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1. 发明申请

WO2016193959A2 IMPROVED PRODUCTIVITY AND BIOPRODUCT FORMATION IN PHOTOTROPIN KNOCK/OUT MUTANTS IN MICROALGAE 审中-公开
标题翻译：在微藻中改善光合作用的生物量和生物量
公开(公告)号：WO2016193959A2
公开(公告)日：2016-12-08
申请号：PCT/IB2016/054466
申请日：2016-07-26
申请人： NMC, INC. , LOS ALAMOS NATIONAL SECURITY, LLC
发明人： NEGI, Sangeeta , SAYRE, Richard Thomas , STARKENBURG, Shawn Robert
IPC分类号： C12P19/04 , C12N1/13
CPC分类号： C12N9/12 , C07K14/405 , C12N1/12 , C12N9/0051 , C12N9/1205 , C12N9/22 , C12N15/01 , C12N15/11 , C12N2310/20 , C12N2800/80 , C12P7/64 , C12P19/04 , C12P23/00 , C12Y108/01009 , C12Y207/11
摘要： Phototropin is a blue light receptor, which mediates a variety of blue-light elicited physiological processes in plants and algae. In higher plants these processes include phototropism, chloroplast movement and stomatal opening. In the green alga Chlamydomonas reinhardtii , phototropin plays a vital role in progression of the sexual life cycle and In the control of the eye spot size and light sensitivity Phototropin is also involved in blue-light mediated changes in the synthesis of chlorophylls, carotenoids, chlorophyll binding proteins. We compared the transcriptome of phototropin knock out (PHOT KO) mutant and wild-type parent to analyze differences in gene expression in high light grown cultures (500 μmol photons m -2 s -1 ). Our results indicate the up-regulation of genes involved in photosynthetic electron transport chain, carbon fixation pathway, starch, lipid, and cell cycle control genes. With respect to photosynthetic electron transport genes, genes encoding proteins of the cytochrome b6f and ATP synthase complex were up regulated potentially facilitating proton-coupled electron transfer. In addition genes involved in limiting steps in the Calvin cycle Ribulose-1,5-blsphosphate carboxylase/oxygenase (RuBisCO), Sidoheptulose 1,7 bisphosphatase (SBPase), Glyceraldehyde-3- phosphate dehydrogenase (3PGDH) and that mediate cell-cycle control (CDK) were also up regulated along with starch synthase and fatty acid biosynthesis genes involved in starch and lipid synthesis, in addition, transmission electron micrographs show increased accumulation of starch granules in PHOT mutant compared to wild type, which is consistent with the higher expression of starch synthase genes. Collectively, the altered patterns of gene expression in the PHOT mutants were associated with a two-fold increase in growth and biomass accumulation compared to wild type when grown in environmental photobioreactors (Phenometrics) that simulate a pond environment. In conclusion, our studies suggest that phototropin may be a master gene regulator that suppresses rapid cell growth and promotes gametogenesis and sexual recombination in wild type strains.
摘要翻译：光吗啉是蓝光受体，其介导植物和藻类中各种蓝光引发的生理过程。在高等植物中，这些过程包括光转动，叶绿体运动和气孔开放。在绿藻藻衣藻中，光滑石在性生活周期的进展中起着至关重要的作用，并且在眼睛斑点大小和光敏感性的控制中。光喜素也参与蓝光介导的叶绿素，类胡萝卜素，叶绿素合成的变化结合蛋白。我们比较了光诱导敲除（PHOT KO）突变体和野生型亲本的转录组，以分析高光生长培养物（500μmol光子m-2s-1）中基因表达的差异。我们的结果表明参与光合电子传递链，碳固定途径，淀粉，脂质和细胞周期控制基因的基因的上调。对于光合电子转运基因，编码细胞色素b6f和ATP合成酶复合物蛋白质的基因上调可能促进质子 - 偶联电子转移。此外，参与限制加尔文周期中的步骤的基因Ribulose-1,5-blsphosphate羧化酶/加氧酶（RuBisCO），Sidoheptulose 1,7二磷酸酶（SBPase），甘油醛-3-磷酸脱氢酶（3PGDH），并介导细胞周期控制（CDK）也与淀粉合成酶和脂肪酸生物合成基因参与淀粉和脂质合成相关，此外，透射电子显微照片显示，与野生型相比，透明电子显微照片显示淀粉颗粒在PHOT突变体中的积累增加，这与更高的表达一致的淀粉合成酶基因。总之，在模拟池塘环境的环境光生物反应器（Phenometrics）中生长时，与野生型相比，PHOT突变体中基因表达的改变模式与生长和生物量积累相比增加了两倍。总之，我们的研究表明光滑素可能是一个主要的基因调节器，抑制快速的细胞生长和促进野生型菌株的配子形成和性重组。

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