Microalgal biorefineries have emerged as significant reservoirs of therapeutic compounds,including pigments and proteins.Facilitating a robust circular bioeconomy necessitates the augmentation of pigment synthesis alo...Microalgal biorefineries have emerged as significant reservoirs of therapeutic compounds,including pigments and proteins.Facilitating a robust circular bioeconomy necessitates the augmentation of pigment synthesis alongside algae biofuel pro-duction.Nevertheless,inherent constraints in ketocarotenoid synthesis exist in naturally fast-growing microalgae strains,such as Chlamydomonas reinhardtii.To address this limitation,we overexpressed two pivotal enzymes in the carotenoid biosynthetic pathway,namelyβ-carotene hydroxylase(crt)andβ-carotene ketolase(bkt),in C.reinhardtii utilizing strong promoters to amplify carotenoid production.The genetically modified(GM)microalgae were validated through PCR,South-ern hybridization,and Western blot assays,confirming the presence and expression of both genes in the C.reinhardtii strains.These GM lines exhibited a substantial enhancement over wild-type(WT)algae,showcasing a remarkable 5.39-fold increase inβ-carotene concentration and twofold increase in total carotenoids compared to the WT microalgae.Notably,the GM microalgae achieved astaxanthin production up to 1.47±0.063 mg/g DCW,a compound absent in WT C.reinhardtii.These findings indicate the successful functionalization of Hematococcus pluvialis genes through nuclear expression in C.reinhardtii,facilitating ketocarotenoid production.This study presents a valuable strategy to boost carotenoid production in microalgae by stable overexpression of two heterologous genes within the nuclear genome of C.reinhardtii.展开更多
MicroRNAs （miRNAs） usually contain 19-24 nucleotides and have been identified as important eukaryotic gene regulators. Applications of various computational approaches have simplified the task by predicting miRNAs f...MicroRNAs （miRNAs） usually contain 19-24 nucleotides and have been identified as important eukaryotic gene regulators. Applications of various computational approaches have simplified the task by predicting miRNAs from available sequence data sources. In this study, we identified a conserved miR414 from a computational analysis of EST sequence data available from Stevia rebaudiana. In addition, we also identified six conserved miRNAs namely miR169, miR319, miR414, miR164, miR167 and miR398 using stem-loop RT-PCR analysis. Hence, miR414 was commonly identified using both methods. The expression analysis of these miRNAs documented their roles in growth and development of Stevia. Furthermore, the detected miRNAs were found to target genes involved in plant growth, development, metabolism and signal transducfion. This is the first study reporting these conserved miRNAs and their expression in Stevia.展开更多
Despite our many advances in deconstructing and understanding cellular and biomolecular machineries,the homochirality of life remains an enigma.Since our earliest ancestors,the central dogma has operated solely on D-DNA
基金supported by DST INSPIRE Ph.D.Fellowship(Grant no.IF190164).
文摘Microalgal biorefineries have emerged as significant reservoirs of therapeutic compounds,including pigments and proteins.Facilitating a robust circular bioeconomy necessitates the augmentation of pigment synthesis alongside algae biofuel pro-duction.Nevertheless,inherent constraints in ketocarotenoid synthesis exist in naturally fast-growing microalgae strains,such as Chlamydomonas reinhardtii.To address this limitation,we overexpressed two pivotal enzymes in the carotenoid biosynthetic pathway,namelyβ-carotene hydroxylase(crt)andβ-carotene ketolase(bkt),in C.reinhardtii utilizing strong promoters to amplify carotenoid production.The genetically modified(GM)microalgae were validated through PCR,South-ern hybridization,and Western blot assays,confirming the presence and expression of both genes in the C.reinhardtii strains.These GM lines exhibited a substantial enhancement over wild-type(WT)algae,showcasing a remarkable 5.39-fold increase inβ-carotene concentration and twofold increase in total carotenoids compared to the WT microalgae.Notably,the GM microalgae achieved astaxanthin production up to 1.47±0.063 mg/g DCW,a compound absent in WT C.reinhardtii.These findings indicate the successful functionalization of Hematococcus pluvialis genes through nuclear expression in C.reinhardtii,facilitating ketocarotenoid production.This study presents a valuable strategy to boost carotenoid production in microalgae by stable overexpression of two heterologous genes within the nuclear genome of C.reinhardtii.
文摘MicroRNAs （miRNAs） usually contain 19-24 nucleotides and have been identified as important eukaryotic gene regulators. Applications of various computational approaches have simplified the task by predicting miRNAs from available sequence data sources. In this study, we identified a conserved miR414 from a computational analysis of EST sequence data available from Stevia rebaudiana. In addition, we also identified six conserved miRNAs namely miR169, miR319, miR414, miR164, miR167 and miR398 using stem-loop RT-PCR analysis. Hence, miR414 was commonly identified using both methods. The expression analysis of these miRNAs documented their roles in growth and development of Stevia. Furthermore, the detected miRNAs were found to target genes involved in plant growth, development, metabolism and signal transducfion. This is the first study reporting these conserved miRNAs and their expression in Stevia.
文摘Despite our many advances in deconstructing and understanding cellular and biomolecular machineries,the homochirality of life remains an enigma.Since our earliest ancestors,the central dogma has operated solely on D-DNA
