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ABSTRACT: Organogenetic buds were induced from hypocotyl and cotyledon explants of oil crop Perilla frutescens in Murashige and Skoog (MS) medium supplemented with 5.7 M indole-3-acetic acid (IAA) and 8.9 – 13.3 M 6-benzylaminopurine (BA). Shoots were rooted on MS medium with 2.9 M IAA and 1.4 M gibberellic acid (GA3) and the regenerated plants flowered and set seeds normally.
Biologia Plantarum 02/2005; 49(1):129-132. · 1.97 Impact Factor
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ABSTRACT: An efficient and reproducible protocol is described for the regeneration of Astragalus melilotoides protoplasts isolated from hypocotyl-derived embryogenic calli. Maximum protoplast yield (11.74 +/- 0.6x10(5)/g FW) and viability (87.07 +/- 2.8%) were achieved using a mixture of 2% (w/v) Cellulase Onozuka R10, 0.5% (w/v) Cellulase Onozuka RS, 0.5% (w/v) Macerozyme R10, 0.5% (w/v) Hemicellulase, and 1% (w/v) Pectinase, all dissolved in a cell protoplast wash (CPW) salt solution with 13% (w/v) sorbitol. First divisions occurred 3-7 days following culture initiation. The highest division frequency (9.86 +/- 0.68%) and plating efficiency (1.68 +/- 0.05%) were obtained in solid-liquid medium (KM8P) supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid, 0.5 mg/l 6-benzylaminopurine (BA), 0.2 mg/l kinetin, 0.2 M glucose, 0.3 M mannitol and 500 mg/l casein hydrolysate. Upon transfer to MS medium with 0.5 mg/l alpha-naphthaleneacetic acid and 1-2 mg/l BA, the protoplast-derived calli produced plantlets via somatic embryogenesis (56.3 +/- 4.1%) and organogenesis (21.6 +/- 0.6%). Somatic embryos or adventitious shoots developed into well-rooted plantlets on MS medium without any plant growth regulators or supplemented with 3.0 mg/l indole-3-butyric acid, respectively. About 81% of the regenerants survived in soil, and all were normal with respect to morphology and growth characters.
Plant Cell Reports 06/2004; 22(10):741-6. · 2.27 Impact Factor
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ABSTRACT: An efficient and reproducible protocol is described for the regeneration of Astragalus melilotoides protoplasts isolated from hypocotyl-derived embryogenic calli. Maximum protoplast yield (11.740.6105/g FW) and viability (87.072.8%) were achieved using a mixture of 2% (w/v) Cellulase OnozukaR10, 0.5% (w/v) Cellulase OnozukaRS, 0.5% (w/v) MacerozymeR10, 0.5% (w/v) Hemicellulase, and 1% (w/v) Pectinase, all dissolved in a cell protoplast wash (CPW) salt solution with 13% (w/v) sorbitol. First divisions occurred 3–7days following culture initiation. The highest division frequency (9.860.68%) and plating efficiency (1.680.05%) were obtained in solid-liquid medium (KM8P) supplemented with 1.0mg/l 2,4-dichlorophenoxyacetic acid, 0.5mg/l 6-benzylaminopurine (BA), 0.2mg/l kinetin, 0.2M glucose, 0.3M mannitol and 500mg/l casein hydrolysate. Upon transfer to MS medium with 0.5mg/l -naphthaleneacetic acid and 1-2 mg/lBA, the protoplast-derived calli produced plantlets via somatic embryogenesis (56.34.1%) and organogenesis (21.60.6%). Somatic embryos or adventitious shoots developed into well-rooted plantlets on MS medium without any plant growth regulators or supplemented with 3.0mg/l indole-3-butyric acid, respectively. About 81% of the regenerants survived in soil, and all were normal with respect to morphology and growth characters.
Plant Cell Reports 04/2004; 22(10):741-746. · 2.27 Impact Factor
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ABSTRACT: Embryogenic callus was obtained only from hypocotyl explants of Astragalus adsurgens and light inhibited the formation of embryogenic callus. A high yield (1.2 x 10(6)/g F. Wt.) of protoplasts with high viability (over 80%) could be isolated from 10-day-old embryogenic callus. Protoplasts were induced to undergo sustained division and to form cell colonies when cultured in agarose-solidified medium (KMP) containing 1/4 strength of mineral salts and supplemented with 1.5 mg/L 2, 4-D, 0.5 mg/L BA and 0.5 mol/L glucose at a plating density of 1.0 x 10(5)mL, where the plating efficinency was 16.8%. Conditioning medium significantly improved the formation of cell colonies. When protoplast-derived colonies were maintained at 4 degrees C for 2 weeks and subsequently transferred onto medium (MS) with 0.1 mg/L NAA and 1.0 mg/L BA, somatic embryogenesis occurred. Frequency of cell colonies producing somatic embryos reached 70%, and the number of somatic embryos per gram cells was over 200. Cultured on hormone-free half-strength MS medium, somatic embryos developed into healthy plantlets with normal chromosome complement.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 02/2000; 16(1):17-21.
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ABSTRACT: Protoplasts from 4-day-old embryogenic cell suspension cultures of Astragalus adsurgens, when cultured in KM8P medium which ammonium concentration was reduced to 2.5 mmol/L and supplemented with 0.5 mg/L NAA, 1.0 mg/L 2, 4-D, 0.7 mg/L BA and 0.4 mol/L glucose, underwent cell sustained divisions and formed cell colonies at a frequency of 16%-20%. Preplasmolysis or low temperature treatment of suspension cells prior to enzyme incubation enhanced colony formation. Following proliferation on MS medium containing 1.0 mg/L 2, 4-D and 0.5 mg/L BA, cell colonies were cultured on MS medium containing 0.1 mg/L NAA and 1.0 mg/L BA, where approximately 40% of colonies produced somatic embryos ranging in number from 20 to 40 per colony. No significant decrease was found in the potential of somatic embryogenesis when protoplast colonies were obtained from long-term cell suspensions. On hormone-free 1/2 MS medium, somatic embryos developed into intact plants, which showed normal morphology and stable chromosome number.
Shi yan sheng wu xue bao 01/2000; 32(4):401-8.
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ABSTRACT: An efficient procedure was developed for inducing callus and plant regeneration using hypocotyl segments of Astragalus adsurgens. The combinations and concentrations of different growth regulators were shown to be critical factors for both the frequency
and the type of callus formation as well as for the potential of callus differentiation. Of the four morphologically distinct
types of calli that were induced, a friable, yellow callus, i.e. type I, induced on MS medium supplemented with 9.0 µM 2,4-dichlorophenoxyacetic acid and 2.2 µM N6-benzylaminopurine (BA), and then transferred to MS medium containing 0.5 µM α-naphthaleneacetic acid and 8.9 µM BA, exhibited the maximum frequency of shoot regeneration (75%). After regenerated shoots were transferred onto half-strength
MS medium without growth regulators, they rooted and complete plants were obtained. Plantlet regeneration from callus cultures
required 7–8 weeks.
Plant Cell Reports 03/1998; 17(6):567-570. · 2.27 Impact Factor
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ABSTRACT: A reproducible release of viable protoplasts was obtained from friable calli of Astragalus adsurgens. Protoplasts underwent sustained divisions and formed cell colonies when cultured in either liquid or agarose-solidified
Kao and Michayluk (1975) protoplast medium (KM8P) supplemented with 1.5 mg/l 2,4-D, 0.5 mg/l BA and 0.5 M glucose. Compared to liquid culture, agarose bead culture improved division frequency effectively, the two culture systems
showing a plating efficiency of 0.8±0.5% and 6.5±0.7%, respectively. Upon transfer to Murashige and Skoog (1962) medium (MS)
with 1–2 mg/l BA, alone or in combination with NAA or 2,4-D at 0.1 mg/l, the protoplast-derived calli produced complete plantlets
through somatic embryogenesis. The maximum percentage of calli producing somatic embryos (52.5± 2.2%) occurred on MS medium
containing 0.1 mg/l NAA and 1 mg/l BA, whereas the maximum number of calli regenerating plantlets (64.7±6.2) was obtained
on MS medium with 0.1 mg/l NAA and 2 mg/l BA.
Plant Cell Reports 01/1998; 17(4):313-317. · 2.27 Impact Factor
