USPP13110P2 - Lippia alba plant named ‘Bhurakshak’ - Google Patents

Lippia alba plant named ‘Bhurakshak’ Download PDF

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USPP13110P2
USPP13110P2 US09/538,341 US53834100V USPP13110P2 US PP13110 P2 USPP13110 P2 US PP13110P2 US 53834100 V US53834100 V US 53834100V US PP13110 P2 USPP13110 P2 US PP13110P2
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plant
bhurakshak
citral
lippia
artificial sequence
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US09/538,341
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Sushil Kumar
Janak Raj Bahl
Ravi Prakash Bansal
Shri Niwas Garg
Arif Ali Naqvi
Suman Preet Singh Khanuja
Ajit Kumar Shasany
Mahendra Pandurang Darokar
Vikram Singh
Sweta Sinha
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Council of Scientific and Industrial Research CSIR
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Council of Scientific and Industrial Research CSIR
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/02—Flowers
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/12—Leaves
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/86—Verbenaceae, e.g. Verbena

Definitions

  • the invention relates to a new and distinct mutant plant of Lippia alba named ‘Bhurakshak’ characterized by its aroma has been developed.
  • the plant possesses a novel aroma combining linalool, citral a and citral b as the major constituents in its oil.
  • the present invention relates to a new and distinct chemotype of Lippia alba , a member of the family Verbenaceae.
  • the genus Lippia is known to consist of about 200 species of herbs, shrubs and small trees, widely distributed in tropical to semi-temperate areas of America, Africa and Asia. Materials in various forms and essential oils of Lippia species have been used in the traditional medicine and control of pest infestations in food grains. Specific strains are recognized as resources of several commercially important terpenoid compounds.
  • the present invention provides a new and distinct mutant plant of Lippia alba called ‘Bhurakshak’, characterized by novel aroma of the oil constituting 7.2% 1,8 cineole, 42.3% linalool, 12.9% citral (b) and 14.2% citral (a) as a major terpenoids, and having a unique RAPD profile as shown in FIG. 1 of the drawings.
  • the new chemotype of Lippia alba named ‘Bhurakshak’ has been developed through a planned mutation breeding program conducted at Central Institute of Medicinal and Aromatic Plants, Lucknow with an aim to create genotypes with novel aroma suitable for cosmetic and soap industry.
  • a spreading shrub variety ‘Kavach’ of Lippia alba which can be grown on waste land, ravines and slopes to check soil erosion, was selected for mutation breeding.
  • a total of 17,700 single nodal stem cuttings were y-irradiated with different doses (1 kR to 5 kR) during July, 1999.
  • Out of the 3641 surviving plants, 1360 were visually selected after smelling and hydrodistilled in clevenger's apparatus during October 1999.
  • a plant with number CIMAP/L-640 possessing a novel aroma has been identified with 1,8 cineole (9.6%), linalool (41.7%), citral b (13.5%), and citral a (15.3%) in its essential oil as the major terpenoids.
  • the GC on oil sample was performed using varian gas chromatograph model CX-3400 under the following conditions carrier gas hydrogen, injector and detector temperatures 220° and 225° C., respectively; capillary column (supelcovax-10, 30 m ⁇ 0.3 mm, film thickness 0.20 ⁇ m); and oven temperature programmed from 80-150° C. @ 5° C./min., then 215° C. @ 7° C./min. The area percentage were obtained on Varian 4400 integrator.
  • This selected plant was multiplied through stem cuttings during the first week of November, 1999. Out of the 14 plants, five randomly selected plants were hydrodistilled in the first week of February 2000 and analyzed for its oil constituents on Gas Chromatograph following similar conditions of GLC. The percentage of major terpenoids in the essential oil were similar to that of the parent mutant plant having 1,8 cineole 7.2%, linalool 42.3%, citral a 14.2% and citral b 12.9% confirming its true breeding behavior.
  • the mutant genotype L-640 has remained stable and uniform for its morphological characters and showed consistency in performance for its oil quality attributes during its evaluation and vegetative multiplication.
  • FIG. 1 shows a RAPD profile of ‘Bhurakshak.’
  • FIG. 2 shows a close up of the flowering shoot of ‘Bhurakshak.’
  • the genotype CIMAP/L-640 (‘Bhurakshak’) possesses a unique novel aroma combining 1,8 cineole (7.2%), linalool (42.3%), citral a (14.2%) and citral b (12.9%) altogether. No other Lippia alba genotype has been reported to possess these terpenoids in this proportion to give a special novel aroma.
  • a typical RAPD profile of ‘Bhurakshak’ generated by 13 primers whose decamer sequences are given in FIG. 1 was generated, which clearly distinguished the genotype from existing accessions of Lippia alba was established and is given in FIG. 1 .
  • Leaf Ovate-oblong or elliptic-oblong, leaf tip acute, margin finely serrate, base truncate, softly strigose, olderleaves dark green and younger ones light or pale green, size: length 3-10 cm and width 2-5 cm. (FIG. 2 ).
  • Inflorescence Flower sessile arranged in axillary dense capitate spikes, corolla light purple, pink or violetish with yellow throat.
  • Citral a ( geranial ).—14.2.
  • the mutant clone possesses a novel aroma with oil constituents which are intermediate between the two other accessions in collection.
  • the mutant genotype CIMAP/L-640 (‘Bhurakshak’) was bred at the Central Institute of Medicinal and Aromatic Plants (CUVIAP), Lucknow, India under the genetic improvement program on Lippia.
  • the mutant genotype was bred from a parent plant christened ‘Kavach’.
  • the parent plant Kavach was released by Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India and is now in the public domain.
  • the plant material of ‘Kavach’ (parent plant) is also made available to the public through National Gene Bank of CIMAP.
  • CIMAP is a non-profit Governmental Organization having the facility for multiplying the plant material and there is no restriction to the access and supply of the plant material of the parent plant.
  • the parent plant Kavach is not patented.
  • the distinct mutant plant of Lippia Alba named “Bhurakshak” is planted during the month of July when the maximum temperature remains between 35-42° C. under hot and sunny conditions and the minimum temperature between 27-32° C.
  • the cuttings after gamma-rays treatment are planted immediately and the cuttings establish in the soil within a week by the growth of roots and the plants grow thereafter.
  • the plants are grown by asexual reproduction through stem cutting, in the farm of Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India.
  • Corolla Five petals, gamopetalous, bilipped(4/1), coronary outgrowth present at the throat of corolla.
  • Androecium Four stamens, didynamous, epipetalous, polyandrous, dorsifixed situated at the throat of corolla and discrete.
  • Gynoccium Bicarpellary, syncarpous, ovary superior, bilocular with one ovule in each locule, axile placenta, terminal style and capitate stigma.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physiology (AREA)
  • Botany (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

A new and distinct mutant plant of Lippia alba called "Bhurakshak,' characterized by novel aroma of the oil constituting 7.2% 1, 8 cineole, 42.3% linalool, 12.9% citral (b) and 14.2% citral (a) as a major terpenoids, and having a unique RAPD profile as shown in FIG. 1 of the drawings.

Description

FIELD OF INVENTION
The invention relates to a new and distinct mutant plant of Lippia alba named ‘Bhurakshak’ characterized by its aroma has been developed. The plant possesses a novel aroma combining linalool, citral a and citral b as the major constituents in its oil.
BACKGROUND OF THE INVENTION
The present invention relates to a new and distinct chemotype of Lippia alba, a member of the family Verbenaceae. The genus Lippia is known to consist of about 200 species of herbs, shrubs and small trees, widely distributed in tropical to semi-temperate areas of America, Africa and Asia. Materials in various forms and essential oils of Lippia species have been used in the traditional medicine and control of pest infestations in food grains. Specific strains are recognized as resources of several commercially important terpenoid compounds.
Certain Lippia alba wild populations and their cultivated clones have been reported to yield essential oils rich in linalool or terpenoids such as citral and cineole closely related to linalool (Terblanche and Kornelius, 1996. J. Essent. Oil. Res., 8: 471-485). A new chemotype is developed through mutation breeding which has all the four constituents in appreciable amounts to give a novel aroma, which can find wider usage in the cosmetic and soap industry.
SUMMARY OF THE INVENTION
The present invention provides a new and distinct mutant plant of Lippia alba called ‘Bhurakshak’, characterized by novel aroma of the oil constituting 7.2% 1,8 cineole, 42.3% linalool, 12.9% citral (b) and 14.2% citral (a) as a major terpenoids, and having a unique RAPD profile as shown in FIG. 1 of the drawings.
DETAILED DESCRIPTION
The new chemotype of Lippia alba named ‘Bhurakshak’ has been developed through a planned mutation breeding program conducted at Central Institute of Medicinal and Aromatic Plants, Lucknow with an aim to create genotypes with novel aroma suitable for cosmetic and soap industry. For this purpose, a spreading shrub variety ‘Kavach’ of Lippia alba which can be grown on waste land, ravines and slopes to check soil erosion, was selected for mutation breeding. A total of 17,700 single nodal stem cuttings were y-irradiated with different doses (1 kR to 5 kR) during July, 1999. Out of the 3641 surviving plants, 1360 were visually selected after smelling and hydrodistilled in clevenger's apparatus during October 1999.
A plant with number CIMAP/L-640 possessing a novel aroma has been identified with 1,8 cineole (9.6%), linalool (41.7%), citral b (13.5%), and citral a (15.3%) in its essential oil as the major terpenoids. The GC on oil sample was performed using varian gas chromatograph model CX-3400 under the following conditions carrier gas hydrogen, injector and detector temperatures 220° and 225° C., respectively; capillary column (supelcovax-10, 30 m×0.3 mm, film thickness 0.20 μm); and oven temperature programmed from 80-150° C. @ 5° C./min., then 215° C. @ 7° C./min. The area percentage were obtained on Varian 4400 integrator. This selected plant was multiplied through stem cuttings during the first week of November, 1999. Out of the 14 plants, five randomly selected plants were hydrodistilled in the first week of February 2000 and analyzed for its oil constituents on Gas Chromatograph following similar conditions of GLC. The percentage of major terpenoids in the essential oil were similar to that of the parent mutant plant having 1,8 cineole 7.2%, linalool 42.3%, citral a 14.2% and citral b 12.9% confirming its true breeding behavior.
EVIDENCE OF UNIFORMITY AND STABILITY
The mutant genotype L-640 has remained stable and uniform for its morphological characters and showed consistency in performance for its oil quality attributes during its evaluation and vegetative multiplication.
TYPE AND FREQUENCY OF VARIANTS DURING REPRODUCTION AND MULTIPLICATION
No variant had been found among the progeny during multiplication through vegetative multiplication.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 shows a RAPD profile of ‘Bhurakshak.’
FIG. 2 shows a close up of the flowering shoot of ‘Bhurakshak.’
STATEMENT OF DISTINCTION
The genotype CIMAP/L-640 (‘Bhurakshak’) possesses a unique novel aroma combining 1,8 cineole (7.2%), linalool (42.3%), citral a (14.2%) and citral b (12.9%) altogether. No other Lippia alba genotype has been reported to possess these terpenoids in this proportion to give a special novel aroma. A typical RAPD profile of ‘Bhurakshak’ generated by 13 primers whose decamer sequences are given in FIG. 1 was generated, which clearly distinguished the genotype from existing accessions of Lippia alba was established and is given in FIG. 1.
OBJECTIVE DESCRIPTION OF THE MUTANT CLONE L-640 (‘BHURAKSHAK’)
1. Genus: Lippia.
2. Species: Alba.
3. Family: Verbenaceae.
4. Plant height (cm): 70-80.
5. Growth: Spreading shrub, much branched.
6. Stem: Hard, brownish green.
7. Stem diameter (cm) (at 5th-6th node): 0.8.
8. Leaf: Ovate-oblong or elliptic-oblong, leaf tip acute, margin finely serrate, base truncate, softly strigose, olderleaves dark green and younger ones light or pale green, size: length 3-10 cm and width 2-5 cm. (FIG. 2).
9. Leaf/stem ratio: 0.38.
10. Inflorescence: Flower sessile arranged in axillary dense capitate spikes, corolla light purple, pink or violetish with yellow throat.
11. Fruits: Globose, dry, splitting into two 1-seeded pyrenes.
12. Oil content in the fresh herbage (%): 0.14.
13. Active constituents in the essential oil (%):
1,8 cineole.—7.2.
Linalool.—42.3.
Terpinen- 4 -ol.—1.1.
Citral b (neral).—12.9.
Citral a (geranial).—14.2.
Geranyl acetate.—1.9.
Geraniol.—1.0.
14. Seed length (mm): 1.9.
15. Seed width (mm): 1.0.
16. Seed color: Green group (137B).
17. Length of flower (cm): 1.43.
18. Diameter of flower: 0.73.
19. No. of leaves/branches: 28.
20. No. of branches/plant: 2.
21. Leaf texture: Rough.
22. Arrangement of leaves: Opposite.
23. Length of branches and stem (cm): 142.3.
24. Internode length (cm): 10.2.
25. Stem:
a. Texture.—Rough.
b. Diameter (cm).—0.8.
26. Petiole length (cm): 0.68.
27. Color of upper surface of leaf: Green group (137C).
28. Color of lower surface of leaf: Green group (138B).
All references to color herein refer to The Royal Horticultural Society Chart. The green foliage, i.e., the leaves and the stem, of the plant are used to obtain the essential oil. Cuttings of the plant establish in soil within a week.
ADDITIONAL DESCRIPTION OF THE STRAIN
The mutant clone possesses a novel aroma with oil constituents which are intermediate between the two other accessions in collection.
TABLE 1
Major terpenoids in the essential oil of available
accessions including parent plant and the mutant clone.
Active constituents LAC-2 New mutant
Sl. No. in the oil LAC-1 Parent clone Clone L-640
1. 1,8 Cineole 0.5 5.0 7.2
2. Linalool 1.3 64.9 42.3
3. Terpinen-4-ol 1.1 2.5 1.1
4. Citral b (neral) 20.3 1.7 12.9
5. Citral a (geranial) 28.2 2.0 14.2
6. Geranyl acetate 0.3 0.6 1.9
7. Geraniol 1.8 3.7 1.0
The mutant genotype CIMAP/L-640 (‘Bhurakshak’) was bred at the Central Institute of Medicinal and Aromatic Plants (CUVIAP), Lucknow, India under the genetic improvement program on Lippia. The mutant genotype was bred from a parent plant christened ‘Kavach’. The parent plant Kavach was released by Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India and is now in the public domain. The plant material of ‘Kavach’ (parent plant) is also made available to the public through National Gene Bank of CIMAP. CIMAP is a non-profit Governmental Organization having the facility for multiplying the plant material and there is no restriction to the access and supply of the plant material of the parent plant. The parent plant Kavach is not patented.
The distinct mutant plant of Lippia Alba named “Bhurakshak” is planted during the month of July when the maximum temperature remains between 35-42° C. under hot and sunny conditions and the minimum temperature between 27-32° C. The cuttings after gamma-rays treatment are planted immediately and the cuttings establish in the soil within a week by the growth of roots and the plants grow thereafter. The plants are grown by asexual reproduction through stem cutting, in the farm of Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India.
Bhurakshak has the following botanical characteristics:
a) Flowers: Bracteate, sessile, complete, zygomorphic, pentamorous, hypogynous and cyclio.
b) Calyx: Five sepals and gamosepalous.
c) Corolla: Five petals, gamopetalous, bilipped(4/1), coronary outgrowth present at the throat of corolla.
d) Androecium: Four stamens, didynamous, epipetalous, polyandrous, dorsifixed situated at the throat of corolla and discrete.
e) Gynoccium: Bicarpellary, syncarpous, ovary superior, bilocular with one ovule in each locule, axile placenta, terminal style and capitate stigma.
13 1 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 1 aaatcggagc 10 2 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 2 tgcgcgatcg 10 3 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 3 aacgtacgcg 10 4 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 4 gcacgccgga 10 5 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 5 cgggatccgc 10 6 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 6 gcgaattccg 10 7 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 7 ccctgcaggc 10 8 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 8 ccaagcttgc 10 9 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 9 aagatagcgg 10 10 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 10 ggatctgaac 10 11 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 11 ttgtctcagg 10 12 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 12 ggactccacg 10 13 10 DNA Artificial Sequence primer for generating random amplified polymorphic DNA profile of claimed plant 13 agcctgacgc 10

Claims (1)

We claim:
1. A new and distinct variety of Lippia alba plant, substantially as illustrated and described
US09/538,341 2000-03-29 2000-03-29 Lippia alba plant named ‘Bhurakshak’ Expired - Lifetime USPP13110P2 (en)

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Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Linalool rich Lippia alba var. Kavach, Kumar, et al Journal of Medicinal and Aromatic Plant Sciences 21 (1999) 1094-1095.

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