Several extraction and measurement methods currently employed in the determination of total sugar and starch contents in plant tissues were investigated with the view to streamline the process of total sugar and starch determination. Depending on the type and source of tissue, total sugar and starch contents estimated from samples extracted with 80% hot ethanol were significantly greater than from samples extracted with a methanol:chloroform:water solution. The residual ethanol did not interfere with the sugar and starch determination, rendering the removal of ethanol from samples unnecessary. The use of phenol−sulfuric acid with a phenol concentration of 2% provided a relatively simple and reliable colorimetric method to quantify the total soluble-sugar concentration. Performing parallel sugar assays with and without phenol was more useful for accounting for the interfering effects of other substances present in plant tissue than using chloroform. For starch determination, an enzyme mixture of 1000 U α-amylase and 5 U amyloglucosidase digested starch in plant tissue samples more rapidly and completely than previously recommended enzyme doses. Dilute sulfuric acid (0.005 N) was less suitable for starch digestion than enzymatic hydrolysis because the acid also broke down structural carbohydrates, resulting in overestimates of starch content. After the enzymatic digestion of starch, the glucose hydrolyzate obtained was measured with a peroxidase−glucose oxidase/o-dianisidine reagent; absorbance being read at 525 nm after the addition of sulfuric acid. With the help of this series of studies, we developed a refined and shortened method suitable for the rapid measurement of total sugar and starch contents in woody plant tissues.

Many forested landscapes around the world are severely altered during mining for their rich mineral and energy reserves. Herein we provide an overview of the challenges inherent in efforts to restore mined landscapes to functioning forest ecosystems and present a synthesis of recent progress using examples from North America, Europe and Australia. We end with recommendations for further elaboration of the Forestry Reclamation Approach emphasizing: (1) Landform reconstruction modelled on natural systems and creation of topographic heterogeneity at a variety of scales; (2) Use and placement of overburden, capping materials and organic amendments to facilitate soil development processes and create a suitable rooting medium for trees; (3) Alignment of landform, topography, overburden, soil and tree species to create a diversity of target ecosystem types; (4) Combining optimization of stock type and planting techniques with early planting of a diversity of tree species; (5) Encouraging natural regeneration as much as possible; (6) Utilizing direct placement of forest floor material combined with seeding of native species to rapidly re-establish native forest understory vegetation; (7) Selective on-going management to encourage development along the desired successional trajectory. Successful restoration of forest ecosystems after severe mining disturbance will be facilitated by a regulatory framework that acknowledges and accepts variation in objectives and outcomes.

Non-structural carbohydrates (NSCs), the stored products of photosynthesis, building blocks for growth and fuel for respiration, are central to plant metabolism, but their measurement is challenging. Differences in methods and procedures among laboratories can cause results to vary widely, limiting our ability to integrate and generalize patterns in plant carbon balance among studies. A recent assessment found that NSC concentrations measured for a common set of samples can vary by an order of magnitude, but sources for this variability were unclear. We measured a common set of nine plant material types, and two synthetic samples with known NSC concentrations, using a common protocol for sugar extraction and starch digestion, and three different sugar quantification methods (ion chromatography, enzyme, acid) in six laboratories. We also tested how sample handling, extraction solvent and centralizing parts of the procedure in one laboratory affected results. Non-structural carbohydrate concentrations measured for synthetic samples were within about 11.5% of known values for all three methods. However, differences among quantification methods were the largest source of variation in NSC measurements for natural plant samples because the three methods quantify different NSCs. The enzyme method quantified only glucose, fructose and sucrose, with ion chromatography we additionally quantified galactose, while the acid method quantified a large range of mono- and oligosaccharides. For some natural samples, sugars quantified with the acid method were two to five times higher than with other methods, demonstrating that trees allocate carbon to a range of sugar molecules. Sample handling had little effect on measurements, while ethanol sugar extraction improved accuracy over water extraction. Our results demonstrate that reasonable accuracy of NSC measurements can be achieved when different methods are used, as long as protocols are robust and standardized. Thus, we provide detailed protocols for the extraction, digestion and quantification of NSCs in plant samples, which should improve the comparability of NSC measurements among laboratories.

Aspen (Populus tremuloides Michx.) is a clonal tree species that commonly regenerates via root suckering after disturbance. This paper reviews the literature and identifies critical gaps in our understanding of the dynamics of aspen root suckering. The role of plant growth regulators (e.g., hormones, carbohydrates), environmental conditions (e.g., soil moisture, temperature, nutrient availability), overstory disturbance (e.g., harvesting, wildfire), ground disturbance (e.g., soil compaction, wounding or severing of roots), vegetation competition, predisturbance stand condition, and clonal (genetic) differences are discussed as they relate to sucker initiation, sucker growth, and (or) patterns of site establishment. The paper presents a series of conceptual figures summarizing our knowledge of the factors controlling suckering dynamics and identifies areas of future research.

In a greenhouse study we quantified the gradual change of gas exchange, water relations and root reserves of aspen (Populus tremuloides Michx.) seedlings growing over a 3-month period of severe water stress. The aim of the study was to quantify the complex interrelationship between growth, water and gas exchange, and root carbon (C) dynamics. Various growth, gas exchange and water relations variables in combination with root reserves were measured periodically on seedlings that had been exposed to a continuous drought treatment over a 12-week period and compared with well-watered seedlings. Although gas exchange and water relations parameters significantly decreased over the drought period in aspen seedlings, root reserves did not mirror this trend. During the course of the experiment roots of aspen seedlings growing under severe water stress showed a two orders of magnitude increase in sugar and starch content, and roots of these seedlings contained more starch relative to sugar than those in non-droughted seedlings. Drought resulted in a switch from growth to root reserves storage which indicates a close interrelationship between growth and physiological variables and the accumulation of root carbohydrate reserves. Although a severe 3-month drought period created physiological symptoms of C limitation, there was no indication of a depletion of root C reserve in aspen seedlings.

Sudden dieback and deterioration of mature aspen stands is commonly observed throughout North America. This dieback process has tremendous ecological and economic importance, yet remains poorly understood. This paper summarizes our understanding of aspen dieback in North America, identifies potential processes that contribute to reduced vigour and dieback of aspen stands, and examines the scales (stand, ecosite, regional) at which these processes operate. Many factors including pathogens, nutrition, or successional changes may be involved in the decline of aspen vigour and thereby contribute to the dieback process. However, insect defoliation, drought, and thaw–freeze events appear to be the most likely factors initiating dieback in mature aspen stands. Further study is clearly needed to elucidate the mechanisms and landscape patterns of dieback. Information needs related to identifying processes and modeling landscape patterns of dieback are indicated.