BIOPESTICIDE FORMULATION AND TREATMENT METHOD THEREOF

Abstract

The present disclosure is related to terrein functioning as a biopesticide formulation in drought resistance and growth promotion of crops. The formulation containing terrein can be treated by soaking or spraying on the crops. It is especially suitable for drought and water shortage conditions, promoting growth of plant root length, seedling height, fresh weight, and dry weight, greatly increasing crop yield, and significantly improving drought resistance of crop plants. Under the condition of seed soaking treatment with 10 g/mL terrein, promotion rates of root length, seedling height, fresh weight, and dry weight of pakchoi are 99.19%, 15.66%, 40.34%, and 49.12%, respectively. The source of terrein is easy to obtain and the cost is low. It has a simple structure and is easily soluble in water. In the actual application process, it only needs to simply prepare an aqueous solution or mix with other pesticide formulations for seed soaking or spraying treatment.

Claims

1. An application of terrein as an active ingredient in preparing a biopesticide formulation for crop drought resistance and growth promotion, comprising: an applied concentration of terrein ranged from 2 g/mL to 10 g/mL; wherein the crops include pakchoi, mung bean, sorghum, or wheat.

2. A treatment method for drought resistance and growth promotion of crops, wherein the crops are treated by a seed soaking treatment or a spraying treatment with a formulation containing terrein, and an applied concentration of terrein ranges from 2 g/mL to 10 g/mL, wherein the crops comprise pakchoi, mung bean, sorghum, or wheat.

3. The treatment method for drought resistance and growth promotion of the crops according to claim 2, wherein the seed soaking treatment comprises soaking the seeds of the crops in a formulation containing 2 g/mL to 10 g/mL terrein for 2 to 8 hours, and then sowing them to plant.

4. The treatment method for drought resistance and growth promotion of the crops according to claim 3, wherein the seed soaking treatment comprises soaking the seeds of the crops in a formulation containing 2 g/mL to 10 g/mL terrein for 6 hours, and then sowing them to plant.

5. The treatment method for drought resistance and growth promotion of crops according to claim 2, wherein the spraying treatment comprises spraying the formulation containing 2 g/mL to 10 g/mL terrein at a seedling stage of the crops by 1 to 3 times.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The disclosure will become more fully understood from the detailed description given herein below for illustration only, and thus not limitative of the disclosure, wherein:

[0024] FIG. 1 shows the effect of terrein on root growth of four potted crops under different treatments according to the present invention;

[0025] FIG. 2 shows the effect of terrein on growth of seedling height of four potted crops under different treatments according to the present invention;

[0026] FIG. 3 shows the effect of terrein on fresh weight of four potted crops under different treatments according to the present invention;

[0027] FIG. 4 shows the effect of terrein on dry weight of four potted crops under different treatments according to the present invention;

[0028] FIG. 5 shows the effect of terrein on growth of pakchoi under different treatments according to the present invention;

[0029] FIG. 6 shows the effect of terrein on growth of mung bean under different treatments according to the present invention;

[0030] FIG. 7 shows the effect of terrein on growth of sorghum under different treatments according to the present invention; and

[0031] FIG. 8 shows the effect of terrein on growth of wheat under different treatments according to the present invention.

DETAILED DESCRIPTION

[0032] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. For the experimental methods that do not specify specific conditions in the embodiments, usually follow conventional conditions or conditions suggested by the manufacturer.

[0033] Terrein, is one of the main secondary metabolites produced by the soil fungus Aspergillus terreus, with a CAS number of 582-46-7. It is a colorless needle-like crystal at room temperature. It is a small molecule polyketide compound with a simple structure, which has various biological activities, including anticancer, anti-inflammatory, antibacterial, insecticidal, etc., and has potential application prospects in the fields of beauty, medicine, and agriculture. However, there is no report about the drought resistance and growth promoting effect of terrein on plants at home and abroad. Terrein has a simple structure. It can not only be fermented and extracted by Aspergillus terreus strains widely present in the soil, with high yield and high purity, but also can be prepared on a large scale by chemical synthesis. The source is easy to obtain, and it has a good prospect for development and utilization.

[0034] The structural formula of the terrein involved is:

##STR00001##

[0035] Soil moisture content calculation formula:

[00001]$\mathrm{Soil}\mathrm{moisture}\mathrm{content}=\frac{\mathrm{original}\mathrm{soil}\mathrm{weight}-\mathrm{dried}\mathrm{soil}\mathrm{weight}}{\mathrm{dried}\mathrm{soil}\mathrm{weight}}100\%$

[0036] In Zhang Dan's Regional Drought Long-Term Forecasting and Agricultural Drought Risk Comprehensive Assessment [D]. Dalian University of Technology, 2011, taking the average moisture content of each layer in each period from 2002 to 2009 at Chaoyang Station in the western part of Liaoning Province as an example, when it is in a drought year (2009), the rainfall from mid-April to early November is only 267 mm, and the highest and lowest average moisture content of the 0-50 cm soil layer are 18.8% and 8.3%, respectively, and the average soil moisture content during this period is 12.9%. When it is in a wet year (2003), the rainfall in the same period is 514 mm, and the highest and lowest average moisture content of the 0-50 cm soil layer are 24.5% and 10.7%, respectively, and the average soil moisture content during this period is 15.7%.

[0037] In addition, soil moisture can be classified into saturated moisture, suitable moisture, yellow moisture and dry soil. Wherein, (1) Saturated moisture (moisture content ranges from 18.5% to 20%), the soil color is dark and black, it can be kneaded into a ball by hand, and it will not disperse by throwing it. It can be twisted into strips. There are obvious water marks on the hands. The saturated moisture is the upper limit of suitable tillage, and the effective moisture content of the soil is the largest. (2) Suitable moisture (moisture content ranges from 15.5% to 18.5%), the soil color is deep and dark, it can be kneaded into a ball by hand, and will be broken when thrown. There are wet marks on the hands. Suitable moisture content is suitable for sowing cultivation, and the effective moisture content is relatively high. (3) Yellow soil (moisture content ranges from 12% to 15%), the soil color is yellow, it can be kneaded into a ball, crumbly, and has a cool feeling in the hand. The yellow soil is suitable for cultivation, the effective moisture content is less, and the emergence of seedlings is uneven, so irrigation is required. (4) Dry soil (moisture content below 8%), the soil color is grayish white, with large lumps of induration, fine soil is loose, and dry soil has no moisture that crops can absorb, so it is not suitable for cultivating and sowing. In the present invention, the soil moisture content of the normal treatment group is 18.33% to 19.67%, which is between suitable moisture and saturated moisture, while the soil moisture content of the drought treatment group is 9.33% to 11.89%, which is between dry soil and yellow moisture.

First Embodiment

[0038] An embodiment of effects of 2 g/mL terrein formulation on the growth of four crops by seed soaking treatment under drought conditions is as follows. Terrein (purity 95%) is prepared as a solution with a concentration of 2 g/mL in distilled water, and pakchoi, mung bean, sorghum, and wheat seeds are soaked in it for 6 hours, and the seeds are also soaked in distilled water as a blank control. Subsequently, the soaked crop seeds are sown into potted plants, and each of the treatment group is replicated five times. When the seedlings grew to about 10 cm, the drought treatment culture is carried out. The room temperature is 25 C., the humidity is 70%, 12 hours of light and 12 hours of darkness each day, and the light intensity is 80-100 mol.Math.m.sup.2.Math.s.sup.1. During the period, the soil was kept in a dry state, and 200 mL is watered every week for 30 days, with a total of 800 mL. It is detected that the moisture content of each soil is maintained at 9.33% to 11.89%, as shown in Table 2. The drought culture conditions described below are all treated in the same way. After 30 days, the root length, seedling height, and fresh weight of pakchoi, mung bean, sorghum, and wheat are measured, and 10 seedlings are selected from each of the treatment groups, with a total of 50 plants (N=50), hereafter, they are dried in an oven and then measured dry weight.

[0039] Data statistics method is as follows. The growth rates of root length, seedling height, fresh weight, and dry weight of pakchoi, mung bean, sorghum, and wheat under 2 g/mL compared to the blank control are first calculated, that is,

[00002]$\mathrm{growth}\mathrm{rate}=\frac{\mathrm{trea}\mathrm{tment}-\mathrm{blank}\mathrm{control}}{\mathrm{blank}\mathrm{control}}$

[0040] Then use one-way analysis of variance to detect whether the difference between the data of each of groups is significant, and then use the One-way analysis of variance (One-way ANOVA) method to analyze the data, and the difference between the groups with different marked letters is significant, where the level of probability value is p<0.05.

[0041] Results are as follows. Under the soaking treatment of 2 g/mL terrein, compared with the blank control, the growth rates of the root length of dicotyledonous pakchoi and mung bean are 76.63% and 44.72%, respectively, and the growth rates of their dry weight are 32.47% and 28.07%, respectively, the effects on seedling height, and fresh weight are not significant. In addition, the promotion rates of wheat fresh weight and dry weight are 52.30% and 21.99%, respectively, and there is no significant effect on the growth of sorghum. It can be seen that the seed soaking treatment at this concentration is more suitable for increasing the yield of dicotyledonous pakchoi and mung bean compared with monocotyledonous sorghum and wheat.

Second Embodiment

[0042] An embodiment of effects of 10 g/mL terrein formulation on the growth of four crops by seed soaking treatment under drought conditions is as follows. Terrein is prepared as a solution with a concentration of 10 g/mL in distilled water, and pakchoi, mung bean, sorghum, and wheat seeds are soaked in it for 6 hours, and the seeds are also soaked in distilled water as a blank control. Subsequently, the soaked crop seeds are sown into potted plants. When the seedlings grew to about 10 cm, the drought treatment culture is carried out. After 30 days, the root length, seedling height, and fresh weight of pakchoi, mung bean, sorghum, and wheat are measured, hereafter, they are dried in an oven and then measured dry weight.

[0043] Data statistics method is as follows. The growth rates of root length, seedling height, fresh weight, and dry weight of pakchoi, mung bean, sorghum, and wheat under 10 g/mL compared to the blank control are first calculated, that is,

[00003]$\mathrm{growth}\mathrm{rate}=\frac{\mathrm{trea}\mathrm{tment}-\mathrm{blank}\mathrm{control}}{\mathrm{blank}\mathrm{control}}$

[0044] Then use One-way ANOVA to detect whether the difference between the data of each of groups is significant, and the difference between the groups with different marked letters is significant, where the level of probability value is p<0.05.

[0045] Results are as follows. Under the soaking treatment of 10 g/mL terrein, terrein had a significant promoting effect on the growth of pakchoi, mung bean, sorghum, and wheat, and the effect strength is better than that of 2 g/mL seed soaking treatment. Compared with the blank control, the growth rates of the root length of pakchoi, mung bean, and wheat are 99.19%, 30.92%, and 25.09%, respectively. The growth rate of seedling height of pakchoi is 15.66%. The growth rates of fresh weight of pakchoi and wheat are 40.34% and 76.39%, respectively. The growth rates of dry weight of pakchoi, mung bean, sorghum, and wheat are 49.12%, 35.65%, 15.42% and 22.35%, respectively. It can be seen that compared with the seed soaking treatment of 2 g/mL, the growth of pakchoi can be further promoted under the condition of seed soaking treatment at this concentration, and it is more suitable as a drought-resistant and growth-promoting agent for pakchoi to increase its yield. At the same time, it can also be applied to increase the yield of mung bean, sorghum, and wheat, but the effect is weaker than that of pakchoi.

Third Embodiment

[0046] An embodiment of effects of 2 g/mL terrein formulation on the growth of four crops by spraying treatment under drought conditions is as follows. Seeds of four tested plants are soaked in distilled water and then sowing. Terrein is prepared as a solution with a concentration of 2 g/mL in distilled water. When the potted pakchoi, mung bean, sorghum, and wheat grow to about 10 cm (15-20 days after sowing), spray them once, with an amount of about 3 mL, and distilled water sprayed used as blank control. Subsequently, the drought treatment culture is carried out. The room temperature is 25 C., the humidity is 70%, 12 hours of light and 12 hours of darkness each day, and the light intensity is 80-100 mol.Math.m.sup.2.Math.s.sup.1. During the period, the soil was kept in a dry state, and 200 mL is watered every week for 30 days, with a total of 800 mL. It is detected that the moisture content of each soil is maintained at 9.33% to 11.89%, as shown in Table 2. After 30 days, the root length, seedling height, and fresh weight of pakchoi, mung bean, sorghum, and wheat are measured, hereafter, they are dried in an oven and then measured dry weight.

[0047] Data statistics method is as follows. The growth rates of root length, seedling height, fresh weight, and dry weight of pakchoi, mung bean, sorghum, and wheat under 2 g/mL compared to the blank control are first calculated, that is,

[00004]$\mathrm{growth}\mathrm{rate}=\frac{\mathrm{trea}\mathrm{tment}-\mathrm{blank}\mathrm{control}}{\mathrm{blank}\mathrm{control}}$

[0048] Then use One-way ANOVA to detect whether the difference between the data of each of groups is significant, and the difference between the groups with different marked letters is significant, where the level of probability value is p<0.05.

[0049] Results are as follows. Under the spraying treatment of 2 g/mL terrein, compared with the blank control, the growth rates of the root length of pakchoi, mung bean, and sorghum are 32.48%, 61.95%, and 18.49%, respectively. The growth rates of fresh weight of sorghum and wheat are 18.92% and 60.98%, respectively. The growth rates of dry weight of sorghum and wheat are 20.80% and 16.29%, respectively. Compared with the seed soaking treatment, the growth-promoting effect on pakchoi under this condition is slightly weaker, but the growth-promoting effect on mung bean, sorghum, and wheat is slightly stronger, and it can be applied to increase the yield of mung bean, sorghum, and wheat.

Fourth Embodiment

[0050] An embodiment of effects of 10 g/mL terrein formulation on the growth of four crops by spraying treatment under drought conditions is as follows. Seeds of four tested plants are soaked in distilled water and then sowing. Terrein is prepared as a solution with a concentration of 10 g/mL in distilled water. When the potted pakchoi, mung bean, sorghum, and wheat grow to about 10 cm (15-20 days after sowing), spray them once, with an amount of about 3 mL, and distilled water sprayed used as blank control. Subsequently, the drought treatment is carried out. After 30 days, the root length, seedling height, and fresh weight of pakchoi, mung bean, sorghum, and wheat are measured, hereafter, they are dried in an oven and then measured dry weight.

[0051] Data statistics method is as follows. The growth rates of root length, seedling height, fresh weight, and dry weight of pakchoi, mung bean, sorghum, and wheat under 10 g/mL compared to the blank control are first calculated, that is,

[00005]$\mathrm{growth}\mathrm{rate}=\frac{\mathrm{trea}\mathrm{tment}-\mathrm{blank}\mathrm{control}}{\mathrm{blank}\mathrm{control}}$

[0052] Then use One-way ANOVA to detect whether the difference between the data of each of groups is significant, and then use the LSD method to analyze the data, and the difference between the groups with different marked letters is significant, where the level of probability value is p<0.05.

[0053] Results are as follows. Under the spraying treatment of 10 g/mL terrein, pakchoi, mung bean, sorghum and wheat showed significant promoting effects. Compared with the blank control, the growth rates of root length are 16.45%, 57.20%, 30.50% and 67.40%, respectively. The growth rates of fresh weight of mung bean, sorghum and wheat are 56.18%, 19.90% and 97.21%, respectively. The growth rates of dry weight of mung beans, sorghum and wheat are 26.98%, 25.19% and 34.79%, respectively. Compared with other groups of treatment, this condition has a slightly weaker growth-promoting effect on pakchoi, but has the strongest growth-promoting effect on mung bean, sorghum and wheat, so the treatment is more suitable as a drought-resistant growth-promoting agent for mung bean, sorghum and wheat to increase their yield. The test results of first embodiment to fourth embodiment are shown in FIG. 1 to FIG. 8, wherein FIG. 1 shows the effect of terrein on root growth of four potted crops under different treatments according to the present invention. FIG. 2 shows the effect of terrein on growth of seedling height of four potted crops under different treatments according to the present invention. FIG. 3 shows the effect of terrein on fresh weight of four potted crops under different treatments according to the present invention. FIG. 4 shows the effect of terrein on dry weight of four potted crops under different treatments according to the present invention. FIG. 5 shows the effect of terrein on growth of pakchoi under different treatments according to the present invention. FIG. 6 shows the effect of terrein on growth of mung bean under different treatments according to the present invention. FIG. 7 shows the effect of terrein on growth of sorghum under different treatments according to the present invention. FIG. 8 shows the effect of terrein on growth of wheat under different treatments according to the present invention. Among them, in FIGS. 5-8, from left to right: CK (control, 0 g/mL), terrein seed soaking 2 g/mL, terrein seed soaking 10 g/mL, terrein spraying 2 g/mL, terrein spraying 10 g/mL mL. The order of marked letters in FIGS. 1-4 is also arranged in this order from left to right. The error bars represent the standard errors of the values, and different lowercase letters (a, b, c, etc.) indicate significant differences according to Fisher's LSD test (p<0.05) in FIGS. 1-4.

[0054] The normal treatment is watering twice a week, 300 mL each time, 8 times in total, and the soil moisture content is shown in Table 1. The drought treatment is watering once a week, 200 mL each time, four times in total. Table 3 shows the effects of terrein on the growth of each of crops under drought treatment.

TABLE-US-00001 TABLE 1 Watering amount and soil moisture content of crops under normal control treatment Normal control treatment pakchoi mung bean sorghum wheat Total watering 2400 2400 2400 2400 amount/mL Soil moisture 18.67% 18.33% 19.67% 19.12% content

TABLE-US-00002 TABLE 2 Watering amount, spraying amount and soil moisture content of crops under drought treatment Drought treatment pakchoi mung bean sorghum wheat Total watering amount/mL 800 800 800 800 Soil Control treatment (0 g/mLterrein) 11.00% 10.00% 9.33% 11.67% moisture 2 g/mL terrein seed soaking 11.89% 10.30% 10.33% 9.33% content treatment 10 g/mL terrein seed soaking 11.33% 10.33% 10.00% 10.00% treatment Control treatment (0 g/mLterrein) 10.25% 11.22% 9.90% 10.50% 2 g/mL terrein spraying treatment 11.33% 10.00% 9.33% 9.67% 10 g/mL terrein spraying treatment 11.89% 10.90% 10.00% 10.33%

TABLE-US-00003 TABLE 3 Effects of terrein on the growth of each of crops under drought treatment 2 10 g/mL g/mL 2 10 Control terrein terrein Control g/mL g/mL treatment seed seed treatment terrein terrein Species (0 g/mL soaking soaking (0 g/mL spraying spraying of crops Growth index terrein) treatment treatment terrein) treatment treatment pakchoi root length (cm) 8.75 15.46 17.43 8.75 11.59 10.19 seedling height (cm) 10.55 11.45 12.21 10.55 11.40 10.73 fresh weight (g) 1.74 1.91 2.45 1.74 1.75 1.85 dry weight (g) 0.12 0.16 0.18 0.12 0.13 0.14 mung bean root length (cm) 9.81 14.19 12.84 9.81 15.88 15.42 seedling height (cm) 23.29 22.54 24.21 23.29 23.99 22.08 fresh weight (g) 0.74 0.98 0.87 0.74 0.92 1.16 dry weight (g) 0.12 0.15 0.16 0.12 0.14 0.15 sorghum root length (cm) 32.19 35.67 33.99 32.19 38.15 42.01 seedling height (cm) 40.99 40.43 39.69 40.99 39.14 38.16 fresh weight (g) 1.61 1.71 1.88 1.61 1.92 1.93 dry weight (g) 0.33 0.37 0.38 0.33 0.40 0.42 wheat root length (cm) 15.16 16.13 18.96 15.16 17.02 25.37 seedling height (cm) 20.30 21.55 20.25 20.30 18.93 20.49 fresh weight (g) 0.72 1.10 1.27 0.72 1.16 1.42 dry weight (g) 0.21 0.26 0.26 0.21 0.25 0.29

[0055] The present invention has done corresponding indoor petri dish bioassay experiments before, with pakchoi under drought treatment conditions as the recipient plant, terrein as the donor, and the concentration of terrein is set to 0, 2, 10, 50 and 250 g/mL. The study found that when the concentration of terrein is 2 g/mL and 10 g/mL, it could significantly promote the growth of pakchoi, among which, the promotion rates of pakchoi root length are 13.51% and 18.82%, respectively. The promotion rates of pakchoi seedling height are 21.70% and 23.26%, respectively. When the concentration increased to 50 g/mL, the inhibition rate of pakchoi root length is 16.01%, and the effect on seedling height is not significant. When the concentration reached 250 g/mL, the inhibitory effect on the growth of pakchoi is further enhanced, and the inhibition rates of its root length and seedling height reached 86.83% and 5.71%, respectively. Therefore, the present invention only selects low concentrations (2 g/mL, 10 g/mL) for experiments to confirm the effect of terrein on drought resistance and growth promotion of crops in outdoor pot experiments.

[0056] Terrein in the present invention has remarkable promoting effect on pakchoi, mung bean, sorghum, and wheat, especially under drought conditions, and the dosage is extremely small, and can be used as a plant growth regulator in agricultural production. Compared with several other crops, pakchoi has relatively strong sensitivity, and a very small amount can significantly increase its biomass, which can be mainly used to increase the yield of pakchoi. In actual production, terrein has a relatively mature formulation method. It is easily soluble in water, so the formulation method is simple and the operation is convenient. Therefore, the development of terrein as a drought-resistant and growth-promoting agent has high practical value in agricultural production.

[0057] Finally, it should be noted that the above embodiments are merely the preferred embodiments of the present disclosure, and are not intended to limit the present disclosure. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present disclosure should be included in the protection of the present disclosure within the scope.