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Micropropagation in banana using high levels of cytokinins does not involve any genetic changes as revealed by RAPD and ISSR markers

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Abstract

Use of high levels of growth regulators during micropropagation results in undesirable clonal variability in important commercial crops such as banana. The present study investigated the effects of high levels of cytokinins on micropropagation in banana (genotype AAB), and the genetic stability of plantlets was assessed using RAPD and ISSR markers. Cytokinins, such as BA and kinetin were added to the routine shoot multiplication medium at concentrations up to 10 mg l−1. After 12 weeks of culture involving three subcultures, the maximum number of shoot buds were produced in cultures receiving either 5 mg l−1 BA (80 shoot buds) or 4 mg l−1 kinetin (62 shoot buds). Certain morphological abnormalities observed during proliferation of shoot buds in vitro were not observed during acclimatization ex vitro. To check the genetic stability, RAPD and ISSR profiles of micropropagated plantlets obtained from different cytokinin-treatments were compared with control microplants maintained on MS medium as well as the field-grown mother plant. A total of 50 RAPD and 12 ISSR primers resulted in 625 distinct and reproducible bands. Thus a total of 17,400 bands were generated showing homogeneous RAPD and ISSR patterns. Band intensity histogram of each gel confirmed their monomorphic nature with no genetic variation in all the plantlets analysed. Based on these results a protocol for high rate shoot multiplication was worked out leading to uniform shoot production.

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Abbreviations

AS:

Adenine sulphate

BA:

N6-benzyladenine

HRMM:

High rate multiplication medium

IBA:

Indole-3-butyric acid

2iP:

N6-(2-isopentenyl)adenine

ISSR:

Inter Simple Sequence Repeats

NAA:

α-naphthaleneacetic acid

NR:

Nanjanagudu Rasabale (AAB)

RH:

Relative Humidity

RAPD:

Random Amplified Polymorphic DNA

PCR:

Polymerase chain reaction

MS:

Murashige and Skoog’s (1962) medium

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Acknowledgements

The first author acknowledges the Council of Scientific and Industrial Research (CSIR), India for financial support in the form of a JRF. Encouragement by Dr. G. A. Ravishankar Head, PCBT and Dr. V. Prakash, Director, CFTRI for research activities are gratefully acknowledged. Authors are extremely grateful to R. Thimmaraju for assisting in data analysis and making graphics, and R. Kaunain-Roohie for her help in maintaining stock cultures of banana

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  1. Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore, Karnataka, 570 020, India

    Lakshmanan Venkatachalam, R. V. Sreedhar & Neelwarne Bhagyalakshmi

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  1. Lakshmanan Venkatachalam
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  2. R. V. Sreedhar
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  3. Neelwarne Bhagyalakshmi
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Correspondence to Neelwarne Bhagyalakshmi.

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Venkatachalam, L., Sreedhar, R.V. & Bhagyalakshmi, N. Micropropagation in banana using high levels of cytokinins does not involve any genetic changes as revealed by RAPD and ISSR markers. Plant Growth Regul 51, 193–205 (2007). https://doi.org/10.1007/s10725-006-9154-y

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