The development of blackcurrant fruit from flower to maturity: a comparative study by 3D nuclear magnetic resonance (NMR) micro-imaging and conventional histology

Abstract

The development of fruits of blackcurrant (Ribes nigrum) cv. Ben Alder from flower to maturity was studied non-invasively by nuclear magnetic resonance (NMR) microscopy, using attached and detached fruits, and the images were compared with those from low temperature scanning electron microscopy (LTSEM) and conventional resin histology. The NMR images derived from 2-D and 3-D datasets showed the previously unreported growth of arillar tissues to the extent that they almost completely occlude the locular cavity, but LTSEM and resin histology revealed that no fusion occurs between the arillar tissues and the gelatinous sheath surrounding each seed, or between the arillar tissues and the endocarp. The discontinuities between these tissues cause magnetic inhomogenities which result in these structures being clearly resolved by gradient echo imaging sequences. During seed maturation the endosperm changed from high (bright) to low (dark) signal intensity as lipid reserves formed and solidified, whereas the gelatinous sheath had high signal intensity throughout maturation. The high lipid concentration in the seed was manifested by chemical shift effects in the images and the increasing viscosity of the endosperm was reflected in the decrease in spin–lattice (T₁) relaxation times. The funiculi, throughout development of seeds, appeared in NMR images with low signal intensity and 3-D surface-rendered reconstructions illustrated the complexity of the spatial array of seeds and funiculi arising from parietal placentas within the loculus. All other vascular tissues in the pericarp and placentas were resolved as a small bright core surrounded by a dark region, within a matrix of moderate signal intensity. Conventional microscopical studies then showed that the bright core discernible by NMR imaging encompassed an entire vascular bundle whereas the darker surrounding region represented small parenchyma cells with pronounced intercellular gas spaces. Other regions of the pericarp which included extremely large parenchyma cells, however, had few intercellular spaces and consequently gave rise to brighter regions of the image.