Holistic, cost effective method for management of huang long bing (HLB), phytophthora gummosis, asian citrus psyllid and other serious infestations in citrus and other crops

Abstract

Method of controlling infestation of citrus comprising the steps of: i) application of nanobiocide component-1 as a soil drench and foliar spray followed by simultaneous application of component-2 and component-4 followed by the application of component-3, as and when the citrus psyllid observed at threshold levels; ii) application of component-6 which is silicone based wetting and spreading agent cum penetrant mixed with components 3 and 5 and water as foliar spray; ii) finally applying component-5from the onset of flowering at 30-45 day intervals.

Claims

1. A method of controlling at least one of bacterial, fungal, viral nematode and insect infestation of a crop comprising the steps of: i) application of a first component as a soil drench and foliar spray followed by simultaneous application of a second component and a fourth component followed by application of a third component, as and when an insect pest is observed at threshold levels; ii) application of a sixth component which is a silicone based wetting and spreading agent and penetrant mixed with the third component and a fifth component and water as foliar spray; and iii) applying the fifth component as a foliar spray from an onset of flowering of the crop at 30-45 day intervals; wherein: the first component includes a water based formulation containing nanoparticles of silver alone in microencapsulated form or in conjunction with either 2.5% hydrogen peroxide or nanoparticles of a metal; the second component includes a consortium of plant growth promoting rhizosphere microbes which occur in nature; the third component includes a plant extract which is an oil extracted from the seed of Karanj (Pongamia glabra/Derris indica); the fourth component includes a slow release, granulated formulation or Soluble Powder (SP) formulation comprising amino acids, sea weed extracts, humic and fulvic acids which arc extracted from compost; the fifth component includes nitrogen fixing bacteria along with folic acid and thiazolidine carboxylic acid; and the sixth component includes a silicone based wetting and spreading agent and penetrant which when mixed with the third and fifth components and water is applied as a foliar spray.

2. The method according to claim 1 wherein the first component further includes citric acid thereby reducing a pH range to between 2 and 3 and reducing volatilization of H.sub.2O.sub.2.

3. The method according to claim 1 wherein nanoparticles of the first component are sized in a range of 1-100 nm.

4. The method according to claim 1 wherein the second component further comprises micropropagules of different species of the endo mycorrhiza Glomus, and wherein the consortium of plant growth promoting rhizosphere microbes fix Nitrogen (N).

5. The method according to claim 4 wherein the micropropagules of different species of the endo mycorrhiza Glomus are selected from the group consisting of G. mosseae, G. intraradices, G. aggregatum, G. microaggregatum, G. fasciculatum and G. etinucatum.

6. The method according to claim 4 wherein the consortium of microbes is used either as a single product or divided into two components containing mixtures of rhizosphere microbes.

7. The method according to claim 4 wherein the rhizosphere microbes in the second component are Pseudomonas fluorescens, Bacillus subtilis, Bacillus mycoides (the fungus) Trichodema viride occurring in nature.

8. The method according to claim 4 wherein the concentration of each of the microbes in the second component is between 110.sup.8 and 110.sup.9 CFU per gram, except the endo mycorrhiza which is present at a concentration in a range 50,000 to 150,000micropropagules/gram.

9. The method according to claim 1 wherein the third component is an oil extracted from seed of Karanj (Pongamia glabra/Derris indica) as an emulsifiable concentrate comprising 2% alkaloids extracted from the oil in semi purified form together with oil ranging from 25-35% formulated with petroleum solvents.

10. The method according to claim 1 wherein the fourth component is a slow release, granulated formulation and slow release of the granulated formulation is ensured by incorporating constituents of the formulation in double roasted bentonite granules with a coating of any natural polysaccharide or natural or synthetic polymer.

11. The method according to claim 1 including repeating the steps i) through iii) three or four times between early February and July.

12. The method according to claim 1 wherein the infestation of the crop is by at least one disease selected from the group consisting of Huanglongbing (HLB), Phytopthora citrophthora, Phytophthora Gummosis, Xanthomonas, Pseudomonas Asian Citrus Psyllid, Diaphorina citri Kuwayama and soil dwelling pathogenic nematodes.

13. The method according to claim 1 wherein the infestation of the crop is caused by pathogens on crops selected from the group consisting of potato, tomatoes, egg plants, bell peppers, oil palm, durian, black pepper, blue berries, capsicum, cotton, rice, wheat, chilies, onions, carrots, cucurbitaceous crops, pulses and forest trees.

14. The method according to claim 1 wherein the infestation of the crop is caused by soil borne pathogens selected from the group consisting of Rhizoctonia, Fusarium, Aspergillus, Macrophomina, Pythium, Scerotinia, Verticillium, Alternaria, Magnoporthe grisea, Peronospora, Puccinia, and Cercospora and the crops selected from the group consisting of strawberry, cole crops, celery, spinach, cucumber, melons, squash, beans and peas.

15. The method according to claim 1 wherein the infestation of the crop is caused by an insect pest or nematode, wherein the insect pest is selected from the group consisting of mites, whiteflies and thrips.

16. The method according to claim 1 wherein the infestation of the crop is caused by a disease selected from the group consisting of yellow vein mosaic and leaf curl virus, and the crop is selected from the group consisting of solanaceous crops, cucurbitaceous crops pulses and cotton.

Description

DETAILED DESCRIPTION

(1) As per the studies, the HLB bacteria initially enter the crop roots and multiply, after which they enter the phloem tissue and spread through the entire plant. At the same time their disruptive activities in the roots that lead to tip dying attract another dangerous pathogenphytophthora which is already present in the soil. Phytophthora resides in soil as a weak saprophyte wherever there has been excessive deposition of phosphates and other alkaline salts resulting in elevation of soil pH to 8 or 9. At these levels the natural enemies of this pathogen are in a weakened state. Phytophthora multiplies rapidly after entering the host tree and then it becomes more difficult to manage. Soil dwelling nematodes can also be significant factors that accelerate root decay and lead to the hastened death of the diseased tree.

(2) Treatment Process

(3) a) The first component of the innovative technique is to loosen the grip of both pathogens on the crop by application of a potent biocide that is effective at extremely low doses. To make this basic ingredient, only naturally occurring substances are used and rely on the principle of the disruptive action of heavy metals on the metabolism and reproductive capability of disease causing microbe.

(4) The principle of microencapsulation and finer particle size (near nano size) is used to achieve the objective of using the lowest possible doses. The concentration of the metals in the formulation is at ppm levels and when it is diluted in water and applied, this will be reduced to ppb levels. On entering the plant tissues it is likely to degrade and further reduced to practically undetectable levels.

(5) The extremely fine particle size offers the advantage of not just low dose but also enable the product to reach inaccessible regions of the plant. This enables action at key sites and a rapid reversal of the infection. Since the phloem tissue is blocked, the deeper set xylem cells enable the product to be translocated rapidly to the growing root and shoot tips, once they reach these sites of infection the effect of their strong contact action begins to be noticed. There is a visible darkening of leaf colour as the yellow mottled leaves start to regain their normal appearance.

(6) These nano particles also appear to have a salutary effect on the curling of young shoots caused by the feeding of the nymphs and adult citrus psyllids and the new leaves that emerge are bright green in colour, very shiny in appearance and bereft of disease symptoms.

(7) The metals that can be used are those with proven biocidal action such as silver, copper, iron and more with combinations also possible.

(8) This biocide is to be applied both as a soil drench as well as foliar spray. It works by preventing germination of fungal spores and preventing spread of the disease. Its effectiveness at low doses suggests some positive impact also on the mechanism of crop resistance which enable crops to withstand the attack of pathogens much better.

(9) The critical reasons of the product appears to work so much better than conventional chemical fungicides are the nano sized particles which are able to pass through plant tissues and cell walls very easily and being metals, they pass relatively unchanged to the site where they encounter the pathogens.

(10) The application as a soil drench enables the particles to reach the sites of infection rapidly through xylem tissue which are not blocked by the pathogens. The phloem and xylem tissue lie in close proximity to each other at the growing tips of the roots and shoots and the nano particles are able to reach the phloem and penetrate the masses of bacteria easily stopping the disease in its tracks.

(11) Regular follow up foliar sprays serve to strengthen this effect and the revival of the tree startlingly quickly. Virtually every twig suffering from die back seems to herald the end of the disease by putting forth new buds just at the point where the disease had killed the tissue and new leaves and branches quickly grow to cover the bald spots. The colour and sheen of the leaves reveals that health has been restored. The old diseased leaves drop off quickly and are replaced by healthy new foliage that transform the appearance of the tree within a few months. There is a stark difference observed in the appearance of the emerging inflorescens. The new buds that emerge are pristine white in colour, compared to off white in the diseased plants. The dehiscing of buds is observable in the case of diseased trees, where relatively fewer buds transform into flowers. Brown specks can be observed on the buds, flowers as well as the empty spots in the inflorescens of the diseased trees.

(12) b) The application of the nano biocide is quickly followed by the application of a microbial formulation containing plant growth promoting rhizosphere bacteria that get associated with crop roots in a symbiotic relationship.

(13) The formulation can contain organisms that attack soil borne pathogens and also microbes that facilitate the availability of vital macro and micro nutrients through their biological action. These microbes are known grow rapidly with roots to increase availability of nutrients and release polysaccharides, antibiotics, fulvic and humic acids and plant growth regulators that increase the ability of the crop to withstand stress and disease. These also serve to reduce the soil pH to normal levels which is less suitable to phytophthora and more congenial to beneficial microbes. This product contains microbes that actively suppress soil nematodes further reducing the stress on the crop.

(14) The formulation can contain microbes from among: Thiobacillus, Trichoderma viride, T. harzianum, Bacillus subtilis, Frauteria aurantia, Azotobacter chroococcum, B. megaterium, Pseudomonas fluorescens Beauvaria bassiana, Metarhizium anisopliae, Zinc solubilising bacteria, and many others jointly or singly.

(15) The organisms are carefully chosen keeping in mind the multiple functions required to be performed right from suppression of phytophthora and nematodes to the HLB causal organism over an extended period to reduce the need for frequent application. The beneficial phytochemicals help to suppress C. liberibacter which can be seen by the sudden cessation of the symptoms of HLB like progressive die back of affected shoots and small yellow mottled leaves. The new foliar growth emerges below each and every dead twig, the buds give rise to fresh and luxuriant growth and within a few months the tree recovers completely with the new growth bereft of disease symptoms.

(16) These organisms also improve soil conditions, by reducing soil pH and increasing agglomeration. This increases the moisture holding capacity of the soil and keeps it cooler enabling the crop to withstand moisture and temperature stress.

(17) Another major benefit they provide is to regenerate the root system and help in the formation of vigorous new feeding roots which help the plants to meet the demand for nutrients and water.

(18) By reducing pH to normal levels and increasing solubility of P, helping fix atmospheric N and increasing bio availability of K these microbes prevent the crop from facing a shortage of nutrients during critical stages of growth (including flowering, fruiting and fruit development) thereby helping its swift recovery.

(19) It is observed that trees treated with this system seem to attract fewer psyllids for feeding and also egg-laying. The leaves appear glossier leading us to the conclusion that they will be less desirable to newly hatched nymphs. Similarly the quick reversal of the yellow green colouration of the leaves to a healthy darker green seems to make the trees less attractive to the psyllid.

(20) This formulation out performs conventional chemical fungicides and offers salvation for the farmers in many ways.

(21) Unlike chemical fungicides which are rapidly degraded after application, the living organisms in the product actually multiply, rapidly doubling in numbers every few minutes, thereby reducing the need for repeat application. In the first year just 2 applications a year is needed, which could reduce to just a single annual application from the second year onwards as populations of the beneficial microbes build up.

(22) These microbes constantly evolve to remain a step ahead of the pathogens they compete against, so there is no problem of resistance and the need to resort to increasingly more expensive patented chemicals. Countless millions of poor farmers all over the world will benefit. As informed before, conventional antibiotics are harmful to the plant and cannot be used, however, the phytochemicals (including antibiotics) produced by the product are not harmful and actually help the crop which is evidenced by the extremely good quality of the leaves and produce. Record yields are achieved by a whole range of crops benefiting from this treatment and not just citrus.

(23) A new variant of this product includes a foliar spray so that any spores of the pathogen that are deposited on the foliage can also be subdued with only a few applications a year.

(24) c) The third component comprises an unique formulation containing plant derived amino acids, cytokinins, humic and fulvic acids together with a N fixing microbe is added simultaneously to make the soil more amenable to the growth of the microbes in the soil. This product formulation has proved highly effective as it also provides the plant with much needed N for production of protein that helps the plant to create amino acids to fight the disease.

(25) This product works in a synergistic manner with the other products mentioned above and encourages the rapid growth of microbes enabling it to outstrip conventional products in terms of performance.

(26) For instance trials carried out with both products applied at the time of sowing in some annual crops show a rapid development of the roots within 15 days after germination compared to crops grown using conventional chemicals alone. Root growth is 2-3 times greater and shoots are stronger and able to withstand high temperatures as well as disease like damping off (caused by Fusarium) much better. A 20% higher germination percentage has been observed and crop stands are much better enabling farmers to reduce seed rates and save money. Leaf area and chlorophyll content is better and crops treated with the disclosed method are able to overhaul crops planted 2 weeks earlier in terms of appearance (root development, height, leaf area etc) within a month after sowing.

(27) d) Further the citrus psyllid is controlled by a fourth component which is formulation based on a plant extract that can contain oils like neem, karanj, soap nut, custard apple, lemon grass, clove oil, citronella, ginger and the like having some insecticide repellent properties either singly or in combination with one another. On application it is observed that the citrus psyllid adults and nymphs stop feeding and die. The product interferes with the moulting of the nymphs causing mortality and failure to advance to their next stage of development.

(28) Another advantage of this application that has been observed is the reduction in sooty mould, which shows that the nymphs and adult psyllids have stopped feeding almost immediately after application.

(29) The application of natural oils such as those used in the product help overcome the problem of resistance to chemical insecticides as they contain a cocktail of natural alkaloids.

(30) e) All sprays are applied along with a silicone based super spreading and penetrating agent presently introduced as component-6. This product is extensively tested by leading Government research organisations of India over several seasons. Its use enables better penetration of the products through the bark as well as better spreading and rain fastness, these result in a more economic use of the product due to better efficacy.

(31) f) Another problem encountered due to disease is an increased dropping of immature fruits due to the weakened state of the crop and its inability to withstand high temperatures in the summer months. The shortage of vital photosynthates in the twigs which provide the much needed energy for the developing fruits is one problem. The second problem is the inability of the root system which too is starved of nutrients (due to blockage of phloem) to grow and provide the moisture and dissolved minerals to the plant. This is due to death of root hairs and growing root tips caused by HLB.

(32) Another problem caused by such stresses is the uncoiling of protein structures which provide strength, such as the stalks of flowers and fruits. When the plants are unable to generate enough proteins and amino acids to sustain their flowers and fruits, these drop to the ground.

(33) The application of a product (component 5) containing an amino acid as well as a precursor for a second, helps overcome these problems by enhancing the activity of the metabolic pathways of the plant that help combat moisture stress and disease. The activation of the proline pathways to increase the concentration of this super solute in the plant sap reduces loss of moisture and helps retain turgidity of cells required for preventing senescence and fruit dropping. This product increases the ability of the plant to withstand temperature stress and resist the uncoiling of protein structures thereby enabling the retention of more flowers and fruits and increasing yields as well as quality of produce.

(34) This product further increases pollination and thereby fruiting. Pollinators can sense the presence of proline in nectar which is required by them to power their flight muscles, so increased presence of this amino acid ensures greater visitation of such flowers by pollinators especially early in the morning for their first daily flights when their energy levels are at their lowest.

(35) Proline is a short chain amino acid, also known as imino acids which contain sulphur, these are also activated by trees when confronted with disease and are responsible for developing natural resistance.

(36) The success of this system lies not just with the 6 carefully selected products but also in their timing and sequence of use. The treatment is started when the citrus plant is naturally at its most vigorous stage of growth and the treatment accelerates these processes resulting in the reversal of the course of this deadly disease and both pathogens as well as the vector.

(37) This method encourages the plant's natural defences and uses the synergism of the microbes to ensure the continuous production of beneficial phytochemicals that further encourage this property.

(38) g) Introduction of a concentrated mycorrhizal formulation with added bio catalysts for increasing efficiency of crop root system (in component 2) is a part of the process which needs to be applied just once a year. Global warming leading to rising temperatures and lower rainfall leads to a strain on the root systems of crops to absorb nutrients and moisture. This means that more energy needs to be expended by crop roots, leading to reduced amounts available for flowering and fruiting. Together with deteriorating soil conditions mentioned earlier, this is another reason why yields have stagnated.

(39) Mycorrhizae also protect plants from attacks by pathogens and mobilise critical nutrients like P and K as well as micronutrients. Significant savings in fertilizer application costs can be achieved by farmers.

(40) By introducing a concentrated formulation of endo mycorrhizae in the disclosed method and kit, the twin objectives of healthy crop and soil is achieved together with increased yields.

(41) These advantages have enabled the disclosed method with crop specific variations to be used with success in many other crops such as capsicum, cotton, rice, wheat, chillies, potatoes, capsicum (bell pepper), tomatoes, onions, carrots, cucurbitaceous crops, pulses and many others with resounding success.

(42) This system has proved useful in controlling not just HLB and Phytophthora but also other fungal organisms like Fusarium, Rhizoctonia, Alternaria, Magnoporthe grisea, Peronospora, Puccinia, Cercospora and many others.

(43) This system has also worked well against bacterial diseases like Xanthomonas, Pseudomonas and others.

(44) The system has also provided significantly superior results in deterring pests like mites, whiteflies and thrips in various crops.

(45) However it has provided unprecedented suppression of viral diseases which cannot be controlled by any chemical pesticide. The various viral diseases against which it has proved successful are yellow vein mosaic and leaf curl virus which affect solanaceous crops, cucurbitaceous crops as well as pulses, cotton and many others.

(46) Silver Nano Product Formulation

(47) The production of nano silver particles is an established science. The product differs in the formulation, where activity is enhanced using hydrogen peroxide at a concentration of 2.5% and using additives and stabilisers for a more stable product. The size of the nano particles is anywhere for 1 nM to 100 nM's but is maintained within a range of 1-10 nMs by regulating the current. The aqueous solution of nano silver particles is concentrated to a level of just 50 ppm-100 ppm and it can be diluted to any concentration desired by adding deionized water. Citric acid has been used to reduce the pH range between 2 and 3 and reduce volatilisation of H.sub.2O.sub.2.

(48) Silver is a powerful germicide, however the nano particles make it much more effective at low doses because these tiny particles can pass very easily through cells and reach the site of infection. The fact that these are effective at infinitesimally low concentrations gives the disclosed method a big advantage over chemical fungicides which are required to be used at much higher concentrations in order to be effective.

(49) What is not commonly known is that the bacteria responsible for HLB resides in the roots for a few years before it builds up its population and then grows onwards through the phloem cells till it infests every emerging shoot or root hair. Phloem cells are critical for passing nutrients (photosynthates) produced by leaves to the growing tips of the plant. Since these delicate and small parts of the plant have relatively few phloem cells and low stored energy, they are the first to succumb to the infection.

(50) Since roots are not visible to the farmer, the first symptom of the disease he sees is the die back of the young shoots and erroneously assumes this to be caused by Phytophthora and commences fungicidal application.

(51) As explained earlier, the dying roots attract secondary infection by the fungus like organismPhytophthora, another deadly plant pathogen. This also grows upwards through the dying tissue and later infects the entire plant. Symptoms are seen in the form of oily secretions emanating from the main stem and branches. This disease can cause death of the tree once it girdles the phloem tissue all along the circumference of the main stem or branches. The HLB bacteria have evolved to attract an insect vector, in this case the Asian Citrus Psyllid which is attracted to yellowish colouration of citrus leaves. It is believed that trees infested by this disease also release chemical signals which are also attractive to these vectors. The HLB bacterium causes the leaves of the infected tree to turn yellow and release the chemical signals which attract the psyllid in large numbers enabling the bacterium to spread to healthy trees.

(52) What is also not realised is the nature of the plant's defence mechanism, which relies on the formation of free radicals like nascent oxygen to kill pathogens and also the surrounding tissue which has been infected. The pathogen requires living tissue to proliferate, by killing the surrounding tissue; the plant is able to limit the infection to a very small area. The nascent oxygen quickly kills the pathogen in this encircled tissue which dries up and drops off, freeing the plant from the clutches of disease. By adding hydrogen peroxide to the nano silver formulation and applying this to the plant, several purposes are served: a. The nano silver particles proliferate through the plant tissues especially the roots where they are drenched, reducing the load of inoculum present so there are fewer bacterial cells available to proliferate. b. They also serve to reduce the intensity of attack of Phytophthora c. Silver nano particles are also carried upwards through xylem tissue till they reach the very tips of the dying shoots where they come into contact with the bacterial cells killing them d. The foliar spray enables silver nano particles to pass through infected leaf tissue where bacterial cells are destroyed. e. The application of hydrogen peroxide in small quantities stimulates the plant's defence mechanism to release more nascent oxygen through a complex physiological reaction involving a series of enzymes. This magnifies the impact of even a single application several fold, enabling the plant to show quick recovery. f. The destruction of bacterial cells and the stimulation of the plant's defence mechanism enable the tree to reverse the symptoms of disease. The leaves quickly turn green reducing its attractiveness to the insect vector thereby relieving the secondary damage caused by the insect.

(53) This formulation is the first measure to be used in the said systematic procedure. Soil drenching with the nano silver product is done after preparing the base of the trees receiving the drench for maximum impact. A light irrigation is given in advance so as to reduce the quantity of product required for drenching, depending on the soil type watering 3-4 days in advance of the activity is generally sufficient. The dampness of soil also facilitates easy weeding which is a prerequisite to remove any space for the pathogens to seek shelter.

(54) The best time for initiation of treatment is before flower initiation. In India this is usually the month of February. This will enable farmers to save their crop in the current season/year. The second best time is after the onset of monsoons usually July as this will enable the plants to recover faster and give a much better crop in the next year. Monsoons are also a time when the plant is geared towards producing more leaves and shoots so at this time the procedure enables the plant to recover faster. The rainy season is also favourable for the growth of the beneficial soil microbes since soil temperatures are lower and moisture is adequate.

(55) On DAY ONE: A circular berm of 6 height is prepared around the tree trunk. The area of this berm should encompass the area shaded by the canopy of the tree for maximum coverage of the roots. All weeds within the circumference are to be uprooted and removed from the site. A light irrigation is given so as to raise soil moisture content.

(56) DAY 4: The nano silver formulation is to be used at 10 ml/liter of water and depending on the size of the tree, 15-20 liters of spray solution is to be used for drenching the root zone of each tree. In effect 150 ml-200 ml of the product will be used for each tree. Soil drenching will be required just once a year, sometimes depending on the observed recovery, a second application might never be required.

(57) DAY 4, 34, 64 & 94: A foliar spray is repeated at monthly intervals in heavily infested citrus orchards for 3-4 months to remove any vestiges of infection. The product is applied at 5 ml/liter of water with the addition of a special wetting agent also developed by us containing a concentrated mix of trisiloxane super spreading and penetrating agents. This enables the product to even penetrate bark. This super spreading and penetrating agent is mixed with the spray solution at a concentration of 0.25-0.4 ml/liter of water. Generally 5-6 liters of spray solution is enough to cover a tree but larger volumes can be used on larger trees for thorough wetting.

(58) The super spreading and wetting agent also helps prevent the product being washed off during rains, rendering the disclosed method flexible enough to be used even if there is a break for 2-3 hours in the rain.

(59) Microbial Consortium

(60) The microbes which are used are commonly available soil dwelling rhizosphere microbes which are commonly available as standalone products and are made by many companies. Since the objective was to keep the cost of treatment low, we have used such microbes that are easily available off the shelf. What is unique about this product is the combination of microbes selected, each with different characteristics serving different purposes that have never before been used together to address the entire set of complex issues that prevent farmers from raising a good crop and making profits.

(61) This product formulation is also unique because of the high concentrations of microbes used. This enables greater efficacy at lower doses and improves the survival chances of microbes under inimical soil conditions.

(62) A dextrose carrier has been used for manufacturing this concentrated formulation instead of talc so as to provide readymade energy for the growing numbers of beneficial microbes.

(63) The product can be modified based on an analysis of the soil in different parts of the country to address specific issues that can come up. A standard product could contain in any combination the following organisms: Azotobacter chroococcum, Frateuria aurantia, Pseudomonas aeruginosa, Bacillus subtilis, Trichoderma viride, Pseudomonas fluorescens, Bacillius megaterium, Bacillus mycoides, Metarhizium anisopliae. The concentration of microbes in the formulation would be between 110.sup.8 to 110.sup.9 cfu's/gram. In addition, the consortium can also include a high concentration of endo mycorrhiza with a formulation strength of 50,000-150,000 infective micropropagules per gram of the product.

(64) Depending on the soil and crop, the products can be formulated containing a single or different multiple combinations of the above microbes for reducing cost of application and increased specificity.

(65) The purpose behind this product is to address the problems posed by HLB, Phytophthora, deteriorating soil conditions caused by the present method of agriculture and nematodes that can aggravate the intensity of disease infection.

(66) HLB is caused by bacteria that cannot be controlled by the application of fungicides, which is the common practice resorted by farmers in India assuming this to be an attack of Phytophthora.

(67) HLB can be controlled by antibiotics which are expensive and require to be injected in the stem which makes their use even more laborious. However the major problem posed by the use of antibiotics is their innate phytotoxicity to citrus. The deep seated nature of HLB necessitates the use of a heavy dose of antibiotics which can injure and even kill the citrus tree.

(68) As explained earlier, secondary infection (through roots injured by HLB) caused by Phytophthora compounds the problem, a tree weakened by HLB is easy prey to Phytophthora.

(69) Plant nematodes are soil dwelling pests which also cause serious damage to roots of citrus and lead to secondary infestation by fungal pathogens. Commonly used chemical nematicides are expensive and highly toxic.

(70) The higher soil pH and excessive phosphates also suppress the availability of micronutrients like Zinc which are essential for crop growth. Plants weakened by the deficiency of micronutrients are more susceptible to disease.

(71) The use of heavy farm machinery for intercultivation destroys soil structure and creates a hard pan at a depth of 10-12 beneath the soil surface. This coupled with the low soil organic content of soil results in periodic waterlogged conditions (during flood irrigation and rain events) that favour Phytophthora and other soil pathogens. Reduced soil moisture holding capacity arising from a destruction of soil structures can accentuate the impact of drought and erosion of this irreplaceable resource.

(72) This is accentuated by global warming which is resulting in periods of heavy rain interspersed with periods of high temperature and near drought conditions.

(73) The disclosed microbial consortium has been evolved after a thorough study of all these problems and offers the following benefits: a. Antagonistic agents of Phytophthora and other soil pathogens have been incorporated which suppress these pathogens by antibiosis, predation and parasitisation. Under favourable conditions, they grow much faster than the pathogen and occupy the soil areas near the roots creating a barrier to pathogens. Chemical signals released by the pathogens encourage these microbes to grow towards their colonies and suppress them. b. They chelate nutrients making them unavailable to pathogens yet available to crop roots, reducing the problem of micro nutrient deficiency. c. In a symbiotic relation with crop roots, they continually release a range of beneficial phytochemicals like humic and fulvic acids, antibiotics, growth regulators, cytokinins, auxin and gibberellins that encourage roots to grow faster. They release polysaccharides that bind soil particles creating air spaces for root aeration. d. This continual release of organic material increases soil organic content and restores soil texture and fertility and also prevents erosion. e. Increased moisture holding capacity enables the soil to remain much cooler which benefits the crop immensely and reduces loss of water and nutrients due to leaching. f. They improve the availability of major nutrients like N, P and K which the tree was unable to access due to damage to the root system caused by the pathogens. These enable the tree to fight the disease. g. Through better root growth, they improve the ability of crops to withstand periods of high temperature and drought much better than conventionally raised crops. h. These microbes also suppress nematodes thereby preventing root damage and secondary infection by pathogens. i. The addition of high concentrations of endo mycorrhizae helps improve the efficiency of roots in drawing nutrients by 10 to 100 times, thereby enabling them to withstand stress conditions far better. This allays the adverse effects of disease as well. j. New root growth enables the infected tree to draw more nutrients and reverse the weakness caused by root decay caused by the pathogens.

(74) This product is applied in the following manner: Day 9: Five days after the application of the nano silver product as a soil drench, the microbial consortium is applied at a dilution of 1.5 to 2 grams in 20 liters of water in the same manner as stated above for the nano silver product.

(75) If the first application is given in February, the response of the citrus trees are observed and a second application given after 150 days in the month of July with the onset of the monsoons. If the response after the first application is good, then a second application may not be required.

(76) Plant Extract

(77) Solvent extracts of Derris Indica also known as Pongamia or Karanj plant were tried out on a small scale against sucking insect pests on a range of crops and found to have beneficial properties. The oil from the seed of this plant seed of Karanj plant contains the highest content of the active principles which are primarily plant alkaloids and flavonoids.

(78) The product is made by a conventional solvent extraction process used for any oil seed. The seed kernel is subject to reaction with organic solvents and the main active ingredient, karanjin is isolated. However in order to retain the advantages of the natural product, 30% by weight of the oil that was extracted is incorporated in the formulation. The active principlekaranjin that was isolated earlier is also incorporated at a concentration of 2%-3%.

(79) This combination of active principle and oil is dissolved in a petroleum solvent and a non ionic wetting agent is added to give a stable formulation containing not just the active ingredientKaranjin, but also other alkaloids and flavonoids present in the natural oil.

(80) The disclosed product offers the following benefits over conventional chemical pesticides: a. Being a natural product, it does not leave harmful residues. b. The presence of a cocktail of natural alkaloids and flavonoids with different modes of action reduces the risk of the development of resistance. c. Though this is a natural product, damage caused by the psyllids and other sucking pests like white flies can be stopped immediately through its anti feedant action. d. It is known that Karanjin also works by preventing moulting. This prevents the insects from growing to their next stage. Psyllid nymphs are unable to transform into adults and this breaks the life cycle of the insect and reduces pest pressure. e. Its gentle healing action reduces stress exhibited by plants with chemical insecticides and leaves remain green unlike the dry yellow appearance they take on after repeated chemical insecticide sprays. f. Unlike most chemical pesticides, this product does not harm predators and parasites of the psyllid making this most amenable for. IPM (Integrated Pest Management). g. Citrus psyllids do not merely infect healthy plants with disease, during the course of their feeding; they release toxins (and possibly viruses) into the growing tips of the citrus trees. This causes the leaves to curl and new shoots remain stunted, this has a direct impact on flowering and fruiting. h. These psyllids and their nymphs also release a secretion called honey dew which covers the leaves and attracts the sooty mould fungus which forms a black mat on the leaf surface, completely blocking sunlight and stopping photosynthesis which weakens the plant further. i. This accentuates the yellowing of leaves and attracts even more psyllids to congregate on such trees magnifying the problem and helping the spread of disease. j. The disclosed system takes cognizance of all these complex variables and delivers benefits rapidly by: Enabling the leaves to regain their lustrous green colour thereby discouraging psyllids from congregating there This return to health and increased chlorophyll content helps the plant to photosynthesize and regain its health rapidly Preventing the formation of the mat of sooty mould and encouraging photosynthesis Neutralises the toxins and viruses injected by these pests and enables new healthy growth to emerge bereft of any disease symptoms Increased flowering and fruit setting is observed whenever the system is implemented in time

(81) This product is dissolved in water at a concentration of 2-2.5 ml per liter of water and the super spreading agent and penetrant is added at 0.25-0.4 ml per liter of solution. This product is applied during periods of peak flush of citrus (when new leaves emerge) since this is when the citrus psyllid is most active, attracted by the pale yellowish green coloration of diseased leaves as well as new flush.

(82) Applications are synchronised with the observation of psyllids in the orchard when their populations are at Economic Threshold Levels (ETL's) these can vary from country to country.

(83) Granular Formulation Containing Beneficial Phytochemicals of Plant Origin

(84) Most agricultural soils contain low organic content due to decades of excessive reliance on chemical inputs and mechanisation. The excessive presence of salts arising from chemical fertilisers and irrigation using ground water having salt content is inimical to the soil ecosystem and result in the problem of soil salinity. Often these salts can result in an increase in soil pH and result in soil alkalinity. Both conditions cause a marked reduction in the presence of beneficial microbes and render such soils unfit for cultivation.

(85) If biological products are used in such soils, the survival of the beneficial organisms contained therein is doubtful; so the results obtained from the use of such products will be far from satisfactory.

(86) This problem has been overcome by incorporating a granular formulation that provides a) humic acid b) amino acids c) cytokinins

(87) These products are made from enzymatic extraction from vermicompost, plants and sea weed respectively. These are available off the shelf as standalone solo products. The unique formulation is the use of all 3 components together in a concentrated form reducing the dosage requirements by 40-50% per acre and result in significant cost reductions for the farmer.

(88) By applying this along with the soil microbes, we are encouraging their growth which in turn helps root growth and a substantial increase in soil exudates. This product effectively kick starts this beneficial cycle rapidly restoring soil conditions and ecological balance.

(89) This product comprises all three basic constituents coated onto double roasted bentonite clay granules with a concentration of 15-20% by weight. Application of these ingredients on to the granules can be by means of spray or mechanical mixing using a drum mixer. The bentonite granules play a crucial role in product efficacy as they absorb these products and release them slowly in the soil in the presence of adequate soil moisture in the root zone, preventing excessive release and leaching.

(90) This product can also be supplied as a soluble powder formulation in concentrated form which is preferred for long distance transportation where logistics costs are a significant factor.

(91) This product is applied along with the microbial consortium mentioned earlier so that soil conditions are rendered more amenable for the growth of the beneficial microbes present therein. Even the standalone products are applied primarily for crop growth and never intended to be used along with microbial consortia.

(92) Foliar Spray Comprising Nitrogen Fixing Bacteria Along with Buffer

(93) The disclosed product contains a high concentration of N fixing bacteria, Azotobacter chroococcum present in a concentration of 110.sup.8 to 110.sup.9. The addition of the buffer enables a rapid proliferation of the beneficial microbes present and improved product efficacy. N is an important macro nutrient that is a constituent of proteins, amino acids, nucleic acids, vitamin and other important building blocks of the plant. Without the presence of N, plants would not be able to grow new leaves. The ability of leaves to photosynthesize is severely impaired in the absence of N. Protein is an important constituent of load bearing structures such as the stalks of flowers and leaves. A shortage of N results in flower and fruit dropping and thereby reduced yields. N applied as a chemical fertilizer is highly susceptible to leaching, this results in an apparent shortage during critical stages of crop growth. This foliar spray mitigates these problems and ensures a steady supply even during critical stages of growth like flowering and fruiting.

(94) In order to improve the efficacy of the N fixing microbes, a buffer is provided which contains carboxylic acids like thiazolidine carboxylic acid (TCA) or carboxylic acids that are acetylation by products of TCA along with folic acid in a ratio of 5%:0.1% respectively.

(95) This product is a unique combination of microbes and buffer and is seen to provide several benefits which include: a. Reduced fruit and flower dropping b. Better quality of produce c. Increase drought tolerance d. Improved growth of microbes present and improved product efficacy e. Increased secretion of amino acids by the tree especially sulphur containing amino acids like proline are believed to help in fighting disease f. Proline also attracts pollinators which improves fruit set g. Higher presence of N in leaves also helps them to turn green faster thereby reducing the attractiveness of the tree for egg laying by the citrus psyllid. This in turn reduces the population of nymphs on the tree and reduces the risk of secondary infection.

(96) The said product is applied during periods of excessive heat and dryness usually between March and June which also coincides with flowering and fruit initiation. Sprays are continued at intervals of 30-45 days depending on agro climatic conditions. A maximum of 3-5 sprays may be given during the course of the year.

(97) If the botanical extract of Karanjin has been applied earlier, it is advised that this product be applied after a gap of 15 days.

(98) Observations

(99) A reversal of symptoms is observed within 3-4 weeks after application of the 2nd step on day 9. The progression of the disease is seen to stop and the branches put forth new buds showing the infected area is regaining its health. Three months after Step 2 and application of all other intermediate steps listed above, it is observed that the bald patches of the tree are getting covered with new branches. Improved flowering and visitations by pollinators is observed Restoration of the environment is seen by increased presence of predators like spiders and beetles which prey on the sucking pests like the psyllid and its nymphs. Shoot growth is seen to be rapid and leaves emerging from the new shoots are healthy and shiny. The chlorophyll content of the leaves appears to be much higher and the leaf size is 2-3 times larger than the diseased older leaves. Increased flowering and pristine white coloration of buds and flowers is observed in contrast with mottled disease infested buds and flowers seen in conventionally raised crops. Flower dropping is visibly reduced and fruit setting is heavy. It is observed that yields reach pre infestation levels in the same year if treatment is done in February and increase further in the 2nd year with a continuation of the treatment.
Trials:

(100) Several field trials have been conducted over the years in North India and 2 examples are given herewith

(101) TRIAL 1 Village Lilawali, District Hanumangarh, Rajasthan.

(102) A severely infested orchard was selected after confirming symptoms of HLB and Phytophthora. The incidence of disease was so severe that the farmer was contemplating uprooting his entire orchard. Income was Rs 30,000 ($517) from the entire orchard for the year.

(103) Number of trees: 600 with 100% infestation.

(104) Trial commenced June Observation by December: Almost 80% recovery with 482 trees having completely recovered and in good fruit bearing condition 95 trees, 15% of the population were on the way to recovery with partially girdled branches still showing disease symptoms and 23 trees or about 4% of the trees were too diseased to recover and had to be uprooted.

(105) Income increased to Rs 3 lacs for the year (approximately $5000)

(106) Diseased branches were pruned and trials continued for a second year

(107) Observation December

(108) Remaining 577 trees appeared to be almost completely recovered and farmer reported increased income to Rs 5 lacs (approximately $8600) Trees started showing fresh growth within 2 months after applying 2.sup.nd STEP Leaf size was much larger and leaves were shiny without mottling symptoms Leaf curling due to psyllid feeding was absent Very few leaves showed traces of sooty mould fungus, indicative of low infestation by citrus psyllids Large number of spiders and webs seen with insects trapped showing a restoration of the ecological balance of the orchard Fruit quality was good and fruits were larger in size.
Trial 2

(109) Village Lakhian (5-O), District Sri Ganganagar

(110) The farmer had grown citrus over 1.25 hectares and his income had decline from Rs 250,000 ($4300) in 2012 to Rs 130,000 ($2240) in 2013. He was assuming further deterioration and the prospect of having to uproot his orchard. All trees showed symptoms of HLB and 80 trees exhibited phytophthora gummosis.

(111) The trial started in February with the farmer agreeing to take up the complete system of treatment. Out of a total of 312 trees, 263 had survived.

(112) The 1.sup.st observations were taken on end March and all trees had recovered remarkably well within a month of the 2.sup.nd STEP. Heavy flowering was observed with increased bee visitation.

(113) Application of STEP 3, 4 and 5 proceeded on schedule and heavy fruit bearing was observed. There was some damage to fruits caused by hailstorms in May leading to some losses but the progress of the orchard was clearly visible.

(114) No HLB symptoms were to be seen on the leaves, there were no bald patches in their crowns (indicative of die back) by end May.

(115) An attractive bid of Rs 4,10,000 ($7,100) was received by the farmer in June for his orchard, an increase of income 215% over the previous year.

(116) Although the foregoing description of the present invention has been shown and described with reference to particular embodiments and applications thereof, it has been presented for purposes of illustration and description and is not intended to be exhaustive or to limit the invention to the particular embodiments and applications disclosed. It will be apparent to those having ordinary skill in the art that a number of changes, modifications, variations, or alterations to the invention as described herein may be made, none of which depart from the spirit or scope of the present invention. The particular embodiments and applications were chosen and described to provide the best illustration of the principles of the invention and its practical application to thereby enable one of ordinary skill in the art to utilize the invention in various embodiments and with various modifications as are suited to the particular use contemplated. All such changes, modifications, variations, and alterations should therefore be seen as being within the scope of the present invention as determined by the appended claims when interpreted in accordance with the breadth to which they are fairly, legally, and equitably entitled.