Abstract Random mutagenesis is still the main strategy for increasing the lycopene yield from Blakeslea trispora because of its complex, unclear genetic regulation. In this study, an effective mutation breeding method was established to obtain lycopene-overproducing mutants, and the transcriptional changes between the mutant and parent strains were then revealed by RNA-seq. After ARTP mutation and preliminary screening on solid plates, 50 color mutants were simultaneously inoculated into a microtiter plate-based system (MTPS) and traditional flasks to estimate the dependability of the high-throughput screening method. As expected, a significant linear correlation (R2 = 0.924) was observed between the absorbance of pigment in the MTPs and lycopene production in flask fermentation. A mutant (WY-239) showing a maximum lycopene concentration of 21.80 ± 1.58 mg/g was obtained, which represented a 56.27 % increment compared with the parent strain. Transcriptomic analysis showed that the carbon metabolism flowed more intensively to acetyl-CoA and then to lycopene in the WY-239 mutant compared with the parent strain. Some primary metabolic and sexual activity may be correlated with lycopene biosynthesis. The mutation breeding method could also be used as a reference for breeding other filamentous fungi, and the findings of RNA-seq can serve as a basis for metabolic engineering.

中文翻译：

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通过结合ARTP突变与微量滴定板培养和RNA-seq揭示的转录变化高通量筛选三孢布拉克斯菌的番茄红素过量突变体

摘要 由于其复杂、不明确的遗传调控，随机诱变仍然是提高三孢布拉克斯菌番茄红素产量的主要策略。本研究建立了一种有效的突变育种方法来获得番茄红素过量产生的突变体，然后通过RNA-seq揭示突变体与亲本菌株之间的转录变化。在固体平板上ARTP突变和初步筛选后，将50个颜色突变体同时接种到基于微量滴定板的系统（MTPS）和传统烧瓶中，以评估高通量筛选方法的可靠性。正如预期的那样，在 MTP 中色素的吸光度与烧瓶发酵中的番茄红素产量之间观察到显着的线性相关性 (R2 = 0.924)。显示最大番茄红素浓度为 21 的突变体 (WY-239)。获得了 80 ± 1.58 mg/g，与亲本菌株相比增加了 56.27%。转录组学分析表明，与亲本菌株相比，WY-239 突变体中碳代谢更强烈地流向乙酰辅酶 A，然后流向番茄红素。一些初级代谢和性活动可能与番茄红素生物合成有关。突变育种方法也可以作为其他丝状真菌育种的参考，RNA-seq的结果可以作为代谢工程的基础。一些初级代谢和性活动可能与番茄红素生物合成有关。突变育种方法也可以作为其他丝状真菌育种的参考，RNA-seq的结果可以作为代谢工程的基础。一些初级代谢和性活动可能与番茄红素生物合成有关。突变育种方法也可以作为其他丝状真菌育种的参考，RNA-seq的结果可以作为代谢工程的基础。