Increasing cellulose production and transgenic plant growth in forest tree species

Abstract

Cellulose is one of many important polymers in plants. Cellulose is made of repeat units of the monomer glucose. Cellulose is a major industrial biopolymer in the forest products, textile, and chemical industries. It also forms a large portion of the biomass useful in the generation of energy. Moreover, cellulose-based biomass is a renewable energy source that can be used for the generation of ethanol as a fuel. Cellulose is synthesized by a variety of living organisms such as plants and algae. It is the major component of plant cell walls with secondary cell walls having a much higher content of cellulose. The relationship between cellulose and lignin biosynthesis is complicated, but it is confirmed that inhibition of lignin biosynthesis in transgenic trees will increase cellulose biosynthesis and plant growth. Cellulose accumulation may be increased by down-regulating 4-coumarate:coenzyme A ligase (4CL, EC 6.2.1.12) as shown in transgenic aspen. There is no similar reports on down-regulating 4CL in transgenic conifers. Based on our establishedAgrobacterium tumefaciens-mediated transformation system in loblolly pine, we are able to produce antisense 4-CL transgenic loblolly pine which is predicted to have increasing cellulose accumulation. The overall objective of this project is to genetically engineer forest tree species such as loblolly pine with reduced amount of lignin and increased cellulose content. The research strategy includes: (1) isolate the 4-coumarate:coenzyme A ligase gene from loblolly pine seedlings by reverse transcription-polymerase chain reaction (RT-PCR) and Rapid Amplification of cDNA Ends-Polymerase Chain Reaction (RACE-PCR) techniques from the cDNA library; (2) construct binary expression vectors with antisense 4CL coding sequences and introduce antisense constructs of the 4-coumarate:coenzyme A ligase gene cloned from loblolly pine into the loblolly pine to down regulate the 4-coumarate:coenzyme A ligase gene expression; (3) study the effect of the antisense transgene expression on lignin content, cellulose accumulation, and loblolly pine biomass; and (4) select fast growth and high cellulose accumulation transgenic loblolly pine lines for future commercial application.

Keywords

Antisense transgene expression / Cellulose biosynthesis / Genetic engineering / Pine / S722.34 / A

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