these studies that the effect of phosphite in controlling the pathogen is determined by the phosphite concentration at the host–pathogen interface. When phosphite concentrations within the roots are low, phosphite interacts with the pathogen at the site of ingress to stimulate host defence enzymes. At high phosphite concentrations, phosphite acts directly on the pathogen to inhibit its growth before it is able to establish an association with the host, and the host defences remain unchanged.     

Tolerance of black, cranberry, kidney, and white bean to cloransulam-methyl

The level of tolerance of various market classes of dry bean to cloransulam-methyl is not known. Three field studies were conducted in Ontario, Canada during 2007 and 2008 to determine the level of tolerance of black, cranberry, kidney, and white bean to the pre-emergence (PRE) and postemergence (POST) application of cloransulam-methyl at 17.5, 35, and 70 g ai ha⁻¹. Cloransulam-methyl applied at 17.5, 35, and 70 g ha⁻¹ caused between 13 and 23% injury in black, cranberry, kidney, and white bean, respectively. Cloransulam-methyl applied at 17.5, 35, and 70 g ha⁻¹ reduced the shoot dry weight by between 16 and 28% compared to the untreated control. Cloransulam-methyl applied PRE reduced the height of black bean by 27% and the height of cranberry bean by 25% at 70 g ha⁻¹ and reduced the height of white bean by 19% at 35 g ha⁻¹ and by 37% at 70 g ha⁻¹. Cloransulam-methyl applied PRE reduced the yield of black bean by 29% at 35 g ha⁻¹ and by 43% at 70 g ha⁻¹, reduced the yield of cranberry bean by 43% at 70 g ha⁻¹, and reduced the yield of white bean by 36% at 35 g ha⁻¹ and by 54% at 70 g ha⁻¹. Based on these results, there is not an adequate margin of crop safety for the PRE and POST application of cloransulam-methyl in black, cranberry, kidney, and white bean at the rates evaluated.