Optimization of Mature Embryo-Based Tissue Culture and Agrobacterium-Mediated Transformation in Model Grass Brachypodium distachyon
Optimization of Mature Embryo-Based Tissue Culture and Agrobacterium-Mediated Transformation in Model Grass Brachypodium distachyon
复制标题
模型草二穗短柄草成熟胚组织培养和农杆菌介导转化的优化
DOI：
10.3390/ijms20215448
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发表时间：
2019-10
影响因子：
5.6
通讯作者：
Wang Kai
中科院分区：
文献类型：
--
作者：
Yu Guangrun;Wang Jianyong;Miao Li;Xi Mengli;Wang Qiongli;Wang Kai
农杆菌介导的遗传转化在模型草 Brachypodium distachyon 中已得到很好的证实。然而，大多数方案都采用未成熟胚胎，因为它们具有更好的再生能力。与未成熟胚系统相关的一个主要问题是它们仅在植物生长的有限时间窗口内可用。在这项研究中，我们开发了一种利用成熟胚胎的优化农杆菌介导的遗传转化方案。我们在愈伤组织诱导和根再生方面分别采用了种子剪切和光合自养生根（PR），在这些方面取得了明显的显着改善。我们还发现新开发的化学诱导剂 Fipexide (FPX) 具有诱导愈伤组织、芽和根的能力。通过比较，我们已经证明，在我们的成熟胚胎系统中，FPX 在芽生成方面表现出比其他常用化学品更高的效率。此外，我们还发现胚性愈伤组织的年龄严重影响转化效率（TE），7周龄的胚性愈伤组织的TE最高达到52.6%，与未成熟胚转化的TE相当。这里报道的新方法将促进短柄草作为草类基因组学新模型系统的开发和利用。
Agrobacterium-mediated genetic transformation is well established in the model grass Brachypodium distachyon. However, most protocols employ immature embryos because of their better regenerative capacity. A major problem associated with the immature embryo system is that they are available only during a limited time window of growing plants. In this study, we have developed an optimized Agrobacterium-mediated genetic transformation protocol that utilizes mature embryos. We have adopted seed shearing and photoautotrophic rooting (PR) in callus induction and root regeneration, respectively, with evident significant improvement in these aspects. We have also revealed that the newly developed chemical inducer Fipexide (FPX) had the ability to induce callus, shoots, and roots. By comparison, we have demonstrated that FPX shows higher efficiency in shoot generation than other frequently used chemicals in our mature embryo-based system. In addition, we demonstrated that the age of embryogenetic callus severely affects the transformation efficiency (TE), with the seven-week-old embryogenetic callus having the highest TE reaching 52.6%, which is comparable with that in immature embryo transformation. The new methodologies reported here will advance the development and utilization of Brachypodium as a new model system for grass genomics.
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影响因子：
--
作者：
Man Bo Lee;W. Jeon;D. Kim;O. Bold;M. Hong;Yong Jin Lee;Jae Han Park;Y. Seo
通讯作者：
Man Bo Lee;W. Jeon;D. Kim;O. Bold;M. Hong;Yong Jin Lee;Jae Han Park;Y. Seo
影响因子：
--
作者：
R. Vunsh
通讯作者：
R. Vunsh
影响因子：
64.8
作者：
Vogel, John P.;Garvin, David F.;Baxter, Ivan
通讯作者：
Baxter, Ivan
影响因子：
3
作者：
Vogel, JP;Garvin, DF;Hayden, DM
通讯作者：
Hayden, DM
DOI：
10.1007/s00468-010-0488-8
发表时间：
2010-09
期刊：
Trees
影响因子：
--
作者：
C. Ragonezi;K. Klimaszewska;Mário Rui da Costa Basílio e Castro;M. Lima;P. Oliveira;M. A. Zavattieri
通讯作者：
C. Ragonezi;K. Klimaszewska;Mário Rui da Costa Basílio e Castro;M. Lima;P. Oliveira;M. A. Zavattieri