Research Centre for Plant Growth and Development, School of Life Sciences, University of KwaZulu-Natal, Private Bag X01, Scottsville, Pietermaritzburg, 3209, South Africa.
Planta. 2012 Dec;236(6):1775-90. doi: 10.1007/s00425-012-1721-z. Epub 2012 Aug 11.
The effect of supplementing either meta-topolin (mT) or N(6)-benzyladenine (BA) requiring cultures with roscovitine (6-benzylamino-2-[1(R)-(hydroxymethyl)propyl]amino-9-isopropylpurine), a cyclin-dependent kinase (CDK) and N-glucosylation inhibitor, and INCYDE (2-chloro-6-(3-methoxyphenyl)aminopurine), an inhibitor of cytokinin (CK) degradation, on the endogenous CK profiles and physiology of banana in vitro was investigated. Growth parameters including multiplication rate and biomass were recorded after 42 days. Endogenous CK levels were quantified using UPLC-MS/MS while the photosynthetic pigment and phenolic contents were evaluated spectrophotometrically. The highest regeneration rate (93 %) was observed in BA + roscovitine while mT + INCYDE plantlets produced most shoots. Treatment with BA + roscovitine had the highest shoot length and biomass. Although not significant, there was a higher proanthocyanidin level in BA + roscovitine treatments compared to the control (BA). The levels of total phenolics and flavonoids were significantly higher in mT + roscovitine treatment than in the mT-treated regenerants. The presence of roscovitine and/or INCYDE had no significant effect on the photosynthetic pigments of the banana plantlets. Forty-seven aromatic and isoprenoid CKs categorized into nine CK-types were detected at varying concentrations. The presence of mT + roscovitine and/or INCYDE increased the levels of O-glucosides while 9-glucosides were higher in the presence of BA. Generally, the underground parts had higher CK levels than the aerial parts; however, the presence of INCYDE increased the level of CK quantified in the aerial parts. From a practical perspective, the use of roscovitine and INCYDE in micropropagation could be crucial in the alleviation of commonly observed in vitro-induced physiological abnormalities.
研究了补充环化依赖激酶（CDK）和 N-糖基化抑制剂罗司维亭（6-苄基氨基-2-[1(R)-(羟甲基)丙基]氨基-9-异丙基嘌呤）和 INCYDE（2-氯-6-(3-甲氧基苯基)氨基嘌呤），一种细胞分裂素（CK）降解抑制剂，对香蕉离体内源 CK 谱和生理的影响。42 天后记录生长参数，包括繁殖率和生物量。使用 UPLC-MS/MS 定量内源性 CK 水平，同时评估光合色素和酚类含量。在 BA+罗司维亭处理中观察到最高的再生率（93%），而 mT+INCYDE 植株产生最多的芽。BA+罗司维亭处理的芽长和生物量最高。尽管不显著，但 BA+罗司维亭处理的原花青素水平高于对照（BA）。mT+罗司维亭处理的总酚和类黄酮水平明显高于 mT 处理的再生植株。罗司维亭和/或 INCYDE 的存在对香蕉苗的光合色素没有显著影响。在不同浓度下检测到 47 种芳香族和异戊二烯 CK，分为九种 CK 类型。mT+罗司维亭和/或 INCYDE 的存在增加了 O-葡萄糖苷的水平，而 BA 的存在则提高了 9-葡萄糖苷的水平。一般来说，地下部分的 CK 水平高于地上部分；然而，INCYDE 的存在增加了空中部分定量的 CK 水平。从实际的角度来看，罗司维亭和 INCYDE 在微繁殖中的使用可能对缓解常见的体外诱导的生理异常至关重要。
