CLIAMS:1. A composition for early flowering, enhanced seed yield and breed cycle reduction in tobacco plants, comprising:
a) 2, 4-D in a concentration ranges from 10 to 80ppm;
b) non-ionic surfactant at a concentration 0.025%;
c) zinc sulphate in a concentration ranges from 20 to 70ppm; and
d) manganese sulphate in a concentration ranges from 5 to 10ppm.
2. The composition as claimed in claim 1, wherein said 2, 4-D is at a concentration of 40ppm.
3. The composition as claimed in claim 1, wherein said non-ionic surfactant is APSA-80.
4. The composition as claimed in claim 1, wherein said zinc sulphate at a concentration of 50ppm.
5. The composition as claimed in claim 1, wherein said manganese sulphate is at a concentration of 5ppm.
6. The composition as claimed in claim 1, wherein said composition is adapted for application to tobacco plants 30 days after transplantation.
7. The composition as claimed in claim 1, wherein said composition is in the form of foliar application using field sprayer.
8. The composition as claimed in claim 1 wherein said tobacco plants are selectively Flue cured virginia (FCV)-K326 variety.
,TagSPECI:Field of the invention
The present invention relates to agricultural compositions which induces early flowering, enhanced seed yield and reduction in breeding cycle in plants. More specifically the present invention relates to agricultural compositions comprising 2, 4-D which induces early flowering, enhanced seed yield and reduction in breeding cycle in tobacco plants, more particularly in Flue cured Virgina (FCV) variety-K326.

Background of the Invention and prior art
Flowering in plants is influenced by many factors. Some plants flower early or late depending on environment they are in. Flowering of plants at right time is important for breeding programs so that generation advancement can be made quickly. In India, tobacco has two growing seasons. One is from February to August (KLS) and second one is from October to March (NLS, SBCS, and SLS) due to which 2 generations can be advanced in a year. In breeding program if 2 generations are take up in a year then breeding cycle can be reduced to half i.e., from 6 to 3 years. This is also applicable for the development of RIL (Recombinant inbred lines) population. As the two seasons in tobacco are back to back and very close, by the time seed sets and capsule gets dried, the sowing time for the next region will be already on. Hence, reducing the flowering time by 20 days can help to take 2 generations in a year more comfortably and there by reducing breeding cycle to half time.
Early flowering in plants can be induced by creating stress in plants with application of chemicals, depriving plant from nutrients or by adverse environmental conditions. Based on literature survey, hormones effecting plant metabolism like auxins, gibberellins, polyamines are applied at varying concentrations to induce flowering.
Auxins are the plant hormones involved in plant growth and development. There are two types of auxins, natural (Indole acetic acid) and synthetic (naphthylacetic acid, 2, 4 Dichloro phenoxy acetic acid). Generally the synthetic plant hormone auxin 2, 4-D is used as weedicide and plant growth regulator. At high concentration auxin kills different types of weeds on the contrary auxin at low concentrations enhances the plant growth and yield.
CN200910061831 relates to the use of plant growth regulators on tobacco, in particular relates to a plant growth regulator to reduce the nicotine content of tobacco-release method. In the method, the plant growth regulator is dissolved in a release medium and is filled into conic caps to prepare release caps; and the release caps are covered at sections of the top of stems of tobacco plants.
WO2010149732 relates to a mixture comprising active compounds which further comprises a herbicide selected from the group of synthetic auxins consisting of (i) Phenoxy-carboxylic-acids selected from clomeprop, 2,4-D, 2,4-DB, dichlorprop, MCPA, MCPB and mecoprop; (ii) Benzoic acids selected from chloramben, dicamba and 2,3,6-TBA; (iii) Pyridine carboxylic acids selected from aminopyralid, clopyralid, fluroxypyr, picloram and triclopyr; (iv) Quinoline carboxylic acids selected from quinclorac and quinmerac; (v) benazolin-ethyl; and (vi) amino-cylopyrachlor; and a fungicide selected from the group of strobilurines consisting of pyraclostrobin, orysastrobin, azoxystrobin, dimoxystrobin, enestroburin, fluox- astrobin, kresoxim-methyl, metominostrobin, picoxystrobin, pyribencarb, triflox- ystrobin, 2-(2-(6-(3-chloro-2-methyl-phenoxy)-5-fluoro-pyrimidin-4-yloxy)-phenyl)- 2-methoxyimino-N-methyl-acetamide, 3-methoxy-2-(2-(N-(4-methoxy-phenyl)- cyclopropane-carboximidoylsulfanylmethyl)-phenyl)-acrylic acid methyl ester, methyl (2-chloro-5-[1 -(3-methylbenzyloxyimino)-ethyl]benzyl)carbamate and 2- (2-(3-(2,6-dichlorophenyl)-1 -methyl-allylideneaminooxymethyl)-phenyl)-2- methoxyimino-N-methyl-acetamide for synergistically increasing the health of a plant. However the prior art uses 2, 4-D as herbicide which kills the unwanted plants (weeds), so that the unwanted plants do not compete with crop plants (maize, soybean etc) for nutrients and water. Thereby the overall growth enhances in crop plants.
WO2013185036 relates to methods for improving plant height and/or yield of crop plants which are resistant to herbicide 2, 4-D by treating the plants with 2, 4-D at application rates which are not harmful to the plants. WO2013185036 provides a method using 2, 4-D to increase yield of crop plants which express AAD-12 gene for 2, 4-D resistance. This method is of particular interest for the treatment of crops plants including maize, soybean, spring and winter oil seed rape (canola), sugar beet, wheat, sunflower, barley, and rice. However the prior art uses 2, 4-D as fungicide to control pathogen infection and enhanced plant growth, but not as an agent for enhancing early flowering and enhanced seed yield.
Further an article titled “The Dual Role of Auxin in Flowering” by Frank B et al. discloses about dual role of auxin. Auxins inhibits flowering in crops like Xanthium, Alaska pea on the contrary it promotes flower initiation in other crop plants like pineapple and Ananas. The dual role action of auxin depends on whether it is a short day or long day plants and concentration of auxin applied (at higher concentration it inhibits flowering irrespective of short day or long day plants). However the dual role of auxin is different for different plant species, there is no generalized response elicited by auxin application. The prior art uses auxin for promoting initiation of flower primordial and elongation of shoot axis but not in advancing flower formation as early as 2-3 weeks.
Further, in tobacco plants “plant growth” means 1) increase in the leaf production where nicotine is stored which has got high economic value and 2) increase in seed production by allowing the plants to produce more flowers. Hence, inducing flowering in tobacco varieties is not a desired choice especially in tobacco industry. Thus the prior art compositions which comprises 2, 4-D relates to use of 2, 4-D to increase the leaf production where nicotine is stored which has got high economic value.
The reduction in flowering time by 20 days can help to take 2 generations in a year more comfortably and there by reducing breeding cycle to half time. Accordingly the present invention provides a composition for early flowering and enhanced seed yield which concedes 20 days early flowering and at least 45% increased seed production.

Objectives of the present invention
It is an object of present invention to overcome the drawbacks of the prior art.
It is another object of present invention to induce early flowering that is 20 days before in tobacco plants.
It a further object of present invention to provide an agricultural composition for foliar application comprising 2, 4 D combined with micronutrients.
It is also an object of present invention to provide a composition to achieve significant reduction in breeding cycle, early flowering and 40-45% enhanced seed yield.

Summary of the present invention
A composition for early flowering, enhanced seed yield and breed cycle reduction in tobacco plants, comprising:
a) 2, 4-D in a concentration ranges from 10 to 80ppm;
b) non-ionic surfactant at a concentration 0.025%;
c) Zinc sulphate in a concentration ranges from 20 to 70ppm; and
d) Manganese sulphate in a concentration ranges from 5 to 10ppm.

Brief description of accompanying drawings
Figure.1 depicts the effect of 2, 4D concentration on tobacco plants.
Figure.2 depicts the effect of 2, 4-D on morphological attributes of tobacco plants.

Detailed description of the present invention
The present invention provides a composition comprising 2, 4-D combined with micronutrients which significant reduces the breeding cycle (time taken from seed sowing to 1st generation seed set), and induces early flowering (seed to flower) and enhances seed yield (seed to seed) in tobacco plants, more particularly in Flue cured Virgina (FCV) variety-K326.
The present inventors have found that the composition of the present invention induces early flowering, wherein the flowering starts at least 20 days early. There is a significant increase in seed quantity along with the change in leaf morphology compared to the control tobacco plants which are not subjected to the composition of the present invention. The induced early flowering and increased seed quantity results in reducing the breeding cycle by taking up two generations in a year (two seasons back to back).
The present inventors have found that the reduction in breeding cycle, inducing early flowering and enhanced seed yield are dependent on the critical parameters such as concentration of 2, 4-D in composition, critical stage of application and tobacco plant variety.
According to the present invention the desired results such as reduction in breeding cycle, early flowering and enhanced seed production is observed when the composition comprises 2, 4-D at a concentration of 40ppm and said composition further comprises a non-ionic surfactants such as APSA-80 at a concentration of 0.025%. The APSA-80 act as an adjuvant used for ease spread and penetration of active molecules into plants
Further, the desired results such as reduction in breeding cycle, early flowering and enhanced seed production is observed only when the composition is applied on 30 days after transplantation (DAT).
The inventors have surprisingly found that there is no effect on the tobacco plants when the composition of the present invention was applied at 25 or 40 days after transplantation. The plants are observed to be similar to control when the composition of the present invention was applied at 25 or 40 days after transplantation.
Further, the present inventors have found that the composition of the present invention showed desired results in FCV tobacco variety K326 only. The composition of the present invention does not have any positive impact towards flowering in other plants.
The composition according to the present invention comprises:
(i) 10 to 80ppm, preferably 40ppm of 2, 4-D;
(ii) 0.025% by weight of non-ionic surfactant;
(iii) 20 to 70ppm, preferably 50ppm of zinc sulphate; and
(iv) 5 to 10ppm, preferably 5ppm of manganese sulphate.
The non-ionic surfactant according to the present invention is APSA-80.
The composition according to the present invention is applied to the foliar parts using field sprayer.
The present invention is now illustrated by way of non-limiting examples. The examples, which are intended to be purely exemplary of the invention, should therefore not be considered to limit the invention in any way.

Example 1
An early flowering and enhanced seed yield composition in accordance with the present invention can have the composition as mentioned in the Table 1.
Table.1 An early flowering and enhanced seed yield composition comprising 2, 4-D and micronutrients
Composition details Amounts
2,4D 40 ppm
APSA-80 (non-ionic surfactants) 0.025%
Zinc sulphate 50ppm
Manganese sulphate 5ppm
The above composition was prepared by dissolving required amount of 2,4-D in minimum amount of ethanol and then this solution was mixed in water. Then zinc sulphate and manganese sulphate were added, followed by addition of APSA-80 just before treatment imposition using field sprayer.

Example 2
Effect of 2, 4-D concentration on tobacco plants
The tobacco plants were treated with three different compositions comprising 2, 4-D at a concentration of 80, 40 and 20ppm along with APSA (0.025%).
The compositions were prepared by dissolving required amount of 2,4-D in minimum amount of ethanol and then the solutions were mixed in water. Then zinc sulphate and manganese sulphate were added, followed by addition of APSA-80 just before treatment imposition using field sprayer. It is observed that the tobacco plants treated with higher concentration (80ppm) become weak and fall down. The tobacco plants treated with 40ppm shows the desired results of early flowering and enhanced seed yield. The effect of 2,4D concentration on tobacco plants is shown in Figure.1.

Example 3
Effect of 2, 4-D on tobacco plant days of flowering and life cycle
A composition comprising 2, 4-D (40ppm); Zinc sulphate (50ppm); Manganese sulphate (5ppm); and APSA 80 (0.025%) was prepared by dissolving required amount of 2,4-D in minimum amount of ethanol and then the solution was mixed in water. Then zinc sulphate and manganese sulphate were added, followed by addition of APSA-80 just before treatment imposition using field sprayer.
The above composition was applied to the tobacco plant in the form of foliar application at a crop stage of 30 DAT (days after transplantation). The results of foliar application of 2, 4-D to the tobacco plants are shown in Table.2.
Table.2 Depicts the effect of 2,4D on tobacco plant life days for flowering and cycle
S.No Treatments (units are in ppm) Days for flowering (seed to flowering) Life cycle(Seed to seed)
1 Control – water 180 200 days
2 2,4D (40ppm) + Zn (50ppm) + Mn (5ppm) + APSA (workable range) 164 180 days

Zn- Zinc sulphate; Mn- Manganese sulphate; APSA – Neutral adjuvant at 0.025%
Numbers indicate concentration of particular chemical in ppm
From Table.2 it is observed that foliar application of 2, 4-D decreases in life cycle by at least 15-20 days and enhanced seed set of about 40-45% compared to the control.
Example 4
Effect of 2, 4-D on morphological attributes of tobacco plants
To study the effect of 2, 4-D on morphological attributes of tobacco plants, sample composition was prepared by dissolving required amount of 2,4-D (40ppm) in minimum amount of ethanol and then this solution was mixed in water. Then zinc sulphate (50ppm) and manganese sulphate (5ppm) were added, followed by addition of APSA-80 (0.025%) just before treatment imposition using field sprayer.
The composition of present invention also contributes towards the morphological changes in the plant such as plant height, leaf area and no. of leaves. The effect of 2, 4-D on morphological attributes of tobacco plants is present in Table.3 and shown in Figure.2.

Table.3 depicts the effect of 2, 4-D on morphological attributes of tobacco plants
Treatments PH NL Avg LA YLD Avg INL Avg SD
Control 159.3 31.0 1936 183.3 6.3 8.6
2,4D 40+zn50+Mn 5 (Test) 125.7 29.3 661 59.3 4.4 6.1
PH- Plant height (cm); NL- No. of leaves; YLD-Leaf yield (g/plant); SD-Stem diameter

Form Table.3 it is observed that the tobacco plants treated with 2, 4-D showed significant changes in phenotypic characters such as leaf area (reduced by 66%), plant height, stem diameter and internodal length apart from early flowering and higher seed yield compared with control (untreated tobacco plants). However the seeds obtained from these treated plants when sown result in healthy plants with leaves which are suitable for tobacco products.

Example 5
Effect of 2, 4-D on seed yield of tobacco plants
To study the effect of 2, 4-D on seed yield of tobacco plants, sample composition was prepared by dissolving required amount of 2,4-D (40ppm) in minimum amount of ethanol and then this solution was mixed in water. Then zinc sulphate (50ppm) and manganese sulphate (5ppm) were added, followed by addition of APSA-80 (0.025%) just before treatment imposition using field sprayer.
Tobacco plants were treated with the above solution.
Seeds were collected from the treated and controlled plants randomly and results are present in Table.4.
Table.4 Depicts the seed yields (g/plant) of 2, 4-D treated and untreated plants
Season 1 Season 2
Sample Control Test Control Test
1 25 40 32 46
2 25 42 34 48
3 31 38 31 43
t test is performed
p value 0.0049 0.0014
1% level of significance

As evident from Table.4 the seed yield advantage of 2, 4-D treated plants is 40-45% over control in the both seasons which appears to be more significant.

Example 6
Criticality in the stage of application
Sample composition was prepared by dissolving required amount of 2,4-D (40ppm) in minimum amount of ethanol and then this solution was mixed in water. Then zinc sulphate (50ppm) and manganese sulphate (5ppm) were added, followed by addition of APSA-80 (0.025%) just before treatment imposition using field sprayer. When tobacco plants reach appropriate stage i.e., 30 days after transplantation (DAT) the present composition was applied into the foliar regions of the plants using field sprayer. It was observed that there occurred reduction in breeding cycle, early flowering and enhanced seed production in the resultant plants.
There was no effect on the tobacco plants when the composition of the present invention was applied at 25 or 40 DAT. Plants were observed to be similar to control.

Example 7
Criticality in selection of plant species
Sample composition was prepared by dissolving required amount of 2,4-D (40ppm) in minimum amount of ethanol and then this solution was mixed in water. Then zinc sulphate (50ppm) and manganese sulphate (5ppm) were added, followed by addition of APSA-80 (0.025%) just before treatment imposition using field sprayer.
Effect of 2,4D on flowering is observed only in FCV tobacco variety K326. However there is no effect (i.e. no flowering) of 2, 4D in other tobacco species / variety such as few Burley varieties which did not flower at all, whereas few FCV varieties like RT (Red Tobacco), Delhi 61, Cubano de las sierra, strain ER, had shown late/delayed flowering (delayed by atleast 10 days when compared with control) upon treatment with 2,4D when compared to control.