Genetic Transformation – Biolistics

Altpeter, Fredy; Sandhu, S. G.; Davey, M. R.; Anthony, Paul

doi:10.1002/9780470686522.ch12

Cited by 19 publications

(8 citation statements)

References 68 publications

(72 reference statements)

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“…CP‐88‐1762 were obtained from the University of Florida, Everglades Research and Education Center in Belle Glade, FL. Plasmids were coprecipitated onto gold particles in a 1:2:2 molar ratio for pJK403NPTII: pJK304605: pJK60iPXAAGP (Figure ) as described by Altpeter and Sandhu ().…”

Section: Methodsmentioning

confidence: 99%

Metabolic engineering of sugarcane to accumulate energy‐dense triacylglycerols in vegetative biomass

Zale

Jung

Kim

et al. 2015

Plant Biotechnology Journal

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143

118

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SummaryElevating the lipid content in vegetative tissues has emerged as a new strategy for increasing energy density and biofuel yield of crops. Storage lipids in contrast to structural and signaling lipids are mainly composed of glycerol esters of fatty acids, also known as triacylglycerol (TAG). TAGs are one of the most energy-rich and abundant forms of reduced carbon available in nature. Therefore, altering the carbon-partitioning balance in favour of TAG in vegetative tissues of sugarcane, one of the highest yielding biomass crops, is expected to drastically increase energy yields. Here we report metabolic engineering to elevate TAG accumulation in vegetative tissues of sugarcane. Constitutive co-expression of WRINKLED1 (WRI1), diacylglycerol acyltransferase1-2 (DGAT1-2) and oleosin1 (OLE1) and simultaneous cosuppression of ADP-glucose pyrophosphorylase (AGPase) and a subunit of the peroxisomal ABC transporter1 (PXA1) in transgenic sugarcane elevated TAG accumulation in leaves or stems by 95-or 43-fold to 1.9% or 0.9% of dry weight (DW), respectively, while expression or suppression of one to three of the target genes increased TAG levels by 1.5-to 9.5-fold. Accumulation of TAG in vegetative progeny plants was consistent with the results from primary transgenics and contributed to a total fatty acid content of up to 4.7% or 1.7% of DW in mature leaves or stems, respectively. Lipid droplets were visible within mesophyll cells of transgenic leaves by confocal fluorescence microscopy. These results provide the basis for optimizations of TAG accumulation in sugarcane and other high yielding biomass grasses and will open new prospects for biofuel applications.

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Section: Methodsmentioning

confidence: 99%

Metabolic engineering of sugarcane to accumulate energy‐dense triacylglycerols in vegetative biomass

Zale

Jung

Kim

et al. 2015

Plant Biotechnology Journal

Self Cite

143

118

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“…For delivery of the minimum linear transgene cassette (MC), the COMT RNAi cassette and npt II expression cassettes were released by restriction enzyme digestion with Xmn I and I‐Sce I, separated by gel electrophoresis and extracted using the QIAquick Gel Extraction kit (Qiagen, Valencia, CA). COMT and npt II – MC DNAs were mixed in a 2 : 1 m ratio, co‐precipitated onto 1‐μm‐diameter gold micro‐carriers (Bio‐Rad) and accelerated into target tissues as described earlier (Altpeter et al. , 2010).…”

Section: Methodsmentioning

confidence: 99%

RNAi suppression of lignin biosynthesis in sugarcane reduces recalcitrance for biofuel production from lignocellulosic biomass

Jung¹,

Fouad²,

Vermerris³

et al. 2012

Plant Biotechnology Journal

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152

120

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SummarySugarcane is a prime bioethanol feedstock. Currently, sugarcane ethanol is produced through fermentation of the sucrose, which can easily be extracted from stem internodes. Processes for production of biofuels from the abundant lignocellulosic sugarcane residues will boost the ethanol output from sugarcane per land area. However, unlocking the vast amount of chemical energy stored in plant cell walls remains expensive primarily because of the intrinsic recalcitrance of lignocellulosic biomass. We report here the successful reduction in lignification in sugarcane by RNA interference, despite the complex and highly polyploid genome of this interspecific hybrid. Down-regulation of the sugarcane caffeic acid O-methyltransferase (COMT) gene by 67% to 97% reduced the lignin content by 3.9% to 13.7%, respectively. The syringyl ⁄ guaiacyl ratio in the lignin was reduced from 1.47 in the wild type to values ranging between 1.27 and 0.79. The yields of directly fermentable glucose from lignocellulosic biomass increased up to 29% without pretreatment. After dilute acid pretreatment, the fermentable glucose yield increased up to 34%. These observations demonstrate that a moderate reduction in lignin (3.9% to 8.4%) can reduce the recalcitrance of sugarcane biomass without compromising plant performance under controlled environmental conditions.

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“…Biolistics is a common method used for sugarcane transformation (Altpeter and Sandhu, 2010) due to its simplicity and applicability to a wide range of tissues and genotypes (Lakshmanan et al, 2005). However, it presents some disadvantages as low reproducibility, integration of a large number of transgene copies (Zhangsun et al, 2007).…”

Section: Engineering Sugarcane For Water Stress Tolerancementioning

confidence: 99%

“…Two main strategies are widely used to produce transgenic plants in sugarcane: direct transformation via microprojectile (biolistics) (Bower and Birch, 1992 ) and indirect transformation mediated by Agrobacterium tumefaciens (Arencibia et al, 1998 ). Biolistics is a common method used for sugarcane transformation (Altpeter and Sandhu, 2010 ) due to its simplicity and applicability to a wide range of tissues and genotypes (Lakshmanan et al, 2005 ). However, it presents some disadvantages as low reproducibility, integration of a large number of transgene copies (Zhangsun et al, 2007 ).…”

Section: Engineering Sugarcane For Water Stress Tolerancementioning

confidence: 99%

Sugarcane Water Stress Tolerance Mechanisms and Its Implications on Developing Biotechnology Solutions

et al. 2017

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Sugarcane is a unique crop with the ability to accumulate high levels of sugar and is a commercially viable source of biomass for bioelectricity and second-generation bioethanol. Water deficit is the single largest abiotic stress affecting sugarcane productivity and the development of water use efficient and drought tolerant cultivars is an imperative for all major sugarcane producing countries. 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 our knowledge on genes and mechanisms underpinning plant response to water stress. We also overview transgenic studies in sugarcane, with an emphasis on the potential strategies to develop superior sugarcane varieties that improve crop productivity in drought-prone environments.

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Genetic Transformation – Biolistics

Cited by 19 publications

References 68 publications

Metabolic engineering of sugarcane to accumulate energy‐dense triacylglycerols in vegetative biomass

Metabolic engineering of sugarcane to accumulate energy‐dense triacylglycerols in vegetative biomass

RNAi suppression of lignin biosynthesis in sugarcane reduces recalcitrance for biofuel production from lignocellulosic biomass

Sugarcane Water Stress Tolerance Mechanisms and Its Implications on Developing Biotechnology Solutions

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