Regeneration of stevia rebaudiana and analysis of somaclonal variation by rapd

被引：12

作者：

Mokladuzzaman, Md. ^{[1
]}

Mahbubur Rahman, S.M. ^{[1
]}

机构：

[1] Biotechnology and Genetic Engineering Discipline, Khulna University

来源：

Biotechnology | 2009年 / 8卷 / 04期

关键词：

Callus; RAPD; Regenerated plants; Root; Shoot; Somaclonal variation; Transplantation;

D O I：

10.3923/biotech.2009.449.455

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The objectives of this experiment were to develop the optimal concentration of auxin and cytokinin for regeneration of Stevia rebaudiana and finally analysis somaclonal variation by RAPD. Various degrees of callus induced from the leaf segments cultured on MS medium supplemented with the different concentrations and combinations of NAA+BA and 2,4-D +BA. Among them. 1.5 mg L-1 NAA with 1.0 mg L-1 BA was the best for callus induction (91.67%) which also produced highest fresh weight (621.7 mg) and dry weight (79.00 mg) of callus. For shoot formation, calli were transferred on to MS medium supplemented with different concentrations and combinations of BA and NAA with control. The highest number of shoots (2.17) and the highest average length of the shoot (3.22 cm) per culture was observed at 1.8 mg L-1 of BA with 0.12 mg L-1 of NAA. The regenerated shoots were then transferred to MS liquid medium supplemented with same concentration of IBA and NAA. All the treatments produced roots and 1.50 mg L-1 IBA produced highest percentage of root (93.33%), but 1.00 mg L-1 NAA produced highest number (no. 7.66) of roots and highest length of roots (13.33 cm) per culture. The regenerated plantlets were successfully transferred into pots containing 75% soil and 25% sand and finally transferred into the field. Apparently somaclonal variations were examined among regenerated plants along with mother plant by RAPD. DNA samples from mother plant and 9 randomly selected regenerated plants were subjected to RAPD analysis. Bands generated through RAPD-PCR were scored according to whether they were present (1) or absent (0) to determine the extent of somaclonal variation. The estimation of genetic similarity coefficient based on RAPD band-sharing data analyzed indicated that some regenerated plants were 100% similar to the mother plants and some were 71, 57 or 14% similar may be due to variation in vitro condition. © 2009 Asian Network for Scientific Information.

引用

页码：449 / 455

页数：6

共 17 条

[1]

Caswell K.L., Leung N., Chibbar R.N., An efficient method for in vitro regeneration from immature inflorescence explanis of Canadian wheat cultivars, Plant Cell Tissue Organ Cult., 60, pp. 69-73, (2000)

[2]

Delporte F., Mostade O., Jacquemin J.M., Plant regeneration through callus initiation from thin mature embryo fragments of wheat, Plant Cell Tissue Organ Cult., 67, pp. 73-80, (2001)

[3]

Hashmi G., Huettel R., Meyer R., Krusberg L., Hammerschlag F., RAPD analysis of somaclonal variants derived from embryo callus cultures of peach, Plant Cell Rep., 16, pp. 624-627, (1997)

[4]

Isabel N., Tremblay L., Michaud M., Tremblay F.M., Bousquet J., RAPDs as an aid to evaluate the genetic integrity of somatic embryogenesis-derived populations of Picea mariana (Mill.) B.S.P. Theor, Applied Genet., 86, pp. 81-87, (1993)

[5]

Karp A., Somaclonal variation as a tool for crop improvement, Euphytica, 85, pp. 295-302, (1995)

[6]

Khanam S., Rajasekharan P.E., Das K., Raman D., Shivananda B.G., In vitro methods for production of stevioside from Stevia rebaudiana, Indian J. Nat. Prod., 21, pp. 14-15, (2005)

[7]

Mitra A., Pal A., In vitro regeneration of Stevia rebaudiana (Bert) from the nodal explant, J. Plant Biochem. Biotechnol., 16, pp. 59-62, (2007)

[8]

Munthali T., Newbury H.J., Ford-Lloyd B.V., The detection of somaclonal variants of beet using RAPD, Plant Cell Rep., 15, pp. 474-478, (1996)

[9]

Nei M., Nei's Original measures of genetic identity and genetic distance, Am. Nat., 106, pp. 283-292, (1972)

[10]

Saharan V., Yadav R.C., Yadav N.R., Chapagain B.P., High frequency plant regeneration from desiccated calli of indica rice (Oryza saliva L.), Afr. J. Biotechnol., 3, pp. 256-259, (2004)

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