Highly Efficient Nanoscale Analysis of Plant Stomata and Cell Surface Using Polyaddition Silicone Rubber.

Yi He,Junliang Fu,Dean Jiang,Kaiyue Zhou,Boxiu Li,Zhemin Wu,Chinho Lin

doi:10.3389/fpls.2019.01569

Abstract

Stomata control gas exchange and water transpiration and are one of the most important physiological apparatuses in higher plants. The regulation of stomatal aperture is closely coordinated with photosynthesis, nutrient uptake, plant growth, development, and so on. With advances in scanning electron microscopy (SEM), high-resolution images of plant stomata and cell surfaces can be obtained from detached plant tissues. However, this method does not allow for rapid analysis of the dynamic variation of plant stomata and cell surfaces in situ under nondestructive conditions. In this study, we demonstrated a novel plant surface impression technique (PSIT, Silagum-Light as correction impression material based on A-silicones for all two-phase impression techniques) that allows for precise analysis of plant stomata aperture and cell surfaces. Using this method, we successfully monitored the dynamic variation of stomata and observed the nanoscale microstructure of soybean leaf trichomes and dragonfly wings. Additionally, compared with the analytical precision and the time used for preparing the observation samples between PSIT and traditional SEM, the results suggested that the analytical precision of PSIT was the same to traditional SEM, but the PSIT was more easy to operate. Thus, our results indicated that PSIT can be widely applied to the plant science field.

Highlights

Stomata are microscopic pores found in large numbers on the epidermal surface of most aerial parts of higher plants, and they have been documented in fossil records as early as the late Silurian era, nearly 411 million years ago (Lawson et al, 2009; Edwards et al, 1998)
SilagumLight is easy to operate for the preparation of molds (Figure 1), which can be stored for extended periods; the precision of the mold would not be affected even when stored at room temperature for several months
There is more surface texture visible in the scanning electron microscopy (SEM) images, whereas the plant surface impression technique (PSIT) images appear to show smoother surfaces (Figure 4D), which might be attributed to not completely dehydrating the soybean leaf trichomes by traditional SEM. These results indicated that the precision of imaging by PSIT can satisfy demand for the study of plant stomata and cell surfaces

Summary

Introduction

Stomata are microscopic pores found in large numbers on the epidermal surface of most aerial parts of higher plants, and they have been documented in fossil records as early as the late Silurian era, nearly 411 million years ago (Lawson et al, 2009; Edwards et al, 1998). Stomata are central to the physiology of land plants, as environmentally induced changes in stomatal development and movements have profound effects on gas exchange between the atmosphere and the leaf (Jiang et al, 2012; Webb and Baker, 2002). Stomata are formed by pairs of guard cells and function as gateways for controlling gas exchange and transpirational water loss. Stomatal opening promotes plant growth by enhancing carbon dioxide uptake and transpirational water loss, which are both essential for photosynthesis and nutrient uptake from the soil to the plant body, respectively (Gray et al, 2000; Murata et al, 2015; Jezek and Blatt, 2017). The number and distribution of stomata affect gas exchange and are closely regulated and coordinated with cell growth and division, while they preserve a level of plasticity to respond to ever-changing environmental conditions (Pilliteri et al, 2012)

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