Title of Invention

A COMPOSITION USEFUL FOR THE PROPOGATION OF PHRAGMITES SPECIES"

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.

Documents:

1709-del-1997-abstract.pdf

1709-del-1997-claims.pdf

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

1709-del-1997-form-1.pdf

1709-del-1997-form-2.pdf


Patent Number 186894
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:
# Inventor's Name Inventor's Address
1 RAJANI SATISH NADGAUDA NATIONAL CHEMICAL LABORATORY, PUNE, MAHARASHTRA, INDIA.
2 MURARI MOHAN JANA NATIONAL CHEMICAL LABORATORY, PUNE, MAHARASHTRA, INDIA.
3 INSIYA SAIFUDDIN POONAWALA NATIONAL CHEMICAL LABORATORY, PUNE, MAHARASHTRA, INDIA.
PCT International Classification Number C02F 5/00
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA