TALEN mediated targeted mutagenesis of the caffeic acid O-methyltransferase in highly polyploid sugarcane improves cell wall composition for production of bioethanol
TLDR
The findings demonstrate that targeted mutagenesis can improve cell wall characteristics for production of lignocellulosic ethanol in crops with highly complex genomes.Abstract:
Sugarcane (Saccharum spp. hybrids) is a prime crop for commercial biofuel production. Advanced conversion technology utilizes both, sucrose accumulating in sugarcane stems as well as cell wall bound sugars for commercial ethanol production. Reduction of lignin content significantly improves the conversion of lignocellulosic biomass into ethanol. Conventional mutagenesis is not expected to confer reduction in lignin content in sugarcane due to its high polyploidy (x = 10-13) and functional redundancy among homo(eo)logs. Here we deploy transcription activator-like effector nuclease (TALEN) to induce mutations in a highly conserved region of the caffeic acid O-methyltransferase (COMT) of sugarcane. Capillary electrophoresis (CE) was validated by pyrosequencing as reliable and inexpensive high throughput method for identification and quantitative characterization of TALEN mediated mutations. Targeted COMT mutations were identified by CE in up to 74 % of the lines. In different events 8-99 % of the wild type COMT were converted to mutant COMT as revealed by pyrosequencing. Mutation frequencies among mutant lines were positively correlated to lignin reduction. Events with a mutation frequency of 99 % displayed a 29-32 % reduction of the lignin content compared to non-transgenic controls along with significantly reduced S subunit content and elevated hemicellulose content. CE analysis displayed similar peak patterns between primary COMT mutants and their vegetative progenies suggesting that TALEN mediated mutations were faithfully transmitted to vegetative progenies. This is the first report on genome editing in sugarcane. The findings demonstrate that targeted mutagenesis can improve cell wall characteristics for production of lignocellulosic ethanol in crops with highly complex genomes.read more
Citations
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Journal ArticleDOI
Applications and potential of genome editing in crop improvement
TL;DR: The current applications of genome editing in plants are described, focusing on its potential for crop improvement in terms of adaptation, resilience, and end-use, and novel breakthroughs that are extending the potential of genome-edited crops and the possibilities of their commercialization are reviewed.
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Plant genome editing with TALEN and CRISPR.
TL;DR: In recent years, transcription activator-like effector nucleases and clustered regularly interspaced palindromic repeats (CRISPR) and CRISpr associated protein 9 or CRISPR from Prevotella and Francisella 1 have emerged as the preferred SSNs for research purposes.
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Ali Razzaq,Fozia Saleem,Mehak Kanwal,Ghulam Mustafa,Sumaira Yousaf,Hafiz Muhammad Imran Arshad,Muhammad Khalid Hameed,Muhammad Sarwar Khan,Faiz Ahmad Joyia +8 more
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Sugarcane Water Stress Tolerance Mechanisms and Its Implications on Developing Biotechnology Solutions.
Thais Helena Silva Ferreira,Max Seiji Tsunada,Denis Bassi,Pedro Araújo,Lucia Mattiello,Giovanna Vieira Guidelli,Germanna Lima Righetto,Vanessa Regina Gonçalves,Prakash Lakshmanan,Marcelo Menossi +9 more
TL;DR: This review summarizes the physiological and molecular studies on water deficit stress in sugarcane, with the aim to help formulate more effective research strategies for advancing knowledge on genes and mechanisms underpinning plant response to water stress.
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Current and future editing reagent delivery systems for plant genome editing
Yidong Ran,Zhen Liang,Caixia Gao +2 more
TL;DR: This review will focus on an update of recent developments in the methodologies of editing reagent delivery, and consider the pros and cons of current delivery systems.
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