Cut and paste: temperature-enhanced cotyledon micrografting for Arabidopsis thaliana seedlings
Cut and paste: temperature-enhanced cotyledon micrografting for Arabidopsis thaliana seedlings
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DOI：
10.1186/s13007-020-0562-1
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发表时间：
2020-02-05
期刊：
影响因子：
5.1
通讯作者：
Quint, Marcel
中科院分区：
文献类型：
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作者：
Bartusch, Kai;Trenner, Jana;Quint, Marcel
背景子叶微嫁接是研究子叶在早期植物发育过程中的核心作用的有用工具，特别是它们在长距离运输方面与其他植物器官的相互作用。虽然下胚轴微嫁接方法已经很成熟，但子叶微嫁接仍然效率低下。通过优化子叶微嫁接，我们的目标是提高模式物种拟南芥的成功率和产量。结果我们在平坦、坚固但潮湿的表面上建立了剪切和粘贴子叶手术程序，这改善了小苗的处理。通过应用特定的切割和连接模式，吞吐量增加至每小时 40 棵幼苗。短日照周期和低光强度的发芽和长日照加上高光强度、升高的温度和嫁接后垂直板定位的组合显着提高了“连接”效率。特别是高温对成功率有有利的影响。总的来说，我们在拟南芥中的嫁接成功率高达 92%。通过脉管系统特异性染料跨接枝部位的运输来证明脉管系统的重新连接。韧皮部和木质部重新连接分别在嫁接后3-4天和4-6天完成，以温度依赖性方式完成。我们观察到，具有嫁接子叶的植物在生物量生产和莲座丛发育方面与具有完整子叶的植物相匹配。结论 这种剪切粘贴子叶到叶柄微嫁接方案简化了手术中植物幼苗的处理，增加了每小时嫁接植物的数量，大大提高了拟南芥幼苗的成功率。所开发的子叶微嫁接方法也适用于其他大小相当的植物物种。
Background Cotyledon micrografting represents a useful tool for studying the central role of cotyledons during early plant development, especially their interplay with other plant organs with regard to long distance transport. While hypocotyl micrografting methods are well-established, cotyledon micrografting is still inefficient. By optimizing cotyledon micrografting, we aim for higher success rates and increased throughput in the model species Arabidopsis thaliana. Results We established a cut and paste cotyledon surgery procedure on a flat and solid but moist surface which improved handling of small seedlings. By applying a specific cutting and joining pattern, throughput was increased up to 40 seedlings per hour. The combination of short-day photoperiods and low light intensities for germination and long days plus high light intensities, elevated temperature and vertical plate positioning after grafting significantly increased 'ligation' efficiency. In particular high temperatures affected success rates favorably. Altogether, we achieved up to 92% grafting success in A. thaliana. Reconnection of vasculature was demonstrated by transport of a vasculature-specific dye across the grafting site. Phloem and xylem reconnection were completed 3-4 and 4-6 days after grafting, respectively, in a temperature-dependent manner. We observed that plants with grafted cotyledons match plants with intact cotyledons in biomass production and rosette development. Conclusions This cut and paste cotyledon-to-petiole micrografting protocol simplifies the handling of plant seedlings in surgery, increases the number of grafted plants per hour and greatly improves success rates for A. thaliana seedlings. The developed cotyledon micrografting method is also suitable for other plant species of comparable size.