| Genomic Edition of Ashbya gossypii Using One-vector CRISPR/Cas9
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Author:
Date:
2020-06-20
[Abstract] The CRISPR/Cas9 system is a novel genetic tool which allows the precise manipulation of virtually any genomic sequence. In this protocol, we use a specific CRISPR/Cas9 system for the manipulation of Ashbya gossypii. The filamentous fungus A. gossypii is currently used for the industrial production of riboflavin (vitamina B2). In addition, A. gossypii produces other high-value compounds such as folic acid, nucleosides and biolipids. A large molecular toolbox is available for the genomic manipulation of this fungus including gene targeting methods, rapid assembly of heterologous expression modules and, recently, a one-vector CRISPR/Cas9 editing system adapted for A. gossypii that allows marker-free engineering strategies to be implemented. The CRISPR/Cas9 ...
[摘要] [摘要] CRISPR/Cas9系统是一种新的基因工具，可以精确地操作几乎所有的基因组序列。在这个协议中，我们使用一个特定的CRISPR/Cas9系统来操纵棉蚜。丝状真菌A.gossypii目前用于核黄素（vitamina B2）的工业生产。此外，棉蚜还产生其他高价值化合物，如叶酸、核苷和生物脂类。一个大型的分子工具箱可用于该真菌的基因组操作，包括基因靶向方法、异源表达模块的快速组装，以及最近一个适用于棉铃虫的单载体CRISPR/Cas9编辑系统，该系统允许实施无标记工程策略。CRISPR/Cas9系统包括RNA引导的DNA内切酶（Cas9）和与基因组靶区互补的引导RNA（gRNA）。Cas9核酸酶需要5′-NGG-3′三核苷酸，称为原间隔基序（PAM），在基因组靶区产生一个双链断裂（DSB），该断裂可以通过同源重组（HR）由合成的致突变供体DNA（dDNA）修复，从而引入一个特定的设计突变。适用于棉铃虫的CRISPR/Cas9系统极大地促进了这种工业真菌的基因组编辑。
[背景] ...
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| Trichoderma harzianum Root Colonization in Arabidopsis
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Author:
Date:
2015-07-05
[Abstract] Trichoderma is a soil-borne fungal genus that includes species with a significant impact on agriculture and industrial processes. In this article we show a detailed protocol of Trichoderma harzianum (T. harzianum) root invasion procedure described by Alonso-Ramírez et al. (2014). Some Trichoderma strains exert beneficial effects in plants through root colonization. They promote growth and development, modify root architecture, facilitate efficient nutrient use, or stimulate defenses against pathogens, although little is known about how this interaction takes place. For this purpose, Trichoderma-Arabidopsis hydroponic cultures were grown inside Phytatray II boxes, using mycelia obtained from spores of T. harzianum and Arabidopsis ...
[摘要] <木霉属是土壤传播的真菌属，包括对农业和工业过程具有显着影响的物种。 在本文中，我们展示了由alonso-ramírez等人（2014年）所描述的哈茨木霉的（哈茨木霉）根侵入程序的详细方案="" ）。="" 一些木霉菌株通过根定植在植物中发挥有益效果。="" 它们促进生长和发育，改变根结构，促进有效的营养物利用或刺激对病原体的防御，尽管关于如何发生这种相互作用了解甚少。="" 为此目的，使用从t孢子获得的菌丝体，在phytatray="" ii盒内培养木霉属拟南芥水培培养物。="" harzianum="" 和拟南芥（拟南芥）幼苗。="">
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