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Slow-growth conservation and clonal fidelity of Tetrastigma hemsleyanum microplants

  • Plant Tissue Culture
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Abstract

Storage under slow growth is one of the most successful techniques for medium-term germplasm conservation and could effectively reduce somaclonal variation during subculture. Tetrastigma hemsleyanum is an endangered medicinal herb with anticancer potential. In the present study, an effective protocol was developed for clonal propagation and medium-term conservation of this endangered medicinal plant. Subculture duration was significantly prolonged to at least 10 mo by using 0.2 mg L−1 of maleic hydrazide (MH) at 8°C. Analyses of inter-simple sequence repeat (ISSR) markers, sequence-related amplified polymorphism (SRAP) markers, total proteins, and peroxidase isozymes together demonstrated the genetic stability of the regenerated germplasm. Total flavonoid content of regenerated microplants also remained stable. These results suggest that MH has potential application in slow-growth conservation of plants.

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References

  • Agarwal T, Gupta AK, Patel AK, Shekhawat NS (2015) Micropropagation and validation of genetic homogeneity of Alhagi maurorum using SCOT, ISSR and RAPD markers. Plant Cell Tissue Organ Cult 120:313–323

    Article  CAS  Google Scholar 

  • Aliyu OM, Awopetu JA (2007) Assessment of genetic diversity in three populations of cashew (Anacardium occidentale L.) using protein-isoenzyme-electrophoretic analysis. Genet Resour Crop Evol 54:1489–1497

    Article  CAS  Google Scholar 

  • Bairu MW, Aremu AO, Staden JV (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173

    Article  CAS  Google Scholar 

  • Devi SP, Kumaria S, Rao SR, Tandon P (2014) Single primer amplification reaction (SPAR) methods reveal subsequent increase in genetic variations in micropropagated plants of Nepenthes khasiana Hook. f. maintained for three consecutive regenerations. Gene 538:23–29

    Article  CAS  PubMed  Google Scholar 

  • Hao YJ, Liu QL, Deng XX (2001) Effect of cryopreservation on apple genetic resources at morphological, chromosomal, and molecular levels. Cryobiology 43:46–53

    Article  CAS  PubMed  Google Scholar 

  • Hao YJ, Wen XP, Deng XX (2004) Genetic and epigenetic evaluations of citrus calluses recovered from slow-growth culture. J Plant Physiol 161:479–484

    Article  CAS  PubMed  Google Scholar 

  • Haque SM, Ghosh B (2013) Field evaluation and genetic stability assessment of regenerated plants produced via direct shoot organogenesis from leaf explant of an endangered ‘asthma plant’ (Tylophora indica) along with their in vitro conservation. Natl Acad Sci Lett 36:551–562

    Article  CAS  Google Scholar 

  • He FG (2010) Research progress in anticancer effect of Tetrastigma hemsleyanum Diels et Gilg and its mechanism. J Oncol 16:75–77 (in Chinese)

    CAS  Google Scholar 

  • Hrahsel L, Basu A, Sahoo L, Thangjam R (2014) In vitro propagation and assessment of the genetic fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers. Appl Biochem Biotechnol 172:1530–1539

    Article  CAS  PubMed  Google Scholar 

  • Jung HJ, Hong Y, Yang HS, Chang HC, Kim HY (2012) Distribution of lactic acid bacteria in garlic (Allium sativum) and green onion (Allium fistulosum) using SDS-PAGE whole cell protein pattern comparison and 16S rRNA gene sequence analysis. Food Sci Biotechnol 21:1457–1462

    Article  Google Scholar 

  • Kaity A, Ashmore SE, Drew RA, Dulloo ME (2008) Assessment of genetic and epigenetic changes following cryopreservation in papaya. Plant Cell Rep 27:1529–1539

    Article  CAS  PubMed  Google Scholar 

  • Largia MJV, Shilpha J, Pothiraj G, Ramesh M (2014) Analysis of nuclear DNA content, genetic stability, Bacoside A quantity and antioxidant potential of long term in vitro grown germplasm lines of Bacopa monnieri (L.). Plant Cell Tissue Organ Cult 120:399–406

    Article  Google Scholar 

  • Lata H, Moraes RM, Bertoni B, Pereira AMS (2010) In vitro germplasm conservation of Podophyllum peltatum L. under slow growth conditions. In Vitro Cell Dev Biol Plant 46:22–27

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nayak SA, Kumar S, Satapathy K, Moharana A, Behera B, Barik DP, Acharya L, Mohapatra PK, Jena PK, Naik SK (2013) In vitro plant regeneration from cotyledonary nodes of Withania somnifera (L.) Dunal and assessment of clonal fidelity using RAPD and ISSR markers. Acta Physiol Plant 35:195–203

    Article  CAS  Google Scholar 

  • Peng X, He JY (2013) The inhibitory effect of Ca2+ on the flavonoid production of Tetrastigma hemsleyanum suspension cells induced by metal elicitors. In Vitro Cell Dev Biol Plant 49:550–559

    Article  CAS  Google Scholar 

  • Peng X, Zhou SL, He JY, Ding L (2013) Influence of rare earth elements on metabolism and related enzyme activity and isozyme expression in Tetrastigma hemsleyanum cell suspension cultures. Biol Trace Elem Res 152:82–90

    Article  CAS  Google Scholar 

  • Peng X, Zhuang DD, Guo QS (2015a) Induction of S phase arrest and apoptosis by ethylacetate extract from Tetrastigma hemsleyanum in human hepatoma HepG2 cells. Tumor Biol 36:2541–2550

    Article  Google Scholar 

  • Peng X, Zhang TT, Zhang J (2015b) Effect of subculture times on genetic fidelity, endogenous hormone level and pharmaceutical potential of Tetrastigma hemsleyanum callus. Plant Cell Tissue Organ Cult 122:67–77

    Article  CAS  Google Scholar 

  • Phulwaria M, Patel AK, Rathore JS, Ram K, Shekhawat NS (2014) An improved micropropagation and assessment of genetic stability of micropropagated Salvadora oleoides using RAPD and ISSR markers. Acta Physiol Plant 36:1115–1122

    Article  CAS  Google Scholar 

  • Rawat JM, Rawat B, Mehrotra S, Chandra A, Nautiyal S (2013) ISSR and RAPD based evaluation of genetic fidelity and active ingredient analysis of regenerated plants of Picrorhiza kurroa. Acta Physiol Plant 35:1797–1805

    Article  CAS  Google Scholar 

  • Refoufi A, Esnault MA (2008) Population genetic diversity in the polyploid complex of wheat grasses using isoenzyme and RAPD data. Biol Plant 52:543–547

    Article  CAS  Google Scholar 

  • Renau-Morata B, Arrillaga I, Segura J (2006) In vitro storage of cedar shoot cultures under minimal growth conditions. Plant Cell Rep 25:636–642

    Article  CAS  PubMed  Google Scholar 

  • Sarkar D, Chakrabarti SK, Naik PS (2001) Slow-growth conservation of potato microplants: efficacy of ancymidol for long-term storage in vitro. Euphytica 117:133–142

    Article  CAS  Google Scholar 

  • Sharma N, Satsangi R, Pandey R, Singh R, Kaushik N, Tyagi RK (2012) In vitro conservation of Bacopa monnieri (L.) using mineral oil. Plant Cell Tissue Organ Cult 111:291–301

    Article  CAS  Google Scholar 

  • Sisunandar RA, Turquay P, Samosir Y, Adkins SW (2010) Cryopreservation of coconut (Cocos nucifera L.) zygotic embryos does not induce morphological, cytological or molecular changes in recovered seedlings. Planta 232:435–447

    Article  CAS  PubMed  Google Scholar 

  • Yadava K, Aggarwalb A, Singha N (2013) Evaluation of genetic fidelity among micropropagated plants of Gloriosa superba L. using DNA-based markers—a potential medicinal plant. Fitoterapia 89:265–270

    Article  Google Scholar 

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Acknowledgments

This work was supported by the public welfare technology research projects of Zhejiang province (Grant No. 2013C32103), the traditional Chinese medical science technology research projects of Zhejiang province (Grant No. 2013ZA119), and the agricultural research projects of Ningbo city (Grant No. 2014C10031).

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Correspondence to Yulan Li.

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Editor: Barbara M. Reed

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Peng, X., Ji, Q., Wu, H. et al. Slow-growth conservation and clonal fidelity of Tetrastigma hemsleyanum microplants. In Vitro Cell.Dev.Biol.-Plant 51, 463–470 (2015). https://doi.org/10.1007/s11627-015-9709-6

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  • DOI: https://doi.org/10.1007/s11627-015-9709-6

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