Gururaj CHALAGERI, Uddagiri Venkanna BABU

In vitro plant regeneration via petiole callus of Viola patrinii and genetic fidelity assessment using RAPD markers

Viola patrinii DC. (Violaceae) petiole explants were used for inducing calli. Significant callus proliferation was observed on MS medium supplemented with 16.12 mM NAA and 13.33 mM BA. Shoot regeneration was achieved upon transferring the green friable calli to MS medium with a 2-fold dilution of potassium dihydrogen phosphate supplemented with 23.25 mM Kn and 2.68 mM NAA. Maximum shoot regeneration was achieved in 4 weeks. Multiple shoots were separated and further cultured in half-strength MS medium supplemented with 9.85 mM IBA with 2% sucrose. Profuse root development was observed after 20-25 days of culturing. Regenerated plants were successfully transferred to soil, showing an 85% survival rate. The genetic fidelity of the micropropagated plants was analysed using random amplified polymorphic DNA (RAPD) markers. The RAPD profile of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrating the homogeneity of the micropropagated plants. This protocol could be successfully used for the mass multiplication and germplasm conservation of this medicinal plant.

Anahtar Kelimeler:

Key words: Viola patrinii, regeneration, micropropagation, leaf explants, RAPD, genetic fidelity

In vitro plant regeneration via petiole callus of Viola patrinii and genetic fidelity assessment using RAPD markers

Keywords:

Key words: Viola patrinii, regeneration, micropropagation, leaf explants, RAPD, genetic fidelity,

PDF

___

Anand PHM, Harikrishnan KN, Martin KP & Hariharan M (1997). In vitro propagation of Kaempferia rotunda L. Indian Crocus A medicinal plant. Phytomorphology 47: 281-286.
Araki H, Shi L & Yakuwa T (1992). Eff ects of auxin, cytokinin and nitrogen concentration on morphogenesis of tissue-cultured shoot apex of Chinese yam (Dioscorea opposita Th unb.). Journal of the Japanese Society for Horticultural Science 60: 851- 857.
Barik DP, Naik SK, Mudgal A & Chand PK (2007). Rapid plant regeneration through in vitro axillary shoot proliferation of butterfl y pea (Clitoria ternatia L.) – a twinning legume. In Vitro Cellular & Developmental Biology-Plant 43: 144-148.
Bhuyan AK, Pattnaik S & Chand PK (1997). Micropropagation of curry leaf tree [Murraya koenigii (L.) Spreng.] by axillary proliferation using intact seedlings Plant Cell Reports 16: 779- 782.
Bouman H, Kuijpers AM & de Klerk GJ (1992). Th e infl uence of tissue culture methods on somaclonal variation in Begonia L. Physiologia Plantarum 85: A45.
Carlson JE, Tulsieram LK, Glaubitz JC, Luk VWK, Kauff eldt C & Rutledge R (1991). Segregation of random amplifi ed DNA markers in F1 progeny of conifers. Th eoretical and Applied Genetics 83: 194-200.
Chopra RN, Chopra I, Kapur LD & Handa KL (1958). Indigenous Drugs of India. Calcutta: Dhur UN and Sons Pvt Ltd.
Council of Scientifi c and Industrial Research (CSIR) (1976). Th e Wealth of India -- A Dictionary of Indian Raw Material and Industrial Products. Vol. X, New Delhi: Publications and Information Directorate, CSIR.
Erişen S, Atalay E & Yorgancılar M (2011). Th e eff ect of thidiazuron on the in vitro shoot development of endemic Astragalus cariensis in Turkey. Turkish Journal of Botany 35: 521-526.
Gupta PK & Varshney RK (1999). Molecular markers for genetic fi delity during micropropagation and germplasm conservation. Current Science 76: 1308-1310.
Han SH, Lee JS & Kwon OC (1990). Callus induction from petiole segment of wild viola (Viola patrinii DC.). Research Reports of Korea Agricultural Technology Research Institute 11: 13-19 (In Korean with English abstract).
Isabel N, Tremblay L, Michaud M, Tremblay FM & Bousquet J (1993). RAPDs as an aid to evaluate the genetic integrity of somatic embryogenesis-derived populations of Picea mariana (Mill.) B.S.P. Th eoretical and Applied Genetics 86: 81-87.
Kirtikar KR & Basu BD (1995). Indian Medicinal Plants. Vol. I, Dehradun, India: International Book Distributions.
Kobayashi H (1982). Application of tissue culture in sweet potato maintenance and multiplication of germplasm. Japanese Journal of Breeding 32(Suppl. 2): 13-14.
Manickam VS, Elango Mathavan R & Antonisamy R (2000). Regeneration of Indian ginseng plantlets from stem callus. Plant Cell Tissue and Organ Culture 62: 181-185.
Martins M, Sarmento D & Oliveira MM (2004). Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers. Plant Cell Reports 23: 492-496.
Matthew KM (1981). Th e Flora of the Tamilnadu Carnatic. Vol. I, Tiruchirapalli, India: Th e Rapinat Herbarium, St.Joseph’s College.
Murashige T & Skoog F (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiologia Plantarum 15: 473-497.
Nahako I & Shizufumi T (1990). Plant regeneration from Bulgarian rose callus. Plant Cell Tissue and Organ Culture 22: 197-199.
Pierik RLM (1991). Horticulture, new technologies and applications. In: Prakash J & Pierik RLM (eds.) Proceedings of the International Seminar on New Frontiers in Horticulture. Dordrecht: Kluwer Academic Publishers, pp. 141-153.
Prakash E, Shavalli Khan PS, Sairam Reddy P & Rao KR (1999). Regeneration of plants from seed-derived callus of Hybanthus enneaspermus L. Muell., a rare ethnobotanical herb. Plant Cell Reports 18: 873-878.
Rani V, Parida A & Raina SN (1995). Random amplifi ed polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh. Plant Cell Reports 14: 459- 462.
Rani V & Raina SN (2000). Genetic fi delity of organized meristem- derived micropropagated plants: A critical reappraisal. In Vitro Cellular & Developmental Biology-Plant 36: 319-330.
Rout GR (2005). Micropropagation of Clitoria ternatea L. (Fabaceae) - An important medicinal plant. In Vitro Cellular & Developmental Biology-Plant 41: 516-519.
Roy A, Frascaria N, MacKay J & Bousquet J (1992). Segregating random amplifi ed polymorphic DNAs (RAPDs) in Betula alleghaniensis. Th eoretical and Applied Genetics 85: 173-180.
Salvi ND, George L & Eapen S (2001). Plant regeneration from leaf base callus of turmeric and random amplifi ed polymorphic DNA analysis of regenerated plants. Plant Cell Tissue and Organ Culture 66: 113-119.
Samantaray S, Rout GR & Das P (1995). An in vitro study on organogenesis in Trema orientalis (Blume) L. Plant Science 105: 87-94.
Sahoo Y, Pattnaik SK & Chand PK (1997). In vitro clonal propagation of an aromatic medicinal herb Ocimum basilicum L. (Sweet basil) by axillary shoot proliferation. In Vitro Cellular & Developmental Biology-Plant 33: 293-296.
Sato T, Kwon OC, Miyake H, Taniguchi T & Maeda E (1995). Regeneration of plantlets from petiole callus of wild viola (Viola patrinii DC.). Plant Cell Reports 14: 768-772.
Sharma N, Chandel KPS & Paul A (1993). In vitro propagation of Gentiana kurroo: an indigenous threatened plant of medicinal importance. Plant Cell Tissue and Organ Culture 34: 307-309.
Sharma TR & Singh BM (1997). High frequency in vitro multiplication of disease free Zingiber offi cinale Rose. Plant Cell Reports 17: 68-72.
Shrivastava N & Rajani M (1999). Multiple shoot regeneration and tissue culture studies on Bacopa monnieri (L.) Pennell. Plant Cell Reports 18: 919-923.
Tripathi YK, Gurha P, Ghosh D, Kumar RV & Prakash V (2007). Determination of phytogenetic relationships among Isoetes species using random primers. Turkish Journal of Botany 31: 367-372.
Usha R & Swami PM (1998). In vitro micropropagation of sweet wormwood (Artemisia annua L.). Phytomorphology 48: 149- 154.
Yao DY & Wang JZ (1987). Jr. Shandong Teachers’s University (Natural Science Ed.) 2: 243-284.