Method For Promoting The Growth Of Corn Under Low Temperature Stress

Abstract

Present invention teaches the method of using a keratin hydrolysis peptide (KHP) solution to improve the growth of corn under low temperature conditions. By selectively choosing specific weights of feathers and water, and treating the mixture to a high-temperature high-pressure hydrolysis process, the resulting solution is confirmed to contain at least 253 peptides and then infused to the soil containing the corn seeds, the solution can also be sprayed to the leaf surface of corn plants at a specific growth stage. Optionally, the KHP solution can be diluted by water, as disclosed in the specification, for applying to the soil and for spraying to leaf surface of corn seedlings/plants.

Claims

1. A method of using a keratin hydrolysis peptide (KHP) solution to promote the growth of corn under low temperature stress, comprising the steps of: a. Preparing the KHP solution by putting 70 kg of feathers whose water content is 46%, without mixing any water, in a sealed container; b. hydrolyzing the mixture in the container with a temperature and pressure setting of 180 C. and 13 kg/cm.sup.2 for a duration of 40 minutes; c. using a mass spectrometer to confirm the combination of peptides in the solution to contain at least 253 peptides as listed in the specification where their molecular masses are between 500 and 4,000 Daltons, and the concentration is in the range of 2.010.sup.54.510.sup.5 ppm; and d. Applying the KHP solution by first diluting with water by volume at the ratio of 1:50-100 before application.

2. The method of using a keratin hydrolysis peptide solution of claim 1 wherein the application of the solution is by infusing into the soil immediately after planting the corn seeds.

3. The method of using a keratin hydrolysis peptide solution of claim 1 wherein the application of the solution is by infusing into the soil at corn's seedling stage.

4. The method of using a keratin hydrolysis peptide solution of claim 1 wherein the application of the solution is by infusing into the soil at corn's V3-V4 growth stage.

5. The method of using a keratin hydrolysis peptide solution of claim 1 wherein the application of the solution is by leaf spraying to the plants at corn's V3-V4 growth stage.

6. A method of using a keratin hydrolysis peptide (KHP) solution to promote the growth of corn under low temperature stress, comprising the steps of: a. Preparing the KHP solution by mixing 50 kg of feathers whose content is 50% water and 40 kg of water in a sealed container; b. hydrolyzing the mixture in the container with a temperature and pressure setting of 185 C. and 12 kg/cm.sup.2 for a duration of 80 minutes; c. using a mass spectrometer to confirm the combination of peptides in the solution to contain at least 253 peptides as listed in the specification where their molecular masses are between 500 and 4,000 Daltons, and the concentration is in the range of 2.010.sup.54.510.sup.5 ppm; and d. Applying the KHP solution by first diluting with water by volume at the ratio of 1:50-500.

7. The method of using a keratin hydrolysis peptide solution of claim 6 wherein the application of the solution is by infusing into the soil immediately after planting the corn seeds.

8. The method of using a keratin hydrolysis peptide solution of claim 6 wherein the application of the solution is by infusing into the soil at corn's seedling stage.

9. The method of using a keratin hydrolysis peptide solution of claim 6 wherein the application of the solution is by infusing into the soil at corn's V3-V4 growth stage.

10. The method of using a keratin hydrolysis peptide solution of claim 6 wherein the application of the solution is by leaf spraying to the plants at corn's V3-V4 growth stage.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0021] The accompanying drawings, figures and tables, which are incorporated in and constitute a part of this specification, illustrate and exemplify the preferred embodiments of the invention. Together with the description, serve to explain the principles of the invention.

[0022] FIG. 1 shows the comparison of underground bio mass weights among the low temperature groups vis--vis the normal temperature group.

[0023] FIG. 2A shows the comparison of plant heights among the low temperature groups vis--vis the normal temperature group.

[0024] FIG. 2B shows the comparison of stem widths among the low temperature groups vis--vis the normal temperature group.

[0025] FIG. 3A shows the comparison of above ground fresh weights among the low temperature groups vis--vis the normal temperature group.

[0026] FIG. 3B shows the comparison of above ground dry weights among the low temperature groups vis--vis the normal temperature group.

[0027] FIG. 4A shows the comparison of leaf surface areas among the low temperature groups vis--vis the normal temperature group.

[0028] FIG. 4B shows the comparison of SPAD reading (chlorophyll) from the third leaves among the low temperature groups vis--vis the normal temperature group.

[0029] FIG. 4C shows the comparison of SPAD reading (chlorophyll) from the fourth leaves among the low temperature groups vis--vis the normal temperature group.

[0030] FIG. 5 shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3-V4 stage, underground dry weights among the low temperature groups vis--vis the normal temperature group.

[0031] FIG. 6A shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3-V4 stage, above ground dry weights among the low temperature groups vis--vis the normal temperature group.

[0032] FIG. 6B shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3-V4 stage, the plant heights among the low temperature groups vis--vis the normal temperature group.

[0033] FIG. 6C shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3-V4 stage, the stem widths among the low temperature groups vis--vis the normal temperature group.

[0034] FIG. 7A shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3V4 stage, the leaf surface areas among the low temperature groups vis--vis the normal temperature group.

[0035] FIG. 7B shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3-V4 stage, the SPAD reading from the 5th leaves among the low temperature groups vis--vis the normal temperature group.

[0036] FIG. 7C shows, given treatment of KHP solutions, both leaf-spray and soil infusion at V3-V4 stage, the SPAD reading from the 6th leaves among the low temperature groups vis--vis the normal temperature group.

DETAILED DESCRIPTION OF THE INVENTION

[0037] The keratin hydrolysis peptide (KHP) solution of present invention is made by a high-temperature and high-pressure process to treat a mixture of water and feathers as shown in the parameters herein.

[0038] The mixture ratio, temperature, pressure and duration parameters are shown herein:

TABLE-US-00001 Water content Feather Water in feather Pressure Temp. Time Mass Concen. KHP # (kg) (kg) (%) (kg/cm.sup.2) ( C.) (min) (Da) (ppm) 1 70 0 46% 13 180 40 705.9~3194.7 381250 2 50 40 50% 12 185 80 593.3~3508.9 301500

[0039] A first embodiment of keratin hydrolysis peptide (KHP) solution, without water, can be made by 70 kg of feathers, with the feathers' water content being 46%, and then treated by the steps of: [0040] a. stirring the feathers in a sealed container; [0041] b. hydrolyzing the feathers in the container with a temperature and pressure setting of 180 C. and 13 kg/cm.sup.2 for a duration of 40 minutes; [0042] c. using a mass spectrometer to confirm the combination of peptides in the solution to contain at least 253 peptides as listed in the specification whereby their molecular masses are between 705.9 and 3,194.7 Dalton, and the concentration is in the range of 2.010.sup.54.510.sup.5 ppm.

[0043] The keratin hydrolysis peptide (KHP) solution of the first embodiment is further filtered and concentrated to 381,250 ppm concentration.

[0044] The KHP solution of the second embodiment takes the steps of: [0045] a. Preparing the KHP solution by mixing 50 kg of feathers whose content is 50% water with 40 kg of water in a sealed container; [0046] b. hydrolyzing the mixture in the container with a temperature and pressure setting of 185 C. and 12 kg/cm.sup.2 for a duration of 80 minutes; [0047] c. using a mass spectrometer to confirm the combination of peptides in the solution to contain at least 253 peptides as listed in the specification where their molecular masses are between 500 and 4,000 Daltons, and the concentration is in the range of 2.010.sup.54.510.sup.5 ppm.

[0048] The KHP solution of the second embodiment is further filtered and concentrated to 301,500 ppm concentration.

[0049] The confirmation of some of the 253 peptides is further done by referencing the BIOPEP-UWM database.

[0050] To test the effectiveness of the method of promoting corn growth by application of KHP solutions under low temperature setting, the YuMeiJeng corn species was selected by the inventors to conduct experiments and tests.

[0051] Healthy seeds are screened and planted into 3.5-inch pots, one seed per pot. The pots are placed in climate-control rooms with 16 hours of day light and 8 hours of dark time. In addition to normal watering, common TaiFei 43 instant fertilizer was applied to the pots in accordance with common practice.

[0052] Corn's nutritional growth period, after germination, are divided into stages: VE (sheath leaf protruding above ground), V1 (first leaf collar visible), V2 (second leaf collar visible), V3 (third leaf collar visible), V4 (fourth leaf collar visible, with stem tip branching tissue underground), V5 (fifth leaf collar visible, with stem tip branching tissue above ground), etc.

[0053] The inventors set KHP solution's application manners of soil infusion (50 ml per pot) or leaf spray (0.8 ml to leaves) based upon the growth stages of the corn as set forth below.

[0054] First, the inventors conducted soil infusion after seeding based upon the conditions detailed in the following table:

TABLE-US-00002 Temp ( C.) Infusion (50 Group Low Temp (light/dark) ml) to soil Dilution ratio Normal Temp 25/23 water and environment Low Temp Ck 1 0-5 days after 18/18 water Low Temp Exp seeding KHP-1 100x 1A Low Temp Exp KHP-1 50x 1B Low Temp Ck 2 6-9 days after 18/18 water Low Temp Exp seeding (VE KHP-2 100x 2A stage) Low Temp Exp KHP-2 50x 2B

[0055] For the normal temperature and environment group, the corns are given day light temperature of 25 C. (light amount of 27,000 Lux) and dark time temperature of 23 C.

[0056] For the corn pots in the low temperature check group 1 (Low Temp Ck 1), immediately after seeding, 50 ml of water was given, and are then given 5 days' low temperature treatment (18 C. for both day light and dark time). After the 5 days, the temperature is reverted back to normal temperature setting.

[0057] The Low Temp Exp groups 1A and 1B are given the same low temperature treatment, but instead of 50 ml of water, they are given KHP-1 solution, diluted with water by volume at ratios of 100 (noted as 100) and 50 (noted as 50) respectively, by infusing into the soil.

[0058] For the low temperature check group (Low Temp Ck 2), the pots are similarly given 50 ml of water after seeding and the first 5 days after germination are given normal temperature. On the 6th to 9th day, during VE stage, they are given the low temperature treatment of 18 C. (light amount 27,000 Lux during day time) for four days, then the temperature is reverted back to normal setting.

[0059] For the Low Temp Exp 2A and Low Temp Exp 2B groups, the pots of corns are given the same temperature and lighting treatment as the Low Temp Ck2 group, but instead of given 50 ml water, the soil is infused with 50 ml's KHP-2 100 and KHP-2 50 solutions.

[0060] On the 13.sup.th day after seeding, the inventors took various measurements to assess the bio mass, stem heights and widths, leaf SPAD value and surface area as noted below.

[0061] The inventors extracted and cleaned the root portions and took pictures. The plant heights and stem widths are measured. They used a digital scale (AP224X, Shimadazu) to measure the underground biomass and tabulated the findings. For the leaf's measurement, the inventors used a analysis tool WinFOLIA Pro LA2400 (Regent) to compute the leaf surface area; the inventors used SPAD 502 plus to compute SPAD values. The findings are tabulated into the figures herein.

[0062] The inventors experimented with the soil infusion of KHP solution when the corn experienced low temperature at two different times: immediately after seeding and immediately after germination.

[0063] In FIG. 1, it can be seen that both low temperature check groups have much lower underground bio mass weights than the normal temperature setting group. But with the application of KHP-1 solutions (at either 100 or 50 dilution ratio), the bio mass weight increased substantially. The increase is between 22% and 32% in the Low Temp Exp 1A and 1B group; the increase is between 13% and 20% in the Low Temp Exp 2A and 2B group. The general observation in FIG. 1 is that lower concentration of KHP solution (the 100 group) performs better in promoting the corn growth in low temperature setting.

[0064] On the 13.sup.th day after seeding, the heights in different groups are measured and noted in FIG. 2A. As shown, low temperature has huge impact on the height development of corn plants at seeding time, but less so in the VE stage. The increases in height in the Low Temp Exp 1A and Low Temp 1B groups are 21% and 19% over that of the Low Temp Ck 1 group respectively. The general observation in FIG. 2A is that KHP solution at lower concentration (the 100 dilution) works better to increase plant heights.

[0065] For stem widths, The Low Temp Exp 1A and Low Temp Exp 1B groups show increase of 13% and 20%, respectively, over that of the Low Temp Ck 1 group as reflected in FIG. 2B. The Low Temp Exp 2A and Low Temp 2B group both show 10% increase over that of the Low Temp Ck 2 group.

[0066] FIG. 3A shows the comparisons of fresh weights, among the two groups of low temperature treatment immediately after seeding and immediately after germination. The Low Temp Exp 1A and 1B groups show that the KHP solution infusion increased the above ground fresh weight by 49% and 53%, respectively, over that of the Low Temp Ck 1 group. The Low Temp Exp 2A and 2B group also show, respectively, 10-20% increase over that of the Low Temp Ck 2 group.

[0067] FIG. 3B shows above ground dry weight comparisons. The soil infusion of KHP solution to counter the low temperature stress showed that the Low Temp Exp 1A and 1B groups have 51% and 49% increase, respectively, over that of the Low Temp Ck 1 group. The Low Temp Exp 2A and 2B show increase of 21% and 9%, respectively, over that of the Low Temp Ck 2 group.

[0068] FIG. 4A shows the comparisons of leaf surface area. By the analysis done via WinFOLIA Pro LA2400 instrument, the inventors found that the leaf surface area of the Low Temp Exp 1A and 1B shows, respectively, 84% and 66%, increase over that of the Low Temp Ck 1 group. The Low Temp Exp 2A and 2B groups also show substantial increase of 25% and 29%, respectively, over that of the Low Temp Ck 2 group.

[0069] The inventors specifically measured the chlorophyll count by obtaining the SPAD values on the third and fourth leaves. The results are tabulated into FIG. 4B, showing the comparisons of SPAD reading for the third leaf and FIG. 4C, showing the comparison of SPAD reading for the fourth leaf.

[0070] In FIG. 4B, the soil infusion of KHP solution increased the third leaf's SPAD reading (reflecting chlorophyll count) over the corresponding low temperature check group by between 7% and 10%.

[0071] In FIG. 4C, the soil infusion of KHP solution increased the fourth leaf's SPAD reading over the corresponding low temperature check group by between 15% and 29%.

[0072] During the V3-V4 stages, the inventors applied to the corn plants by leaf spray, using 0.8 ml of KHP solutions to spray on the leaves, and also using 50 ml of KHP solution for infusing into the soil containing the corn plants. The conditions of the low temperature treatment and KHP application are noted herein:

TABLE-US-00003 Temp ( C.) Application at Dilution Group Light/dark V3-V4 Applied with Ratio Normal Temp 25/23 Soil infusion 50 ml water setting Low Temp 18/18 Soil infusion 50 ml water Ck3 (V3-V4) Low Temp Leaf spray 0.8 ml KHP-1 500x Exp 3A Low Temp Leaf spray 08 ml KHP-2 500x Exp 3B Low Temp Soil Infusion 50 ml KHP-1 100x Exp 3C

[0073] Corns in all group are given the general NPK fertilizer (20-20-20) by infusion to the soil during the early growth period of between the 7.sup.th-14.sup.th days.

[0074] On the 14.sup.th day after the low temperature treatment, measurements are obtained regarding biomass weights, plant heights, stem widths, leaf SPAD reading and leaf surface area, etc. The results are noted and tabulated into the figures as further discussed herein.

[0075] FIG. 5 shows the Low Temp Ck3 group's underground biomass weight is severely impacted by the low temperature. Such stunted growth, however, is greatly relieved by the method of KHP applications shown in the 3 groups with KHP treatments: Low Temp Exp 3A (leaf spray), Low Temp Exp 3B (leaf spray) and Low Temp Exp 3C (soil infusion).

[0076] FIG. 6A shows the boost result of KHP solution to counter the low temperature stress, as evidenced in the above ground bio mass weights. The Low Temp Exp 3C group, with infusion of 50 ml KHP-1 100 solution, shows a whopping 88% increase over that of the Low Temp Ck3 group.

[0077] FIG. 6B shows the improvement of plant heights. All three KHP treatment groups show increase of between 12% and 25% over that of the Low Temp Ck3 group.

[0078] FIG. 6C shows the improvement of stem widths. All three KHP treatment group show increase of between 13% and 23% over that of the Low Temp Ck3 group.

[0079] The inventors measured the leaf development status of the corns in different groups and compared the leaf surface area, and SPAD readings from the 5th and the 6th leaves. The results are tabulated in the figures below.

[0080] FIG. 7A shows the extremely poor leaf surface area in the Low Temp Ck3 group. With the application of KHP solutions as taught herein, the three treatment groups show increase of between 31% and 60% over that of the Low Temp Ck3 group.

[0081] FIG. 7B shows the comparison of SPAD readings from the 5th leaf among different groups. The soil infusion of 50 ml KHP-1 100 solution in the Low Temp Exp 3C group shows vastly improved result over that of the Low Temp Ck3 group, while the two other KHP treatment groups did not register positive result.

[0082] FIG. 7C shows the comparison of SPAD readings from the 6th leaf among different groups. Similar to the result in FIG. 7B, only the soil infusion of 50 ml KHP-1 100 solution in the Low Temp Exp 3C group shows substantial increase, by 60%, over that of the Low Temp Ck3 group, while the other two group did not register obvious positive result.

[0083] As has been proven by the tests conducted and verified via the scientific analysis/measurement done by the inventors, the method of creating the KHP solution and the method of application will promote the growth of corn under adversity of low temperature, and to reach the health conditions comparable to corns growing in normal temperature settings.

[0084] While the disclosure herein gave limited teachings and embodiment examples, it should be noted that the description and disclosure made herein illustrated the preferred embodiments of the invention and are not meant to limit the scope of the applicant's rights. Variations and alterations may be employed for yet additional embodiments without departing from the scope of the invention herein.