CA1224055A - Colletotrichum coccodes spore production, spore preparation, formulation and agricultural process - Google Patents
Colletotrichum coccodes spore production, spore preparation, formulation and agricultural processInfo
- Publication number
- CA1224055A CA1224055A CA000452069A CA452069A CA1224055A CA 1224055 A CA1224055 A CA 1224055A CA 000452069 A CA000452069 A CA 000452069A CA 452069 A CA452069 A CA 452069A CA 1224055 A CA1224055 A CA 1224055A
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Abstract
PROCESS FOR CONTROLLING UNDESIRABLE VEGETATION
WITH COLLETOTRICHUM COCCODES SPORES
Abstract of the Disclosure There is disclosed a process for controlling undesirable vegetation, especially velvetleaf by apply-ing Colletotrichum coccodes spores onto the undesirable vegetation or onto the situs of the undesirable vege-tation. The composition including the fungus spores in an agricultural carrier as well as the growing of the fungus on solid or liquid media are also disclosed.
The spores germinate, infect the undesirable vegetation and result in a pathogenic disease which kills or reduces the growth of this undesirable plant.
WITH COLLETOTRICHUM COCCODES SPORES
Abstract of the Disclosure There is disclosed a process for controlling undesirable vegetation, especially velvetleaf by apply-ing Colletotrichum coccodes spores onto the undesirable vegetation or onto the situs of the undesirable vege-tation. The composition including the fungus spores in an agricultural carrier as well as the growing of the fungus on solid or liquid media are also disclosed.
The spores germinate, infect the undesirable vegetation and result in a pathogenic disease which kills or reduces the growth of this undesirable plant.
Description
This invention relates to a process and a composition for controlling undesirable vegetation.
More particularly, the present invention is directed to a process for controlling the growth of velvetleaf by applying Colletotrichum spores thereto.
This invention also relates to the growing of a fungus, Colletotrichum coccodes (Wallr.) Hughes, collecting spores, concentrating the spores and for-mulating the spores for application over geographic areas to control undesirable vegetation, namely velvet-leaf (Abutilon theophrasti Medic.). According to the invention, after application the spores germinate9 infect the velvetlea-f and result in a pathogenic disease which terminates or reduces the growth of the undesirable plant.
Velvetleaf, often called button-weed or elephant-ear, is an erect annual broadleaf plant species from 0.3 to 3m in height, but generally grows to just above thecrop in which it is growing. It is a prolific seed producer with 17~000 seeds per plant being produced on average. The seeds can persist in the soil for up to 50 years and are unaffected by pas-sage through animals or by ensilement.
Velvetleaf is widely distributed -throughout most of the United States and much of southeastern Canada.
It is considered as one of the most common and costly weeds in the North Central region of the United States.
- 1 - ~
~2~ 5S
This weed species occurs in waste places, vacant lots, gardens and cultivated fields. Velvetleaf is a serious weed problem in corn7 soybeans, cotton, wheat, oats, barley, sugarbeets, sunflower, tomato and other culti-vated crops. Infestations of this weed presently cause significant yield reductions in these crops and the problem is increasing due to the prolific seed production of velvetleaf and the difficulty in the control thereof.
In most cropping systems~ chemical herbicides have been developed to provide acceptable to excellent control of many grass and broadleaf weeds. However, widespread and continuous use of certain herbicides has favored the establishment of problem, difficult to control weeds such as velvetleaf. A cornbination of chemical herbicides and cultivation are presently used to control velvetleaf populations in various crops, but these methods are inadequate for the control of velvet-leaf as infestations of this weed are continuing to increase in size and in nurnber. Problems with cultiva-tion for the control of velvetleaf includeo 1. appropriate timing to avoid injury to the crop plants is difficult,
More particularly, the present invention is directed to a process for controlling the growth of velvetleaf by applying Colletotrichum spores thereto.
This invention also relates to the growing of a fungus, Colletotrichum coccodes (Wallr.) Hughes, collecting spores, concentrating the spores and for-mulating the spores for application over geographic areas to control undesirable vegetation, namely velvet-leaf (Abutilon theophrasti Medic.). According to the invention, after application the spores germinate9 infect the velvetlea-f and result in a pathogenic disease which terminates or reduces the growth of the undesirable plant.
Velvetleaf, often called button-weed or elephant-ear, is an erect annual broadleaf plant species from 0.3 to 3m in height, but generally grows to just above thecrop in which it is growing. It is a prolific seed producer with 17~000 seeds per plant being produced on average. The seeds can persist in the soil for up to 50 years and are unaffected by pas-sage through animals or by ensilement.
Velvetleaf is widely distributed -throughout most of the United States and much of southeastern Canada.
It is considered as one of the most common and costly weeds in the North Central region of the United States.
- 1 - ~
~2~ 5S
This weed species occurs in waste places, vacant lots, gardens and cultivated fields. Velvetleaf is a serious weed problem in corn7 soybeans, cotton, wheat, oats, barley, sugarbeets, sunflower, tomato and other culti-vated crops. Infestations of this weed presently cause significant yield reductions in these crops and the problem is increasing due to the prolific seed production of velvetleaf and the difficulty in the control thereof.
In most cropping systems~ chemical herbicides have been developed to provide acceptable to excellent control of many grass and broadleaf weeds. However, widespread and continuous use of certain herbicides has favored the establishment of problem, difficult to control weeds such as velvetleaf. A cornbination of chemical herbicides and cultivation are presently used to control velvetleaf populations in various crops, but these methods are inadequate for the control of velvet-leaf as infestations of this weed are continuing to increase in size and in nurnber. Problems with cultiva-tion for the control of velvetleaf includeo 1. appropriate timing to avoid injury to the crop plants is difficult,
2. lack of control of late germinating plants,
3. it cannot be used in new cropping systerns such as solid seeded soybeans (SSS).
The problems associated with the present chemical herbicides for velvetleaf control include:
1. lack of soil persistence resulting in no control of late germinating ve~vetleaf 20 most chernical herbicides cause some det-rimental effects to the growth of the crop, 3. chemical herbicides may adversely affect non-target organisms,
The problems associated with the present chemical herbicides for velvetleaf control include:
1. lack of soil persistence resulting in no control of late germinating ve~vetleaf 20 most chernical herbicides cause some det-rimental effects to the growth of the crop, 3. chemical herbicides may adversely affect non-target organisms,
4. toxic residues of chemical herbicides may persist in the environment or contaminate our food.
Dinqley, J.M. and Gilmour, J.W., New Zeal. J.
For. Sci. 2(2): 192 (1972) discloses the C. coccodes (Wallr.) Hughes fungus in order to distinguish it from the C. acutatum, to which the studies refer. Stevenson, W R., Evans, G.E~, and Barksdale, T.H., P1. Dis. Rept.
62(11): 937 (1978) discloses the losses in fresh market tomato crops produced by anthracnose disease caused by C. coccodes (Wallr.) Hughes among others, and the develop-ment of tomato cultivars with genetic resistance to the disease.
Although many patents deal with the fungal control of vegetation, none of them disclose the use of Colletotrichum coccodes (Wallr.) Hughes for this treat-ment. Such patents include the following:
U.S. Patents: 3,087,865 Drake et al 3,300,390 Tiner et al 3,308,038 Rhodes et al 3,150,062 Greenberg et al 3,357,895 Che-rry 3,361,555 Herschler 3,999,973 Templeton 4 9 061,488 Mann 4,162,912 Charudattan 4,390,360 walker
Dinqley, J.M. and Gilmour, J.W., New Zeal. J.
For. Sci. 2(2): 192 (1972) discloses the C. coccodes (Wallr.) Hughes fungus in order to distinguish it from the C. acutatum, to which the studies refer. Stevenson, W R., Evans, G.E~, and Barksdale, T.H., P1. Dis. Rept.
62(11): 937 (1978) discloses the losses in fresh market tomato crops produced by anthracnose disease caused by C. coccodes (Wallr.) Hughes among others, and the develop-ment of tomato cultivars with genetic resistance to the disease.
Although many patents deal with the fungal control of vegetation, none of them disclose the use of Colletotrichum coccodes (Wallr.) Hughes for this treat-ment. Such patents include the following:
U.S. Patents: 3,087,865 Drake et al 3,300,390 Tiner et al 3,308,038 Rhodes et al 3,150,062 Greenberg et al 3,357,895 Che-rry 3,361,555 Herschler 3,999,973 Templeton 4 9 061,488 Mann 4,162,912 Charudattan 4,390,360 walker
5~
In a broad aspect, -the present invention relates to a process for controlling undesirable vegetation by appling Colletotrichum cocco~es spores onto the undesirable vegetation or onto the situs of the undesirable vegetation.
The preferred composition which is used in the process according to the invention comprises Colletotrichum coccodes spores in an agricultural carrier preferably having a spore concentration of 1 x 107 to 1 x 109 spores per ml of carrier.
The process according to the invention is especially useful for the control of velvetleaf (Abutilon theophrasti) which involves the application of a spore suspension of a fungus, Colletotrichum coccodes which results in a debilitating disease of velvetleaf. The fungal spore application provides a unique narrow spectrum herbicide for the control of a single noxiousweed species, velvetleaf in soybeans, corn and other crops.
Ihe preferred agricultural carrier comprises water, with or without 1% by weight gelatin, the spores being uniforrnly dispersed in the carrier.
The spore containing composition is preferably applied at the cotyledon to 6-leaf stage of velvetleaf, and under warm and moist conditions. m e composition may be applied onto the leaf surface of the velvetleaf.
The organism, C. coccodes has been deposited and can be obtained from the permanent collections of ~22~55 the Biosystematic Research Institute Agriculture Canada, Ottawa, Canada where it has been deposited under an unrestricted deposit as C. coccodes D~OM 183088.
The fungus produces necrotic spots (lesions) on the leaves of velvetleaf which coalesce and cause the death of leaves and stems. When the fungus infects young velvetleaf plants, a devastating blight develops and most diseased plants are killed~
C.coccodes can be grown on solid or in liquid media for spore production. Media can be potato dextrose, malt extract, V-8, or beef extract. For pro-duction of larger quantities of spores liquid media is used. For example:
Formula 1 - Modified Beef Extract dextrose10.67g nutrient broth 10.69g peptone 0.44g distilled water (to make)1000 ml Formula II - Richard's Modified V-8 Medium sucrose 10.00g KNO3 10.00g g 4 2 2.50g 2 4 g FeC13 0.02g V-8 juice150 ml distilled water (to make)1000 ml Liter flasks containing 500 ml of the liquid media are inoculated with spores and incubated at 28C
with occasional agitation for 5 to 10 days. The contents of the flasks are then filtered to remove fungal mycelium.
The remaining liquid containing the Eungal spores is centrifuged in a refrigerant centrifuge. The supernatant is discarded and the spore concentrate is collected and stored at 4C prior to application.
The spore concentrate is mixed with an agri-culturally acceptable diluent or carrier for application to the target weed, such as velvetleaf. The preferred carrier is water containing 1% gelatin and the spore concentrate is uniformly dispersed in the carrier. The spore suspension (formulation) is sprayed onto the leaf surface of the undesirable vegetation at a preferred rate of 107 to 109 spores per m2 in 47 to 187 ml of water per m2.
The velvetleaf isolate of C.coccodes does not damage soybean, corn, and other crop plants and can be used in these crops to control velvetleaf. me preferred application is at the cotyledon to 6~1eaf stage of velvet-leaf with the associated environmental conditions being warm and moist. Repeat applications of the spore formul-ation may be required and control is improved if the growth of the associated crop is vigorous.
The invention will now be illus-trated by means of the following non-limitating examples.
Example 1. Growth Chamber Studies Cultures of C. coccodes are grown on malt agar in petri dishes and when the cul-tures are sporu-lating, small portions of the agar (about 1 cm3) are removed and used for inoculation of culture flasks.
250 ml flasks containing 75 ml of modified beef extract media are aseptically inoculated with an agar cube.
Flasks are incubated on a rotary shaker (250 rpm) at 20 to 25C for 5 to 10 days. The cultures are then filtered to remove mycelia, the remaining liquid con-taining the spores is centrifuged in an IEC B20A
refrigerated centrifuge (870 rotor) at 7000 rpm for 20 minu-tes. The spore concentrate is resuspended in dis-tilled water to reach a final spore concentration of 1 x 107 spores per ml. Velvetleaf plants growing within controlled environment chambers are inoculated by spraying the spore suspension onto the plant surface until runoff occurs.
Optimum conditions for spore germination, infection and disease development wexe determined.
Warm temperatures and free moisture are required for spore germination and infection cf host plant material.
These processes occur best when temperatures are 24 to 30C and when a saturated atmosphere is maintained for 8 to 24 hours. After infection occurs higher tempera-tures accelerate disease development with 30C night and 35C day temperatures being optimum. All growth stages tested (cotyledon to the 12-leaf stage) were equally susceptible to infection.
Example 2. Field Studies A. Spore preparation Cultures of C coccodes are grown on PDA
plates for 1 to 2 weeks. When cultures are sporulating, spores are aseptically suspended in 10 ml o-f sterile water by swirling. Spore suspension is then transferred to 1 L flasks containing 500 ml of Richard's Modified V-8 medium. Cultures are then incubated at 28C for
In a broad aspect, -the present invention relates to a process for controlling undesirable vegetation by appling Colletotrichum cocco~es spores onto the undesirable vegetation or onto the situs of the undesirable vegetation.
The preferred composition which is used in the process according to the invention comprises Colletotrichum coccodes spores in an agricultural carrier preferably having a spore concentration of 1 x 107 to 1 x 109 spores per ml of carrier.
The process according to the invention is especially useful for the control of velvetleaf (Abutilon theophrasti) which involves the application of a spore suspension of a fungus, Colletotrichum coccodes which results in a debilitating disease of velvetleaf. The fungal spore application provides a unique narrow spectrum herbicide for the control of a single noxiousweed species, velvetleaf in soybeans, corn and other crops.
Ihe preferred agricultural carrier comprises water, with or without 1% by weight gelatin, the spores being uniforrnly dispersed in the carrier.
The spore containing composition is preferably applied at the cotyledon to 6-leaf stage of velvetleaf, and under warm and moist conditions. m e composition may be applied onto the leaf surface of the velvetleaf.
The organism, C. coccodes has been deposited and can be obtained from the permanent collections of ~22~55 the Biosystematic Research Institute Agriculture Canada, Ottawa, Canada where it has been deposited under an unrestricted deposit as C. coccodes D~OM 183088.
The fungus produces necrotic spots (lesions) on the leaves of velvetleaf which coalesce and cause the death of leaves and stems. When the fungus infects young velvetleaf plants, a devastating blight develops and most diseased plants are killed~
C.coccodes can be grown on solid or in liquid media for spore production. Media can be potato dextrose, malt extract, V-8, or beef extract. For pro-duction of larger quantities of spores liquid media is used. For example:
Formula 1 - Modified Beef Extract dextrose10.67g nutrient broth 10.69g peptone 0.44g distilled water (to make)1000 ml Formula II - Richard's Modified V-8 Medium sucrose 10.00g KNO3 10.00g g 4 2 2.50g 2 4 g FeC13 0.02g V-8 juice150 ml distilled water (to make)1000 ml Liter flasks containing 500 ml of the liquid media are inoculated with spores and incubated at 28C
with occasional agitation for 5 to 10 days. The contents of the flasks are then filtered to remove fungal mycelium.
The remaining liquid containing the Eungal spores is centrifuged in a refrigerant centrifuge. The supernatant is discarded and the spore concentrate is collected and stored at 4C prior to application.
The spore concentrate is mixed with an agri-culturally acceptable diluent or carrier for application to the target weed, such as velvetleaf. The preferred carrier is water containing 1% gelatin and the spore concentrate is uniformly dispersed in the carrier. The spore suspension (formulation) is sprayed onto the leaf surface of the undesirable vegetation at a preferred rate of 107 to 109 spores per m2 in 47 to 187 ml of water per m2.
The velvetleaf isolate of C.coccodes does not damage soybean, corn, and other crop plants and can be used in these crops to control velvetleaf. me preferred application is at the cotyledon to 6~1eaf stage of velvet-leaf with the associated environmental conditions being warm and moist. Repeat applications of the spore formul-ation may be required and control is improved if the growth of the associated crop is vigorous.
The invention will now be illus-trated by means of the following non-limitating examples.
Example 1. Growth Chamber Studies Cultures of C. coccodes are grown on malt agar in petri dishes and when the cul-tures are sporu-lating, small portions of the agar (about 1 cm3) are removed and used for inoculation of culture flasks.
250 ml flasks containing 75 ml of modified beef extract media are aseptically inoculated with an agar cube.
Flasks are incubated on a rotary shaker (250 rpm) at 20 to 25C for 5 to 10 days. The cultures are then filtered to remove mycelia, the remaining liquid con-taining the spores is centrifuged in an IEC B20A
refrigerated centrifuge (870 rotor) at 7000 rpm for 20 minu-tes. The spore concentrate is resuspended in dis-tilled water to reach a final spore concentration of 1 x 107 spores per ml. Velvetleaf plants growing within controlled environment chambers are inoculated by spraying the spore suspension onto the plant surface until runoff occurs.
Optimum conditions for spore germination, infection and disease development wexe determined.
Warm temperatures and free moisture are required for spore germination and infection cf host plant material.
These processes occur best when temperatures are 24 to 30C and when a saturated atmosphere is maintained for 8 to 24 hours. After infection occurs higher tempera-tures accelerate disease development with 30C night and 35C day temperatures being optimum. All growth stages tested (cotyledon to the 12-leaf stage) were equally susceptible to infection.
Example 2. Field Studies A. Spore preparation Cultures of C coccodes are grown on PDA
plates for 1 to 2 weeks. When cultures are sporulating, spores are aseptically suspended in 10 ml o-f sterile water by swirling. Spore suspension is then transferred to 1 L flasks containing 500 ml of Richard's Modified V-8 medium. Cultures are then incubated at 28C for
6-9 days with occasional swirling. Spores are harvested by filtration through 2 layers of cheesecloth followed by centrifugation in 250 ml bottles in a Beckman JA-14 rotor at 4,000 RPM for 15 minutes at 15C. Pellets of spores are resuspended in distilled water and rinsed twice with distilled water followed by centrifugation each time. Spores are then stored at 4C. Yields exceed 6 x 10 spores per liter of growth media used.
B. Formulation and application Spores are resuspended in 1% gelatin concen-tration and are adjusted to apply 107 or 109 spores per m2 in 47, 94, or 187 ml per m2~
C. Fl d control for velvetleaf When one application of spores is applied under warm field conditions, velvetleaf biomass is reduced by 85% as compared to unsprayed control within 3 weeks after spraying. The fungus spreads from infected foliage to surviving foliage or new growth and suppresses velvet-leaf growth for the rest of the growing season.
B. Formulation and application Spores are resuspended in 1% gelatin concen-tration and are adjusted to apply 107 or 109 spores per m2 in 47, 94, or 187 ml per m2~
C. Fl d control for velvetleaf When one application of spores is applied under warm field conditions, velvetleaf biomass is reduced by 85% as compared to unsprayed control within 3 weeks after spraying. The fungus spreads from infected foliage to surviving foliage or new growth and suppresses velvet-leaf growth for the rest of the growing season.
Claims (15)
1. The process for controlling velvet leaf (Abutilon theophrasti),comprising applying an effective amount of Colletotrichum coccodes spores on to the said velvet leaf or onto the situs of said velvet leaf.
2. A process according to claim 1, which comprises applying onto said velvet leaf or onto the situs of said velvet leaf a composition comprising Colletotrichum coccodes spores in an agricultural carrier having a spore concentration of 1 x 107 to 1 x 109 spores per ml of carrier.
3. A process according to claim 2, wherein said agricultural carrier comprises water, said spores being uniformly dispersed in said carrier.
4. A process according to claim 2, wherein said agricultural carrier comprises water containing 1% by weight gelatin, said spores being uniformly dispersed in said carrier.
5. A process according to claim 2, wherein said spore containing composition is applied at the cotyledon to 6-leaf stage of velvet leaf.
6. A process according to claim 5, wherein the application is carried out under warm and moist conditions.
7. A process according to claim 4, which comprises spraying said spore containing composition onto the leaf surface of said velvet leaf.
8. A process according to claim 7, which comprises spraying said composition at a rate of 107 to 109 spores per m2 in 47 to 187 ml of water per m2.
9. A process which comprises growing the fungus Colletotrichum coccodes (Wallr.) Hughes, on solid or liquid media, thereby obtaining spores of said fungus.
10. Process according to claim 9, wherein said media are selected from the group consisting of potato dextrose, malt extract, V-8TM and beef extract.
11. Process according to claim 10, wherein the medium is a modified beef extract comprising dextrose, nutrient broth, peptone and distilled water.
12. Process according to claim 11, wherein the medium is a modified V-8TM medium comprising sucrose, KNO3, MgSO4, H2O, KH2PO4, FeCl3, V-8TM juice and distilled water.
13. An agricultural composition for controlling undesirable vegetation comprising Colletotrichum coccodes spores which are dispersed in an agricultural carrier, at a spore concentration of 1 x 107 to 1 x 109 spores per ml of carrier.
14. A composition according to claim 13, wherein said agricultural carrier comprises water, said spores being uniformly dispersed in said carrier.
15. A composition according to claim 13, wherein said agricultural carrier comprises water containing 1% by weight gelatin, said spores being uniformly dispersed in said carrier.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US57360984A | 1984-01-25 | 1984-01-25 | |
| US573,609 | 1990-08-27 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1224055A true CA1224055A (en) | 1987-07-14 |
Family
ID=24292684
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000452069A Expired CA1224055A (en) | 1984-01-25 | 1984-04-16 | Colletotrichum coccodes spore production, spore preparation, formulation and agricultural process |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1224055A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5208026A (en) * | 1990-02-06 | 1993-05-04 | The Royal Institution For The Advancement Of Learning | Composition for biocontrol of plantain |
| CN112941006A (en) * | 2021-02-07 | 2021-06-11 | 广东省科学院生物工程研究所 | Preparation method of rice blast bacterium spores |
-
1984
- 1984-04-16 CA CA000452069A patent/CA1224055A/en not_active Expired
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5208026A (en) * | 1990-02-06 | 1993-05-04 | The Royal Institution For The Advancement Of Learning | Composition for biocontrol of plantain |
| CN112941006A (en) * | 2021-02-07 | 2021-06-11 | 广东省科学院生物工程研究所 | Preparation method of rice blast bacterium spores |
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