Title of Invention | A COMPOSITION USEFUL FOR THE PROPOGATION OF PHRAGMITES SPECIES" |
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Abstract | A composition useful Eor the propogation of Phraqmites species which comprises mixture of the following ingredients in the specified ranges mentioned below: a) Major elements Concentration mg/L In the range of K"+ 250 to 1090 NH+ 125 to 365 Mg++ 22 to 41 Ca++ 4 0 to 125 NO3- 893 to 2800 CI 80 to 241 SO4-- 72 to 159 PO4- 82 to 147 |
Full Text | This invention is related to composition useful for the propagation of PHRAGMITES spp. More particularly it is related to nutrient media which could be used for the mass propagation by shoot sprouting & plantlet production of the said species using Tissue culture techniques. The Phraqmites spp. has an important application in effluent treatment of industries. Hence its large scale propagation assumes great significance. The tissue culture method has been successfully employed for production of this species. In recent years, due to rapid industrialization and population, the degree of pollution is increasing at a very high rate. Treatment of waste water has become a global concern. Now a days, major efforts have been undertaken to develop waste water treatment techniques which are effective at removing both organic and inorganic contaminants so that waste water discharge and water quality standards can be met. In many cases tertiary or advanced waste water treatment processes are needed to meet these water quality goals. However, advanced waste water treatment systems are generally energy intensive, expensive and relatively ineffective for the removal of many trace contaminants. Innovative waste water treatment systems are therefore needed to amend this situation, which should be cost effective as well. One class of treatment system which has recently been proposed for advanced waste water treatment is aquatic treatment or "Rootzone system". This is a low cost, low energy demanding solution to waste water treatment. Aquatic treatment systems are ones in which wastewater is applied to natural or man-made wetlands or to basins containing free floating or submerged aquatic macrophytes. These systems not only remove organic matter but also the other nutrients (nitrogen and phosphorus), heavy metals and traced organelles. They are also biomass generaters which may generate plant material for anaerobic digestion and methane production. In these systems, treatment occurs through a combination of biological, physical and chemical processes which are similar to those occuring in conventional secondary treatment processes. The aquatic macrophytes play an essential role in aquatic treatment systems. The roots, stems and leaves of the plants in the water column provide a surface on which bacteria can grow and provide a media for degradation and adsorption of solids. They also provide oxygen to the bacteria through photosynthesis. The stems and leaves at or above the water surface help to attenuate sunlight and thus prevent growth of suspended algae and reduce effects of wind on the water, thus minimizing roiling of sediments. So also, heavy metals and nutrients are assimilated by these plants. The extensive 'root zone of many species of the genus Phragmites viz. P-- australis , P- communis, P. karka etc. are used for the treatment of wastewater. With the growing demand of efficient treatment and large quantity of water coming out from various industries, rapid propagation of this plant species is very much needed for quick application and also to meet the increasing demand of today's industrialization. Use of naturally occuring species like Phragmites in such treatments in Industry are also envi-ronmment friendly. One of the ways to acheive mass propagation of the species needed for the treatment, is the use of tissue culture techniques. with this background in mind, the inventors in the present invention initiated work on developing a tissue cluture medium for the mass propagation of the genus Phragmites. Phraqmites Trin. is a genus of tall perennial grasses of the family Gramineae. Many species of this genus have thick weeping rhizome and are found on the banks of lakes and streams. The species are used in the manufacture of pulps for rayon and paper, employed for thatching and making mats, baskets, chairs, fences , fish traps etc. The culms are used for 'huk-aah' pipes, flutes and pens. The panicles are used for brooms. The seed when young is eaten by cattle. The roots are reported to be used in medicines, especially in East Asia , they are reported to be employed in rheumatic complaints and diabetes and are used for fractured bones. Many compositions for propogation of various plants by tissue culture techniques are available. However the same compositions are not found applicable to Phraqmites. The main object of the present invention therefore is to develop composition (medium) which will stimulate shoot and plantlet production utilizing explants from known clumps. As a result, new compositions (media) for culturing Phragmites species inducing multiple shoots and rooting has been developed and also complete plantlets have been successfully transferred to field. The development of such media also ensures a steady supply of plants throughout the year without having any dependence on. the availability of any new material. This composition developed by the present invention has been used for various species of Phragmites like P^ australis , P^ karka P. communis. Compositions for the said Phragmites spp. bas been developed for the first time and hence there is no prior compositions. A basic composition (medium) containing inorganic salts added at macro and micro levels and other organic constituents like sucrose, amino acids etc and formulated by Murashige and Skoog (MS) in 1962, is described in composition I. This basal medium alongwith different concentration of harmones have been used for tissue culture of tomato, teak, eucalyptus, rose wood, ferns, gerberas etc. However the same could not be used for Phragmites species. Composition I Concentration mg/L a) Major elements K++ 734.91 NH4+ 365.65 Mg*+ 36.48 Ca++ 119.95 NO3" 2422.00 CI 212.21 SO4-- 144.20 PO4-- 118.63 b) Minor elements B 1.08 Mn 5.49 Fe [As a chelate with 3.78 ethylene diamine tetra acetic acid ] Co 0.006 Cu 0.006 Zn 2.46 Mo 0.1 I 0.63 c) Organic components Sucrose 20,000 Myo-inositol 100 Thiamine hydrochloride 0.1 Nicotinic acid 0.5 Pyridoxine hydrochloride 0.5 Glycine 2 . 0 Edamin 1,000 Accordingly, the present invention provides a composition useful for the propogation of Phragmites species which comprises mixture of the following ingredients in the specified ranges mentioned below: a) Major elements Concentration mg/L In the range of K+ 250 to 1090 NH4+ 125 to 365 Mg++ 22 to 41 Ca+ 40 to 125 NO- 893 to 2800 CI 80 to 241 SO4-- 72 to 159 PO4- 82 to 147 b) Minor elements In the range of B 0.52 to 1.32 Mn 2.90 6.65 Fe [As a chelate with 2.20 5.20 ethylene diamine tetra acetic acid ] Co 0.002 0.008 Cu 0.0018 0.0075 Zn 0.91 2.81 Mo 0.07 0.17 I 0.45 0.82 c) Organic components In the range of Sucrose 15000 50000 Glucose 15000 50000 Mannitol 15000 50000 Fructose 15000 50000 Myo-inositol 75 200 Thiamine hydrochloride 0.07 0.13 Nicotinic acid 0.30 0.80 Pyridoxine hydrochloride 0.25 0.75 Glycine 0.80 3.00 Kinetin 0.1 0.5 Benzyl adenine 0.1 0.5 Malt extract 25 200 Yeast extract 25 200 Coconut milk 2 0 70 Agar agar 4000 5000 for solidification •* - added as mL/L The stimulatory action of this composition in enhancing bud sprouting, multiplication and root initiation is attributed to the synergistic action of the different combinations of ingredients added in the composition in a particular manner. These could cause a higher photosynthetic activity and/ or a modification in cellular structure permitting uptake of nutrients due to increased permeability of the cell membrane. The levels of the mineral salts and other organic growth substances also play a prominent role in this synergistic effect, which results in the unexpected physiological response. The composition can be prepared by any of the standard tissue culture methods. Stock solutions of major and minor elements were prepared as a ten times concentrated solution initially and was diluted to give the required strength. To this final diluted solution, other organic constituents can be added, individually or in combination at different concentrations specified above. The composition of the present invention works successfully for sprouting, multiplication and root induction. In order to elongate roots and shoots, the multiple shoots with root primordia obtained on this Composition have to be transferred to other known composition for root and shoot elongation which comprises the following ingredients. Composition III Concentration mg/L in the range of a) Major elements K+ 100 to 900 NH4+ 45 275 Mg++ 10 35 Ca++ 18 100 NO3" 388 1800 CI 40 150 SO4 - 3 0 93 PO4-- 35 100 b) Minor elements B 0.20 1.00 Mn 1.25 3.65 Fe [As a chelate with ethylene diamine tetra acetic acid ] 1.00 3.05 Co 0.001 0.004 Cu 0.0008 0.0042 Zn 0.40 1.41 Mo 0.028 0.09 I 0.20 0.50 c) Organic components Sucrose 10000 25000 Glucose 10000 25000 Mannitol 10000 25000 Fructose 10000 25000 Myo-inositol 40 100 Thiamine 0.035 0.07 hydrochloride Nicotinic acid 0.20 0.50 Pyridoxine 0.15 0.4 0 hydrochloride Glycine 0.40 1.50 The compositions developed in the present invention are described herein below as examples which are illustrative only and should not be construed to limit the scope of the invention. COMPOSITION FOR SPROUTING, MULTIPLICATION AND ROOT INITIATION OF THE SHOOTS Example I Example II Concentration mg/L Concentration mg/L a) Major elements K+ 300 890 NH4+ 225 125 Mg*+ 30 35 Ca++ 55 100 NO3" 1935 1443 CI 150 120 SO4" 72 125 PO4" 100 82 b) Minor elements B 0.92 0.82 Mn 4.25 5.65 Fe [As a chelate with ethylene diamine tetra acetic acid ] ' 3.40 4.20 Co 0.005 0.006 Cu 0.0062 0.0075 Zn 1.91 2.50 Mo 0.10 0.15 I 0.65 0.45 c) Organic components Sucrose 30000 20000 Mannitol - 15000 Myo-inositol 100 150 Thiamine 0.13 0.09 hydrochloride Nicotinic acid 0.63 0.42 Pyridoxine 0.50 0.67 hydrochloride Glycine 1.90 2.5 0 Kinetin 0.15 0.20 Benzyl adenine 0.10 0.2 0 Malt extract - 75 Coconut milk 50 Agar Agar 4000 4000 for solidification * - added as mL/L Nodal buds were collected from mature clumps, cleaned and sterilized with ethanol , a detergent antiseptic solution and lastly with HgCl2 (0.05 - 1.5% w/v), The buds were then inoculated on sprouting medium solidified with agar. After getting sufficient sprouts at 25± 2°C unde 500-1500 lux light intensity with 16h light and 8h dark photoperiod, the sprouted buds were transferred in the liquid medium and kept for incubation till sufficient multiples and root initiation was observed. Subculturing within 18-25 days was then carried out on the same medium. After getting the sprouting, multiplication androotinduction in the composition described in examples I and II, the shoots were separated and transferred on the composition described in examples III and IV for shoot and root elongation. Example - III Example IV Concentration mg/L Concentration mg/L a) Major elements K+ 100 500 NH+ 150 200 Mg4+ 25 10 Ca++ 50 25 NO3- 438 1000 CI 122 88 SO4- 40 62 PO4 - 60 70 b) Minor elements B 0.50 1.00 Mn 1.65 2.60 Fe [As a chelate with EDTA ] 2.00 1.00 Co 0.002 0.004 Cu 0.003 0.0009 Zn 0.80 1.2 0 Mo 0.080 0.050 I 0.30 0.20 c) Organic components Sucrose 10000 20000 Glucose 10000 Myo-inositol 80 50 Thiamine 0.06 0.05 hydrochloride Nicotinic acid 0.30 0.50 Pyridoxine 0.25 0.35 hydrochloride Glycine 0.80 1.00 Incubation was done with the photoperiod and temperature as described for sprouts. After root and shoot elongation (15-25 days) on this composition, the plantlets were transferred to a soil : sand (1:1) mixture. The results of the present invention are very striking resulting in prolific shoot multiplication on subculture. This immediately opens up the possibility for around the year production of the plants which can be used to cover a larger area of wet lands for wastewater treatment as and when needed. We claim : 1} A composition useful for the propogation of Phragmites species which comprises mixture of the following ingredients in the specified ranges mentioned below: a) Major elements Concentration mg/L In the range of K+ 250 to 1090 NH4+ 125 to 365 Mg4+ 22 to 41 Ca++ 40 to 125 NO3- 893 to 2800 CI 80 to 241 SO4- 72 to 159 PO4-- 82 to 147 b) Minor elements In the range of B 0.52 to 1.32 Mn 2.90 6.65 Fe [As a chelate with 2.20 5.20 ethylene diamine tetra acetic acid ] Co 0.002 0.008 Cu 0.0018 0.0075 Zn 0.91 2.81 Mo 0.07 0.17 I 0.45 0.82 c) Organic components In the range of Sucrose 15000 50000 Glucose 15000 50000 Mannitol 15000 50000 Fructose 15000 50000 Myo-inositol 75 200 Thiamine hydrochloride 0.07 0.13 Nicotinic acid 0.30 0.80 Pyridoxine hydrochloride 0.25 0.75 Glycine 0.80 3.00 Kinetin 0.1 0.5 Benzyl adenine 0.1 0.5 Malt extrat 25 200 Yeast extract 25 200 Coconut milk 20 70 Agar agar 4000 5000 for solidification * - added as mL/L 2) Compositions useful for the propogation of Phragmites species as substantially descreibed hereinbefore with reference to examples. |
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1709-del-1997-complete specification (granted).pdf
1709-del-1997-correspondence-others.pdf
1709-del-1997-correspondence-po.pdf
1709-del-1997-description (complete).pdf
Patent Number | 186894 | ||||||||||||
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Indian Patent Application Number | 1709/DEL/1997 | ||||||||||||
PG Journal Number | 48/2001 | ||||||||||||
Publication Date | 01-Dec-2001 | ||||||||||||
Grant Date | 26-Jul-2002 | ||||||||||||
Date of Filing | 24-Jun-1997 | ||||||||||||
Name of Patentee | COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH | ||||||||||||
Applicant Address | RAFI MARG, NEW DELHI-110001, INDIA. | ||||||||||||
Inventors:
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PCT International Classification Number | C02F 5/00 | ||||||||||||
PCT International Application Number | N/A | ||||||||||||
PCT International Filing date | |||||||||||||
PCT Conventions:
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