Basal stem cluster bud induction and efficient regeneration for the Tibetan endemic medicinal plant Swertia conaensis
Keywords:adventitious buds, basal stem cluster buds, callus, proliferation coefficient, Swertia conaensis
The artificial rapid propagation system for Swertia conaensis T. N. Ho et S. W. Liu was explored to screen the appropriate plant regeneration method and to provide an efficient propagation mode, useful for artificial breeding technology or for further research and development of the Tibetan endemic medicinal plant. In this study, the most suitable explant and hormone were chosen according to single factor test. Next, the effects of different hormone combinations on basal stem cluster bud induction, callus induction, adventitious bud occurrence and plant regeneration were investigated by using complete combination and orthogonal experiment. The obtained results showed that the explants suitable for in vitro of S. conaensis were stem tips with leaves, which were regenerated through the method of basal stem cluster bud occurrence in the MS medium with 2.0 mg∙L-1 6-BA, 0.5 mg∙L-1 NAA, but the proliferation coefficient was low, only 3.16 after 40 days of culture. Subsequently, the proliferation coefficient failed to improve, irrespective of change of the concentration ratio of 6-BA and NAA. Therefore, in the orthogonal experiment of adding ZT, the MS medium with 1.0 mg∙L-1 ZT, 0.5 mg∙L-1 NAA and 2.5 mg∙L-1 6-BA induced a large number of callus green and compact, with 86.30% callus occurrence rate. After 40 days of culture, the rate of adventitious bud occurrence was 96.55% and the proliferation coefficient was high (10.37). The rooting rate was 100% in the 1/2MS medium with 0.5 mg∙L-1 NAA. The survival rate of regenerated plants was more than 95%. Indirect organogenesis was more efficient than direct organogenesis in in vitro culture of S. conaensis. In this study, the efficient and stable regeneration system of S. conaensis was achieved through the method of explant to callus to adventitious buds, which provided an effective way to an endangered species.
Afshan N, Anwar S, Mohammad A (2014). Effect of adenine sulphate interaction on growth and development of shoot regeneration and inhibition of shoot tip necrosis under in vitro condition in adult Syzygium cumini L. -a multipurpose tree. Applied Biochemistry and Biotechnology 173:90-102. https://doi.org/10.1007/s12010-014-0797-2
Ahluwalia V, Elumalai S, Kumar V, Kumar S, Sangwan RS (2018). Nano silver particle synthesis using Swertia paniculata herbal extract and its antimicrobial activity. Microbial Pathogenesis 114:402-408. https://doi.org/10.1016/j.micpath.2017.11.052
Cao TW, Geng CA, Ma YB, Zhang XM, Zhou J, Tao YD, Chen JJ (2015). Chemical constituents of Swertia mussotii and their anti-hepatitis B virus activity. Fitoterapia 102:15-22. https://doi.org/10.1016/j.fitote.2015.01.020
Chen YS (2014). Six new species of Saussurea (Asteraceae) from eastern Himalaya. Phytotaxa 177(4):191-206. https://doi.org/10.11646/phytotaxa.177.4.1
Chinese Academy of Sciences, China flora editorial board (1988). Flora of China (Volume 62). Science Press, Beijing, China.
Debnath SC (2005). Micropropagation of lingonberry: influence of genotype, explant orientation, and overcoming TDZ-induced inhibition of shoot elongation using zeatin. HortScience 40(1):185-188. https://doi.org/10.21273/HORTSCI.40.1.185
Debnath SC (2008). Zeatin-induced one-step in vitro cloning affects the vegetative growth of cranberry (Vaccinium macrocarpon Ait.) micropropagules over stem cuttings. Plant Cell, Tissue and Organ Culture 93(2):231-240. https://doi.org/10.1007/s11240-008-9366-0
Debnath SC, Mcrae KB (2002). An efficient adventitious shoot regeneration system on excised leaves of micropropagated lingonberry (Vaccinium vitis-idaea L.). Journal of Horticultural Science & Biotechnology 77:744-752. https://doi.org/10.1080/14620316.2002.11511567
He TN, Xue CY, Wang W (1994). The origin, dispersal and formation of the distribution pattern of Swertia L. (gentianaceae). Acta Phytotaxonomica Sinica 32(6): 525-537. (in Chinese with English abstract)
Huang HY (2005). Tissue culture of medicinal plant Swertia mileensis. Chinese Traditional and Herbal Drugs 36(2):261-265. (in Chinese with English abstract)
Huang HY, Huang J, Wang MR, Ma XN, Xi YK (2016). Comparison on progeny regeneration capacity among different mating patterns and optimization of cultivation condition in Swertia mileensis. Chinese Traditional and Herbal Drugs 47(3):480-487 (in Chinese with English abstract). https://doi.org/10.7501/j.issn.0253-2670.2016.03.022
Khanal S, Shakya N, Thapa K, Pant DR (2015). Phytochemical investigation of crude methanol extracts of different species of Swertia from Nepal. BMC Research Notes 8(1):1-9. https://doi.org/10.1186/s13104-015-1753-0
Kishore KC, Arifullah M, Gayathri D, Rama GG (2011). In vitro shoot regeneration and control of shoot tip necrosis in tissue cultures of Soymida febrifuga (Roxb.) Plant Tissue Culture and Biotechnology 21(1):11-25. https://doi.org/10.3329/ptcb.v21i1.9559
Kshirsagar P, Chavan J, Nimbalkar M, Yadav S, Dixit G, Gaikwad N (2015a). Phytochemical composition, antioxidant activity and HPLC profiles of Swertia species from Western Ghats. Natural Product Research 29(8):780-784. https://doi.org/10.1080/14786419.2014.986124
Kshirsagar PR, Chavan JJ, Umdale SD, Nimbalkar MS, Dixit GB, Gaikwad NB (2015b). Highly efficient in vitro regeneration, establishment of callus and cell suspension cultures and RAPD analysis of regenerants of Swertia lawii Burkill. Biotechnology Reports 6:79-84. https://doi.org/10.1016/j.btre.2015.03.003
Li N, Huang HY, Zeng B (2020). Cluster bud induction of base stem and establishment of high efficiency regeneration system of Lycium ruthenicum. Chinese Traditional and Herbal Drugs 51(13):3545-3553 (in Chinese with English abstract). https://doi.org/10.7501/j.issn.0253-2670.2020.13.02
Li ZJ, Jiao PP, Zhou ZL, Li Q, Li JQ (2012). Morphological and anatomical features of root sucker propagation of Populus pruinosa. Chinese Bulletin of Botany 47(2):133-140 (in Chinese with English abstract). https://doi.org/10.3724/SP.J.1259.2012.0013
Lin Q, Li Z, Zhang L, Tan XF, Long HX, Wu LL (2016). High-efficiency regeneration of seedlings from hypocotyl explants of tung tree (Vernicia fordii). International Journal of Agriculture and Biology 18:370-376. https://doi.org/10.17957/IJAB/15.0097
Liu C, Allow P, Rowland LJ, Hancock JF, Song GQ (2010). Adventitious shoot regeneration from leaf explants of southern highbush blueberry cultivars. Plant Cell, Tissue and Organ Culture 103:137-144. https://doi.org/10.1007/s11240-010-9755-z
Mahendran G, Bai VN (2017). Plant regeneration through direct somatic embryogenesis, antioxidant properties and metabolite profiles of Swertia corymbosa (Griseb.) Wight ex C.B. Clarke. Plant Biosystems 151(1):39-49. https://doi.org/10.1080/11263504.2015.1064043
Matt A, Jehle JA (2005). In vitro plant regeneration from leaves and internode sections of sweet cherry cultivars (Prunus avium L.). Plant Cell Reports 24: 468-476. https://doi.org/10.1007/s00299-005-0964-6
Nagalakshmi M, Vishwanath S, Viswanath S (2014). Adventitious shoot regeneration from hypocotyls of Wrightia arborea (Dennst.) Mabb.: an endangered toy wood species. Journal Cell and Tissue Research 14:4339-4344.
Preece JE, Huetteman CA, Ashby WC, Roth PL (1991). Micro and cutting propagation of silver maple. I. Results with adult and juvenile propagules. Journal of the American Society for Horticultural Science 116:142-148. https://doi.org/10.21273/JASHS.116.1.142
Song JY, Mattson NS, Jeong BR (2011). Efficiency of shoot regeneration from leaf, stem, petiole and petal explants of six cultivars of Chrysanthemum morifolium. Plant Cell, Tissue and Organ Culture 107(2):295-304. https://doi.org/10.1007/s11240-011-9980-0
Takahashi W, Sugawara F, Yamamoto N, Bando E, Matsushita J, Tanaka O (2004). Plant regeneration in Actinidia polygama Miq. by leaf, stem, and petiole culture with zeatin, and from stem-derived calli on low-sucrose medium. Journal of Forest Research 9(1):85-88. https://doi.org/10.1007/s10310-003-0053-z
Wang QQ, Wang YZ, Huang HY, Guo FM (2018). Rapid propagation system for tissue culture of Codonopsis bulleyana. Journal of Chinese Medicinal Materials 41(6):1262-1266 (in Chinese with English abstract). https://doi.org/10.13863/j. issn1001-4454. 2018. 06.00
Xi YK, Wang Y, Zeng B, Huang HY, Yang WD (2020). Callus induction and adventitious bud differentiation of Cyclocodon lancifolius (Roxb.) Kurz. Botanical Sciences 98(4):534-544. https://doi.org/10.17129/botsci.2609
Xiang FN, Xing MQ, Xia GM, Hu FZ, Li Y (1999). Tissue culture and antihepatitis constituent in calli of Swertia franchetiana H. Smith. Bulletin of Botanical Research 19(2):172-178 (in Chinese with English abstract).
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