Effect of cytokinins and light quality on adventitious shoot regeneration from leaflet explants of peanut (Arachis hypogaea)

Juvenal Assou, Hans Bethge, D. Wamhoff, T. Winkelmann
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Abstract

ABSTRACT Plant tissue culture and novel breeding techniques such as genome editing in the economically important crop Arachis hypogaea constitute potential for genetic improvement. Therefore, in this study, high-frequency in vitro regeneration via formation of adventitious shoots was described for different A. hypogaea lines. The leaflets of the primary leaves of 5-day-old seedlings were cultured for 16 weeks on Murashige and Skoog’s medium supplemented with equimolar concentrations (22.19 µM) of the cytokinins 6-benzylaminopurine (BAP); thidiazuron (TDZ); zeatin (Zea) or meta-topolin (MT), which was reduced to 7.40 µM after eight weeks plus 2.3 µM kinetin. Highest shoot regeneration percentages of 89.1% and 81.3% for BAP and TDZ containing media, respectively, were achieved in the line ’Jimmy’s pride’ with the maximum shoot number obtained on BAP. The regeneration percentages for three further lines were lower than 50%. Furthermore, the effect of different light qualities, applied using LEDs, and compared to fluorescent lamps was investigated. Shoot regeneration percentages did not improve significantly under the different LEDs, instead a pronounced effect on the regeneration efficiency of the culture temperature was observed which differed among the light treatments. However, fluorescent lamps can be replaced by red or blue LEDs without negative effects on shoot regeneration.
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细胞分裂素和光照质量对花生小叶外植体不定芽再生的影响
植物组织培养和基因组编辑等新育种技术为重要经济作物花生的遗传改良提供了潜力。因此,在本研究中,描述了通过形成不定芽的高频离体再生对不同的山竹系。将5日龄幼苗初生叶的小叶在添加等摩尔浓度(22.19µM)细胞分裂素6-苄氨基嘌呤(BAP)的Murashige和Skoog培养基上培养16周;物质(TDZ);玉米素(Zea)或元topolin (MT), 8周后减少到7.40µM,加上2.3µM激动素。含BAP和TDZ培养基的“Jimmy’s pride”植株再生率最高,分别为89.1%和81.3%。另外3个品系的再生率均低于50%。此外,还研究了不同光质量对发光二极管的影响,并与荧光灯进行了比较。在不同的led下,芽再生率没有显著提高,相反,培养温度对再生效率有显著的影响,不同的光处理之间存在差异。然而,可以用红色或蓝色led代替荧光灯,而不会对茎部再生产生负面影响。
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