Agrobacterium-Mediated Genetic Transformation of Taiwanese Isolates of Lemna aequinoctialis

Wang, Kuang-Teng; Hong, Ming‐Chang; Wu, Yu‐Sheng; Wu, Tzong‐Ming

doi:10.3390/plants10081576

Cited by 5 publications

(3 citation statements)

References 44 publications

(78 reference statements)

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“…In addition to reliable genome information, efficient stable transformation protocols are needed for the generation of transgenic lines. Protocols based on Agrobacterium ‐mediated transformation of callus cultures have been reported for multiple duckweed species (Boehm et al ., 2001 ; Geng et al ., 2018 ; Vunsh et al ., 2007 ; Wang et al ., 2021 ; Yamamoto et al ., 2001 ), but the transformation of L. japonica clone 8627 is the most efficient by far, so we chose this species in these studies (Cantó‐Pastor et al ., 2015 ). This clone was previously misclassified as L. minor (Braglia et al ., 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Engineering triacylglycerol accumulation in duckweed (Lemna japonica)

Liang

Anaokar

et al. 2022

Plant Biotechnology Journal

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Duckweeds are amongst the fastest growing of higher plants, making them attractive highbiomass targets for biofuel feedstock production. Their fronds have high rates of fatty acid synthesis to meet the demand for new membranes, but triacylglycerols (TAG) only accumulate to very low levels. Here we report the engineering of Lemna japonica for the synthesis and accumulation of TAG in its fronds. This was achieved by expression of an estradiol-inducible cyan fluorescent protein-Arabidopsis WRINKLED1 fusion protein (CFP-AtWRI1), strong constitutive expression of a mouse diacylglycerol:acyl-CoA acyltransferase2 (MmDGAT), and a sesame oleosin variant (SiOLE(*)). Individual expression of each gene increased TAG accumulation by 1-to 7-fold relative to controls, while expression of pairs of these genes increased TAG by 7-to 45-fold. In uninduced transgenics containing all three genes, TAG accumulation increased by 45fold to 3.6% of dry weight (DW) without severely impacting growth, and by 108-fold to 8.7% of DW after incubation on medium containing 100 μM estradiol for four days. TAG accumulation was accompanied by an increase in total fatty acids of up to 3-fold to approximately 15% of DW. Lipid droplets from fronds of all transgenic lines were visible by confocal microscopy of BODIPY-stained fronds. At a conservative 12 tonnes (dry matter) per acre and 10% (DW) TAG, duckweed could produce 350 gallons of oil/acre/year, approximately 7-fold the yield of soybean, and similar to that

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Section: Introductionmentioning

confidence: 99%

Engineering triacylglycerol accumulation in duckweed (Lemna japonica)

Liang

Anaokar

et al. 2022

Plant Biotechnology Journal

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“…Additionally, PGRs play an important role in inducing buds and roots with different explants. For example, callus tissues of duckweed ( Lemna aequinoctialis ) in Schenk and Hildebrandt (SH) medium containing 1 μmol/L 6-BA were differentiated into adventitious bud with 100% [ 26 ]; 2 mg/L 6-BA and 0.1 mg/L NAA in MS could induce buds with leaves of Taraxacum officinale [ 23 ]; 25 μmol/L thidiazuron (TDZ) and 10 μmol/L IAA in MS or B5 medium effectively induced adventitious buds with whole or half-leaf attached petiolar explants of Paulownia elongata [ 27 ]. The combination of IAA 0.05 mg/L and KT 2 mg/L efficiently induced cotyledonary nodes to differentiate adventitious in our study ( Fig 1 ).…”

Section: Discussionmentioning

confidence: 99%

Tissue culture and Agrobacterium-mediated genetic transformation of the oil crop sunflower

Chen,

Zeng,

Cheng

et al. 2024

PLoS ONE

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Sunflower is one of the four major oil crops in the world. ‘Zaoaidatou’ (ZADT), the main variety of oil sunflower in the northwest of China, has a short growth cycle, high yield, and high resistance to abiotic stress. However, the ability to tolerate adervesity is limited. Therefore, in this study, we used the retention line of backbone parent ZADT as material to establish its tissue culture and genetic transformation system for new variety cultivating to enhance resistance and yields by molecular breeding. The combination of 0.05 mg/L IAA and 2 mg/L KT in MS was more suitable for direct induction of adventitious buds with cotyledon nodes and the addition of 0.9 mg/L IBA to MS was for adventitious rooting. On this basis, an efficient Agrobacterium tumefaciens-mediated genetic transformation system for ZADT was developed by the screening of kanamycin and optimization of transformation conditions. The rate of positive seedlings reached 8.0%, as determined by polymerase chain reaction (PCR), under the condition of 45 mg/L kanamycin, bacterial density of OD600 0.8, infection time of 30 min, and co-cultivation of three days. These efficient regeneration and genetic transformation platforms are very useful for accelerating the molecular breeding process on sunflower.

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“…Duckweed has 37 species in the world, belonging to five genera including Lemna, Spirodela, Wolffia, Wolffiella , and Landoltia ( Wang et al., 2021 ). Lemna aequinoctialis is a common and widely distributed aquatic plant in China, which has been extensively used in the research of plant biology, aquatic ecotoxicology, and water pollution remediation due to its rapid reproduction, high yield and rich nutritional value ( Kummerová et al., 2016 ; Ekperusi et al., 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Physiological responses and antibiotic-degradation capacity of duckweed (Lemna aequinoctialis) exposed to streptomycin

et al. 2022

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Aquatic plants are constantly exposed to various water environmental pollutants. Few data on how antibiotics affect duckweed health and its removal ability. The aim of this study was to investigate the impact of streptomycin on the physiological change and uptake capability in duckweed (Lemna aequinoctialis) after exposure at different time points (0, 5, 10, 15 and 20 days). Duckweeds were exposed to streptomycin at a range of concentrations (0.1-10 mM). Results indicated that the high streptomycin concentrations (≥1 mM) resulted in a lower duckweed biomass (21.5-41.5%), RGR (0.258-0.336 g d−1), decrease in total Chl and increase in carotenoids. Antioxidative enzymes, including CAT (18-42.88 U mg protein-1), APX (0.41-0.76 U mg protein-1), and SOD (0.52-0.71 U mg protein-1) were found to accumulate in the streptomycin groups in comparison to the control group. The significant reduction (72-82%) in streptomycin content at 20 d compared to the control (40-55%) suggested that duckweed has a high ability in removing streptomycin. Transcriptome analysis showed that the secondary metabolic pathways including phenylpropanoid biosynthesis and flavonoid biosynthesis were significantly upregulated in the streptomycin setup compared to the control. Therefore, our findings suggested that duckweed can contribute to the streptomycin degradation, which should be highly recommended to the treatment of aquaculture wastewater and domestic sewage.

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Agrobacterium-Mediated Genetic Transformation of Taiwanese Isolates of Lemna aequinoctialis

Cited by 5 publications

References 44 publications

Engineering triacylglycerol accumulation in duckweed (Lemna japonica)

Engineering triacylglycerol accumulation in duckweed (Lemna japonica)

Tissue culture and Agrobacterium-mediated genetic transformation of the oil crop sunflower

Physiological responses and antibiotic-degradation capacity of duckweed (Lemna aequinoctialis) exposed to streptomycin

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