Morphogene-assisted transformation of Sorghum bicolor allows more efficient genome editing

第一作者：Kiflom Aregawi

第一单位：加州大学伯克利分校

通讯作者：Peggy G. Lemaux

 Abstarct 

背景回顾：Sorghum bicolor (L.) Moench, the fifth most important cereal worldwide, is a multi-use crop for feed, food, forage and fuel. To enhance the sorghum and other important crop plants, establishing gene function is essential for their improvement. For sorghum, identifying genes associated with its notable abiotic stress tolerances requires a detailed molecular understanding of the genes associated with those traits. The limits of this knowledge became evident from our earlier in-depth sorghum transcriptome study showing that over 40% of its transcriptome had not been annotated. 

高粱是全球第五大谷类作物，可作为饲料、粮食、饲草和燃料等多种用途。对基因功能的鉴定是对高粱和其他重要作物植物进行遗传改良的重要基础。对于高粱来说，鉴别与其显著的非生物胁迫耐受性相关的基因，需要对与这些性状相关的基因进行详细的分子鉴定。早期对高粱转录组的深入研究显示超过40%的高粱转录组没有被功能注释，表明目前对于高粱很多基因的功能还是未知的。

主要研究：Here, we describe a full spectrum of tools to engineer, edit, annotate and characterize sorghum’s genes. 

本文中，作者建立了一套技术体系可以用来对高粱的基因进行基因工程和编辑，从而对其进行功能鉴定。

结果1-MAT方法促进不依赖于基因型的高校遗传转化：Efforts to develop those tools began with a morphogene-assisted transformation (MAT) method that led to accelerated transformation times, nearly half the time required with classical callus-based, non-MAT approaches. These efforts also led to expanded numbers of amenable genotypes, including several not previously transformed or historically recalcitrant. 

该技术体系的基础是一个叫做形态基因辅助转化（MAT）的方法，该方法能够显著加快高粱的遗传转化时间：与传统的、基于愈伤的非MAT方法相比，MAT方法介导的高粱遗传转化能够加速一倍的时间。该方法同样能够作用于不同的高粱基因型，包括了先前从未转化成功或者对转化较为顽拗的基因型。

结果2-利他转化方法：Another transformation advance, termed altruistic, involved introducing a gene of interest in a separate Agrobacterium strain from the one with morphogenes, leading to plants with the gene of interest but without morphogenes. 

此外，由于形态基因ZmBBM/WUS2辅助的遗传转化会导致再生植株的异常表型，因此发展出了一种称之为“利他转化”的改良版转化方法，该方法通过用携带不同载体的农杆菌菌株侵染受体材料，一种农杆菌携带形态基因，另外一种农杆菌携带有目的基因，从而能够获得只转化目的基因但是没有形态基因的再生植株。

结果3-MAT方法用于基因编辑：The MAT approach was also successfully used to edit a target exemplary gene, phytoene desaturase. 

作者进一步利用MAT方法进行了基因编辑的测试，成功编辑掉了PDS基因。

结果4-单拷贝转化植株高效鉴定：To identify single-copy transformed plants, we adapted a high-throughput technique and also developed a novel method to determine transgene independent integration. 

为了鉴定单拷贝的转化植株，作者开发了一个高通量技术，同时还开发了一个新的方法来检测转基因的独立整合。

结论：These efforts led to an efficient method to determine gene function, expediting research in numerous genotypes of this widely grown, multi-use crop.

综上，作者开发的技术能够快速鉴定基因功能，从而加速了高粱多种基因型的育种研究。

** Peggy G. Lemaux **

doi: https://doi.org/10.1111/pbi.13754

Journal: Plant Biotechnology Journal

Published date: November 26, 2021