Biocontrol of Sclerotinia sclerotiorum infection of cabbage by Coniothyrium minitans and Trichoderma spp.

Nine fungal isolates [Clonostachys rosea (1), Coniothyrium minitans (1), Trichoderma crassum (1), T. hamatum (4), T. rossicum (1) and T. virens (1)] were tested in two bioassays for their ability to degrade sclerotia and reduce apothecial production and carpogenic infection of cabbage seedlings. C. minitans LU112 reduced apothecial production in both bioassays, with T. virens LU556 significantly reducing apothecial production in the sclerotial parasitism assay. Both isolates also reduced sclerotial viability in this assay to 5% for C. minitans and 22% for T. virens. C. minitans LU112 and T. virens LU556 reduced the infection of cabbage seedlings in the pot bioassay 126 days after sowing but not after 147 days, partly due to ascospore cross-infection between treatments. C. minitans LU112, T. virens LU556 and T. hamatum LU593 as maizemeal-perlite (MP) soil incorporation and transplant potting mix incorporation were evaluated for their ability to control Sclerotinia sclerotiorum disease of cabbage in field experiments. S. sclerotiorum infection of cabbage was reduced by 46–52% and 31–57% by both C. minitans LU112 and T. hamatum LU593 as MP soil incorporations, respectively, in the two field experiments. T. virens LU556 MP soil incorporation and C. minitans LU112 and T. hamatum LU593 transplant potting mix incorporations reduced S. sclerotiorum disease in the first experiment but not in the second experiment. A commercial C. minitans LU112 formulation, C. Mins LU112 WG, also significantly reduced S. sclerotiorum disease by 59%. Soil incorporation of C. minitans and T. hamatum was shown to have potential to control S. sclerotiorum disease in cabbage.     

Sensitivity of Clonostachys rosea and Trichoderma spp. as Potential Biocontrol Agents to Pesticides

Clonostachys rosea 47 (CR47), Trichoderma atroviride 59 (TA59), T. atroviride 312 (TA312), Trichoderma harzianum 24 (TH24), Trichoderma longibrachiatum 9 (TL9), T. longibrachiatum 144 (TL144) and Trichoderma viride 15 (TV15) were tested to evaluate their in vitro sensitivity towards five fungicides (carboxin, guazatine, prochloraz, thiram and triticonazole) and four herbicides (chlorsulfuron, chlorotoluron, flufenacet and pendimethalin). All antagonists showed low sensitivity to carboxin and thiram and high sensitivity to prochloraz. For mycelial radial growth, TV15 was highly sensitive to guazatine, prochloraz and triticonazole and TH24 moderately insensitive to carboxin, guazatine and thiram. For conidial germination TL144 was the most sensitive to the fungicides, for mycelial radial growth and conidial germination CR47 was the least sensitive. None of the antagonists showed any mycelial radial growth inhibition in presence of the herbicides at field dose, except for TL144. Most antagonists did not show any conidial germination inhibition by the herbicides. The in vitro toxicity of prochloraz, guazatine and triticonazole towards the antagonists was confirmed by light and scanning electron microscope showing hyphal disruptions and extrusion of cytoplasmic content. A mixture of CR47 and/or TA312 with carboxin, thiram and triticonazole, applied to wheat seeds, was able to control Fusarium culmorum artificially inoculated to wheat seedlings in growth chambers. In the field, the antagonists applied along with triticonazole or thiram, at 1/10 of the field dose to seeds naturally infected by F. culmorum, gave a disease control comparable to that induced by triticonazole at full field dose. Our results demonstrate how an integration of microorganisms with pesticides makes the control of wheat foot rot possible.     

Effect of opportunistic fungi on the life cycle of the root-knot nematode ( Meloidogyne javanica) on brinjal

The effect of four opportunistic fungi viz., Paecilomyces lilacinus, Cladosporium oxysporum, Gliocladium virens and Talaromyces flavus on the life cycle of the root-knot nematode, Meloidogyne javanica, on brinjal was evaluated under glasshouse conditions. The results revealed that these fungi affected the penetration and development of M. javanica. The life cycle of M. javanica was delayed by 10, 7, 4 and 2 days in the presence of P. lilacinus, C. oxysporum, G. virens and T. flavus respectively. Fecundity, number of eggs per eggmass and number of larvae was also reduced in the presence of these opportunistic fungi. However, the number of males increased in the presence of opportunistic fungi.     

Does seasonal reproduction occur at the optimal time for fertilization in the polychaetes Arenicola marina L. and Nereis virens Sars?

Summary

Many marine invertebrates exhibit highly seasonal and synchronised reproduction, with offspring production often being confined to just two or three days each year. Several models have been proposed to explain the fitness benefits of this reproductive pattern, many of which assume enhanced offspring survival due to temperature constraints placed on fertilization and development at other times of the year. In this investigation the temperature limits and optimum for fertilization were determined for two polychaete species, Arenicola marina and Nereis virens. These two polychaete species are exposed to the same environmental conditions throughout the year, yet breed at very different times. Other seasonal impacts on fertilization, i.e., reduced salinity due to rainfall and the effect of sub-zero temperatures on sperm of A. marina, were also investigated. In both A. marina and N. virens fertilization success was significantly influenced by temperature, with the maximum success recorded at 15–18°C. The ambient seawater temperature at the time of natural spawning for both worms is around 10°C, which means that both species are spawning right at the lower limit for maximum fertilization. Salinity and exposure of A. marina sperm to sub-zero temperatures were also found to influence fertilization success, but only at levels that would not be experienced by these polychaetes under natural conditions at the time of spawning. These results suggest there must be additional selective pressures acting on the fitness of these two polychaetes causing A. marina to breed later than, and N. virens to breed earlier than, the optimum time for fertilization. A. marina apparently waits as late as possible to maximise adult fecundity and survival. N. virens breeds as soon as it can achieve high fertilization to maximise larval and juvenile competitiveness.     

Effects of copper and other metals on fertilization,embryo development,larval survival and settlement of the polychaete Nereis (Neanthes) virens

Summary

Nectochaete larvae of the ecologically and economically important ragworm, Nereis virens, were exposed to cadmium, chromium, copper, lead and zinc dissolved in seawater to nominal concentrations ranging from 0 to 5000 μg l^?1. Copper was the most toxic (mean LC50 of 76.5 μg l^?1 ± 95% CI 73.8–79.2 after 96 h exposure) and so was used for subsequent experiments. Exposure of gametes to greater than 500 μg l^?1 copper for 2 or 4 h at 10°C prior to fertilization, or a 10 min exposure during fertilization, significantly reduced embryo developmental success. The effect of copper on larval settlement was also assessed using sediment spiked to a range of concentrations (0, 50, 250, 500, 1000 mg kg!1 dry weight). Significantly fewer larvae were found in sediment of $250 mg kg!1 in comparison to the control or the 50 mg kg!1 treatment. Assessment of living larvae also confirmed a significant reduction in settlement, but in all treatments compared to the control, although the number of dead larvae also increased as the concentrations increased. These effects may have important implications for reproductive success and recruitment of N. virens to polluted sediments.     

Is the Cape White-eye Zosterops virens or Zosterops capensis?

The recent scientific literature employs three binomial names for the southern African endemic bird known as the Cape White-eye: Zosterops capensis, Z. pallidus and Z. virens. This unacceptable inconsistency reflects the contention regarding white-eye systematics. Recent molecular work by Oatley and colleagues led to the suggestion that Z. virens and Z. capensis should both fall under the name Z. virens. We urge ornithologists to adopt this convention, and we encourage further molecular stud ies in order to clarify the taxonomy of southern African white-eyes.     

Different mechanisms of Trichoderma virens‐mediated resistance in tomato against Fusarium wilt involve the jasmonic and salicylic acid pathways

In the present study, we investigated the role of Trichoderma virens (TriV_JSB100) spores or cell‐free culture filtrate in the regulation of growth and activation of the defence responses of tomato (Solanum lycopersicum) plants against Fusarium oxysporum f. sp. lycopersici by the development of a biocontrol–plant–pathogen interaction system. Two‐week‐old tomato seedlings primed with TriV_JSB100 spores cultured on barley grains (BGS) or with cell‐free culture filtrate (CF) were inoculated with Fusarium pathogen under glasshouse conditions; this resulted in significantly lower disease incidence in tomato Oogata‐Fukuju plants treated with BGS than in those treated with CF. To dissect the pathways associated with this response, jasmonic acid (JA) and salicylic acid (SA) signalling in BGS‐ and CF‐induced resistance was evaluated using JA‐ and SA‐impaired tomato lines. We observed that JA‐deficient mutant def1 plants were susceptible to Fusarium pathogen when they were treated with BGS. However, wild‐type (WT) BGS‐treated tomato plants showed a higher JA level and significantly lower disease incidence. SA‐deficient mutant NahG plants treated with CF were also found to be susceptible to Fusarium pathogen and displayed low SA levels, whereas WT CF‐treated tomato plants exhibited moderately lower disease levels and substantially higher SA levels. Expression of the JA‐responsive defensin gene PDF1 was induced in WT tomato plants treated with BGS, whereas the SA‐inducible pathogenesis‐related protein 1 acidic (PR1a) gene was up‐regulated in WT tomato plants treated with CF. These results suggest that TriV_JSB100 BGS and CF differentially induce JA and SA signalling cascades for the elicitation of Fusarium oxysporum resistance in tomato.     

Evaluation of seed dressing and soil application formulations of Trichoderma species for integrated management of dry root rot of chickpea

The efficacy of the newly developed seed dressing and soil application formulations of Trichoderma viride, T. virens and T. harzianum were evaluated individually and in combinations under pot and field experiments for the management of dry root rot (Rhizoctonia bataticola) of chickpea (Cicer arientinum). In pot experiments, T. harzianum based seed dressing formulation, Pusa 5SD, and soil application formulations, Pusa Biogranule 5 (PBG 5) and Pusa Biopellet 10G (PBP 10G), were found to be effective in reducing dry root rot incidence in chickpea and increasing the seed germination, shoot and root lengths of the crop. Under field experiments, a combination of soil application of T. harzianum based PBP 10G and seed treatment with Pusa 5SD+carboxin was found to be the best by providing the highest seed germination, shoot and root lengths and grain yield and the lowest dry root rot incidence in chickpea.     

Morphogenesis in the Heterotrich Ciliate Climacostomum virens. I. Oral Development During Cell Division

The principal characteristics of stomatogenesis during division in Climacostomum virens are: (A) Kinetosomal proliferation on the left side of a variable number of kineties in the ventral somatic cortex forms an oral primordium consisting of several kinetosomal fields, which then fuse to form a single anarchic field. (B) A constant topographical relationship exists between the primordium and a well defined cortical pattern, the zone of discontinuity. (C) The anarchic field primordium divides into 2 unequal parts—to the left, the AZM primordium, and to the right, the paroral primordium, which differentiates into the apical membranelles, the peristomial field, and the buccal tube. (D) Preoral and oblique kineties of the somatic cortex form along the right side of the paroral primordium. (E) Parental oral structures are partially dedifferentiated. Stomatogenesis in C. virens and other heterotrichs is compared.     

Electronic nose evaluation of onion headspace volatiles and bulb quality as affected by nitrogen, sulphur and soil type

Edaphic factors affect the quality of onions (Allium cepa). Two experiments were carried out in the field and glasshouse to investigate the effects of N (field: 0,120 kg ha⁻¹; glasshouse: 0,108 kg ha⁻¹), S (field: 0, 20 kg ha⁻¹; glasshouse: 0, 4.35 kg ha⁻¹) and soil type (clay, sandy loam) on onion quality. A conducting polymer sensor electronic nose (E-nose) was used to classify onion headspace volatiles. Relative changes in the E-nose sensor resistance ratio (%dR/R) were reduced following N and S fertilisation. A 2D Principal Component Analysis (PCA) of the E-nose data sets accounted for c. 100% of the variations in onion headspace volatiles in both experiments. For the field experiment, E-nose data set clusters for headspace volatiles for no N-added onions overlapped (D²= 1.0) irrespective of S treatment. Headspace volatiles of N-fertilised onions for the glasshouse sandy loam also overlapped (D²=1.1) irrespective of S treatment as compared with distinct separations among clusters for the clay soil. N fertilisation significantly (P < 0.01) reduced onion bulb pyruvic acid concentration (flavour) in both experiments. S fertilisation increased pyruvic acid concentration significantly (P < 0.01) in the glasshouse experiment, especially for the clay soil, but had no effect on pyruvic acid concentration in the field. N and S fertilisation significantly (P < 0.01) increased lachrymatory potency (pungency), but reduced total soluble solids (TSS) content in the field experiment. In the glasshouse experiment, N and S had no effect on TSS. TSS content was increased on the clay by 1.2-fold as compared with the sandy loam. Onion tissue N: water-soluble SO₄²⁻ ratios of between five and eight were associated with greater %dR/R and pyruvic acid concentration values. N did not affect inner bulb tissue microbial load. In contrast, S fertilisation reduced inner bulb tissue microbial load by 80% in the field experiment and between 27% (sandy loam) and 92% (clay) in the glasshouse experiment. Overall, onion bulb quality discriminated by the E-nose responded to N, S and soil type treatments, and reflected their interactions. However, the conventional analytical and sensory measures of onion quality did not correlate with %dR/R.     

Management Fusarium wilt on melon and watermelon by Penicillium oxalicum

The potential of the biological control fungus Penicillium oxalicum to suppress wilt caused by Fusarium oxysporum f. sp. melonis and F. oxysporum f. sp. niveum on melon and watermelon, respectively, was tested under different growth conditions. The area under disease progress curve of F. oxysporum f. sp. melonis infected melon plants was significantly reduced in growth chamber and field experiments. In glasshouse experiments, it was necessary to apply P. oxalicum and dazomet in order to reduce Fusarium wilt severity in melons caused by F. oxysporum f. sp. melonis. For watermelons, we found that P. oxalicum alone reduced the area under the disease progress curve by 58% in the growth chamber experiments and 54% in the glasshouse experiments. From these results, we suggested that P. oxalicum may be effective for the management of Fusarium wilt in melon and watermelon plants.     

UVI_02019870, a Puptive Effector from <i>Ustilaginoidea virens</i>, Interacts with a Chloroplastic-Like Protein OsCPL1

Ustilaginoidea virens, which causes rice false smut (RFS), is one of the most detrimental rice fungal diseases and poses a severe threat to rice production and quality. Effectors in U. virens often act as a group of essential virulence factors that play crucial roles in the interaction between host and the pathogen. Thus, the functions of individual effectors in U. virens need to be further explored. Here, we found a small secreted hypersensitive response-inducing protein UVI_02019870 was highly conserved in fungi. Furthermore, we performed Y2H and BiFC assay to demonstrated UVI_02019870 interacted with OsCPL1, which was predicted as a chloroplast precursor to regulate chloroplast signaling pathways. Our data provide a theory for gaining an insight into the molecular mechanisms underlying the UVI_02019870 virulence function.     

Effect of Burkholderia (Pseudomonas) cepacia and soil type on the control of crown rot in wheat

A rifampicin-resistant isolate of Burkholderia (Pseudomonas) cepacia (A3R) reduced crown rot (Fusarium graminearum Group 1) symptoms significantly (P 0.05) in wheat in glasshouse and field experiments and increased grain yield significantly (P 0.05) in one of two field experiments. In glasshouse experiments, applying the bacteria as a soil drench (2.5 × 109 cfu/g soil) was more effective than coating the bacteria on wheat seed (3.4 × 107cfu/seed). In field experiments, the bacteria were applied as a soil drench at the rate of 1.8 x 10¹⁰ cfu/m row. In both the glasshouse and the field, disease severity in the bacteria-inoculated treatments was significantly less in a silt loam than in a sandy loam. The silt loam had a large proportion of fine clay and silt particles (51.7%), which may have favoured the biocontrol activity and survival of the introduced B. cepacia. In a glasshouse experiment, control by B. cepacia was significantly greater in the silt loam than in the sandy loam, which in turn was greater than in a loamy sand. The loamy sand appeared to favour crown rot development but not the activity or survival of the bacterial antagonist. The latter was reflected by the relative populations of the rifampicin-resistant bacteria re-isolated from the various soils during a 5-week period after application of the bacteria (silt loam > sandy loam > loamy sand). This study further confirms that soil type can influence the populations and the level of biocontrol activity of some bacterial antagonists.     

Cloning and sequencing of endochitinase genes from gliocladium virens and trichoderma species

Nucleotide sequences containing the genetic determinants of the endochitinase of Gliocladiurn virences as well as those of Trichoderma harzianum, T. hamatum, T. viride and T. pseudokoningii were determined. The ORFs of 1287 or 1293 nucleotides were interrupted by three introns. The deduced amino acid sequences of endochitinase of G. virens were highly homologous to those of Trichoderma species (82–97% identity), although the culture filtrates from G. virens showed a much stronger inhibitory effect on growth of Helicobasidium mompa than those from Trichoderma species. Phylogenic analysis showed that these sequences could be divided into three clusters regardless of genus or species.     

In vitro screening of biological and chemical agents together on the growth of Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. causing inflorescence die back in arecanut

Colletotrichum gloeosporioides is a destructive pathogen of many crop species causing diseases in many annual, biennial and perennial plants. A study was undertaken to find out the effect of biological and chemical agents together on the growth of C. gloeosporioides causing inflorescence die back in arecanut at the Department of Plant Pathology, CPCRI, Kasaragod. To reduce the release of chemical pesticides to the environment, integrated control strategies have been adopted extensively by combining both bioagents and chemical agents. So in the present study in vitro experiments were conducted with two compatible Trichoderma sp. viz., Trichoderma virens and Trichoderma viride and fungicides viz. Blitox 50 W and Mixol 72. The results indicated that all the treatments had a significant inhibitory effect on the growth of C. gloeosporioides and reduced its colony diameter. High percent inhibition was found when 0.05% of Mixol 72 was used with T. virens (87.61%). The least inhibition was shown by T. virens+0.05% Blitox 50 W (80.95%). It is concluded that the combination of bioagents with fungicides provided higher disease suppression than achieved with fungicides and bioagents when used alone.     

The essential effector SCRE1 in Ustilaginoidea virens suppresses rice immunity via a small peptide region

The biotrophic fungal pathogen Ustilaginoidea virens causes rice false smut, a newly emerging plant disease that has become epidemic worldwide in recent years. The U. virens genome encodes many putative effector proteins that, based on the study of other pathosystems, could play an essential role in fungal virulence. However, few studies have been reported on virulence functions of individual U. virens effectors. Here, we report our identification and characterization of the secreted cysteine-rich protein SCRE1, which is an essential virulence effector in U. virens. When SCRE1 was heterologously expressed in Magnaporthe oryzae, the protein was secreted and translocated into plant cells during infection. SCRE1 suppresses the immunity-associated hypersensitive response in the nonhost plant Nicotiana benthamiana. Induced expression of SCRE1 in rice also inhibits pattern-triggered immunity and enhances disease susceptibility to rice bacterial and fungal pathogens. The immunosuppressive activity is localized to a small peptide region that contains an important ‘cysteine-proline-alanine-arginine-serine’ motif. Furthermore, the scre1 knockout mutant generated using the CRISPR/Cas9 system is attenuated in U. virens virulence to rice, which is greatly complemented by the full-length SCRE1 gene. Collectively, this study indicates that the effector SCRE1 is able to inhibit host immunity and is required for full virulence of U. virens.     

Biological characteristics of Streptomyces albospinus CT205 and its biocontrol potential against cucumber Fusarium wilt

In a dual-culture assay, Streptomyces albospinus CT205 inhibited the growth of Fusarium oxysporum in vitro, the casual agent of widespread Fusarium wilt, probably due to the production of chitinase, β-glucanase, and a heat-resistant antagonistic substance. Pot and field experiments were performed to investigate the effects of S. albospinus CT205 alone or combined with organic fertiliser (BOF-CT205) on control of Fusarium wilt. Pot experiments showed that BOF-CT205 treatments obtained the lowest disease index (23.2), compared with organic fertiliser (OF, 55.3), strain CT205 (66.2), and control (72.5) treatments. In the field experiment, BOF-CT205 treatment yielded significantly lower disease incidences than the control, with reductions of ca. 55%. Cucumber yields in the same treatments were significantly higher than in the control, with increases of approximately 30% in the OF treatment. The cucumber yield was maximised in the BOF-CT205 in field experiments, reaching 8.3 × 104 kg ha^?1. The application of BOF-CT205 reduced the presence of the pathogen in the cucumber rhizosphere. In conclusion, S. albospinus CT205 was demonstrated to be a promising biocontrol agent for cucumber wilt, and the most effective treatment was the application of BOF-CT205.     

Induction of defence-related enzymes in onion by combined application of fungal and bacterial biocontrol agents against Fusarium oxysporum f. sp. cepae

Basal rot disease of onion is a major problem in different onion growing regions of Tamil Nadu, India. Fungal and bacterial cultures were isolated and tested their efficiency against Fusarium oxysporum f. sp. cepae under in vitro conditions. Effective bacterial and fungal antagonists were tested alone and in combinations for the control of F. oxysporum f. sp. cepae in glasshouse experiments. Defence-related enzymes such as peroxidase, polyphenol oxidase and phenylalanine ammonia-lyase were induced and accumulated in onion treated with fungal and bacterial antagonists. Defence-related enzymes were significantly higher in onion pretreated with consortial formulation of Pf12 + Pf27 + TH3 at 5 days after the challenge inoculation with F. oxysporum f. sp. cepae and gave resistance to onion against basal rot disease.     

Biocontrol of Sclerotinia lettuce drop by Coniothyrium minitans and Trichoderma hamatum

Fungal isolates, with known activity against Sclerotinia spp. in laboratory assays, were tested for their ability to control Sclerotinia minor in four field experiments (1998–2000). In the first experiment, eight fungal isolates (Trichoderma hamatum LU595, LU593, LU592, Trichoderma virens LU555 and LU556, Coniothyrium minitans LU112, Clonostachys rosea LU115 and Trichoderma rossicum LU596) were evaluated by incorporating spore suspensions into transplant potting mix and planting lettuce seedlings into a S. minor infested field site. At harvest, Trichoderma hamatum LU595, LU593, T. virens LU555 and C. minitans LU112 reduced disease by 30–50% compared with the untreated control under very high disease pressure (100%). In further field experiments C. minitans LU112 and T. hamatum LU593, applied as maizemeal–perlite soil amendments or incorporated into the potting mix, reduced S. minor disease over a range of disease pressures (29–91%). Disease control was equivalent or greater than that achieved with the standard carbendazim fungicide treatment. Both isolates were shown to effectively colonize the lettuce rhizosphere and surrounding soil and this colonization may have protected the roots from infection by S. minor. Multiple applications of C. minitans LU112 or T. hamatum LU593 formulations gave no added disease control compared with a single application at planting. Commercial formulations of both C. minitans LU112 and T. hamatum LU593 applied as transplant treatments, solid substrate soil amendments or as a spore drench gave consistent disease control and are currently being developed further.