caddis
05-05-2007, 06:51 PM
Effect of Aerated Compost Tea on the Development and Growth of Botrytis cinerea
Transform is researching the ability of aerated compost tea made with Transform Plant Product’s
Worm Castings to prevent Botrytis cinerea (Gray Mold) from establishing and colonizing plants. The findings are astonishing and extremely promising.
Botrytis is a very common fungal foliar disease of plants in greenhouses, nurseries and gardens in regions of high relative humidity. Spread by wind, rain or plant movement, it is difficult to control and almost impossible to eradicate. “Estimated losses for vineyards [from Botrytis cinerea] in France amount to 15-40% of the harvest, depending on the climatic conditions” ( www.cns.fr/externe/English/Projets/Projet_LN/organisme_LN.html)
Fig. 3: Botrytis cinerea inoculum
Figure 1 and 2 show the dramatic difference between the control and compost tea treatment
seven days after inoculation with the leaf shown in figure 3.
Figure 4 shows the change in average number of colonies per treatment per leaf five and six days after inoculation. Clearly, there is a dramatic difference between the tea treatments and control.
After carefully examining the changes in total number of colonies per treatment at day 5 and 6, several important observations have been made.
Because of the various concentrations of bacterial and fungal tea used, an arbitrary number of 10 units of bacteria and 10 units of fungi were allocated to the 1:0 Fungal tea. Because the 1:1 fungal tea had a 50% dilution, there are 5 units of fungi and 5 units of bacteria. For the 1:0 Bacterial tea, all the fungi were filtered out, hence the concentration of bacteria increases to 20
units, and for the 1:1 bacterial tea, there are 10 units of bacteria. Using Microsoft Excel’s Correlation analysis function, the correlation between bacterial and fungal concentration and colony formation was determined for day 5 and the change between day 5 and 6. A logarithmic trend line was used since it is assumed the bacteria would reproduce at a logarithmic rate.
Five days after inoculation, the higher bacterial dominated teas (Fig. 5) suppress colony formation better than fungal dominated tea (Fig. 6). However, between day 5 and 6, the fungal dominated teas suppressed the colony formation better as shown in figure 7 and 8.
Figures 5 through 8 suggest that bacteria are more important in preventing Botrytis cinerea initially,
while fungi are more important later on. Figure 5 has a strong correlation between increasing
concentrations of bacteria and decreasing number of established, fruiting colonies of Botrytis. One theory is that over time, the established Botrytis colonies continue to lyse new tissue and larger percentages of the leaf become necrotic. As a result, the bacteria, which are feeding on leaf exudates and/or living in micro-niches on the leaf surface, have less food resources and/or habitat available. Between day 5 and 6, so many bacteria could no longer find any food, that the bacterial population was severely reduced, and with no competition, the Botrytis could develop faster. The fungi however inhabit a different niche on the leaf surface and consequently may be favored by the decaying leaf tissue catalyzed by the Botrytis infection. As a result, as more leaf tissue become necrotic, the fungi grows faster, and competes more directly for resources with the Botrytis.
During the first 5 days, there was no major difference in total number of colonies between the
1:0 bacteria (which have no fungi) and the 1:0 fungi (which have both fungi and bacteria). This
suggests that a limited amount of bacteria are required for adequate control.
The graph above shows the exact differences between the various treatments after seven days.
Columns with the same letter are not statistically significantly different from each other. Hence,
the suppression of Botrytis by compost tea is statistically significant when α = 0.05. There is no
significant difference between bacterial and fungal teas, nor between a 50% water dilution.
Experimental Methodology
50 leaves where cut from 10 seed geranium plants. The leaves where all surface sterilized by submersing and gently agitating in a 1% detergent solution for 3 minutes, rinsed with water,
than transferred to a 10% bleach solution for 10 minutes with occasional agitation. The leaves
were all thoroughly rinsed with water prior to the compost tea application.
The fungal compost tea is Transform’s standard compost tea. For the bacterial treatment, the tea
was filtered through two layers of coffee filter to remove all fungal hyphae and most protozoa.
Each leaf was dipped in the tea for approximately 2 seconds and allowed to air dry.
Botrytis inoculation occurred by building a spore settling tower consisting of a 24” long 4” PVC pipe caped on top and a 1/4” hole bored diagonally through the pipe near the cap. The pipe was fixed
through a sheet of plywood. Three heavily infested leaves where taped spore side out (towards the middle of the pipe) inside the pipe. All geranium leaves were randomized at the base of a garbage can using a 2” numbered grid at the base of the garbage can. Placing the pvc pipe fixed to the plywood on top of the garbage can, a 1.5 minute blast of compressed air from a commercial air compressor was used to dislodge the Botrytis conidia from the infested leaves and distribute through the garbage can. Because of the cyclonic action inside the pvc pipe, the spores settled relatively consistently at the bottom of the garbage container. Spores where allowed to settle for approximately 8 hours. The individual treatments where placed in a Ziploc freezer bags, each leaf placed on top of a 2x2” square of paper towel. The squares of paper towel were placed on another piece of plastic that slid into the Ziploc bag, facilitating the removal of specimens for observation. The paper towel squares were wetted down with approximately the same amount of water, and the bags were sealed.
Five and six days after leaf inoculation, the number of flowering Botrytis colonies was counted. If two colonies were growing together but could distinctly be seen as two independent colonies, they were counted as such.
After 7 days, a 5 mm grid printed on a cellophane sheet was used to determine the degree of leaf
infestation by counting the number of times Botrytis fruiting structures where observed directly
below a cross between the horizontal and vertical lines of the grid. The total leaf area was measured using the same method. A one-way ANOVA was performed using Microsoft Excel.
Going forward, Transform intends to repeat the experiment with several other variables to determine the suppressive properties of compost tea. Also, outdoor experiments will likely be
conducted to determine the effect of compost tea under standard agronomic practices.
http://www.transformcompost.com/tf%20web%20other%20pdf/Botrytis%20Experiment2.pdf
Transform is researching the ability of aerated compost tea made with Transform Plant Product’s
Worm Castings to prevent Botrytis cinerea (Gray Mold) from establishing and colonizing plants. The findings are astonishing and extremely promising.
Botrytis is a very common fungal foliar disease of plants in greenhouses, nurseries and gardens in regions of high relative humidity. Spread by wind, rain or plant movement, it is difficult to control and almost impossible to eradicate. “Estimated losses for vineyards [from Botrytis cinerea] in France amount to 15-40% of the harvest, depending on the climatic conditions” ( www.cns.fr/externe/English/Projets/Projet_LN/organisme_LN.html)
Fig. 3: Botrytis cinerea inoculum
Figure 1 and 2 show the dramatic difference between the control and compost tea treatment
seven days after inoculation with the leaf shown in figure 3.
Figure 4 shows the change in average number of colonies per treatment per leaf five and six days after inoculation. Clearly, there is a dramatic difference between the tea treatments and control.
After carefully examining the changes in total number of colonies per treatment at day 5 and 6, several important observations have been made.
Because of the various concentrations of bacterial and fungal tea used, an arbitrary number of 10 units of bacteria and 10 units of fungi were allocated to the 1:0 Fungal tea. Because the 1:1 fungal tea had a 50% dilution, there are 5 units of fungi and 5 units of bacteria. For the 1:0 Bacterial tea, all the fungi were filtered out, hence the concentration of bacteria increases to 20
units, and for the 1:1 bacterial tea, there are 10 units of bacteria. Using Microsoft Excel’s Correlation analysis function, the correlation between bacterial and fungal concentration and colony formation was determined for day 5 and the change between day 5 and 6. A logarithmic trend line was used since it is assumed the bacteria would reproduce at a logarithmic rate.
Five days after inoculation, the higher bacterial dominated teas (Fig. 5) suppress colony formation better than fungal dominated tea (Fig. 6). However, between day 5 and 6, the fungal dominated teas suppressed the colony formation better as shown in figure 7 and 8.
Figures 5 through 8 suggest that bacteria are more important in preventing Botrytis cinerea initially,
while fungi are more important later on. Figure 5 has a strong correlation between increasing
concentrations of bacteria and decreasing number of established, fruiting colonies of Botrytis. One theory is that over time, the established Botrytis colonies continue to lyse new tissue and larger percentages of the leaf become necrotic. As a result, the bacteria, which are feeding on leaf exudates and/or living in micro-niches on the leaf surface, have less food resources and/or habitat available. Between day 5 and 6, so many bacteria could no longer find any food, that the bacterial population was severely reduced, and with no competition, the Botrytis could develop faster. The fungi however inhabit a different niche on the leaf surface and consequently may be favored by the decaying leaf tissue catalyzed by the Botrytis infection. As a result, as more leaf tissue become necrotic, the fungi grows faster, and competes more directly for resources with the Botrytis.
During the first 5 days, there was no major difference in total number of colonies between the
1:0 bacteria (which have no fungi) and the 1:0 fungi (which have both fungi and bacteria). This
suggests that a limited amount of bacteria are required for adequate control.
The graph above shows the exact differences between the various treatments after seven days.
Columns with the same letter are not statistically significantly different from each other. Hence,
the suppression of Botrytis by compost tea is statistically significant when α = 0.05. There is no
significant difference between bacterial and fungal teas, nor between a 50% water dilution.
Experimental Methodology
50 leaves where cut from 10 seed geranium plants. The leaves where all surface sterilized by submersing and gently agitating in a 1% detergent solution for 3 minutes, rinsed with water,
than transferred to a 10% bleach solution for 10 minutes with occasional agitation. The leaves
were all thoroughly rinsed with water prior to the compost tea application.
The fungal compost tea is Transform’s standard compost tea. For the bacterial treatment, the tea
was filtered through two layers of coffee filter to remove all fungal hyphae and most protozoa.
Each leaf was dipped in the tea for approximately 2 seconds and allowed to air dry.
Botrytis inoculation occurred by building a spore settling tower consisting of a 24” long 4” PVC pipe caped on top and a 1/4” hole bored diagonally through the pipe near the cap. The pipe was fixed
through a sheet of plywood. Three heavily infested leaves where taped spore side out (towards the middle of the pipe) inside the pipe. All geranium leaves were randomized at the base of a garbage can using a 2” numbered grid at the base of the garbage can. Placing the pvc pipe fixed to the plywood on top of the garbage can, a 1.5 minute blast of compressed air from a commercial air compressor was used to dislodge the Botrytis conidia from the infested leaves and distribute through the garbage can. Because of the cyclonic action inside the pvc pipe, the spores settled relatively consistently at the bottom of the garbage container. Spores where allowed to settle for approximately 8 hours. The individual treatments where placed in a Ziploc freezer bags, each leaf placed on top of a 2x2” square of paper towel. The squares of paper towel were placed on another piece of plastic that slid into the Ziploc bag, facilitating the removal of specimens for observation. The paper towel squares were wetted down with approximately the same amount of water, and the bags were sealed.
Five and six days after leaf inoculation, the number of flowering Botrytis colonies was counted. If two colonies were growing together but could distinctly be seen as two independent colonies, they were counted as such.
After 7 days, a 5 mm grid printed on a cellophane sheet was used to determine the degree of leaf
infestation by counting the number of times Botrytis fruiting structures where observed directly
below a cross between the horizontal and vertical lines of the grid. The total leaf area was measured using the same method. A one-way ANOVA was performed using Microsoft Excel.
Going forward, Transform intends to repeat the experiment with several other variables to determine the suppressive properties of compost tea. Also, outdoor experiments will likely be
conducted to determine the effect of compost tea under standard agronomic practices.
http://www.transformcompost.com/tf%20web%20other%20pdf/Botrytis%20Experiment2.pdf