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Confocal Raman microspectroscopy (CRM) has emerged as a powerful approach to visualize the compositional distribution in lignocellulosic biomass of cell walls. In this work, the applicability of CRM for imaging the topochemical correlation between lignin and hydroxycinnamic acids (HCA) in the Miscanthus sinensis internode was explored. Model compound [p-coumaric acid (PCA) and ferulic acid (FA)] analysis indicated that the band region from 1,152 to 1,197 cm−1 can be used to characterize the distribution of HCA. Raman images calculated by integrating over the area intensity of characteristic spectral regions showed heterogeneous distribution of lignin and HCA at cellular and sub-cellular level. When overlaying the Raman image of lignin and HCA distribution, it was found that these two polymers were co-located in the middle lamella and secondary wall of corresponding cells. Raman images for the band intensity ratio (1,173 cm−1/1,603 cm−1) indicated a clear association between lignin and HCA distribution within morphologically distinct cell wall layers of sclerenchyma fibers and the parenchyma. This is the first time that the spatial correlation between lignin and HCA concentration has been illustrated by a microspectroscopy imaging approach. The results are of importance in extending the current understanding of lignin and aromatics topochemistry in herbaceous biomass.
In this study we investigated the importance of two species of fruit bat (Rousettus leschenaulti and Cynopterus sphinx) as seed dispersers for a species of fruit tree (Syzygium oblatum) found in the Xishuangbanna Tropical Botanical Garden in South-West China. We found that although R. leschenaulti and C. sphinx were the two primary seed dispersers of S. oblatum over half of the fruit produced by the tree (65%) fell to the ground. Out of the fruit collected, R. leschenaulti and C. sphinx were able to disperse seeds up to 73 m from the parent tree with the highest density of feeding roosts occurring at 21.3 m (SE = 5.2 m). We found no signs that either species of bat used the parent tree as a feeding roost, instead choosing specific trees that were at lower densities compared with other trees in the forest that were not used. When comparing the viability of seeds in three different habitats (under parent tree, in forest gap, under feeding roost) survival analysis revealed that seedling survival was significantly higher in the forest gap (91.7% ± 4.41%) than under the parent tree (78.3% ± 1.67%), but was not significantly different to seedling survival underneath feeding roosts (86.7 ± 1.67%). Further work also showed that the seeds did not have to be removed from the fruit or ingested by the bat in order to germinate. We conclude that although S. oblatum is not dependent on R. leschenaulti and C. sphinx for successful germination of its seeds, these two species of bat are important seed dispersers and can move seeds to areas where there is a greater chance of germination success and survival.
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