To ensure rhizosphere competence, the distal root isolation procedure, as described in Example 1, was used to identify subisolates. As noted elsewhere, these have some instability and are sectored.
Several different sectors were isolated and tested. The number system for the strains is important. The first letter or letters indicates the specific protoplast fusion that was conducted and the number immediately following described the order in which the specific progeny was picked.
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Each different set of letters indicates a different set of parents. While the original species were known, these indicators now are largely meaningless. At the time of U. Patent No. If the letter is a capital, this means that the selection was made at the time the isolates were first obtained, and the small letter designations are those that were picked in the most recent set of selections. For example, RRl 7Bc indicates that the strain arose from fusion RR, was the 17 th isolate picked, and that in the original selection, sector B was isolated.
For these experiments, seeds were treated with a Trichoderma preparation containing about 1 xlO 10 colony forming units cfu per g. Treated or untreated seeds were planted in a standard peat-vermiculite planting mix in 4 inch plastic pots. There were five seeds planted per pot and each pot was considered a replicate. The results are shown in Figure 1. Similarly, strains were also tested for their abilities to increase resistance to disease.
The model system was powdery mildew on cucumber. Figure 2 shows the reduction of powdery mildew on cucumber foliage.
Based on this testing, Fl IBab was added to list of elite strains. Suboptimal light conditions may even be sufficient to give greater differential growth in the presence of the endophytic strains that are the subject of the present invention than under optimal light conditions. Thus, even with supplemental lighting, variable results may be seen at different times of the year or other environmental variables. As the wheat plant grows, it forms tillers that each provides an additional head of grain. Thus, for example, if the wheat plant sequesters more carbon, this ought to be expressed as additional tillering.
These experiments, which measured the number of tillers Figure 3 , were conducted in a greenhouse using seeds of spring wheat variety Glenn which were treated following the protocol described in Example 1. Assuming that tiller lengths are similar across treatments, the total cumulative tiller length ought to be greater in strains that increase tiller number.
Gary E. Harman (Author of Trichoderma and Gliocladium Vol 2)
This data is shown in Figure 4. Data is shown with the standard deviation. As can be seen, all of the strains tested increased total tiller length relative to the control. These data indicate that a the new strains will increase yields of wheat, b that rooting will be enhanced along with shoot growth, thus permitting lower nitrogen fertilizer usage rates without yield reduction and c that carbon dioxide must be sequestered at a greater level to account for the increase in stem length and weight.
Experiments were conducted as nitrogen rate ranging trials in large pots 4 L volume to avoid root binding in smaller pots. This indicates that the Trichoderma strains were increasing N uptake, and that high levels of N were toxic. Since the toxicity occurred at lower concentrations in the presence of the Trichoderma strains, this data suggests that the strains were indeed increasing uptake and that toxic levels were reached at lower N concentrations in the presence than in the absence of Trichoderma.
This has importance in other examples that follow. However, by 49 days, at the time of emergence of heads, there was a difference in plants that grew from untreated or the different Trichoderma strains in soils to which N at 40 Figure 5A or b of N Figure 5B were added.
The plant heights were first measured, where the differences were significant at the two lower N rates. Plants were then destructively sampled from one replicate of the trials, and total plant weight, N levels, as well as levels of other nutrients was measured. Figure 6 provides the regression between N content of the wheat and plant weight. Since plant weights are tightly correlated with N levels it is likely that one principal effect of the enhancement in plant growth is due to enhanced N uptake. However, even when N is sufficient in soil, there may still be an increase in plant growth due to the enhanced N uptake level since the greater efficiency may increase N levels to higher levels than otherwise.
At 40 Ib N, there was about 2X as much N in plants in the presence of WW10TC4 than occurred with no Trichoderma seed treatment; 2 that much of the enhanced growth of wheat by Trichoderma, and probably other crops, is due to this enhanced nitrogen level in crops, which results from more efficient N fertilizer uptake; 3 seed treatment with the best strain, T. The trial was conducted with lowland rice. The plots were conducted in a randomized complete block design with four replicates. Fertilizer was added at the normal rate of nitrogen or at the half rate of nitrogen; a no fertilizer check was also included with the beneficial organisms.
At the full rate of N, there was no difference between the yields when Trichoderma was applied or not applied, and sometimes, the yields where Trichoderma was applied were lower. Yields with the strains at the half rate were numerically higher than the yields with the same treatment at the full rate of N.
During the course of the trial, after plants emerged, the dike broke and the plants were left without water. The plants without Fl IBab all died from drought and had no harvest, but a normal harvest was obtained in the presence of the beneficial fungus. October, The seeds were treated with the strains described in the present invention or not treated, in a randomized replicated field plot.
In November, stands in three feet of row were counted in each replicate of each treatment and visual vigor ratings were made on scale, with 10 being highly vigorous. The results are shown in Table 2. Numbers in the brackets are standard deviations around the mean. This is the first requirement for increased plant stands. Ref document number : Country of ref document : EP. Kind code of ref document : A1. Country of ref document : CA. Ref country code : DE. Country of ref document : US. A method for enhancing growth of plants comprising contacting a Trichoderma strain with the plant or a plant seed under conditions effective for the Trichoderma strain to colonize the roots of the plant or a plant grown from the plant seed, thereby creating a plant- Trichoderma system.
The plant or plant seed is grown under conditions effective to sustain the plant-Tnchoderma system in a planting medium and to enhance plant growth. Methods of enhancing resistance of plants to abiotic stress, increasing nitrogen use efficacy in plants, reducing nitrous oxide emissions in air, reducing leaching of nitrates into soil and water, and enhancing sequestration of carbon from air are disclosed. Provisional Patent Application Serial No. Wiley and Sons: New York , which is hereby incorporated by reference in its entirety or other deleterious soil microflora Bakker et al, "Microbial Cyanide Production in the Rhizosphere in Relation to Potato Yield Reduction and Ps eudomonas spp-Mediated Plant Growth-Stimulation," Soil Biol Bio- chem , which is hereby incorporated by reference in its entirety.
Example 2 - Selection of Improved Strains.
Example 3 - Increase in Growth of Wheat. The method according to claim 1 further comprising: adding a supplemental source of nutrients to the plant- Trichoderma system. The method according to claim 1, wherein the Trichoderma strain is provided as a granule, dust, powder, slurry, film, liquid suspension, or combinations thereof.
The method according to claim 1, wherein said contacting is carried out by broadcast application, liquid or dry in-furrow application, spray application, irrigation, injection, dusting, pelleting, or coating of the plant or the plant seed or the planting medium with Trichoderma strain.http://iqahyjokaw.cf
Trichoderma And Gliocladium, Volume 2
The method according to claim 1, wherein said contacting is carried out before the plant or the plant seed is introduced to the planting medium. The method according to claim 1, wherein said contacting is carried out after the plant or the plant seed is introduced to the planting medium. The method according to claim 1 , wherein the plant or plant seed is a crop plant selected from the group consisting of alfalfa, rice, wheat, barley, oats, rye, cotton, sorghum, sunflower, peanut, potato, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, soybean, tobacco, tomato, maize, clover, and sugarcane.
The method according to claim 1 , wherein the plant or plant seed is an ornamental plant selected from the group consisting of Arabidopsis thaliana, Saintpaulia, petunia, pelargonium, poinsettia, chrysanthemum, carnation, zinnia, roses, snapdragon, geranium, zinnia, lily, daylily, Echinacea, dahlia, hosta, tulip, daffodil, peony, phlox, herbs, ornamental shrubs, ornamental grasses, switchgrass, and turfgrass.
The method according to claim 1, wherein the plant growth enhancement is in the form of greater root mass, greater depth of rooting, greater shoot mass, greater length of shoots, increased leaf greenness, increased yields, and improved stand and vigor. A method for increasing nitrogen use efficacy in plants comprising: contacting a Trichoderma strain with the plant or a plant seed under conditions effective for the Trichoderma strain to colonize the roots of the plant or a plant grown from the plant seed, thereby creating a piant-Trichoderma system and growing the plant or plant seed under conditions effective to sustain the plant- Trichoderma system in a planting medium and to increase nitrogen use efficacy in plants, wherein the Trichoderma strain is selected from the group consisting of Trichoderma atroviride strain WW10TC4 ATCC accession number PTA , Trichoderma harzianum strain RRl 7Bc ATCC accession number PTA , Trichoderma harzianum strain Fl 1 Bab ATCC accession number PTA , and combinations thereof.
The isolated Trichoderma harzianum strain RRl 7Bc according to claim 20 which is biologically pure. The isolated Trichoderma harzianum strain Fl IBab according to claim 22 which is biologically pure. USP true EP EPB1 en US AOf-International.