Smith R H, Price H J, Thaxton J B
In Vitro. 1977 May;13(5):329-34. doi: 10.1007/BF02616179.
Defined in vitro conditions for callus initiation by Gossypium arboreum L. were determined, and different tissues were evaluated as explant sources, Environmental conditions tested included light versus dark, and low light versus high light. Different nutrient media as well as carbohydrate sources were examined. Our data show that hypocotyl tissue was superior to cotyledon or leaf tissue as the explant source for callus proliferation; the Murashige-Skoog inorganic formulation with (in mg per 1) 100 myo-inositol, 0.4 thiamine-HCl, 2 indoleacetic acid (IAA), 1 kinetin, and 3% glucose solidifield by agar was the best medium to initiate callus. Cultures with sucrose as a carbohydrate source browned rapidly. Callus proliferation was superior under high light (8000 to 9000 lux) conditions at 20 +/- 1 degree C. Various combinations of auxins and cytokinins were tested for their ability to improve callus proliferation and subsequent growth of subcultures. Although the MS medium containing IAA and kinetin was found superior for obtaining rapid proliferation of callus from hypocotyl explants, a second medium containing 2 mg per 1 naphthaleneacetic acid (NAA) and 0.5 to 1 mg per 1 benzyladenine (BA) was found necessary for vigorous growth of subcultured callus. A MS medium with 5 to 10 mg per 1 N6-[delta2-isopentenyl]-adenine (2ip) and 1 mg per 1 NAA was also favorable for continued subculturing.
确定了陆地棉愈伤组织诱导的体外条件，并评估了不同组织作为外植体来源，测试的环境条件包括光照与黑暗、低光照与高光照。研究了不同的营养培养基以及碳水化合物来源。我们的数据表明，下胚轴组织作为愈伤组织增殖的外植体来源优于子叶或叶片组织；含有（每升毫克数）100肌醇、0.4盐酸硫胺素、2吲哚乙酸（IAA）、1激动素和3%葡萄糖并由琼脂固化的Murashige-Skoog无机配方是诱导愈伤组织的最佳培养基。以蔗糖作为碳水化合物来源的培养物迅速褐变。在20±1摄氏度的高光（8000至9000勒克斯）条件下，愈伤组织增殖更佳。测试了各种生长素和细胞分裂素组合促进愈伤组织增殖及后续继代培养生长的能力。虽然发现含有IAA和激动素的MS培养基对于从下胚轴外植体获得快速增殖的愈伤组织更具优势，但发现第二种含有每升2毫克萘乙酸（NAA）和每升0.5至1毫克苄基腺嘌呤（BA）的培养基对于继代培养的愈伤组织旺盛生长是必要的。含有每升5至10毫克N6-[δ2-异戊烯基]-腺嘌呤（2ip）和每升1毫克NAA的MS培养基对于持续继代培养也很有利。
