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Journal of Tropical Agriculture 50 (1-2) : 72-75, 2012


Short communication

Vegetative propagation of stevia [ Stevia rebaudiana (Bertoni) Hemsl.] through stem cuttings
G.R. Smitha1* and K. Umesha2

Directorate of Medicinal and Aromatic Plants Sciences, Boriavi, Anand 387 310, Gujarat, India; 2Division of Horticulture, Gandhi Krishi Vignana Kendra, University of Agricultural Sciences, Bengaluru 560 065, Karnataka, India

Received 6 December 2011; received in revised form 30 March 2012; accepted 30 March 2012.

Abstract Stevia rebaudiana (Bertoni) Hemsl. is a non-caloric bio-sweetener wherein seed propagation is difficult. Propagation of stevia through stem cuttings was evaluated under two environmental conditions (natural shade and greenhouse with intermittent mist) and four combinations of sand, soil, FYM, vermicompost, and coir dust, with growth regulator (IBA), bio-inoculants (Trichoderma viride), and cow’s urine treatments. Stem cuttings treated with T. viride, placed in soil, sand, and vermicompost rooting media (1:1:1) under greenhouse conditions recorded better shoot and root growth parameters, besides better field establishment (90%), over other treatment combinations. Keywords: Bio-inoculants, Environmental conditions, Growth regulators, Rooting media

Stevia [(Stevia rebaudiana (Bertoni) Hemsl.], commonly known as “sweet weed”, “sweet leaf”, “sweet herb” and “honey leaf”, is a perennial herb belonging to the family Asteraceae. Leaves of stevia contain around 10 sweetening glycosides, of which stevioside (3–10%), rebaudiside-A (13%), and rebaudiside-B, C, D are important (Yoshida, 1986). With a huge share of the Indian population being diabetic (Siegel et al., 2008), non-caloric, natural sweeteners safe to diabetics will receive greater focus in future. Although there is greater interest in this plant now as a natural alternative to artificial sweeteners like saccharin, aspartame, asulfam-K, progress towards large scale commercialization of stevia sweetners has been rather slow, largely due to difficulties in propagating the crop. Stevia is propagated either through seeds or cuttings. However, low and erratic seed germination (owing to its

small seed size and related bottlenecks in nutrition) and slow establishment of seedlings warrant its propagation through vegetative means (Randi, 1980). Among different vegetative means, stem cutting is a cheaper and better alternative although tissue culture methods have been standardized. Some basic work on vegetative propagation of stevia regarding use of polythene cover, sand: perlite (1:3) rooting media, and use of growth regulators particularly IBA and NAA to enhance rooting of stevia cuttings (Rajashekara, 2004; Ingle and Venugopal, 2009) also have been carried out. However, information on naturally available, cost effective rooting media, bio-inoculants, and their interaction with the growing conditions for multiplication of stevia has been scarce. Hence, an effort was made to standardize the effects of rooting media, growth regulators, bio-incoulants, and growing con-

*Author for correspondences : Phone +912692271605/606 Ext. 222; Mobile+91-9974303326; Email <>.

Semi-hardwood cuttings with three nodes were prepared using matured basal portions of current year stem. Observations on shoot parameters were recorded from five labelled plants in each replication at regular intervals up to 60 days after planting.21c 14. IBA 500 ppm.79b 19.96c 1. The media were prepared as per the treatment protocol and filled in the seed pan leaving a head space of 2 cm.35 a 20.G.88 c 21.44a 2.64a 30.17b 2. average temperature 30ºC.11c 2.22a 64.42a 33.44b 32.17c 86. IBA 1000 ppm.29 a 21. highest dry weight of the shoot (3.20b 81.19c 34.) and greenhouse with intermittent mist.52 b 20.92C 1. and thickness were recorded by destructive sampling (uprooting five labelled cuttings in each replication at 60 days after planting).29c 22.47 20.65b 79.06a 18.73b 23. and Relative humidity 85%). soil + sand + vermicompost (1:1:1) and soil+sand+coirdust (1:1:1).27 2.R. soil+sand+FYM (1:1:1).87a 30. 930 m altitude) during 2009–’10. and cow’s urine). and six growth regulators and bio-inoculants (control. Basal portion of the cuttings (1 to 2 cm) was dipped in growth regulator solution for 10 seconds.21b 25.29 b 20.14a 1.89 21. Bengaluru (12o58" N. Influence of environmental conditions.02C 1.12b 1. early sprouting (20 days).04d 76. Trichoderma viride.98a 74.06b 0.59b 1. The cuttings were kept under the shade of Singapore cherry and in the medium cost polyhouse with intermittent misting (LDPE with 150 ȝM thickness.12b 26.16a 1. IBA 2000 ppm.90a 20.37 2.08 b 70.06a 18.70b Means with the same superscript do not differ significantly (5%).81b 76.79 c 21.25b Length of Girth of Number longest sprout sprout of leaves (cm) (mm) per rooted cutting 13. highest number of leaves per rooted cutting (27). Rooting parameters such as rooting percentage. The trial was laid out in a factorial controlled randomized design (CRD) with two environmental conditions: natural shade of Singapore cherry (Muntingia calubura L.24a 1.94b 23.88b 71. rooting media and growth regulators on shoot parameters of stevia cuttings.07c 2. .64 21. maximum sprout length (23 cm). root length. Smitha and K. The data were statistically analyzed using ANOVA. Umesha 73 ditions on propagation of stevia through stem cuttings.09b 1.27a 31. Treatments Days taken for sprouting Sprouting percentage Number of sprouts/ rooted cutting 2. Trichoderma viride was prepared by mixing 0.05b 0.63a 29.25 b 19. An experiment was carried out at Sanjeevini Vatika.5 L water to form a uniform slurry.56a 2.04 g). four media: soil+sand (1:1).76a 22.98b 19.92b Environmental conditions Natural shade Greenhouse Media Soil+sand (1:1) Soil+sand +FYM (1:1:1) Soil+sand+vermicompost (1:1:1) Soil+sand+coirdust (1:1:1) Growth regulators Control IBA 500 ppm IBA 1000 ppm IBA 2000 ppm Trichoderma viride Cow’s urine 21.46c 33.12a 15.39 2. Among the two environmental conditions evaluated. highest rooting Table 1.18a 73. as per the treatment protocol. The cuttings were planted in seed pans after treating with growth regulators or bioinoculants with one basal node buried inside the medium.5 kg of culture in 0.54a 2.72 20. 77o55" E.20 2. Fresh cow urine was collected and diluted with water in 1:10 ratio.35c 73.80b 70.31b 2.18b 16.26a 1.93c 19. maximum bud sprout (79%).

Control G2. Higher biomass production in vermicompost media may be due to increased nutrient uptake and enhanced availability of nutrients and growth promoting substances (Thankamani et al. Rooting in stevia cuttings as influenced by different environmental conditions.1). Fig. more roots and higher dry weights for shoot and root (Tables 1 and 2).67) and maximum root length (12.Natural shade E2. maximum root girth (0.13%). sand. Quicker and better sprouting under greenhouse conditions might be attributed to the warmer temperature (30ºC) and higher relative humidity (85%) prevailing under it compared to that of natural shade. maximum number of roots (14).IBA 500 ppm G3-IBA 1000 ppm G4-IBA 2000 ppm G5-Trichoderma viride G6-Cow’s urine (1:10) . rooting media and growth regulators. soil + sand + vermicompost media produced longer and thicker sprouts. As regards to growth media. viride (91.Greenhouse with intermittent mist Media (M) M1-Soil+Sand (1:1) M2. Legends: Environment (E) E1. Similarly.Vegetative propagation of stevia through stem cuttings 74 (77%).37 g) were recorded for the greenhouse treatment (Tables 1 and 2.7 mm) and maximum root dry weight (0.. viride was clearly superior.. 2004). and vermicompost (1:1:1) kept in the greenhouse showed better shoot and root parameters Figure 1..29 cm). Among the interaction effects. Among the six growth regulators and bio-inoculants tried. which accelerated sprouting of the dormant buds (Umesha et al. cuttings treated with T. longest root length (8.Soil+Sand+Vermicompost (1:1:1) M4. the highest field establishment of the rooted cuttings (Table 2) was recorded in greenhouse conditions (90.79%) for cuttings dipped in T. Overall. 2005).Soil+Sand+Coir pith (1:1:1) Growth Regulators (G) G1.Soil+Sand+FYM (1:1:1) M3. 2011). Trichoderma is also capable of increasing nutrient uptake by secreting enzymes that solubilise soil nutrients (Harman et al.02 cm). cuttings treated with Trichoderma viride and planted in a media containing soil. viride planted in soil+sand+vermicompost media and kept in greenhouse gave maximum number of leaves (49. T.

K.20 6.66b 0.R. R.00b 90. and Smitha..37 a 0.83 c 89. Trichoderma sps.18 b 13.25a 6.66 0. University of Agricultural Sciences.66b 0. Environmental conditions and type of cuttings on rooting and growth of vanilla (Vanilla planifolia Andrews). Crops.38 a 0.63c 7.73 a 59. Thankamani.09b 76. Murthy. Spices Arom. I. A.86b 7. Opportunistic. Randi.39a 9. J.63b 66.41b 90. 27: 1077–1090.37 7. 2009. Jap.V. Rajashekara..K. C. K.93 a 70. 1986. Simple determination of sweet glucosides in Stevia plant by thin layer chromato-scanner and their accumulation patterns with plant growth.27 c 0. K. Microbiol . Health Aff.23b 8.26b 75.. 49 (1–2): 121–123.) thesis . Univeridade Estadual de Campinas. Karnataka J.92c 0.79 a 89. . J. Sc. 1980.14b 13.63 c 0.55a 11.32 b 0. 2008. Agric. 2: 43–56. 2004.38a 11. M. 55(2): 189– 195.92 74.64 0. References Harman. M.02a 7. Germinacho de Stevia rebaudiana Bert.M. Sci.. M.35 a 0.22 88. Nature Rev. Homell. 2011.56b 0.19b 91.75 a 0.53 7.R.67 0. and Kinra. 2004. S. 22(2): 460–461. G. and is appropriate for commercial propagation of stevia. Crop Sci.65 0.31b 12.24 c 0. (Millwood).83b 79.29 b 15. A. C. Standardization of vegetative propagation of stevia (Stevia rebaudiana Bertoni) through stem cuttings.68b 8. pp. S. J. K. Sreekala.. Umesha.68b Means with the same superscript do not differ significantly (5%).00 c 70. Smitha and K.67 84. (Hort. Viterbo. Finding a policy solution to India’s diabetes epidemic..18 b 0. Campinas..82 c 90..13 a 90.G. Effect of environmental conditions. avirulenmt plant symbionts. and Venugopal. Treatments Rooting percent Number of roots per rooted cutting 11.33b 67. Effect of Pseudomonas fluorescens (IISR-6) and Trichoderma harzianum (P-26) on growth of black pepper (Piper nigrum L. Bengaluru. G. K.58b 7.03a 16.M. Narayan..33 b 0. Siegel.76 a 0.80 c Length of Girth longest of the roots root (cm) (mm) 6.39 90. M.63 c Dry weight Field of roots survival (g/plant) percentage 0..R.49b 7..20 d 0.50 90. Trop. M. Ingle. and Lorito. and Anandaraj.30 7. Thesis... C.13 e 0. Umesha 75 Table 2. Effect of different growth regulators on rooting of stevia (Stevia rebaudiana Bertoni) cuttings.) in the nursery.60d 0.84a 13. 28–36.17 d 89.61 a 70. Yoshida. 2005. Chet. 14(2): 112–116. Studies on the production of sweet substances in Stevia rebaudiana: I.37 a 0..65 a Environment Natural shade Greenhouse Media Soil+sand (1:1) Soil+sand +FYM (1:1:1) Soil+sand+vermicompost (1:1:1) Soil+sand+coirdust (1:1:1) Growth regulators Control IBA 500 ppm IBA 1000 ppm IBA 2000 ppm Trichoderma viride Cow’s urine 67. rooting media and growth regulators on root parameters of stevia cuttings. G.30 90.39d 12. India. Agric. Sc.