Visualizing water conducting system of woody plants is a critical parameter for estimating water alteration (Wullschleger et al. 2001, Kumagai et al. 2005), and anatomical changes caused by implications promoting seedling growth into the reforestation ecosystems. The xylem, which is the main part of the water conducting system supported experimental evidences in plants, regulates water transport regulated by the differentiation in water potential between the air surrounding leaves and soil surrounding the roots (Myburg and Sederoff, 2001).
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'Visualizing Water Conducting System of Woody Plants'
Several studies have been conducted to visualize water pathways thought xylem in different methods including dye injection (Umebayashi et al. 2007; Maton and Gartner 2005), radioactive dye injection (Postlethwait and Rogers 1958), and water observation in plants with NMR imagining (Utsuzawa et al. 2005). However, these methods are difficult and costly and to implement to small seedlings. In addition, different dye solutions were used to visualize water conducting system (Jupa et al. 2015). Acid fuchsion (Maton and Gartner, 2005) and safranin o (Schulte and Brooks, 2003) dye stains were used for visualizing water conducting system in pine seedlings.
However, protocols belonging to those dye stains were not clearly detailed and operable for having clear visual observation to measure hydraulically active xylem diameter. Hydraulically active xylem diameter and percentage may be one of the best indicators, which may be anatomically improved by the implementations in the nursery growth phase, show and determine seedling survival and growing performance in harsh planting sites.
We have little knowledge about how environmental stresses like drought affect xylem hydraulics in conifers; especially in Pinus ponderosa. Because water-conducting tissues and hydraulics are critical to understand how producing adapted seedlings to planting sites is important to improve post planting success. Morphology of ponderosa pine seedlings such as enhance seedlings height and root collar diameter was promoted by initial nursery practices. Xylem diameter and hydraulically activity in total xylem amount was predicted to be changed with initial nursery practices.
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Thus, to test that prediction, cristal violet staning was used to visualize hydraulically activity and diameter of ponderosa pine seedlings in chapter 1 resulting in a failure of visualizing the xylem diameter. The objective of this study is to develop the best dye staining combination using acid fuchsinand safranin o in three concentrations (0.01%, 0.1%, and 0.5%), three dye exposing time periods (10 min, 20 min, and 30 min), and additional two time series for keeping seedlings in distilled water (0 min, and 5 min). These dye stained combinations including the time series were compared by using light microscopic images based on ranking procedure in terms of clarity of hydraulically active xylem of ponderosa pine seedlings.
