Trichoderma species are currently used as biocontrol agents for crop diseases caused by a number of fungal plant pathogens. However, their biocontrol performance in the field can be unreliable and it is likely that mo...Trichoderma species are currently used as biocontrol agents for crop diseases caused by a number of fungal plant pathogens. However, their biocontrol performance in the field can be unreliable and it is likely that more consistent performance could be achieved through knowledge and manipulation of the genes involved. For example, induction of the genes could be optimised for variable environmental and physiological conditions, superior strains could be selected more effectively and novel strains could be created. One method by which Trichoderma species accomplish biocontrol is mycoparasitism. Several genes involved in the mycoparasitic interaction have previously been characterised, however these consist predominantly of those that encode enzymes that degrade fungal cell walls. In the current study subtractive hybridisation was used to target genes expressed when Trichoderma hamatum and the plant pathogen Sclerotinia sclerotiorum were cultured together, subtracting genes expressed when each are grown individually. This experimental design has the potential to yield T. hamatum genes involved in mycoparasitism of S. sclerotiorum, and S. sclerotiorum genes up-regulated in host defence. The cDNA fragments yielded by the subtraction were characterised with respect to expression, sequence and species of origin. A number of novel T. hamatum genes which were up-regulated during mycoparasitism were identified.展开更多
A 10 year research programme at Lincoln University, investigating the use of Trichoderma species for biological control of soil-borne diseases of vegetable crops, has resulted in the development of two commercial prod...A 10 year research programme at Lincoln University, investigating the use of Trichoderma species for biological control of soil-borne diseases of vegetable crops, has resulted in the development of two commercial products. Trichodry TM. 6S and Trichoflow TM. 6S based upon Trichoderma hamatum isolate 6SR4, are used to control Sclerotinia lettuce drop disease. The Trichodry 6S product is formulated as a dry flake, which is incorporated into nursery seedling mix and the Trichoflow 6S is a wettable powder which is used as a top-up drench before planting. The treatment stimulates seedling establishment and vigour and protects the developing seedling from Sclerotinia minor infection after transplanting in the field.The second commercial product is Trichopel TM. Ali 52, based upon Trichoderma atroviride isolate C52, which is used to control Sclerotium cepivorum, the causal agent of Allium white rot disease. The product is formulated as a granule and applied into the furrow at planting time. The fungus proliferates in the rhizosphere region and protects the growing seedling from pathogen attack by a combination of nutrient competition, antibiosis and mycoparasitism. The use of Trichopel Ali 52 under low to medium disease pressure in Pukekohe, the main vegetable growing region of New Zealand, gave a three fold cost benefit through yield increases in the 2003-2004 season. Current field development work involves the use of a wettable powder formulation of T. atroviride distributed via a T-tape irrigation system to target mid-season applications of the product to the onion roots. Both products perform well under low to moderate disease pressure but, when there is high disease pressure, an integrated programme is required to give satisfactory control. Current research is focused on gaining a greater understanding of the biotic and abiotic factors, which influence biocontrol activity under field conditions as a means to enhance integrated control approaches. For example, T. atroviride C52, when applied to the planting furrow, is compatible with procymidone, benomyl and captan but not thiram when applied as onion seed treatments. It is also compatible with the majority of other fungicides applied to the onion crop to protect against foliar diseases such as Botrytis and downy mildew. The product can be integrated with the use of the germination stimulant diallyl disulphide (DADS) as long as the Trichoderma product is applied at least two weeks after DADS application. The product is not compatible with the use of nitrogen fertilisers due to the sensitivity of the Trichoderma mycelium to high N, thus, care must be taken to separate fertilizer application from that of the biocontrol product. Expansion of the range of crop diseases targeted by the biocontrol products is currently under investigation with promising results obtained against a number of Botrytis diseases.展开更多
With increased imports of foreign microbes either as commercial biocontrol produ cts or for the purposes of research, there is potentially an increased threat to indigenous beneficial microflora. In the present study,...With increased imports of foreign microbes either as commercial biocontrol produ cts or for the purposes of research, there is potentially an increased threat to indigenous beneficial microflora. In the present study, indigenous species of t he fungal genus Hypocrea/Trichoderma are being used as a model system to d etermine the impact of foreign microbes on the native microflora of New Zealand. In order to protect such microflora, one has to first be aware of what is curre ntly present and what sites, if any, are most vulnerable. A preliminary survey f or the presence and diversity of species of Hypocrea/Trichoderma is curren tly underway in New Zealand and samples are being assessed from forest soils, ag ricultural soils, orchards, garden soils, sclerotia of various plant pathogens a nd pasture land. To date 238 isolates have been identified using both morphologi cal characters and DNA sequence data from the ITS regions of the ribosomal gene cluster (ITS1 & ITS2) and, in some instances, sequence of the elongation facto r gene (EF1-α) . Isolates were found to represent 16 known species plus three s pec ies as yet undescribed. In forest soils T. harzianum /T. inhamatum (31%) and T. viride (29%) followed by T. fertile (13%), were clearly th e most abundant species and the remaining five species found in forests (T. a troviride, T. koningii, T. aureoviride, H. cf. flavovirens anamorph and one u nknown) each accounting for <8% of the total. Dominance by the species T. h arzianum/inhamatum is consistent with studies done in South-East Asia, a mid -E uropean primeval floodplain-forest and Moscow. In contrast, when isolations wer e conducted with a bias for biocontrol capabilities it was found that the species T. atroviride (29%), T. koningii (17 %), T. harzianum (1 5%) and T. viride (12%) dominated respectively. This survey is currently on go ing in New Zealand. Future studies will monitor indigenous species and strains f ollowing inoculation of specific microbes to assess the impact of the introduced microbe on the natural ecosystem.展开更多
Achieving a balance between vegetative growth and spore production is essential for successful biocontrol by fungi. Low sporulation rates in the field can result in poor establishment and survival, whereas failure of ...Achieving a balance between vegetative growth and spore production is essential for successful biocontrol by fungi. Low sporulation rates in the field can result in poor establishment and survival, whereas failure of conidia to recognise hosts can lead to persistence without efficacy. Commercial biocontrol products involve bulk preparations of conidia, however considerable variability in conidiation rates exists between biocontrol agents, which can restrict choice of strain for production. The majority of studies on Trichoderma conidiation have focused on the species T. viride and T. atroviride. These species form conidia in response to blue and near-UV light and/or nutrient deprivation and conidiation proceeds in a highly co-ordinated fashion, however relatively little is known on the genetic basis of Trichoderma conidiation. In addition, whilst photoconidiation appears to be a general response detailed studies in other Trichoderma species are absent. In this study, conidiation in the lesser known biocontrol species T. hamatum is being investigated using a combined morphological and molecular approach. In contrast to T. atroviride, conidiation in response to blue-light was weaker and variable and suggested that additional triggers may be required for the T. hamatum photoresponse. A series of comparative photoconidiation assays are currently being undertaken investigating the effect of inoculum type and abiotic factors on timing and intensity of the response. Results will be discussed in relation to the current knowledge on conidial morphogenesis in Trichoderma. In addition to these morphological assays, a selection of genes implicated in sporulation and the blue-light responses are currently being isolated and characterised from T. hamatum. Two genes, phr1 and cmp1, which were isolated previously from T. atroviride will be used as early and late markers of gene expression during the photoresponse in T. hamatum in order to define time points for harvesting comparable stage-specific RNA from T. hamatum and T. atroviride. Using degenerate PCR putative sporulation gene orthologues have also been identified in T. hamatum. Work is currently underway to isolate genomic clones of these genes from T. hamatum and T. atroviride. Sequence and expression analysis of orthologues, including expression in response to abiotic factors will be presented and discussed in relation to the current knowledge of the molecular basis of conidiation in Trichoderma and other filamentous fungi.展开更多
文摘Trichoderma species are currently used as biocontrol agents for crop diseases caused by a number of fungal plant pathogens. However, their biocontrol performance in the field can be unreliable and it is likely that more consistent performance could be achieved through knowledge and manipulation of the genes involved. For example, induction of the genes could be optimised for variable environmental and physiological conditions, superior strains could be selected more effectively and novel strains could be created. One method by which Trichoderma species accomplish biocontrol is mycoparasitism. Several genes involved in the mycoparasitic interaction have previously been characterised, however these consist predominantly of those that encode enzymes that degrade fungal cell walls. In the current study subtractive hybridisation was used to target genes expressed when Trichoderma hamatum and the plant pathogen Sclerotinia sclerotiorum were cultured together, subtracting genes expressed when each are grown individually. This experimental design has the potential to yield T. hamatum genes involved in mycoparasitism of S. sclerotiorum, and S. sclerotiorum genes up-regulated in host defence. The cDNA fragments yielded by the subtraction were characterised with respect to expression, sequence and species of origin. A number of novel T. hamatum genes which were up-regulated during mycoparasitism were identified.
文摘A 10 year research programme at Lincoln University, investigating the use of Trichoderma species for biological control of soil-borne diseases of vegetable crops, has resulted in the development of two commercial products. Trichodry TM. 6S and Trichoflow TM. 6S based upon Trichoderma hamatum isolate 6SR4, are used to control Sclerotinia lettuce drop disease. The Trichodry 6S product is formulated as a dry flake, which is incorporated into nursery seedling mix and the Trichoflow 6S is a wettable powder which is used as a top-up drench before planting. The treatment stimulates seedling establishment and vigour and protects the developing seedling from Sclerotinia minor infection after transplanting in the field.The second commercial product is Trichopel TM. Ali 52, based upon Trichoderma atroviride isolate C52, which is used to control Sclerotium cepivorum, the causal agent of Allium white rot disease. The product is formulated as a granule and applied into the furrow at planting time. The fungus proliferates in the rhizosphere region and protects the growing seedling from pathogen attack by a combination of nutrient competition, antibiosis and mycoparasitism. The use of Trichopel Ali 52 under low to medium disease pressure in Pukekohe, the main vegetable growing region of New Zealand, gave a three fold cost benefit through yield increases in the 2003-2004 season. Current field development work involves the use of a wettable powder formulation of T. atroviride distributed via a T-tape irrigation system to target mid-season applications of the product to the onion roots. Both products perform well under low to moderate disease pressure but, when there is high disease pressure, an integrated programme is required to give satisfactory control. Current research is focused on gaining a greater understanding of the biotic and abiotic factors, which influence biocontrol activity under field conditions as a means to enhance integrated control approaches. For example, T. atroviride C52, when applied to the planting furrow, is compatible with procymidone, benomyl and captan but not thiram when applied as onion seed treatments. It is also compatible with the majority of other fungicides applied to the onion crop to protect against foliar diseases such as Botrytis and downy mildew. The product can be integrated with the use of the germination stimulant diallyl disulphide (DADS) as long as the Trichoderma product is applied at least two weeks after DADS application. The product is not compatible with the use of nitrogen fertilisers due to the sensitivity of the Trichoderma mycelium to high N, thus, care must be taken to separate fertilizer application from that of the biocontrol product. Expansion of the range of crop diseases targeted by the biocontrol products is currently under investigation with promising results obtained against a number of Botrytis diseases.
文摘With increased imports of foreign microbes either as commercial biocontrol produ cts or for the purposes of research, there is potentially an increased threat to indigenous beneficial microflora. In the present study, indigenous species of t he fungal genus Hypocrea/Trichoderma are being used as a model system to d etermine the impact of foreign microbes on the native microflora of New Zealand. In order to protect such microflora, one has to first be aware of what is curre ntly present and what sites, if any, are most vulnerable. A preliminary survey f or the presence and diversity of species of Hypocrea/Trichoderma is curren tly underway in New Zealand and samples are being assessed from forest soils, ag ricultural soils, orchards, garden soils, sclerotia of various plant pathogens a nd pasture land. To date 238 isolates have been identified using both morphologi cal characters and DNA sequence data from the ITS regions of the ribosomal gene cluster (ITS1 & ITS2) and, in some instances, sequence of the elongation facto r gene (EF1-α) . Isolates were found to represent 16 known species plus three s pec ies as yet undescribed. In forest soils T. harzianum /T. inhamatum (31%) and T. viride (29%) followed by T. fertile (13%), were clearly th e most abundant species and the remaining five species found in forests (T. a troviride, T. koningii, T. aureoviride, H. cf. flavovirens anamorph and one u nknown) each accounting for <8% of the total. Dominance by the species T. h arzianum/inhamatum is consistent with studies done in South-East Asia, a mid -E uropean primeval floodplain-forest and Moscow. In contrast, when isolations wer e conducted with a bias for biocontrol capabilities it was found that the species T. atroviride (29%), T. koningii (17 %), T. harzianum (1 5%) and T. viride (12%) dominated respectively. This survey is currently on go ing in New Zealand. Future studies will monitor indigenous species and strains f ollowing inoculation of specific microbes to assess the impact of the introduced microbe on the natural ecosystem.
文摘Achieving a balance between vegetative growth and spore production is essential for successful biocontrol by fungi. Low sporulation rates in the field can result in poor establishment and survival, whereas failure of conidia to recognise hosts can lead to persistence without efficacy. Commercial biocontrol products involve bulk preparations of conidia, however considerable variability in conidiation rates exists between biocontrol agents, which can restrict choice of strain for production. The majority of studies on Trichoderma conidiation have focused on the species T. viride and T. atroviride. These species form conidia in response to blue and near-UV light and/or nutrient deprivation and conidiation proceeds in a highly co-ordinated fashion, however relatively little is known on the genetic basis of Trichoderma conidiation. In addition, whilst photoconidiation appears to be a general response detailed studies in other Trichoderma species are absent. In this study, conidiation in the lesser known biocontrol species T. hamatum is being investigated using a combined morphological and molecular approach. In contrast to T. atroviride, conidiation in response to blue-light was weaker and variable and suggested that additional triggers may be required for the T. hamatum photoresponse. A series of comparative photoconidiation assays are currently being undertaken investigating the effect of inoculum type and abiotic factors on timing and intensity of the response. Results will be discussed in relation to the current knowledge on conidial morphogenesis in Trichoderma. In addition to these morphological assays, a selection of genes implicated in sporulation and the blue-light responses are currently being isolated and characterised from T. hamatum. Two genes, phr1 and cmp1, which were isolated previously from T. atroviride will be used as early and late markers of gene expression during the photoresponse in T. hamatum in order to define time points for harvesting comparable stage-specific RNA from T. hamatum and T. atroviride. Using degenerate PCR putative sporulation gene orthologues have also been identified in T. hamatum. Work is currently underway to isolate genomic clones of these genes from T. hamatum and T. atroviride. Sequence and expression analysis of orthologues, including expression in response to abiotic factors will be presented and discussed in relation to the current knowledge of the molecular basis of conidiation in Trichoderma and other filamentous fungi.
