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Int.J.Curr.Microbiol.App.Sci (2020) 9(10): 1803-1808 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 9 Number 10 (2020) Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2020.910.219 Effect of Pre-sowing Seed Treatments on Aristolochia tagala Cham: A Threatened Medicinal Plant of South India Rajani Bhat1*, G. Raviraja Shetty1, P. E. Rajasekharan2, D. A. Pooja1, M. Ganapathi1 and Sadhashiv Nadukeri1 1 College of Horticulture, Mudigere-577132, University of Agricultural and Horticultural Sciences, Shivamogga, Karnataka, India 2 ICAR- Indian Institute of Horticultural Research, Bengaluru, Karnataka, India *Corresponding author ABSTRACT Keywords Aristolochia tagala, Seed germination, Water soaking, Thiourea Article Info Accepted: 18 September 2020 Available Online: 10 October 2020 An experiment was conducted to standardize the seed germination technique in Aristolochia tagala Cham. at the Division of Floriculture and medicinal crops, ICARIndian Institute of Horticulture, Bengaluru during 2019-20. Fresh seeds of A. tagalawere collected from the Field Gene Bank of RET medicinal plants, IIHR Bengaluru and different treatments such as GA3, KNO3, Thiourea and water soaking was imposed on it. The maximum germination percent (70.57%) was obtained from treatment with thiourea (2.0 %), which was on par (67.33%) with water soaking treatment. The maximum shoot length (15.13 cm), root length (6.40 cm), plant height (21.53 cm), fresh weight of the seedling (726.33 mg/plant), dry weight of seedling (72.90 mg/plant) and seedling vigour index I (1433.52) was recorded in water soaking for 30 minutes. Introduction Aristolochia tagala Cham. is a threatened medicinal plant of Aristolochiaceae family. It is generally called “Nallaisvara”, “Nallayisvari” or “Nallaeeshvara” in Telugu, “Doddaeeshwariballi” or “Gattadaeeshvari” in Kannada, “Aadutheendapalai” in Tamil and “Valia arayan” in Malayalam. It is a sunloving climber found in forests and open lowland thickets, climbing over bushes and trees. It is distributed throughout tropical and subtropical countries (Murugan et al., 2006.). It is found to grow at 1000 to 1400 m above sea level elevation and occurs along the Western Ghats in India (Nayar et al., 1976). This plant is found to have a significant role in the tribal medicinal system in its natural habitat. It is mainly used to treat snake bites and other poisonous bites. Apart from this, roots are also used to treat bone fracture, malaria, indigestion, toothache, rheumatism and various dermatological conditions by Kani tribes, Thiruvananthapuram and Tirunelveli hills (Ravikumar and Ved, 2000). Due to indiscriminate use of its roots and 1803 Int.J.Curr.Microbiol.App.Sci (2020) 9(10): 1803-1808 deforestation, its population in reducing in its natural habitat. The threatened status was assigned to this species under the red data list of south Indian medicinal plants (Sarvalingam and Rajendran, 2016). Hence the conservation of this species is crucial. One way in conservation is to standardize the propagation technique for its commercial cultivation and ex-situ conservation. But in A. tagala, seed germination is low due to the presence of insufficient endosperm, which leads to low seed viability (Biswas et al., 2007). Therefore, the present study aims to improve seed germination and the effect of different pre-soaking seed treatment on seed germination and seedling growth in Aristolochia tagala Cham. each. The pre-treatments include GA3 at 100, 150, 200 ppm, KNO3 at 1%, 1.5%, 2.0%, Thiourea at 1%, 1.5%, 2.0% and water soaking along with control. Apart from control, the seeds were soaked in the above treatments for 30 minutes and then air-dried. The pre-treated seeds sown in the port rays filled with cocopeat. The experiment was carried out in a poly house. The observations for parameters like germination percentage, rate of germination, days have taken to initiate germination, number of leaves, seedling height, fresh weight of shoot and root, dry weight of shoot and root were recorded. Statistical analysis The experimental data were analyzed using Completely Randomized Design (CRD) with three replications. The data were analyzed statistically as per the method suggested by Panse and Sukhatme (1967). Materials and Methods Experimental site The experiment was carried out at the ICARIndian Institute of Horticultural Research (ICAR-IIHR), Hesaraghatta Lake Post, Bengaluru. It is located in the eastern dry zone of Karnataka state at13O58l North latitude and 78OEast longitude. It is situated at an altitude of 890 m above mean sea level (MSL). Seed collection The fresh seeds of Aristolochia tagala Cham. were collected freshly from the plant maintained in the FGB of RET medicinal plants at ICAR-Indian Institute of Horticultural Research (IIHR), Bengaluru. The seeds were harvested in October 2019. Experimental description The experiment was carried out with eleven different pre-sowing treatments consisting of three replications having twenty-five seeds Results and Discussion Effect of pre-sowing treatments on seed germination of Aristolochia tagala Cham The data of seed germination as an influence by pre-sowing treatments in A. tagala Cham. is presented in Table 1. All the treatments were significantly superior over the control. The seed treated with thiourea (2.0 %) recorded the highest germination percent (70.57%), which was on par (67.33%) with water soaking treatment. The minimum time for complete germination (39.67 days)was taken by thiourea (2.0 %) and was followed by water soaking. The rate of germination (0.93) was maximum in thiourea (2.0 %). This may be due to a strong neutralizing effect of thiourea on inhibitor in the seed or increase in cytokinin activity by thiourea to overcome the seed coat inhibiting effect (Hore and Sin, 1994). A similar result was 1804 Int.J.Curr.Microbiol.App.Sci (2020) 9(10): 1803-1808 also obtained in Hippophae salicifolia seeds which showed the highest germination rate (76%–83%) when treated with thiourea (100 mM) (Airi et al., 2009). Khan (2015) reported that the seed germination rate (76% ± 0.6%) of Boswellia serrata Roxb. improved by thiourea 1 % for 30 minutes. Nandi et al., (2000) obtained a higher germination rate and percentage in Aconitum heterophyllum Wall. and A. balfourii Stap f. by thiourea treatment. Table.1 Effect of pre-sowing treatments on seed germination of Aristolochia tagala Cham Treatment T1 - Control T2 - GA3 100 ppm T3 - GA3 150 ppm T4 - GA3 200 ppm T5 - KNO3 1% T6 - KNO3 1.5% T7 - KNO3 2% T8 - Thiourea 1% T9 - Thiourea 1.5% T10 -Thiourea 2% T11 – Water soaking Mean S. Em ± CD@5% Days taken for complete germination 51.78 44.67 44.11 41.89 44.22 44.11 43.55 44.44 44.00 39.67 41.33 43.98 0.548 1.617 Germination percent 46.24 51.19 52.38 62.10 52.38 53.57 54.76 52.57 57.90 70.57 67.33 56.45 1.412 4.167 Rate of germination 0.56 0.64 0.64 0.65 0.63 0.65 0.67 0.75 0.77 0.93 0.78 0.70 0.02 0.06 Table.2 Effect of pre-sowing treatments on seedling parameters of Aristolochia tagala Cham. at 75 days after sowing Treatment T1 - Control T2 - GA3 100 ppm T3 - GA3 150 ppm T4 - GA3 200 ppm T5 - KNO3 1% T6 - KNO3 1.5% T7 - KNO3 2% T8 - Thiourea 1% T9 - Thiourea 1.5% T10 -Thiourea 2% T11 – Water soaking Mean S. Em ± CD@5% Shoot length (cm) 7.68 8.37 10.15 12.33 8.47 10.53 11.50 10.30 10.93 13.50 15.13 10.81 0.239 0.705 Root length (cm) 3.53 3.87 4.82 5.23 4.10 5.07 5.20 4.32 5.15 5.72 6.40 4.85 0.091 0.269 1805 Seedling height (cm) 11.21 12.24 14.97 17.56 12.57 15.60 16.70 14.62 16.08 19.22 21.53 15.66 0.276 0.815 Int.J.Curr.Microbiol.App.Sci (2020) 9(10): 1803-1808 Table.3 Effect of pre-sowing treatments on the fresh and dry weight of shoot and root of Aristolochia tagala Cham. at 75 days after sowing Treatment Fresh weight of seedling (mg/plant) Dry weight of seedling (mg/plant) Seedling vigour index I T1 - Control T2 - GA3 100 ppm T3 - GA3 150 ppm T4 - GA3 200 ppm T5 - KNO3 1% T6 - KNO3 1.5% T7 - KNO3 2% T8 - Thiourea 1% T9 - Thiourea 1.5% T10 -Thiourea 2% T11 – Water soaking Mean S. Em ± CD@5% 191.33 218.00 277.00 317.33 253.33 291.67 425.67 367.33 385.67 532.67 726.33 362.39 7.74 22.64 22.97 36.57 38.80 40.10 38.10 45.43 53.33 45.33 50.57 60.07 72.90 45.83 0.90 2.63 518.86 626.07 783.33 1090.64 658.33 836.55 916.37 768.50 931.21 1356.68 1433.52 901.82 31.332 92.487 Effect of pre-sowing treatments on seedling parameters of Aristolochia tagala Cham All growth parameters influenced by presowing seed treatments of A. tagala are presented in Tables 2 and 3. There was a significant influence of the treatments on the various seedling parameters. Among the different treatment, the maximum shoot length (15.13 cm), root length (6.40 cm), plant height (21.53 cm), fresh weight of the seedling (726.33 mg/plant), dry weight of seedling (72.90 mg/plant) and seedling vigour index I (1433.52) was recorded in the treatment water soaking for 30 minutes. In general, water soaking of seeds at room temperature helps to soften the seed coat, remove inhibitors, and reduce the time required for germination and increase the germination percentage. The water imbibition by seeds increases the fresh weight of the seeds and water uptake drives the emergence of radicle and enhances germination (Yashaswini, 2009). In Aristolochia tagala, the seed's embryo size is small and at the time of seed maturity they are not developed fully (Baskin and Baskin, 1998). The embryo development in A. tagala occurs after water absorption (Biswas et al., 2007). Due to which water soaking helped in improving seed germination in A. tagala Cham. The effect of water soaking on other growth parameters may be due to its germination stimulating effect. The results recorded by Krishnan et al., (2019) showed that increased germination per cent of 52% in soil within 32 days in A. tagala was obtained in seeds soaked in warm water 500C for 5min.Debi and Prakash (2015) reported that maximum germination percentage (100%) and mean germination time were obtained in the seeds treated with normal water (24 hours) in Oroxylum indicum. Similar results were also obtained in Pterocarpus santalinus (Naidu and Mastan, 2001), Rauvolfia serpentina (Gupta, 2003), Gloriosa superba (Anandhi et al., 2016) and Terminalia bellirica (Vikas, 2018). 1806 Int.J.Curr.Microbiol.App.Sci (2020) 9(10): 1803-1808 In conclusion among different pre-sowing treatments, water soaking for 30 minutes was the best treatment with respect to seed germination and other growth parameter and the next best treatment was thiourea 2%. Though the germination percent was highest in thiourea (2.0 %), it was on par with the water soaking. Other growth parameters such as seedling height, root length, shoot length, dry weight, fresh weight and seedling vigour index I were maximum in water soaking. The result of the present investigation will help large scale seedling production conservation of this threatened species. References Airi, S., Bhatt, I. D., Bhatt, A., Rawal, R. S. and Dhar, U., 2009, Variations in seed germination of Hippophae salicifolia with different pre-soaking treatments. J. For. Res., 20(1): 27-30. Anandhi, S. and Rajamani, K., 2012, Studies on seed germination and growth in Gloriosa superba L. Glob. J. Res. Med. Plants Indig. Med., 1(10): 524-528. Baskin, C. C. and Baskin, J. M., 1998, Seeds: Ecology, biogeography and evolution of dormancy and germination. Academic press, San Diego Biswas, A., Bari, M. A., Roy, M.and Bhadra S. K., 2007, In-vitro regeneration of Aristolochia tagala Champ. – A rare medicinal plant of Chittagong hill tracts. J. Biosci., 15: 63-67. Debi, C. and Prakash, V., 2015, Seed sources and habitat variation affect seed germination in Oroxylum indicum (L.) Benth. Ex. An important threatened medicinal tree. Int. J. Life Sci. Technol., 8(1):1-9. Gupta, V., 2003, Seed germination and dormancy breaking techniques for indigenous medicinal and aromatic plants. J. Med. Arom. Plant Sci., 23(2): 402-407. Hore, J. K. and Sen, S. K., 1994, Role of presowing seed treatment on germination, seedling growth and longevity of ber (Ziziphus mauritiana L.) seeds. Indian J. agric. Res., 28(4): 285-284. Khan, M. R., 2015, Studies on seed germination of a threatened, endangered medicinal plant species Boswellia Serrata Roxb. Weekly Science, 2(39): 1-5. Krishnan, S. G., Dan, M. and Kumar, E. S. S., 2019, Studies on the seed germination and seedling morphology of Aristolochia tagala Cham. (Aristolochiaceae) from the Western Ghats, India. Int. J. Res. Anal. Rev.,6(2): 620-625. Murugan, R., Shivanna, K. R. and Rao, R. R., 2006, Pollination biology of Aristolochia tagala, a rare species of medicinal importance. Curr. Sci., 91(6):795-798. Naidu, C. V. and Mastan, M., 2001, Seed pretreatment methods to improve germination in Pterocarpus santalinus Linn. Indian J. For., 24(3): 342-343. Nandi, S. K., Pandey, H., Nadeem, M. and Palni, L. M., S., 2000, Chemical stimulation of seed germination in Aconitum heterophyllum Wall. and A. balfourii Stapf.: Important Himalayan species of medicinal value. Seed Sci. Technol.,28(1): 39-48. Nayar, R. C., Mary, Z., Yoganarasimhan, S. N. and Sharma, A. R., 1976, Pharmacognostical studies on the root of Aristolochia tagala Cham. (Aristolochiaceae). Proc. Natl. Acad. Sci. India B., 84: 90-94. Panse, V. G. And Sukhatme, P. V., 1967, Statistical methods for agricultural workers, ICAR, New Delhi, p.381. Ravikumar, K. and Ved, D. K., 2000, Illustrated field guide-100 Red listed medicinal plants of conservation concern in Southern India. Foundation 1807 Int.J.Curr.Microbiol.App.Sci (2020) 9(10): 1803-1808 for revitalization of local health traditions, Bangalore, Pp. 60-62. Sarvalingam, A. and Rajendran, A., 2016, Rare, endangered and threatened (RET) climbers of Southern Western Ghats, India. Rev. Chil. Hist. Nat., 89(9): 1-5. Vikas, K., 2016, Effect of pre-sowing seed treatment on germination and seedling growth of Terminalia bellirica (Gaertn.) Roxb. Indian J. Ecol., 43(1): 233-238. Yashaswini, S., 2009, Propagation studies in selected ret (Rare, Endangered and Threatened) medicinal plant species. MSc. Thesis, Uni. Agri. Sci., Dharwad (India). How to cite this article: Rajani Bhat, G. Raviraja Shetty, P. E. Rajasekharan, D. A. Pooja, M. Ganapathi and Sadhashiv Nadukeri. 2020. Effect of Pre-sowing Seed Treatments on Aristolochia tagala Cham: A Threatened Medicinal Plant of South India. Int.J.Curr.Microbiol.App.Sci. 9(10): 1803-1808. doi: https://doi.org/10.20546/ijcmas.2020.910.219 1808