Comparative Genomic Analysis of the Mutant Rhodotorula mucilaginosa JH-R23 Provides Insight into the High-Yield Carotenoid Mechanism

Jingyao Huang, Sujing Yang, Huali Jian
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Abstract

In this study, the wild-type Rhodotorula mucilaginosa GDMCC 2.30 and its high carotenoid-producing mutant JH-R23, which was screened from the space mutation breeding treated wild type, were used as materials. Through whole-genome sequencing and resequencing analysis, the carotenoid metabolic pathway and mechanism of high carotenoid production in the mutant were explored. The R. mucilaginosa GDMCC 2.30 genome comprised 18 scaffolds and one circular mitochondrial genome with a total size of 20.31 Mb, a GC content of 60.52%, and encoding 7128 genes. The mitochondrial genome comprised 40,152 bp with a GC content of 40.59%. Based on functional annotations in the GO, KEGG, and other protein databases, nine candidate genes associated with carotenoid metabolic pathways, and candidate genes of the CrtS and CrtR homologous gene families were identified. The carotenoid metabolic pathway was inferred to start from sugar metabolism to the mevalonate pathway, as is common to most fungi, and the final product of the mevalonate pathway, geranylgeranyl diphosphate, is a precursor for various carotenoids, including β-carotene, lycopene, astaxanthin, and torularhodin, formed through the activity of crucial enzymes encoded by genes such as CrtI, CrtYB, CrtS, and CrtR. Resequencing analysis of the mutant JH-R23 detected mutations in the exons of four genes, including those encoding Gal83, 3-oxoacyl-reductase, p24 proteins, and GTPase. These mutations are interpreted to have an important impact on carotenoid synthesis by JH-R23.
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对突变体粘质红豆杉 JH-R23 的基因组比较分析有助于深入了解类胡萝卜素的高产机制
本研究以野生型Rhodotorula mucilaginosa GDMCC 2.30及其高产类胡萝卜素突变体JH-R23为材料,该突变体是从空间突变育种处理的野生型中筛选出来的。通过全基因组测序和重测序分析,探索了突变体类胡萝卜素代谢途径和高产类胡萝卜素的机理。R. mucilaginosa GDMCC 2.30基因组由18个支架和一个环形线粒体基因组组成,总大小为20.31 Mb,GC含量为60.52%,编码7128个基因。线粒体基因组由 40,152 bp 组成,GC 含量为 40.59%。根据 GO、KEGG 和其他蛋白质数据库的功能注释,确定了 9 个与类胡萝卜素代谢途径相关的候选基因,以及 CrtS 和 CrtR 同源基因家族的候选基因。推断类胡萝卜素代谢途径与大多数真菌一样,是从糖代谢到甲羟戊酸途径开始的,而甲羟戊酸途径的最终产物--geranylgeranyl diphosphate是多种类胡萝卜素的前体,包括β-胡萝卜素、番茄红素、虾青素和torularhodin,这些类胡萝卜素是通过CrtI、CrtYB、CrtS和CrtR等基因编码的关键酶的活性形成的。对突变体 JH-R23 的重测序分析发现了四个基因外显子的突变,包括编码 Gal83、3-氧代乙酰还原酶、p24 蛋白和 GTPase 的基因。据解释,这些突变对 JH-R23 合成类胡萝卜素有重要影响。
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