Achieving hybridisation between Miscanthus species: Commercially-scalable methods to manipulate flowering synchronisation and maximise seed yield

Miscanthus is a high-yielding lignocellulosic perennial biomass crop. The low multiplication rate of clonal rhizome propagation is a bottleneck to upscaling plantation areas of feedstock needed to supply and expand the bioeconomy. Novel seeded Miscanthus hybrids are currently being developed to overcome this bottleneck by increasing annual multiplication rates from approximately 10 to over 1000 times. We describe a series of field experiments in southern Italy using agronomic methods to optimise multiplication rates through (i) planting configurations and densities, (ii) ratio of seed parents to pollen parents (iii) supplemental pre-dawn misting to increase humidity during pollination. In these trials the seed-bearing M. sinensis started flowering 2–3 weeks earlier than the M. sacchariflorus pollen parent. Earlier experiments indicated that flowering in M. sacchariflorus was mostly determined by photoperiod while in M. sinensis it was modulated by endogenous signals. Consequently, a second set of experiments were conducted to delay flowering time in M. sinensis: (iv) mid-season stem cut back, (v) oversupply of nitrogen, and (vi) undersupply of water. Across all treatments and years, the multiplication rates varied from 140 to 1300 seeds m². Reducing the proportion of the pollen parent plants (M. sacchariflorus) from 50 % to 25 % did not reduce seed yield per plant. This therefore increases the seed yield per m² and reduces seed production upscaling costs. Flowering time and duration in M. sinensis was significantly impacted by mid-season cutting and water stress, but not by nitrogen supply rates. Mid-season shoot cutting reduced number of flowers per plant (77 %), seed quantity (47 %), seed size (46 %), and resulted in seeds with a low germination rate of 39 %. High M. sinensis planting densities produced higher seed yields in the first year. However, in subsequent years higher density plots were more susceptible to autumn lodging lowering seed production by loss of panicles. Pre-dawn misting to prolong pollen life and stigma receptivity had no significant effects on seed production. This study demonstrates the importance of flowering time synchronization in the open field for commercial seed production. The limited effect of agronomic efforts to reduce the interspecies flowering time gap emphasises the importance of genetic factors in controlling flowering time. The most impactful intervention to change flowering time and improve parental synchronisation was mid-season cutting, while this method reduced seed production when applied to the seed parent it could be ideal for pollen parents.