Culturing of immature inflorescences and Agrobacterium-mediated transformation of foxtail millet (Setaria italica)
Culturing of immature inflorescences and Agrobacterium-mediated transformation of foxtail millet (Setaria italica)
复制标题
谷子（Setaria italica）未成熟花序的培养和农杆菌介导的转化
DOI：
10.5897/ajb10.2330
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发表时间：
2011-11-21
影响因子：
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通讯作者：
Yu, Jing-Juan
中科院分区：
文献类型：
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作者：
Wang, Mei-Zhen;Pan, Yan-Lin;Yu, Jing-Juan
在以前的报道中，我们建立了一个农杆菌介导的谷子遗传转化体系。在这里，我们报告了通过提高再生系统效率和优化基因递送条件来优化系统。谷子未成熟花序外植体。以冀谷11号为材料，在改良MS培养基上进行愈伤组织诱导和再生，其长度分别为0.5 ~ 1.0，1.1 ~ 1.5，1.6 ~ 2.0和> 2.0cm。以0.5 ~ 1.0cm长的幼穗为外植体，胚性愈伤组织的诱导率最高（90.72%），0.5 ~ 1.0cm长的幼穗诱导25 d龄的愈伤组织，分化率最高（90.93%）。此外，影响T-DNA传递的因素进行了检查瞬时β-葡萄糖醛酸酶（GUS）的表达。愈伤组织诱导从年轻的外植体（0.5至1.0厘米未成熟的花序）是最佳的。根癌农杆菌菌株LBA 4404的表现显著优于EHA 105。在感染液和共培养基中于22 ℃与0.15 g/l二硫苏糖醇（DTT）共培养，与其他处理相比，GUS瞬时表达效率更高。利用此优化方法，将马铃薯富含赖氨酸蛋白基因SBgLR转化谷子品种Foxtail Millet cv.转化效率为5.5%。本文所述的方法将有助于谷子的遗传改良。
In previous reports, we developed an Agrobacterium-mediated transformation system for foxtail millet. Here, we report optimization of the system through improvement of the regeneration system efficiency and optimization of conditions for gene delivery. Immature inflorescences explants of foxtail millet cv. Jigu 11 varying in length (0.5 to 1.0, 1.1 to 1.5, 1.6 to 2.0 and >2.0 cm) were cultured on modified MS medium for callus induction and regeneration. The highest embryogenic callus-formation efficiency (90.72%) was achieved with 0.5 to 1.0 cm long inflorescences and 25 days old calli induced from 0.5 to 1.0 cm long immature inflorescences gave rise to the highest differentiation frequency (90.93%). In addition, factors affecting T-DNA delivery were examined by transient beta-glucuronidase (GUS) expression. Calli induced from younger explants (0.5 to 1.0 cm immature inflorescences) were optimal. Agrobacterium tumefaciens strain LBA4404 performed significantly better than EHA105. Co-cultivation at 22 degrees C with 0.15 g/l dithiothreitol (DTT) in the infection solution and co-cultivation medium led to higher GUS transient expression efficiency than with other treatments. Using this optimized procedure, the lysine-rich protein encoding gene SBgLR from potato was transformed into foxtail millet cv. Jigu 11 with 5.5% transformation efficiency. The procedure described here will be useful for genetic improvement of foxtail millet.