Patent Publication Number: US-PP14400-P3

Title: Rose scented geranium pelargonium graveolenes plant ‘Safal’

Description:
Latin name of the plant claimed: The plant claimed is  Pelargonium graveolens  var. ‘Safal’. 
     FIELD OF INVENTION 
     The present invention is related to the development of a novel high essential oil producing plant obtained through a unique method of progeny screening from the seeds obtained from the plant cultivar ‘Bipuli’ (unpatented) of  Pelargonium graveolens,  from which a novel, unique and commercially viable plant of rose scented geranium  Pelargonium graveolens  with high quantity and quality of essential oil yield was screened out. Further, the invention relates to the development of a high essential oil yielding hybrid named as ‘Safal’ through a planned selection and analysis of the ‘Bipuli’ seedling progenies. The essential oil of the plant is rich in rhodinol (89%) which includes the constituents geraniol, citronellyl, geranyl acetate, citronellyl acetate, geranyl formate, citronellyl formate, phenyl ethyl alcohols, cis and trans rose oxides, linalool and likewise. The hybrid of the invention can be planted and maintained for commercial cultivation through vegetative propagation using the stem cuttings. 
     BACKGROUND OF THE INVENTION 
     The members of the genus, Pelargonium commonly known as geraniums, are common ornamental plants. Some of the species such as  P. graveolens  are used as the source of rose scented geranium essential oil of considerable commercial importance. Geranium oil is one of the expensive essential oils used in perfumery, cosmetics and soap industries. On account of its antimicrobial and pesticidal activities, the use of geranium essential oil for technical applications is expanding. There is need to expand the geranium cultivation in diverse geographical areas with suitable agroclimates to meet the increasing industrial demand for the geranium essential oil. Traditionally, the geranium cultivation is confined to areas having semi-temperate to temperate climates where geranium is planted as perennial crop, maintained in the field for 4-5 years. The shoot portion of the perennial plantations of geranium is harvested 1 to 3 times each year to distill the oil. The spread of geranium cultivation has been narrow because the prevalent cultivars of  Pelargonium graveolens  are highly susceptible to water logging in soil, white ants and several bacterial and fungal diseases. Also there is paucity of genetic resources in Pelargonium species cultivated for essential oils. The available varieties suffer from high degree of male and female sterility. Breeding programmes based on cross hybridization have been scarce among essential oil yielding geraniums. So it is important to generate more genotypes with varied characters which can yield high quantity and quality of essential oil. 
     OBJECTS OF THE INVENTION 
     The main object of the present invention is to develop a new and distinct plant of  Pelargonium graveolens  ‘Safal’, through progeny screening from the seeds obtained from the plant cultivar ‘Bipuli’ of  Pelargonium graveolens,  said plant capable of producing higher quantity and quality of essential oil. 
     Another object of the present invention is to develop a new plant, which produces less of isomenthone, menthone and other sesqui-terpenes but rich in total rhodinol with a citroniol:geraniol ratio of 1:1. 
     DESCRIPTION OF THE INVENTION 
     Accordingly, in the present invention we have developed a novel, distinct, unique, and high essential oil yielding plant of geranium ( Pelargonium graveolens ) ‘Safal’, through progeny screening from the seeds obtained from the plant cultivar ‘Bipuli’ of  Pelargonlum graveolens,  possessing the following combination of characters: 
     a. the plant is a hybrid between cultivar ‘Bipuli’ and cultivar ‘Hemanti’ (unpatented) as the pollen donor as indicated by the co-dominance RAPD pattern obtained by the random primer 5′AACGTACGCG3′ [SEQ ID NO:5] 
     b. possessing very large, hairy, soft, yellow-green (upper surface—144A; lower surface—147B) leaves, dark pink (78D) petals in the flowers, 
     c. possessing vigorous and rapid vegetative growth with higher plant height of up to 82±6 cm, higher canopy size of up to 1.42±0.4 m 2 , higher herb yield of up to 11.1±0.9 kg per plant, higher leaf area of up to 101±16 cm 2 , higher oil content up to 0.35 to 0.40% and higher oil yield of up to 41 g/plant, 
     d. producing an essential oil with the following composition; Citronellol 30.6±3.0, Geraniol 28.7±6.0, Isomenthone 8.4±0.5, Linalool 4.7±0.1, Cis rose oxide 0.4±0.1, trans rose oxide 0.2±0.1, Menthone 0.2±0.1, Citronellyl formate 6.6±0.5, Geranyl formate 2.9±0.2, 10-epi-γ-eudesmol 5.4±0.3, 6,9-guaiadiene 0.1±0.1, Decanoic acid 0.1±0.1, Phenyl ethyl tiglate 0.8±0.1, which may not be construed to be limited to these values, 
     e. with distinct molecular profile by random amplified polymorphic DNA (RAPD) using 10 random primers (AAATCGGAGC [SEQ ID NO:1], GTCCTACTCG [SEQ ID NO:2], TGCGCGATCG [SEQ ID NO:4], AACGTACGCG [SEQ ID NO:5], CGGGATCCGC [SEQ ID NO:9], GCGAATTCCG [SEQ ID NO:10], CCCTGCAGGC [SEQ ID NO:11], CCAAGCTTGC [SEQ ID NO:12], AAGATAGCGG [SEQ ID NO:15], GGATCTGAAC [SEQ ID NO:16]) distinguishing the plant from the other existing varieties known to us, 
     f. producing highest herbage, oil yield per plant as compared to any other existing varieties, and 
     g. possessing the following botanical details. 
     Stem shape: Rounded. 
     Stem habit: Ramified. 
     Number of nodes: 
     ( i )  Primary nodes.— 4. 
     ( ii )  Secondary nodes.— 25 to 30. 
     ( iii )  Tertiary nodes.— 7. 
     Average length of primary internode: 3.0 cm. 
     Leaf apex shape: Mucronate (rounded), cuspidate. 
     Leaf shape: Palmately lobed, cordate. 
     Leaf lamina base shape: Hastate. 
     Petiole shape: Long, hairy. 
     Petiole color: Yellow-green 147B. 
     Color of upper leaf surface: Yellow green (144A). 
     Color of lower leaf surface: Yellow green (147B). 
     Leaf length: 10.0 cm. 
     Leaf width: 13 cm. 
     Number of trichomes: 3.5/mm 2 . 
     Trichome ratio (lower leaf/upper leaf): 2:1. 
     Peduncle: 
       Length.— 5.5. 
       Color.— Yellow green (147B). 
     Time for flowering: February end. 
     Lastingness of bloom: Starts from February lasts till April. 
     Flower shape: Tubular. 
     Pedicel length: 2.5 mm. 
     Pedicel color: Yellow green (147B). 
     Calyx diameter: 5 mm. 
     Calyx: 
       Color  ( both surfaces ).—Yellow green (146A). 
       Sepal number.— 5. 
       Shape.— Triangular. 
       Apex.— Acute. 
       Base.— Concave. 
       Margin.— Entire, hairy. 
       Length.— 0.7 cm. 
     Corolla: 
       Petal number.— 5. 
       Shape.— Tubular bi-lipped free, zygomorphic. 
       Apex.— Slightly notched. 
       Base.— Slightly ligulate. 
       Margin.— Entire. 
       Length  ( upper lip ).—1.0 cm. 
       Length  ( lower lip ).—0.9 cm. 
     Corolla color (both surfaces): Purple group 78D. 
     Pubescence of corolla: Absent. 
     Reproductive organs: 
       Androecium.— 10 stamens, filament sub-equal, united at base, anthers 7, dithoecous, versatile. 
       Gynoceium.— Pentacarpellary syncarpous superior ovary. 
     Color of stigma: Magenta (red purple) (78B). 
     Umbel: 
       Inflorescence/plant.— 23. 
       Flowers/inflorescence.— 7. 
       Umbel shape.— Umbrella. 
     Mature plant: 
       Height.— 82 cm. 
       Width.— 140 cm. 
     Susceptibility to disease: Susceptible to stem and root rot by fungus  Fusarium oxysporum, Rhyzoctonia solanii  and Pythium spp. and to termite attack. 
     Age of plants when described: 2 years (Lucknow, India). 
     Conditions of culture: Subtropical climate, soil sandy loam, alkaline (pH 7.8), low in available nitrogen (N 155 kg/ha), medium in available phosphorus (P 2 O 5  30.4 kg/ha) and exchangeable potassium (K 2 O 120 kg/ha). Field was dressed with manure @ 1 ton/ha and chlorphyriphos 1%/ha before planting of cuttings. 
     Asexual reproduction: Stem cuttings of about 10-20 cm length, with 4-5 nodes and a terminal bud, were planted in nursery beds or pots containing course sand/sphagnum moss. These rooted within 60 days and were planted in the field beds, without disturbing the root system, at 60×60 cm spacing. 
     Fruit/seed: Fruit setting did not take place in subtropical climatic conditions of Lucknow. 
     Time to produce finished plant: One month from rooted cutting to flowering. 
     The plant ‘Safal’ can yield more essential oil with less of isomenthone, menthone and other sesqui-terpenes but rich in total rhodinol with a citroniol:geraniol ratio of 1:1, than the prevailing cultivars now grown in India. 
     While observing large populations of the ‘Bipuli’, ‘Hemanti’ and ‘Kunti’ (unpatented) cultivars of rose scented geranium  P. graveolens  growing at the Kodaikanal field station Tamil Nadu, India of this Institute, in the temperate climate of Southern hills, formation of fruits bearing seeds at low frequency was observed in the populations of the ‘Bipuli’ cultivar of  P. graveolens.  It was realized that the seeds obtained may be the product of rare self fertilization within ‘Bipuli’ cultivar or cross pollination of ‘Bipuli’ gynoecia by fertile pollen grains formed on ‘Hemanti’ or ‘Kunti’ cultivar. The present work was carried out to reveal new genetic variation if any present, among the plants that could be raised from the spontaneous seeds borne on the plants of the ‘Bipuli’ cultivar. Comparison of the plants produced from seeds, collected from ‘Bipuli’ cultivar plants growing amongst those of ‘Hemanti’ and ‘Kunti’ cultivar, has shown that the ‘Bipuli’ seed progeny plants indeed differed from the plants of the three cultivars of  P. graveolens.  Out of all progenies, one (BSP-4) named as ‘Safal’ was found to be high yielding with essential oil containing the oil constituents in desirable proportions, novel and was thus the plant of this invention. As used herein, the accession number “BSP-4” and variety name ‘Safal’ are used interchangeably. 
     All the color grouping given in the description are made as per The International Royal Horticultural Society Colour chart. 
    
    
     BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS 
     FIG. 1 shows Co-dominance pattern of RAPD profile for the plant ‘Safal’ with Primer 5′AACGTACGCG3′ [SEQ ID NO:5]. 
     FIG. 2 shows Unique RAPD profile of ‘Safal’. 
     FIG. 3 is a Photograph of plant ‘Safal’. 
    
    
     BREEDING HISTORY 
     The plants of ‘Bipuli’, ‘Kunti’ and ‘Hemanti’ varieties of  P. graveolens  were planted in beds of 20 m 2  size arranged randomly in a field at the experimental farm of Central Institute of Medicinal and Aromatic Plants (CIMAP), located at Kodaikanal at 101° N latitude, 78° E longitude and 1800 m above sea level in the state of Tamil Nadu, India, having temperate agroclimatic features. The distinguishing characteristics of the three accessions (varieties) are given in the Table 1. Table 2 gives the detailed botanical description of the newly developed plant ‘Safal’. The flowers of all the three varieties were generally sterile, however a small number of fruits bearing seed were found to have come up in the year 1997 on the plants of the ‘Bipuli’ cultivar. The seeds were separated from the fruits, soaked in 100 ppm GA 3  solution in water for 24 hours and sown in earthen trays containing 1:1 ratio of soil and farmyard manure in a glasshouse at Lucknow, India (26.5° N 80.5° E and of 120 m altitude, in subtropical north Indian plains), India. Altogether four seedlings were recovered from three separate sowings of 10 seeds each. The seedlings were transplanted individually into pots carrying soil and sand mixture and were continued to be maintained in the glasshouse under 16 hour light and 8 hour dark conditions. Cuttings from these were used to multiply the new accessions called BSP-1 (unpatented), BSP-2 (unpatented), BSP-3 (unpatented) and BSP-4 (‘Safal,’ the subject of the present application) (BSP=Bipuli seed progeny). 
     
       
         
           
               
             
               
                 TABLE 1 
               
               
                   
               
               
                 Distinguishing features of the cultivars Bipuli, Hemanti 
               
               
                 and Kunti compared to the Bipuli seed progenies.  
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 SI. 
                  Cultivar accessions 
               
            
           
           
               
               
               
               
               
            
               
                 No. 
                 Character 
                  cv. Bipuli 
                 cv. Hemanti 
                 cv. Kunti  
               
               
                   
               
               
                 1. 
                 Habit 
                 Semi-erect 
                 Prostrate 
                 Erect 
               
               
                 2. 
                 Canopy 
                 Spread has 100 
                 Spread has 80 
                 Spread has 60 
               
               
                   
                   
                 cm diameter 
                 cm diameter 
                 cm diameter 
               
               
                   
                   
                 and 50 cm 
                 and 55 cm 
                 and 40 cm 
               
               
                   
                   
                 height 
                 height 
                 height 
               
               
                 3. 
                 Stem 
                 Moderately 
                 Highly hairy, 
                 Poorly haired, 
               
               
                   
                   
                 hairy, strong, 
                 weak, 2-3 
                 sturdy usually 
               
               
                   
                   
                 4-5 primary 
                 primary 
                 1-2 primary 
               
               
                   
                   
                 branches which 
                 branches which 
                 branches which 
               
               
                   
                   
                 give out 20-25 
                 are highly 
                 bear 15-20 
               
               
                   
                   
                 secondary 
                 branched into 
                 secondary 
               
               
                   
                   
                 branches and 
                 25-30 secondary 
                 branches and 
               
               
                   
                   
                 5-7 tertiary 
                 branches, 5 
                 3-5 tertiary 
               
               
                   
                   
                 branches 
                 tertiary branches 
                 branches  
               
               
                 4. 
                 Leaf 
                 Medium size, 
                 Long, thin, 
                 Small, stout, 
               
               
                   
                 petiole 
                 thin rough dark 
                 soft, light 
                 rough, 
               
               
                   
                   
                 pink at base 
                 pink at base 
                 anthocyanin 
               
               
                   
                   
                   
                   
                 pigment absent 
               
               
                 5. 
                 Leaf lamina 
                 Small (5 cm 
                 Large (8 cm. 
                 very large (9 
               
               
                   
                   
                 long and 6 cm 
                 long, 12 cm 
                 cm long and 14 
               
               
                   
                   
                 wide), lamina 
                 wide), lamina 
                 cm wide), 
               
               
                   
                   
                 has about 75 
                 has about 60 
                 lamina has 45 
               
               
                   
                   
                 lobes, hairy, 
                 lobes, very 
                 lobes, leathery, 
               
               
                   
                   
                 yellowish green 
                 hairy, dark 
                 yellowish green 
               
               
                   
                   
                 146A a   
                 green 137B b   
                 146A a   
               
               
                 6. 
                 Leaf 
                 Medium (400  
                 Long (800 μm), 
                 Short (300 
               
               
                   
                 trichomes 
                 μm), thin 
                 thin 
                 μm), stout 
               
               
                 7. 
                 Flower 
                 Medium size 
                 Big size (1.5- 
                 small size (1.0 
               
               
                   
                   
                 ((1.5 cm), dark 
                 2.0 cm), pink 
                 cm), petals, 
               
               
                   
                   
                 pink petals 
                 petals (80B d ), 
                 light pink 
               
               
                   
                   
                 (78C c ), yellow 
                 anthers. 
                 (74D e ), pink. 
               
               
                   
                   
                 fertile 
                 incompletely 
                 fertile 
               
               
                   
                   
                 anthers, seeds 
                 developed in 
                 anthers, seeds 
               
               
                   
                   
                 formed 
                 the form of 
                 formed 
               
               
                   
                   
                 occasionally 
                 staminodes 
               
               
                 8. 
                 Shoot 
                 0.20-0.25% 
                 0.10-0.15% 
                 0.25-0.30% 
               
               
                   
                 essential 
               
               
                   
                 oil content 
               
               
                 9. 
                 Physical 
                 Bright yellow 
                 Pale yellow 
                 Greenish 
               
               
                   
                 appearance 
                   
                 greenish yellow 
                 yellow 
               
               
                   
                 of oil 
               
               
                 10. 
                 Citronellol: 
                 1:1 like in 
                 3-4:1 like that 
                 1:5 
               
               
                   
                 geraniol 
                 Bourbon type 
                 in commercial 
               
               
                   
                 ratio 
                 commercial oil 
                 oil of Chinese 
               
               
                   
                   
                 of Reunion 
                 origin 
               
               
                   
                   
                 Island origin  
               
               
                   
               
            
           
           
               
               
            
               
                 SI. 
                  Bipuli seed progeny accessions 
               
            
           
           
               
               
               
               
            
               
                 No. 
                 Character 
                 BSP-1 
                 BSP-2  
               
               
                   
               
               
                 1. 
                 Habit 
                 Erect 
                 Erect 
               
               
                 2. 
                 Canopy 
                 Spread has 42 
                 Spread has 40 
               
               
                   
                   
                 cm diameter and 
                 cm diameter and 
               
               
                   
                   
                 48 cm height 
                 45 cm height 
               
               
                 3. 
                 Stem 
                 Poorly haired, sturdy, 
                 Poorly haired, sturdy, 
               
               
                   
                   
                 3 primary branches 
                 3-4 primary branches 
               
               
                   
                   
                 which give out 15-20 
                 which give out 10-50 
               
               
                   
                   
                 secondary and 3 
                 secondary branches and 
               
               
                   
                   
                 tertiary branches 
                 3-4 tertiary branches 
               
               
                 4. 
                 Leaf 
                 Short, stout, rough, 
                 Very short, stout, 
               
               
                   
                 petiole 
                 faint, anthocyanin 
                 rough, light pink 
               
               
                   
                   
                 present at the base 
                 at the base 
               
               
                 5. 
                 Leaf lamina 
                 Large (9 cm 
                 Large (9 cm 
               
               
                   
                   
                 long and 12 cm 
                 long and 12 cm 
               
               
                   
                   
                 wide), lamina 
                 wide), lamina 
               
               
                   
                   
                 has 75 lobes, leathery, 
                 has 80 lobes, leathery, 
               
               
                   
                   
                 yellowish green 146A a   
                 yellowish green 
               
               
                 6. 
                 Leaf 
                 Very short (250 μm), 
                 Very short (200 μm), 
               
               
                   
                 trichomes 
                 stout 
                 stout 
               
               
                 7. 
                 Flower 
                 Medium size 1.3 cm), 
                 Medium size (1.3 cm), 
               
               
                   
                   
                 pink petals (74B e ), 
                 purple petals 
               
               
                   
                   
                 pink fertile, 
                 (77C c ), pink fertile 
               
               
                   
                   
                 anthers, seeds formed 
                 anthers, seeds formed 
               
               
                 8. 
                 Shoot 
                 0.31-0.35% 
                 0.35-0.40% 
               
               
                   
                 essential 
               
               
                   
                 oil content 
               
               
                 9. 
                 Physical 
                 Greenish yellow 
                 Greenish yellow 
               
               
                   
                 appearance 
               
               
                   
                 of oil 
               
               
                 10. 
                 Citronellol: 
                 1:3 
                 1:2.5 
               
               
                   
                 geraniol 
               
               
                   
                 ratio  
               
               
                   
               
            
           
           
               
               
            
               
                 SI. 
                  Bipuli seed progeny accessions 
               
            
           
           
               
               
               
               
            
               
                 No. 
                 Character 
                 BSP-4 
                 BSP-4 (‘Safal’)  
               
               
                   
               
               
                 1. 
                 Habit 
                 Erect 
                 Semi-erect 
               
               
                 2. 
                 Canopy 
                 Spread has 68 
                 Spread has 140 
               
               
                   
                   
                 cm diameter and 
                 cm diameter and 
               
               
                   
                   
                 52 cm height 
                 82 cm height 
               
               
                 3. 
                 Stem 
                 Moderately hairy, 
                 Moderately hairy, sturdy, 
               
               
                   
                   
                 weak, 3 primary 
                 5 primary branches 
               
               
                   
                   
                 branches which give 
                 which are highly 
               
               
                   
                   
                 out 15-20 secondary 
                 branched into 35-40 
               
               
                   
                   
                 branches and 3- 
                 secondary branches and 
               
               
                   
                   
                 5 tertiary branches 
                 5-7   tertiary branches 
               
               
                 4. 
                 Leaf 
                 Long, thin 
                 Very long, soft 
               
               
                   
                 petiole 
                 soft, without 
                 pink at the base 
               
               
                   
                   
                 any pigmentation 
               
               
                   
                   
                 at base 
               
               
                 5. 
                 Leaf lamina 
                 Medium (8 cm 
                 Very large (10 
               
               
                   
                   
                 long and 10 cm 
                 cm long and 13 
               
               
                   
                   
                 wide) lamina 
                 cm wide), 
               
               
                   
                   
                 has 60 lobes, 
                 lamina has 60 
               
               
                   
                   
                 soft, yellowish 
                 lobes, hairy, 
               
               
                   
                   
                 green 144A a   
                 soft, green 137C b   
               
               
                 6. 
                 Leaf 
                 Short (300 μm), 
                 Medium (350 
               
               
                   
                 trichomes 
                 thin 
                 μm), thin 
               
               
                 7. 
                 Flower 
                 Small size (1 
                 Medium size (1.3 cm) 
               
               
                   
                   
                 cm), light pink 
                 dark pink petals 
               
               
                   
                   
                 petals (74D e ) , 
                 (78D c ), yellow 
               
               
                   
                   
                 yellow anthers, 
                 anthers seeds 
               
               
                   
                   
                 seeds not formed 
                 occasionally formed 
               
               
                 8. 
                 Shoot 
                 0.25-0.30% 
                 0.35-0.40% 
               
               
                   
                 essential 
               
               
                   
                 oil content 
               
               
                 9. 
                 Physical 
                 Bright yellow 
                 Pale yellow 
               
               
                   
                 appearance 
               
               
                   
                 of oil 
               
               
                 10. 
                 Citronellol: 
                 1.5:1 
                 1:1 
               
               
                   
                 geraniol 
               
               
                   
                 ratio  
               
               
                   
               
               
                   a yellow green group;  
               
               
                   b green group;  
               
               
                   c purple group;  
               
               
                   d purple violet group;  
               
               
                   e red purple group;  
               
               
                 *(all color groupings were made as per The Royal Horticultural Society Colour Chart)  
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
               
                   
               
               
                 Detailed Botanical Description of the plant ‘Safal’:  
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                  1. 
                 Stem shape: 
                 ROUNDED 
               
               
                  2. 
                 Stem habit: 
                 RAMIFIED 
               
               
                  3. 
                 Number of nodes 
               
               
                   
                 (i) Primary Nodes: 
                 4 
               
               
                   
                 (ii) Secondary Nodes: 
                 25 to 30 
               
               
                   
                 (iii) Tertiary Nodes: 
                 7 
               
               
                  4. 
                 Average length of 
                 3.0 cm 
               
               
                   
                 primary internode: 
               
               
                  5. 
                 Leaf apex shape: 
                 MUCRONATE (rounded), 
               
               
                   
                 CUSPIDATE 
               
               
                  6. 
                 Leaf shape: 
                 PALMATELY LOBED, CORDATE 
               
               
                  7. 
                 Leaf lamina base shape: 
                 HASTATE 
               
               
                  8. 
                 Petiole shape: 
                 LONG, HAIRY 
               
               
                  9. 
                 Petiole color: 
                 YELLOW-GREEN 147B 
               
               
                   
                 Color of Upper leaf surface: 
                 YELLOW GREEN (144A) 
               
               
                 10. 
                 Color of lower leaf surface: 
                 YELLOW GREEN (147B) 
               
               
                 11. 
                 Leaf length: 
                 10.0 cm 
               
               
                 12. 
                 Leaf width: 
                 13 cm 
               
               
                 13. 
                 Number of trichomes: 
                 3.5/mm 2   
               
               
                 14. 
                 Trichome ratio: 
                 2:1 
               
               
                   
                 (lower leaf/upper leaf) 
               
               
                 15. 
                 Peduncle 
               
               
                   
                 length: 
                 5.5 
               
               
                   
                 color: 
                 YELLOW GREEN (147B) 
               
               
                 16. 
                 Time for flowering: 
                 FEBRUARY END 
               
               
                 17. 
                 Lastingness of bloom: 
                 Starts from February 
               
               
                   
                   
                 lasts till April 
               
               
                 18. 
                 Flower shape: 
                 TUBULAR 
               
               
                 19. 
                 Pedicel length: 
                 2.5 mm 
               
               
                 20. 
                 Pedicel color: 
                 YELLOW GREEN (147B) 
               
               
                 21. 
                 Calyx diameter: 
                 5 mm 
               
               
                 22. 
                 Calyx 
               
               
                   
                 (i) color (both surfaces): 
                 YELLOW GREEN (146A) 
               
               
                   
                 (ii) sepal number: 
                 5 
               
               
                   
                 (iii) shape: 
                 triangular 
               
               
                   
                 (iv) apex: 
                 acute 
               
               
                   
                 (v) base: 
                 concave 
               
               
                   
                 (vi) margin: 
                 entire, hairy. 
               
               
                   
                 (vii) length: 
                 0.7 cm 
               
               
                 23. 
                 Corolla 
               
               
                   
                 (i) petal number: 
                 5 
               
               
                   
                 (ii) shape: 
                 tubular bi-lipped free, zygomorphic 
               
               
                   
                 (iii) apex: 
                 slightly notched 
               
               
                   
                 (iv) base: 
                 slightly ligulate 
               
               
                   
                 (v) margin: 
                 entire 
               
               
                   
                 (vi) length (upper lip): 
                 1.0 cm 
               
               
                   
                 (vii) length (lower lip): 
                 0.9 cm 
               
               
                 24. 
                 Corolla color (both surfaces): 
                 Purple group 78D 
               
               
                 25. 
                 Pubescence of corolla: 
                 ABSENT 
               
               
                 26. 
                 Reproductive organs 
               
               
                   
                 (i) androecium: 
                 10 stamens, filament sub-equal, 
               
               
                   
                   
                 united at base, anthers 7, dithoecous, 
               
               
                   
                   
                 versatile 
               
               
                   
                 (ii) gynoecium: 
                 pentacarpellary 
               
               
                   
                   
                 syncarpous superior ovary 
               
               
                 27. 
                 Color of stigma: 
                 MAGENTA (RED PURPLE) (78B) 
               
               
                 28. 
                 Umbel 
               
               
                   
                 (i) inflorescence/plant: 
                 23 
               
               
                   
                 (ii) flowers/inflorescence: 
                 7 
               
               
                   
                 (iii) umbel shape: 
                 umbrella 
               
               
                 29. 
                 Mature plant 
               
               
                   
                 (i) height: 
                 82 cm 
               
               
                   
                 (ii) width: 
                 140 cm 
               
               
                 30. 
                 Susceptibility to disease: 
                 susceptable to stem and root rot by 
               
               
                   
                   
                 fungus  Fusarium oxysporum , 
               
               
                   
                   
                   Rhyzoctonia solanii  and Pythium spp. 
               
               
                   
                   
                 and to termite attack  
               
               
                   
               
            
           
         
       
     
     Field Experiments 
     Out of these 4 progenies obtained, preliminary analysis revealed encouraging results for BSP-4, (‘Safal’), as the plant showed high vigor in biomass production and higher oil yield. So the plant ‘Safal’ was taken to the field for further evaluation. To variously compare the ‘Safal’ accession with the cultivar accessions ‘Bipuli’, ‘Hemanti’ and ‘Kunti’, field plot experiments were carried out over two winter-summer cropping seasons (1998-1999) and (1999-2000) at the experimental farm of Central Institute of Medicinal and Aromatic Plants, at Lucknow India (26.5° N 80.5° E and of 120 m altitude, in subtropical north Indian plains). The soil in the field used was sandy loam in texture, alkaline in reaction (pH, 7.8), low in available N (155 kg/ha) and medium in available phosphorous (30.4 kg P 2 O 5 /ha) and exchangeable potassium (120 kg K 2 O/ha). In October 1998, six glasshouse-grown plants of each accession were transferred to the field trial. The cuttings drawn from each of the 4 accessions were planted in randomized blocks replicated three times, in January 1999. The field blocks used were given a uniform dose of 100 kg/ha P 2 O 5  and 60 kg/ha K 2 O. Nitrogen was applied in three splits while phosphorus and potassium were applied at the time of planting. A light irrigation was given just after the planting of cuttings and plots were manually weeded and irrigated at regular interval of two weeks throughout the cropping season. The crops were allowed to grow until June when observations on the morphology of the plants were recorded and crops harvested to estimate the yield of herbage as well as yield and quality of essential oil. About 300 g sample of herbage harvested from each plot was distilled to determine its oil content. The oil samples were analyzed by GC, GC-MS procedures. The 4 accessions were similarly grown and assessed in the 1999-2000 winter-summer cropping season at Lucknow. The parameters on which observations were recorded were averaged for the two seasons for presentation in Tables 3 and 4. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 The variation observed in the expression of the essential oil yield 
               
               
                 related traits among 4 accessions in the rose scented geranium 
               
               
                 
                   Pelargonium graveolens 
                 
               
            
           
           
               
               
            
               
                 Acc. No. 
                 Essential oil yield parameter  
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Plant 
                   
                   
                   
               
               
                   
                 height 
                 Canopy 
                 Herb Yield 
                 Number of 
               
               
                   
                 (cm) 
                 Size (m 2 ) 
                 (kg/plant) 
                 branches/plant  
               
               
                   
               
               
                 Bipuli 
                 50 ± 2 
                 0.81 ± 0.1  
                 3.3 ± 0.1 
                 25 ±  2 
               
               
                 Hemanti 
                 55 ± 2 
                 0.80 ± 0.2  
                 3.9 ± 0.5 
                 30 ±  4 
               
               
                 Kunti 
                 40 ± 4 
                 0.58 ± 0.2  
                 2.0 ± 0.1 
                 22 ±  1 
               
               
                 BSP-4 (‘Safal’) 
                 82 ± 6 
                 1.42 ± 0.4  
                 11.1 ± 0.9  
                 47 ±  4 
               
               
                 Mean ± (SEM) 
                 53 ± 5 
                 0.73 ± 0.14 
                 3.9 ± 1.3 
                 27 ±  4 
               
               
                 CD at 5% 
                 5 
                 0.30 
                 0.6 
                 4  
               
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Number of 
                 Leaf/stem 
                 Leaf petiole 
               
               
                   
                 leaves/plant 
                 ratio (cm) 
                 lemgth (cm)  
               
               
                   
               
               
                 Bipuli 
                 1437 ± 100 
                 0.7 ± 0.1 
                  9.7 ± 1.8 
               
               
                 Hemanti 
                 1801 ± 167 
                 0.9 ± 0.1 
                 11.1 ± 1.4 
               
               
                 Kunti 
                 1363 ± 177 
                 1.2 ± 0.1 
                  9.1 ± 1.1 
               
               
                 BSP-4 (‘Safal’) 
                 4632 ± 201 
                 0.9 ± 0.5 
                 24.6 ± 2.8 
               
               
                 Mean ± (SEM) 
                 2445 ± 500 
                  1.0 ± 0.07 
                 11.7 ± 2.3 
               
               
                 CD at 5% 
                 697 
                 0.2 
                 3.8  
               
               
                   
               
               
                   
                 Leaf Area 
                   
                 Oil Yield/ 
               
               
                   
                 (cm 2 ) 
                 Oil Content % 
                 plant (g/plant)  
               
               
                   
               
               
                 Bipuli 
                 70 ± 7 
                 0.25 ± 0.1 
                  8.2 ± 0.1 
               
               
                 Hemanti 
                 68 ± 2 
                 0.20 ± 0.1 
                  7.8 ± 0.2 
               
               
                 Kunti 
                 108 ± 5  
                  0.3 ± 0.1 
                  6.0 ± 0.5 
               
               
                 BSP-4 (‘Safal’) 
                 101 ± 16 
                 0.37 ± 0.1 
                 41.0 ± 0.9 
               
               
                 Mean ± (SEM) 
                 83 ± 6 
                 0.29 ± 0.2 
                 19.3 ± 8.0 
               
               
                 CD at 5% 
                 24 
                 0.08 
                 11.7  
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
               
                   
               
               
                 The variation in the expression of essential oil quality among 4 accessions, 
               
               
                 in the rose scented geranium  Pelargonium graveolens   
               
               
                 % content of terpenoid in essential oil  
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                 Iso- 
                   
                 Cis 
               
               
                 Accession 
                 Citronellol 
                 Geraniol 
                 menthone 
                 Linalool 
                 rose oxide  
               
               
                   
               
               
                 Bipuli 
                 34.5 ± 3.4 
                 21.8 ± 3.3 
                  7.7 ± 0.6 
                 4.3 ± 1.6 
                 0.6 ± 0.3 
               
               
                 Hemanti 
                 50.6 ± 1.4 
                  1.2 ± 0.3 
                 12.4 ± 0.7 
                 1.1 ± 0.1 
                 0.8 ± 0.1 
               
               
                 Kunti 
                 13.0 ± 1.0 
                 43.7 ± 1.4 
                 10.5 ± 0.7 
                 6.6 ± 0.1 
                 0.2 ± 0.1 
               
               
                 BSP-4 
                 30.6 ± 3.0 
                 28.7 ± 6.0 
                  8.4 ± 0.5 
                 4.7 ± 0.1 
                 0.4 ± 0.1 
               
               
                 (‘Safal’) 
               
               
                 Means ± 
                 26.2 ± 5.6 
                 27.2 ± 6.2 
                  9.5 ± 1.0 
                 6.2 ± 1.3 
                 0.3 ± 0.1 
               
               
                 SEM 
               
               
                 CD at 5% 
                 6.5 
                 4.3 
                 1.9 
                 2.7 
                 0.3  
               
               
                   
               
               
                   
                 Trans 
                   
                 Citronellyl 
                 Geranyl 
                 10-epi-γ- 
               
               
                 Accession 
                 rose oxide 
                 Menthone 
                 formate 
                 formate 
                 eudesmol  
               
               
                   
               
               
                 Bipuli 
                  0.3 ± 0.1 
                  0.1 ± 0.1 
                  7.8 ± 0.3 
                 2.1 ± 0.2 
                 5.7 ± 0.3 
               
               
                 Hemanti 
                  0.5 ± 0.1 
                  0.1 ± 0.1 
                 13.8 ± 0.8 
                 0.2 ± 0.1 
                 2.4 ± 0.2 
               
               
                 Kunti 
                  0.1 ± 0.1 
                  0.2 ± 0.1 
                  0.6 ± 0.1 
                 0.2 ± 0.1 
                 4.1 ± 0.1 
               
               
                 BSP-4 
                  0.2 ± 0.1 
                  0.2 ± 0.1 
                  6.6 ± 0.5 
                 2.9 ± 0.2 
                 5.4 ± 0.3 
               
               
                 (‘Safal’) 
               
               
                 Means ± 
                  0.2 ± 0.1 
                  0.2 ± 0.1 
                  4.5 ± 0.4 
                 0.9 ± 0.4 
                 3.7 ± 0.8 
               
               
                 SEM 
               
               
                 CD at 5% 
                 0.1 
                 0.1 
                 1.0 
                 0.5 
                 0.5  
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Decanoic 
                   
                 Phenyl ethyl 
               
               
                 Accession 
                 6,9-guaiadiene 
                 acid 
                 Isodecanoic 
                 tiglate  
               
               
                   
               
               
                 Bipuli 
                  0.1 ± 0.1 
                 ND a   
                 ND 
                 0.8 ± 0.3 
               
               
                 Hemanti 
                  0.4 ± 0.1 
                 ND 
                 ND 
                 0.8 ± 0.2 
               
               
                 Kunti 
                  2.0 ± 0.1 
                 2.8 ± 0.1 
                 0.6 ± 0.2 
                 0.9 ± 0.1 
               
               
                 BSP-4 
                  0.1 ± 0.1 
                 0.1 ± 0.1 
                 ND 
                 0.8 ± 0.1 
               
               
                 (‘Safal’) 
               
               
                 Means ± 
                  1.8 ± 0.7 
                 1.5 ± 0.7 
                 0.4 ± 0.2 
                 1.0 ± 0.6 
               
               
                 SEM 
               
               
                 CD at 5% 
                 0.1 
                 0.1 
                 0.1 
                 0.5  
               
               
                   
               
               
                   a ND = not detected  
               
            
           
         
       
     
     GC and GC-MS Analysis 
     GC analysis of the essential oils was performed on a Perkin-Elmer gas chromatograph 8500 equipped with FID, using two fused silica capillary columns BP-1 coated with dimethyl siloxane (30 m×0.25 mm×0.25 μm film thickness) and BP-20 coated with carbowax 20M (20 m×0.25 mm×0.25 μm thickness), carrier gas nitrogen at 10 psi inlet pressure and temperature programmed to 60-220° C. at 5° C./min. For BP-20 column and split ratio of 1:80. GC-MS was performed on Schimadzu QP-2000 instrument using ULBON. HR-1 fused silica column (50 m×0.25 mm×0.25 μm film thickness), temperature programmed to 100°-250° C. at 10° C./min, carrier gas helium at 2 ml/min, MS conditions of El mode 70 eV and ion source temperature of 250° C. 
     Identification of the Compounds 
     Compounds were identified by comparing the retention indices (relative to C8-C21 alkanes) with those reported in literature by peak enrichment on coinjection with standards wherever possible and by comparison of mass spectra of the peak with those of compounds reported in literature (Jennings, W. &amp; T. Shibamoto, 1980. Qualitative analysis of flavour and fragrance volatile by capillary GC, Academic Press Inc., New York.; Adams, R. P., 1990. Identification of essential oils by ion trap mass spectroscopy. Academic Press, San Diego, Calif.). Relative amounts of individual components were estimated based on peak areas on BP-1 column without FID response correction. 
     Molecular Analysis of Hybrids 
     DNA was isolated from young leaves (1 g) taken from mature plants following the reported protocol (Khanuja, S. P. S., Shasany, A. K., Darokar, M. P., and Kumar, S., 1999, Rapid isolation of PCR amplifible DNA from dry and fresh samples of plant producing large amounts of secondary metabolites and essential oil by modified CTAB procedure. Plant Molecular Biology, 17:74.) and was digested with EcoRI restriction endonucleases. A set of twenty decanucleotide primers (M/S Bangalore Genie, India) were used for PCR amplification. Polymerase chain reaction (PCR) was carried out in 25 μl reaction volume, containing 20-40 ng of plant genomic DNA, 125 μM Of MgCl 2  buffer, 100, μM of each dNTP, 5 p motes of primer and 0.2 units of Taq DNA polymerase. Amplification was carried out in DNA Engine PTC 200 (M J Research, USA) thermal cycler programmed for 45 cycles of 1 min at 94° C., 1 at 36° C. and 2 min at 72° C. The amplification cycle was concluded with final extension at 72° C. for 5 min. Amplification products were electrophoresed in 1.2% (w/v) agarose gel, visualized by ethidium bromide (0.5 μgml −1 ) staining. The pictures of the gel were scanned for the presence of polymorphic fragments which were scored for the presence (+) or absence (−) of bands. The data so generated was used for calculating the index of genetic similarity using Nei and Li&#39;s matching co-efficient method (Nei, M. and Li, W. H., 1979. Mathematical model for studying genetic variation in terms of restriction endonuclease. Proc. Natl. Acad. Sci. U.S.A.,74 : 5267-5273.). The similarity indices were calculated by using the formula: Number of similar bands between the two accessions/total number of bands in the two accessions×2 and present in Table 5. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Average similarity indices of ‘Safal’ with the 
               
               
                 cultivar accessions ‘Bipuli’, ‘Hemanti’  and ‘Kunti’ 
               
               
                 in rose scented geranium  Pelargonium graveolens   
               
            
           
           
               
               
               
               
               
            
               
                 Accession 
                 Bipuli 
                 Hemanti 
                 Kunti 
                 Safal  
               
               
                   
               
               
                 Bipuli 
                 1.00 
                   
                   
                   
               
               
                 Hemanti 
                 0.81 
                 1.00 
               
               
                 Kunti 
                 0.83 
                 0.78 
                 1.00 
               
               
                 Safal 
                 0.76 
                 0.75 
                 0.67 
                 1.00  
               
               
                   
               
            
           
         
       
     
     The similarity indices arrived at using 20 decanucleotide primers (MAP01 to MAP20: AAATCGGAGC [SEQ ID NO:1], GTCCTACTCG [SEQ ID NO:2], GTCCTTAGCG [SEQ ID NO:3], TGCGCGATCG [SEQ ID NO:4], AACGTACGCG [SEQ ID NO:5], GCACGCCGGA [SEQ ID NO:6], CACCCTGCGC [SEQ ID NO:7], CTATCGCCGC [SEQ ID NO:8], CGGGATCCGC [SEQ ID NO:9], GCGAATTCCG [SEQ ID NO:10], CCCTGCAGGC [SEQ ID NO:11], CCAAGCTTGC [SEQ ID NO:12], GTGCAATGAG [SEQ ID NO:13], AGGATACGTG [SEQ ID NO:14], AAGATAGCGG [SEQ ID NO:15], GGATCTGAAC [SEQ ID NO:16], TTGTCTCAGG [SEQ ID NO:17], CATCCCGAAC [SEQ ID NO:19], GGACTCCACG [SEQ ID NO:19], AGC;CTGACGC [SEQ ID NO:20], respectively) in the RAPD analysis are given in the Table 4. Table 4 indicated that the accession ‘Safal’ was more similar to the parent varieties ‘Bipuli’ and ‘Hemanti’ (more than 75%) than ‘Kunti’ (67%). Further the RAPD profile of ‘Safal’ showed co-dominance inheritance from ‘Bipuli’ and ‘Hemanti’ (FIG.  1 ). 
     Distinctiveness of Safal 
     The accession ‘Safal’ expressed most of the characters at much higher levels than the corresponding measurements in the rest of the accessions. In ‘Safal’, the leaf/stem ratio and leaf area measurements fell within the ranges covered for these characters by the cultivar accessions. As compared to ‘Bipuli’, ‘Safal’ was 1.6 fold taller in height, had 1.7 fold larger canopy, yielded 3.5 fold more herbage, had 2 fold more branches, 3 fold more leaves, 3.5 fold longer leaf petiole, 1.4 fold more oil content in herbage and gave 5 fold more oil yield (Table 2). Apparently the accession ‘Safal’ demonstrated the kind of vigour associated with hybrids or transgressive segregants. The variation observed for the essential oil quality parameters among the seven accessions in terms of the 14 terpenoid components that could be identified are summarized in the Table 3. The oil of the accession ‘Safal’ had geraniol to citronellol proportion as 1:1 and had other parameters widely different from those observed for the oils of the other accessions. The cis and trans rose oxides and citronellyl fonnate contents in the oil of ‘Safal’ accession were lower than in the oils of the accessions ‘Bipuli’ and ‘Hemanti’. The essential oil of the accession ‘Safal’ had low concentrations of 6,9-guaiadiene, decanoic acid and, isodecanoic acid, like in the oils of the accessions ‘Bipuli’ and ‘Hemanti’. These observations described above have demonstrated that the accession ‘Safal’ of  P. graveolens  differed from the cultivar accessions ‘Bipuli’, ‘Kunti’ and ‘Hemanti’ not only in some of the morphological and essential oil characteristics but also in their DNA profiles. 
     The accession ‘Safal’ demonstrated the expression of essential oil yield related characters at much higher levels than by all the accessions studied. The terpenoid profile of the essential oil of the accession ‘Safal’ also appeared to be unique, in that it had equally high concentrations of citronellol and geraniol and very high concentration of total rhodinols. Considering all the essential oil yield related characters together with hierarchical relationships arrived at by DNA fingerprinting, it is possible to surmise that the accession ‘Safal’ may be a hybrid between the accession ‘Bipuli’ and ‘Hemanti’. The accession ‘Safal’ is novel, unique and has highly useful combination of yield, essential oil quality related characteristics, and this can be used for commercial cultivation to extract high value essential oil having utility for industrial and pharmaceutical purposes. Finally, the new plant was selected for its high quality essential oil and the genotype can be used in the future for plant improvement. The accession BSP-4 is the plant of this invention and was named as ‘Safal’. 
     Randomly Amplified Polymorphic DNA Analysis 
     The RAPD profiles of the plant ‘Safal’ were unambiguously able to establish its distinct identity as completely different from the parent plant ‘Bipuli’ as well as the known varieties ‘Kunti’ and ‘Hemanti’. The plant of the present invention was developed by screening the ‘Bipuli’ seed progenies and differentiated as distinct, unique and novel at DNA level. The plant is having desirable morphological and economical traits in a rare unmatchable combination and is available only with us in CIMAP. The primers with the sequence AAATCGGAGC [SEQ ID NO:1], GTCCTACTCG [SEQ ID NO:2], TGCGCGATCG [SEQ ID NO:4], AACGTACGCG [SEQ ID NO:5], CGGGATCCGC [SEQ ID NO:9], GCGAATTCCG [SEQ ID NO:10], CCCTGCAGGC [SEQ ID NO:11], CCAAGCTTGC [SEQ ID NO:12], AAGATAGCGG [SEQ ID NO:15], GGATCTGAAC [SEQ ID NO:16], were used to develop a unique and distinct RAPD profile of the Plant (FIG.  2 ). The whole plant ‘Safal’ has been shown in FIG. 3 wherein the canopy and the shape of the leaves are quite apparent. 
     
       
         
           
             20 
           
           
             1 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            1
aaatcggagc                                                            10
 
           
             2 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            2
gtcctactcg                                                            10
 
           
             3 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            3
gtccttagcg                                                            10
 
           
             4 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            4
tgcgcgatcg                                                            10
 
           
             5 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            5
aacgtacgcg                                                            10
 
           
             6 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            6
gcacgccgga                                                            10
 
           
             7 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            7
caccctgcgc                                                            10
 
           
             8 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            8
ctatcgccgc                                                            10
 
           
             9 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            9
cgggatccgc                                                            10
 
           
             10 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            10
gcgaattccg                                                            10
 
           
             11 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            11
ccctgcaggc                                                            10
 
           
             12 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            12
ccaagcttgc                                                            10
 
           
             13 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            13
gtgcaatgag                                                            10
 
           
             14 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            14
aggatacgtg                                                            10
 
           
             15 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            15
aagatagcgg                                                            10
 
           
             16 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            16
ggatctgaac                                                            10
 
           
             17 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            17
ttgtctcagg                                                            10
 
           
             18 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            18
catcccgaac                                                            10
 
           
             19 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            19
ggactccacg                                                            10
 
           
             20 
             10 
             DNA 
             Artificial Sequence 
             
               Primer 
             
           
            20
agcctgacgc                                                            10