Culture and regeneration of mesophyll-derived protoplasts of sorghum [Sorghum bicolor (L.) Moench]

R. V. Sairam; N. Seetharama; P. S. Devi; A. Verma; U. R. Murthy; I. Potrykus

doi:10.1007/s002990050693

Culture and regeneration of mesophyll-derived protoplasts of sorghum [Sorghum bicolor (L.) Moench]

R. V. Sairam, N. Seetharama, P. S. Devi, A. Verma, U. R. Murthy, I. Potrykus

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Abstract

A protocol for plant regeneration from mesophyll/protoplasts of sorghum [Sorghum bicolor (L.) Moench] was developed. The yield of intact protoplasts, their subsequent divisions and regeneration were genotype-dependent. The genotype 296B was always more responsive than IS 32266. For 296B, the sixth leaf from 18-day-old plants kept in dark for 2 days before harvesting was found to be the most suitable source of viable protoplasts. The first division was observed 10-12 days after plating, and the second division after 12-14 days. The maximum plating efficiency was 4.8% in 296 B, followed by 2.48% in IS 32266. Microcolonies were visible after 25-30 days, and microcalli after 60-75 days. Whole plants were obtained after 6-8 weeks of culture of microcalli on MS medium containing 0.2 mg l^-1 kinetin and 2 mg l^-1 BAP. The frequency of regeneration in 296B and IS 32266 was 12.80% and 10.58%, respectively. Ten plants transferred to pots in the glasshouse established well. The seeds collected from glasshouse-grown plants were sown in the field where plants were grown to maturity.

Original language	English (US)
Pages (from-to)	972-977
Number of pages	6
Journal	Plant Cell Reports
Volume	18
Issue number	12
DOIs	https://doi.org/10.1007/s002990050693
State	Published - Sep 1999

All Science Journal Classification (ASJC) codes

Agronomy and Crop Science
Plant Science

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10.1007/s002990050693

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@article{c85e06ddc42d424ab512dd6dc564e112,

title = "Culture and regeneration of mesophyll-derived protoplasts of sorghum [Sorghum bicolor (L.) Moench]",

abstract = "A protocol for plant regeneration from mesophyll/protoplasts of sorghum [Sorghum bicolor (L.) Moench] was developed. The yield of intact protoplasts, their subsequent divisions and regeneration were genotype-dependent. The genotype 296B was always more responsive than IS 32266. For 296B, the sixth leaf from 18-day-old plants kept in dark for 2 days before harvesting was found to be the most suitable source of viable protoplasts. The first division was observed 10-12 days after plating, and the second division after 12-14 days. The maximum plating efficiency was 4.8% in 296 B, followed by 2.48% in IS 32266. Microcolonies were visible after 25-30 days, and microcalli after 60-75 days. Whole plants were obtained after 6-8 weeks of culture of microcalli on MS medium containing 0.2 mg l-1 kinetin and 2 mg l-1 BAP. The frequency of regeneration in 296B and IS 32266 was 12.80% and 10.58%, respectively. Ten plants transferred to pots in the glasshouse established well. The seeds collected from glasshouse-grown plants were sown in the field where plants were grown to maturity.",

author = "Sairam, {R. V.} and N. Seetharama and Devi, {P. S.} and A. Verma and Murthy, {U. R.} and I. Potrykus",

note = "Funding Information: Acknowledgements The first author is grateful to Professors Mariam B. Sticklen, P.B. Kavikishor and K.R. Shivanna for critically reviewing the manuscript. We are also thankful to Dr. D.E. Byth and Dr. B.S. Rana for taking a keen interest in this work and to C. Laxminarayana for technical assistance. The author acknowledges financial support in the form of a grant from ACIAR and the AP-Netherlands Biotechnology projects. The author is also grateful to the Former President, ETH, Dr. Jacob N{\"u}esch, for supporting this work with a Young Scientist Award.",

year = "1999",

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doi = "10.1007/s002990050693",

language = "English (US)",

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T1 - Culture and regeneration of mesophyll-derived protoplasts of sorghum [Sorghum bicolor (L.) Moench]

AU - Sairam, R. V.

AU - Seetharama, N.

AU - Devi, P. S.

AU - Verma, A.

AU - Murthy, U. R.

AU - Potrykus, I.

N1 - Funding Information: Acknowledgements The first author is grateful to Professors Mariam B. Sticklen, P.B. Kavikishor and K.R. Shivanna for critically reviewing the manuscript. We are also thankful to Dr. D.E. Byth and Dr. B.S. Rana for taking a keen interest in this work and to C. Laxminarayana for technical assistance. The author acknowledges financial support in the form of a grant from ACIAR and the AP-Netherlands Biotechnology projects. The author is also grateful to the Former President, ETH, Dr. Jacob Nüesch, for supporting this work with a Young Scientist Award.

PY - 1999/9

Y1 - 1999/9

N2 - A protocol for plant regeneration from mesophyll/protoplasts of sorghum [Sorghum bicolor (L.) Moench] was developed. The yield of intact protoplasts, their subsequent divisions and regeneration were genotype-dependent. The genotype 296B was always more responsive than IS 32266. For 296B, the sixth leaf from 18-day-old plants kept in dark for 2 days before harvesting was found to be the most suitable source of viable protoplasts. The first division was observed 10-12 days after plating, and the second division after 12-14 days. The maximum plating efficiency was 4.8% in 296 B, followed by 2.48% in IS 32266. Microcolonies were visible after 25-30 days, and microcalli after 60-75 days. Whole plants were obtained after 6-8 weeks of culture of microcalli on MS medium containing 0.2 mg l-1 kinetin and 2 mg l-1 BAP. The frequency of regeneration in 296B and IS 32266 was 12.80% and 10.58%, respectively. Ten plants transferred to pots in the glasshouse established well. The seeds collected from glasshouse-grown plants were sown in the field where plants were grown to maturity.

AB - A protocol for plant regeneration from mesophyll/protoplasts of sorghum [Sorghum bicolor (L.) Moench] was developed. The yield of intact protoplasts, their subsequent divisions and regeneration were genotype-dependent. The genotype 296B was always more responsive than IS 32266. For 296B, the sixth leaf from 18-day-old plants kept in dark for 2 days before harvesting was found to be the most suitable source of viable protoplasts. The first division was observed 10-12 days after plating, and the second division after 12-14 days. The maximum plating efficiency was 4.8% in 296 B, followed by 2.48% in IS 32266. Microcolonies were visible after 25-30 days, and microcalli after 60-75 days. Whole plants were obtained after 6-8 weeks of culture of microcalli on MS medium containing 0.2 mg l-1 kinetin and 2 mg l-1 BAP. The frequency of regeneration in 296B and IS 32266 was 12.80% and 10.58%, respectively. Ten plants transferred to pots in the glasshouse established well. The seeds collected from glasshouse-grown plants were sown in the field where plants were grown to maturity.

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Culture and regeneration of mesophyll-derived protoplasts of sorghum [Sorghum bicolor (L.) Moench]

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