浮萍 (Lemna japonica) 中的工程三酰甘油积累,Plant Biotechnology Journal

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浮萍 (Lemna japonica) 中的工程三酰甘油积累
Plant Biotechnology Journal ( IF 10.1 ) Pub Date : 2022-10-09 , DOI: 10.1111/pbi.13943
Yuanxue Liang ₁ , Xiao-Hong Yu ₁ , Sanket Anaokar ₁ , Hai Shi ₁ , William B Dahl ₂ , Yingqi Cai ₁ , Guangbin Luo ₃ , Jin Chai ₁ , Yuanheng Cai ₁ , Almudena Mollá-Morales ₂ , Fredy Altpeter ₃ , Evan Ernst _{2,

4} , Jorg Schwender ₁ , Robert A Martienssen _{2,

4} , John Shanklin ₁

Affiliation

浮萍是生长最快的高等植物之一，这使它们成为生物燃料原料生产的有吸引力的高生物量目标。它们的叶状体具有很高的脂肪酸合成率，可以满足对新膜的需求，但三酰甘油 (TAG) 只能积累到非常低的水平。在这里，我们报告了Lemna japonica在其叶中合成和积累 TAG的工程。这是通过表达一种雌二醇诱导的青色荧光蛋白-拟南芥 WRINKLED1融合蛋白 ( CFP-AtWRI1 )、小鼠二酰基甘油:酰基辅酶 A 酰基转移酶 2 ( MmDGAT ) 和芝麻油质蛋白变体 ( SiOLE(*))的强组成型表达来实现的). 相对于对照，每个基因的单独表达使 TAG 积累增加 1 到 7 倍，而这些基因对的表达使 TAG 增加 7 到 45 倍。在包含所有三种基因的未诱导转基因中，TAG 积累增加了 45 倍至干重 (DW) 的 3.6% 而没有严重影响生长，并且在含有 100 μ m的培养基上孵育后增加了 108 倍至干重的 8.7%雌二醇 4 天。TAG 积累伴随着总脂肪酸的增加高达三倍，达到 DW 的大约 15%。通过 BODIPY 染色的叶状体的共聚焦显微镜可以看到来自所有转基因品系叶状体的脂滴。以每英亩 12 吨（干物质）和 10% (DW) TAG 的保守值计算，浮萍每年每英亩可产 350 加仑油，大约是大豆产量的七倍，与油棕的产量相似。这些发现为优化浮萍中的 TAG 积累提供了基础，并为生产生物燃料和脂质生物产品提供了新的机会。

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Engineering triacylglycerol accumulation in duckweed (Lemna japonica)

Duckweeds are amongst the fastest growing of higher plants, making them attractive high-biomass 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 on 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 45-fold 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 4 days. TAG accumulation was accompanied by an increase in total fatty acids of up to three-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 seven-fold the yield of soybean, and similar to that of oil palm. These findings provide the foundation for optimizing TAG accumulation in duckweed and present a new opportunity for producing biofuels and lipidic bioproducts.

更新日期：2022-10-09

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