Novel genetic engineering technology which increases leaf lipid content modifies the ensiling properties of perennial ryegrass : a thesis presented in partial fulfillment of the requirements for the degree of Masters in Agricultural Science at Massey University, Palmerston North

Abstract

A novel strategy to increase the metabolisable energy (ME) yield of pastures has been the development of a genetic engineering technology which increases the leaf lipid content and biomass production of perennial ryegrass (PRG). Outdoor plot/feeding trials of genetically engineered crops are prohibited under the current New Zealand (NZ) regulatory framework. However, this high lipid PRG may become available to farmers and eventually be used to make silage, which could fulfill an important role as a high ME, inexpensive, supplementary feed for livestock. Ensiling preserves a crop’s nutrients at a high moisture content and at a low pH, by microbial fermentation of plant sugars into lactic acid under anaerobic conditions. In a preliminary investigation into the ensiling biochemistry of this high lipid PRG, glasshouse-grown materials were wilted and inoculated, and then ensiled on a miniature scale. A series of method development ensiling experiments revealed that non-transgenic PRG grown in glasshouse conditions during the NZ spring/summer was very difficult to ensile naturally, due to its low water soluble sugar to buffering capacity ratio. In order to generate well-preserved silage in the main experiment, glucose was added (post-harvest) to a non-transgenic PRG genotype (WT) and two transgenic PRG genotypes containing ‘medium’ and ‘high’ leaf lipid levels (ML and HL). The HL plants produced 51% more dry biomass than WT during the regrowth period. Pre-ensiled HL had 31% higher fatty acid content, 70% higher nitrate content and a 17% lower water soluble sugar to crude protein ratio than WT. ML was intermediate. The glasshouse growth environment resulted in an atypical overall PRG nutritional composition. WT, ML and HL underwent a similar fermentation, and nutrients were well-preserved. The nutritional differences in the ensiled material largely reflected those in their fresh counterparts, although a longer wilt caused greater overall digestible nutrient losses in HL. In an in vitro rumen incubation experiment the fatty acids in HL silage exhibited less complete biohydrogenation than in fresh and ensiled WT. Experiments using a range of high lipid PRG lines grown in a range of environments will be needed to validate these results.