Valizadeh J, Valizadeh M
Department of Biology, College of Science, University of Sistan and Baluchestan, Zahedan, Iran.
Pak J Biol Sci. 2009 Nov 1;12(21):1415-9. doi: 10.3923/pjbs.2009.1415.1419.
Leaf and internode explants of Withania coagulans Dunal were used to evaluate the effect of growth regulators on the in vitro callus induction and plant regeneration. Two independent experiments were performed to establish callus and plant regeneration. In the first experiment, leaf segments were cultured on MS basal medium fortified with 2, 4-dichlorophenoxyacetic acid (2, 4-D, 2-4 mg L(-1)) with combination of 6-benzyladenine (BA, 0.5-1 mg L(-1)) or kinetin (Kin, 0.5-1 mg L(-1)). Callus initiation was observed best in all media after 14-16 days (100%). Highest callus growth in terms of dry weight (76 +/- 5.34 mg) was observed in MS medium fortified with 2 mg L(-1) 2, 4-D and 0.5 mg L(-1) Kin. In the second experiment, callusing also obtained from internode segments cultured on MS medium with the presence of 2, 4-D (2-4 mg L(-1)) in association with BA (0.25-0.5 mg L(-1)). Best callusing rates (42%) and dry weight (86 +/- 3.68 mg) occurred in the presence of 4 mg L(-1) 2, 4-D with combination of 0.25 mg L(-1) BA. Shoot induction was obtained just from callus induced from internode explants on MS medium containing BA (2 mg L(-1)) with IBA (0.5 mg L(-1)), but the yield of shoot regeneration was unsatisfactory (18-33%). Regenerated shoots were rooted best (100%) on half strength MS medium containing IBA (2 mg L(-1)) and produced 23 roots with an average root length of 6.5 +/- 0.41 cm. Rooted plantlets transferred to soil and sand mixture (2:1) showed 75% survival when transferred to outdoor. The procedure reported here in vitro culture of W. coagulans for first time.
使用凝乳茄（Withania coagulans Dunal）的叶片和节间外植体来评估生长调节剂对体外愈伤组织诱导和植株再生的影响。进行了两项独立实验以建立愈伤组织和植株再生体系。在第一个实验中，将叶片切段培养在添加了2,4 - 二氯苯氧乙酸（2,4 - D，2 - 4 mg L(-1)）与6 - 苄基腺嘌呤（BA，0.5 - 1 mg L(-1)）或激动素（Kin，0.5 - 1 mg L(-1)）组合的MS基本培养基上。14 - 16天后在所有培养基中均观察到最佳愈伤组织起始情况（100%）。在添加2 mg L(-1) 2,4 - D和0.5 mg L(-1) Kin的MS培养基中观察到以干重计的最高愈伤组织生长量（76±5.34 mg）。在第二个实验中，在含有2,4 - D（2 - 4 mg L(-1)）并与BA（0.25 - 0.5 mg L(-1)）组合的MS培养基上培养节间切段也获得了愈伤组织。在4 mg L(-1) 2,4 - D与0.25 mg L(-1) BA组合的情况下出现了最佳愈伤组织诱导率（42%）和干重（86±3.68 mg）。仅从节间外植体诱导的愈伤组织在含有BA（2 mg L(-1)）和IBA（0.5 mg L(-1)）的MS培养基上获得了芽诱导，但芽再生产量不令人满意（18 - 33%）。再生芽在含有IBA（2 mg L(-1)）的半强度MS培养基上生根最佳（100%），并产生23条根，平均根长为6.5±0.41 cm。生根的小植株转移到土壤和沙子混合物（2:1）中，转移到室外时显示出75%的存活率。本文首次报道了凝乳茄的体外培养方法。
