Abstract
Protoplasts were isolated from immature cotyledons of Glycine max L. Merr. cv. Clark 63 and cultured in liquid or in agarose-gelled modified KP8 medium. Plating efficiencies of 45–50% were obtained in liquid medium and 55–60% in 1.2% (w/v) agarose beads. Upon regular dilution with K8 medium rapidly growing green microcalli (1–2 mm in size) were obtained in 5–6 weeks, which upon transfer to MSB medium with 0.5 mg 1−1 each of 2,4-D, BA, Kn and 500 mg 1−1 CH produced compact green calli in 4–6 weeks. After 3–4 regular subcultures of 14 days each on MSB medium containing 0.5 mg 1−1 each of BA, Kn, ZT, 0.1 mg 1−1 NAA and 500 mg 1−1 CH, about 21% of the compact calli formed multiple shoots. Addition of glutamine, asparagine and GA3 enhanced shoot regeneration up to 30%. Shoots of 0.5–1.0 cm length were transferred to 1/2 MS medium with 0.01 mg 1−1 TH and 0.5 mg 1−1 GA3 for elongation. In 2 to 3 weeks, approximately 60% of the shoots were 2–3 cm in length. These shoots were rooted on 1/2 MS with 1% sucrose and 0.2 mg 1−1 IBA or 0.5 mg 1−1 NAA. So far, twenty six plants have been transferred to the greenhouse, where they all have set seed.
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Abbreviations
- BA:
-
6-benzyladenine
- CH:
-
casein hydrolysate
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- FDA:
-
fluorescein diacetate
- GA3 :
-
gibberellic acid
- IBA:
-
indole-3-butyric acid
- Kn:
-
kinetin
- MES:
-
2[N-morpholino] ethane sulfonic acid
- NAA:
-
naphthaleneacetic acid
- TH:
-
thidiazuron
- ZT:
-
zeatin
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Communicated by E. D. Earle
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Dhir, S.K., Dhir, S. & Widholm, J.M. Plantlet regeneration from immature cotyledon protoplasts of soybean (Glycine max L.). Plant Cell Reports 10, 39–43 (1991). https://doi.org/10.1007/BF00233030
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DOI: https://doi.org/10.1007/BF00233030