Wheat with greatly reduced accumulation of free asparagine in the grain, produced by CRISPR/Cas9 editing of asparagine synthetase gene TaASN2.
Wheat with greatly reduced accumulation of free asparagine in the grain, produced by CRISPR/Cas9 editing of asparagine synthetase gene TaASN2.
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DOI：
10.1111/pbi.13573
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发表时间：
2021-08
影响因子：
13.8
通讯作者：
Halford NG
中科院分区：
文献类型：
--
作者：
Raffan S;Sparks C;Huttly A;Hyde L;Martignago D;Mead A;Hanley SJ;Wilkinson PA;Barker G;Edwards KJ;Curtis TY;Usher S;Kosik O;Halford NG
游离天冬酰胺是丙烯酰胺的前体，在小麦食品的烘焙、烘烤和高温加工过程中形成。在这项研究中，CRISPR/Cas9被用来敲除小麦（Triticum aestivum）cv.华彩乐段通过粒子轰击将4-gRNA多顺反子基因导入小麦胚中并再生植物。来自14株T0植物中的11株的T1植物显示携带编辑。大多数编辑是删除（多达173个碱基对），但也有一些单碱基对插入和取代。在T1代之后继续编辑。与野生型相比，在所有六个TaASN 2等位基因（每个基因组中的两个等位基因）中携带编辑的植物的谷粒中的游离天冬酰胺浓度显著降低，其中一株植物在T2种子中显示出超过90%的降低。仅在A基因组等位基因中含有编辑的植物显示出谷粒中游离天冬酰胺浓度的较小降低，但浓度仍低于野生型。编辑的植物中的游离天冬酰胺浓度也作为游离氨基酸库的比例降低。T3种子中的游离天冬酰胺浓度在编辑品系中保持显著低于野生型，尽管其高于T2种子，可能是由于胁迫。相比之下，编辑品系中游离谷氨酰胺、谷氨酸和天冬氨酸的浓度均高于野生型。低天冬酰胺的种子表现出不良的发芽，但这可以克服外源天冬酰胺的应用。
Free asparagine is the precursor for acrylamide, which forms during the baking, toasting and high‐temperature processing of foods made from wheat. In this study, CRISPR/Cas9 was used to knock out the asparagine synthetase gene, TaASN2, of wheat (Triticum aestivum) cv. Cadenza. A 4‐gRNA polycistronic gene was introduced into wheat embryos by particle bombardment and plants were regenerated. T1 plants derived from 11 of 14 T0 plants were shown to carry edits. Most edits were deletions (up to 173 base pairs), but there were also some single base pair insertions and substitutions. Editing continued beyond the T1 generation. Free asparagine concentrations in the grain of plants carrying edits in all six TaASN2 alleles (both alleles in each genome) were substantially reduced compared with wildtype, with one plant showing a more than 90 % reduction in the T2 seeds. A plant containing edits only in the A genome alleles showed a smaller reduction in free asparagine concentration in the grain, but the concentration was still lower than in wildtype. Free asparagine concentration in the edited plants was also reduced as a proportion of the free amino acid pool. Free asparagine concentration in the T3 seeds remained substantially lower in the edited lines than wildtype, although it was higher than in the T2 seeds, possibly due to stress. In contrast, the concentrations of free glutamine, glutamate and aspartate were all higher in the edited lines than wildtype. Low asparagine seeds showed poor germination but this could be overcome by exogenous application of asparagine.
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影响因子：
5.8
作者：
Park, Jeongbin;Bae, Sangsu;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
3.8
作者：
Gao R;Curtis TY;Powers SJ;Xu H;Huang J;Halford NG
通讯作者：
Halford NG