Abstract
Pinellia ternate (Thunb.) Breit (Araceae) is an important Chinese medicinal herb. In order to obtain the hexadecaploid plants of P. ternate with improved medicinal qualities, the shoot tips from in vitro-grown plants were treated with colchicine at various concentrations for different time to double the chromosome number. We obtained highest (42.5%) frequency of morphological variation when the tips were treated with 0.1% colchicine for 48 h. The putative hexadecaploid plants showed characteristics such as larger and thicker leaves, deeper green leaf color, stouter petiole, larger stomata, lower density of stomata across the lower epidermis of leaves and increased numbers of chloroplasts per stomata guard cell. Further analysis by flow cytometry and chromosome counting confirmed seven out of 129 were hexadecaploids. Besides, the growth rate (GR) of PLBs in hexadecaploid plants was 10.49% (increase rate in 1 day), 1.6 times higher than that of octoploid plants and the alkaloid content of PLBs in hexadecaploid plants was 0.0794%, 1.8 times higher than that of octoploid plant. The results indicated that we have established a successful procedure for development and propagation of hexadecaploid of P. ternate with higher GR and alkaloid content.
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Abbreviations
- PLBs:
-
Protocorm-like bodies
- ZJTCM:
-
Zhejiang Chinese Medical University
- 6-BA:
-
6-Benzylaminopurine
- MS:
-
Murashige and Skoog (1962) medium
- NAA:
-
1-Naphthaleneacetic acid
- AC:
-
Activated carbon
- IBA:
-
Indole-3-butyric acid
References
Berkov S (2001) Size and alkaloid content of seeds in induced auto-tetraploids of Datura innoxia, Datura stramonium and Hyoscyamus niger. Pharm Biol 39:329–331
Chang L, Xu YM, Xue JP (2007) Formation of microtubers from the petiole of Pinellia ternate (Thunb.) Breit.-in vitro and change of endogenous hormones content (Chinese). J Huazhong Agric Univ 26(5):612–615
Chen LL, Gao SL (2007) In vitro tetraploid induction and generation of tetraploids from mixoploids in Astragalus membranaceus. Sci Hortic 112:339–344
Chen FQ, Fu Y, Wang DL, Gao X, Wang L (2007) The effect of plant growth regulators and sucrose on the micropropagation and microtuberization of Dioscorea nipponica Makino. J Plant Growth Regul 26(1):38–45
Dahanayake N, Chen XL, Zhao FC, Yang YS, Wu H (2009) Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L. J Biomed Biotechnol 7:1–7
Dhooghe E, Grunewald W, Leus L, Van Labeke MC (2009) In vitro polyploidisation of Helleborus species. Euphytica 165:89–95
Fan MH, Zhou JY, Peng Y, Liu SN (2005) Effect of induction and proliferation of callus of Pinellia ternate (Chinese). J Shandong Chin Med 24(3):168–171
Gu XF, Yang AF, Meng H, Zhang JR (2005) In vitro induction of tetraploid plants from diploid Zizyphus jujube Mill. cv. Zhanhua. Plant Cell Rep 24:671–676
Hamill SD, Smith MK, Dodd WA (1992) In vitro induction of banana autotetraploids by colchicine treatment of micropropagated diploids. Aust J Bot 40(6):887–896
Liu GF, Li ZN, Bao MZ (2007) Colchicine-induced chromosome doubling in Platanus acerifolia and its effect on plant morphology. Euphytica 157:145–154
Liu JF, Gao SL, Huang HP, Liu L, Li XY (2009) Allotetraploid induction and identification of Salvia miltiorrhiza Bunge and its hybridization breeding. Pharm Biotechnol 16(3):260–264
Liu YH, Liang ZS, Zhang YJ (2010) Induction and in vitro alkaloid yield of calluses and protocorm-like bodies (PLBs) from Pinellia ternate. In Vitro Cell Dev Pl 46:239–245
Mao ZC, Peng ZS (2003) Progress on research of rapid propagation system of Pinellia ternate (Chinese). Chin J Chin Mater Med 28(3):193–195
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue culture. Physiol Plantarum 15:473–479
Omidbaigia R, Mirzaeea M, Hassanib ME, Sedghi Moghadam M (2010) Induction and identification of polyploidy in basil (Ocimum basilicum L.) medicinal plant by colchicine treatment. Int J Plant Prod 4(2):87–98
The State Pharmacopoeia Commission of China (2010) Pharmacopoeia of the people’s republic of China. China Press of Traditional Chinese Medicine, Beijing
Ye YM, Tong J, Shi XP, Yuan W, Li GR (2010) Morphological and cytological studies of diploid and colchicine-induced tetraploid lines of crape myrtle (Lagerstroemia indica L.). Sci Hortic Amsterdam 124:95–101
Zhang KW, Wu H, Li W (2005) Determination of inosine and guanosine in Rhizoma Pinellia ternate. Chin J Pharm Anal 25:487–489
Zhang XW, Wang XF, Zhang XC (2006) Research advances on Pinellia ternate (Chinese). Chin Med J Res Prac 20(6):57–61
Zhang ZH, Dai HY, Xiao M, Liu X (2008) In vitro induction of tetraploids in Phlox subulata L. Euphytica 159:59–65
Acknowledgments
The authors are grateful for the financial support of Zhejiang Key Innovation Team (Traditional Chinese Medicine Clinical Innovation Team) Research Fund Project (2009R50042), 2011 Zhejiang Provincial Science and Technology Agency Nonprofit Technology Research Project and 2010 Zhejiang Provincial University Students Science and Technology Innovation Program (Xinmiao Talents Scheme) (2010R410041).
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Zhishan Ding contributed equally to this work.
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He, L., Ding, Z., Jiang, F. et al. Induction and identification of hexadecaploid of Pinellia ternate . Euphytica 186, 479–488 (2012). https://doi.org/10.1007/s10681-012-0642-z
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DOI: https://doi.org/10.1007/s10681-012-0642-z