DESC:FIELD OF THE INVENTION

The present invention is related to agricultural field and more particularly, the present invention relates to a neem oil-based composition for treating seed and a process of preparation thereof wherein the composition consisting of neem oil microemulsion for obtaining improved growth of seedling and inhabiting the growth of harmful pests.

BACKGROUND OF THE INVENTION

The ever-increasing concern for improvement of well-being and hygiene in our daily life owing to improved living standards throughout the world attracted the overall global research attention. In such times, investigation on botanical field has emerged as one of the primary research fields. Target applications are agriculture items, biomedical waste containers etc. where colonization of microbes and cross-infection probability is a serious concern. A major challenge of agriculture is to increase food production to meet the needs of the growing world population, without damaging the environment. Therefore, the farmers have been treating pests on the field using certain chemical agents so that you can acquire your fruits, vegetables and other foods at the local market on time. Where Seed treatment refers to the application of fungicide, insecticide, or a combination of both, to seeds so as to disinfect and disinfect them from seed-borne or soil-borne pathogenic organisms and storage insects. It also refers to the subjecting of seeds to solar energy exposure, immersion in conditioned water, etc. In current agricultural practices, the control of pests is often accomplished by means of the excessive use of agrochemicals, which can result in environmental pollution and the development of resistant pests. In this context, biopesticides can offer a better alternative to synthetic pesticides, enabling safer control of pest populations.

In agriculture and horticulture, seed treatment is a chemical treatment, typically antimicrobial or fungicidal treatment, with which seeds are treated prior to planting. In current scenario, captan and thiram have been used in chemical treatment which is effective but harmful. Certain chemical pesticides and insecticides like thiram and captan are used widely in seed coating and seed treatment process to prevent soil borne or seed borne pathogenic organisms. The effects of the fungicides captan and thiram on the survival and phenotypic characteristics of Rhizobium leguminosarum bv. viceae, strain C1. Captan and thiram significantly reduced the numbers of rhizobia recovered from seed. However, only the highest concentrations of captan affected nodulation and plant growth. Contact with some seed-applied fungicides can significantly alter phenotypic characteristics of rhizobia, but these changes might be offset by the presence of host plants.

Seed treatments is chemical treatment by way of using pesticides and insecticides which has limitations of short shelf life, photosensitivity, and volatilization, make it difficult to use them on a large scale. A seed coating is a thicker form of covering of seed and may contain fertilizer, growth promoters and or seed treatment as well as an inert carrier and a polymer outer shell. It is also used to refer to the process of removing chaff, weed seeds and straw from a seed stock.

However, the treated seeds should not be left exposed on the soil surface as it can be detrimental and toxic to birds, mammals, fish as well as aquatic invertebrates. All dispersed seeds should be collected or covered immediately. Thiram-treated seeds can result in delayed egg-laying, decreased clutch size, and effect egg size and eggshell thickness. Consequences of thiram exposure to workers or animals include long term hepatotoxicity, liver damage by enlargement and dysfunction, focal necrosis, and degenerative changes. In humans, acute dermal exposure to captan may lead to conjunctivitis and dermatitis. Captan pesticide is also toxic to fish. Erosion of captan from the treated regions to nearby aquatic region can cause adverse effects to aquatic species.

There is a requirement for a better and more simplistic solution to overcome the limitation of the existing in the art. Therefore, the present invention provides a process for treating a seed wherein the composition consisting of neem oil microemulsion for obtaining improved growth of seedling and inhabiting the growth of harmful pests which is chemical free. This process controls the harmful effects of chemical insecticide and pesticide on different living organisms by considering the gaps and obstacles associated with the development of sustainable agriculture in the not too distant future.

OBJECTS OF THE INVENTION

The main object of the present invention is to provide a process for treating a seed through neem oil microemulsion which protects against seed and soil borne diseases and enhances the efficiency in the growth at the stage of germination of the crop.

SUMMARY OF THE INVENTION

The present invention is related to a process for treating a seed with neem oil microemulsion for obtaining improved growth of seedling and inhabiting the growth of harmful pests. In one aspect, the major ingredient is neem oil which is treating the seed to replace agrochemicals in agricultural pest control. This method chemical free which is efficient, soil melioration, easily degraded, no residue, without pesticide resistance, good stability, be applicable to the feature of directly spraying. Furthermore, the process for the preparation of treating a seed it is advantageous to select natural insecticide and pesticide for seed treatment.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system and/or method, and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates.

Although the present invention has been described in its preferred embodiment. Embodiments of the techniques introduced here include various steps and operations, which have been described below. The main concept of guiding the present invention is to treat a seed through neem oil microemulsion for replacing agrochemicals in agricultural pest control. The main component of the invention is neem oil microemulsion which is treated on the seeds. Neem oil acts as a natural insecticide and pesticide which inhibits growth of harmful pests.

Neem oil is an essential oil derived from the Azadirachta indica plant and is known to source of phytochemicals for use in human health and pest. It also possesses antimicrobial and insect repellent properties. Azadirachta is a fast-growing small-to-medium sized evergreen tree, with wide and spreading branches. It can tolerate high temperatures as well as poor or degraded soil. Neem oil contains at least 100 biologically active compounds. Among them, the major constituents are triterpenes known as limonoids, the most important being azadirachtin, which appears to cause 90% of the effect on most pests. The compound has a melting point of 160°C and molecular weight of 720 g/mol. Other components include meliantriol, nimbin, nimbidin, nimbinin, nimbolides, fatty acids (oleic, stearic, and palmitic), and salannin. The main neem product is the oil extracted from the seeds by different techniques. The other parts of the neem tree contain less azadirachtin, but are also used for oil extraction. It has been suggested that the content of azadirachtin in the seeds can be increased by artificial infection with arbuscular mycorrhiza. The use of neem oil as an insect repellent, pesticide and fungicide, and that the same is a much safer alternative to harmful organic compounds present in other mosquito and insect repellents is well known in the art. This offers an opportunity for using neem oil in insect repellent and related applications. However, neem oil is volatile in nature.

A process for producing a neem oil includes the steps of: Removing of husk from the neem seeds to get neem seed kernel; de-oiling by cold pressing neem seeds kernel to remove a first proportion of the oil content thereof; extracting the so called de-oiled neem seeds cake with water at near neutral pH; separating the liquid phase to obtain clarified aqueous solution of azadirachtin along with polysaccharides and proteins and the remaining oil.

A neem oil-based composition for treating seed is alike neem oil microemulsion liquid which contains a sufficient amount of a neem oil and a surfactant component to obtain the improved growth of seedling and to inhibit and/ or to prevent a biological infestation. Further, neem oil contains azadirachtin, limonoids and polysaccharide-protein matrix and is extracted from seed kernel with different techniques. And, the surfactant component contains one polysorbate component. Where the weight ratio is 0.25% to 75% by weight of the neem oil and from 0.7% to 75% by weight of polysorbate component to obtain the improved growth of seedling and to inhibit and/ or to prevent a biological infestation. And, the biological infestation comprises a fungal species, an algal species, a bacterial species, or any combination thereof.

A process for treating seed by a neem oil-based composition comprising the steps of: promoting (i) growth of a plant, and/or (ii) germination of a seed of said plant, the method comprising contacting said plant and/or its seed and/or its growth substrate with the with the neem oil-based composition, in an amount effective to promote said growth of said plant and/or germination of said seed.

As, shown in the figure in the process of treating a seed, seeds are treated in the following manner. The following process includes: Adding 1 ml neem oil microemulsion dropwise through pipette in the small jar containing 5-7 gm of seeds; stirring constantly for 15-17 minutes till the seeds are coated with the microemulsion neem oil; drying the treated sees for 24 hours in sterilized box; packing the seeds respectively; sowing the treated seeds after 2-3 days; germinating the treated seeds with organic fertilizers; and spraying with the mixture of water-soluble neem oil and water on crops at regular intervals till the fruits are obtained.

Normally, the plants are from different group like thallophyta, bryophyta, pteridophyta, gymnosperms, and angiosperms. And, the seeds are from different group like Grains, pulses, and oilseeds. All of them includes strawberry, squash, cucumber, tomato, rose, pepper eggplant, grapevine, cotton, onion, garlic, wheat, soy, corn and rice.

Analysis of the result

The analysis of variance (ANOVA) is a statistical method for identifying if the means of two or more groups differ significantly.
To determine whether any of the differences between the means are statistically significant, the p-value obtained from ANOVA test is compared to the significance level to assess the null hypothesis. The null hypothesis states that the population means are all equal. Usually, a significance level (denoted as a or alpha) of 0.05 works well.
P-value = a: The differences between some of the means are statistically significant.
P-value > a: The differences between the means are not statistically significant

To support our experimental data we hypothesize,
H0 (null hypothesis) = Neem oil treated seeds does not affect the plant growth positively
HA (alternate hypothesis)= Neem oil treated seeds affect the plant growth positively

Example 1:

Effect of microemulsion neem oil on chili seeds after seedling under vitro conditions: The data has been recorded at harvesting stage of control and microemulsion neem oil treated seed plants. Physical characteristics of the plants such as plant height, shoot weight, no. of leaves, no. of primary branches, no. fruit per plant, single fruit weight, root weight, root length and fruit length was recorded.

CHILI SEEDS GROWTH TRAITS PH SW NOL RL RW NOPB NOF SFW FL
CONTROL 50.1 18.3 20.2 12.45 4.5 2.2 1.4 2 4
NEEM OIL TREATED 55.1 31 24.2 16.2 6.75 2.8 4.2 4.2 7.15

Table 1: Average data of 10 samples of control and neem oil treated chili seeds

Figure 1: Average data of 10 samples of control and neem oil treated chili seeds representing 9 growth traits
Growth traits of chili plant p-value (Control and neem oil treated chili seeds)
PH 0.250
SW 0.002**
NOL 0.017**
NOPB 0.048*
NOF 0.001**
SFW 0.034*
FL 0.070
RL 0.012**
RW 0.035*
**= p = 0.01, *= p = 0.05
Table 2: P- values obtained from ANOVA for growth traits of chili plant.

Example 2:
Effect of microemulsion neem oil on tomato seeds after seedling under vitro conditions: The data has been recorded at harvesting stage of control and microemulsion neem oil treated seed plants. Physical characteristics of the plants such as plant height, no. of primary branches, no. fruit per plant, single fruit weight was recorded.

Result of tomato seeds

PH SFW NOF NOPB
Control 145.7 52.9 9 6.25
Neem oil 170.4 59.5 11.8 7.35

Table 3: Average data of 10 samples of control and neem oil treated tomato seeds

Figure 2: Average data of 10 samples of control and neem oil treated tomato seeds representing 9 growth traits

Growth related traits of tomato plant p-value (Control and neem oil treated tomato seeds)
PH 0.01 **
NOF 0.05*
SFW 0.06
NOPB 0.04*
**= p = 0.01, *= p = 0.05
Table 4: P- values obtained from ANOVA for growth traits of tomato plants

Analysis
Nine growth traits of 10 chili plants for control and neem oil treated seeds were measured namely, Plant height (PH), Shoot weight (SW), No. of leaves (NOL), No. of primary branches per plant (NOPB), No. of fruits per plant (NOF), Single fruit weight (SFW), Fruit length (FL), Root length (RL), Root weight (RW).

Four growth traits of 10 tomato plants for control and neem oil treated seeds were measured namely, Plant height (PH), No. of fruits per plant (NOF), Single fruit weight (SFW), No. of primary branches per plant (NOPB).

The graphs representing average values of all growth traits as shown in figure 1 & figure 2 indicate higher average values for neem oil treated chili and tomato seeds. This indicates that overall strength of plants grown using neem oil treated seeds is better than control seeds.

To support alternate hypothesis ANOVA: single factor was performed for control and neem oil treated seeds for all the growth traits.

For chili plants:
Statistical analysis depicts significant differences between the neem treated seed compared with control except plant height (PL) and fruit length (FL).
For plant height (PH) (p-value: 0.25), the probability of H0 being true is 25%, but an increase of 5cm was observed in the mean values of control and neem oil treated seed plants. There was no significant difference observed for fruit length (FL) (p-value: 0.07).
Shoot weight (SW) (p-value: 0.002), No of leaves (NOL) (p-value: 0.017), No of fruits (NOF) (p-value: 0.001) and Root length (RL) (p-value: 0.012) shows highly significant p-values which strongly rejects the null hypothesis.
Whereas other parameters depict a significant (p=0.05) value as shown in table-2

For tomato plants:
Plant height (PH) showcase highly significant (p-value: 0.01) to reject the null hypothesis, whereas No of leaves (NOL) and No of primary branches (NOPB) shows (p-value: 0.05) and (p-value: 0.04) respectively.
Single fruit weight (SFW) showed slightly insignificant (p-value: 0.06), giving the probability of 6% for the null hypothesis to be true.

Furthermore, the statistical analysis strongly indicates the changes for growth related traits in control and neem oil treated seeds grown plants showcasing significant p-values except Plant height (PH) and fruit length (FL) in chili plants and single fruit weight (SFW) in tomato plants showcasing p-values=0.05 which can be affected by smaller sample size. Other growth traits show significant difference between control and neem oil treated seeds which determines that neem oil treated seeds affect the plant growth positively.

The forgoing disclosure is not intended to limit the present disclosure to the precise forms or particular fields of use disclosed. As such, it is contemplated that various alternate embodiments and/ or modifications to the present disclosure, whether explicitly described or implied herein, are possible in light of the disclosure. Having thus described embodiments of the present disclosure, persons of ordinary skill in the art will recognize that changes may be made in form and detail without departing from the scope of the present disclosure. For example, orders of processes described herein may be changed and are not limited to the manner described herein.
,CLAIMS:1. A neem oil-based composition for treating seed comprising neem oil microemulsion liquid comprising a sufficient amount of a neem oil and a surfactant component to obtain the improved growth of seedling and to inhibit and/ or to prevent a biological infestation;
wherein neem oil contains azadirachtin, limonoids and polysaccharide-protein matrix and is extracted from seed kernel with different techniques;
wherein the surfactant component at contains at least one selected from polysorbate;
wherein the weight ratio is 0.25% to 75% by weight of the neem oil and from 0.7% to 75% by weight of polysorbate and;
wherein the biological infestation comprises a fungal species, an algal species, a bacterial species, or any combination thereof.

2. The neem oil-based composition as claimed in claim 1, wherein said azadirachtin is present in an amount from 150 - 1500 ppm which has pesticidal or plant growth promoting activity.

3. A process for producing a neem oil as claimed in claim 1 comprising the steps of:
Removing of husk from the neem seeds to get neem seed kernel;
de-oiling by cold pressing neem seeds kernel to remove a first proportion of the oil content thereof;
extracting the so called de-oiled neem seeds cake with water at near neutral pH;
separating the liquid phase to obtain clarified aqueous solution of azadirachtin along with polysaccharides and proteins and the remaining oil.

4. A process for treating seed by a neem oil-based composition as claimed in claim 1 comprising the steps of: promoting (i) growth of a plant, and/or (ii) germination of a seed of said plant, the method comprising contacting said plant and/or its seed and/or its growth substrate with the neem oil-based composition, in an amount effective to promote said growth of said plant and/or germination of said seed.

5. The method as claimed in claim 4, wherein said plant is selected from the group consisting of strawberry, squash, cucumber, tomato, rose, pepper eggplant, grapevine, cotton, onion, garlic, wheat, soy, corn and rice.

6. A process for treating seed by a neem oil-based composition comprising the steps of:
Adding 1 ml neem oil microemulsion dropwise through pipette in the small jar containing 5-7 gm of seeds;
Stirring constantly for 15-17 minutes till the seeds are coated with the water-soluble neem oil;
Drying the treated seeds for 24 hours in sterilized box;
Packing the seeds respectively;
Sowing the treated seeds after 2-3 days;
Germinating the treated seeds with organic fertilizers; and
Spraying with the mixture of water-soluble neem oil and water on crops at regular intervals till the fruits are obtained.