Because lignin limits the use of wood for fiber, chemical, and energy production, strategies for its downregulation are of considerable interest. We have produced transgenic aspen (Populus tremuloides Michx.) trees in which expression of a lignin biosynthetic pathway gene Pt4CL1 encoding 4-coumarate:coenzyme A ligase (4CL) has been downregulated by antisense inhibition. Trees with suppressed Pt4CL1 expression exhibited up to a 45% reduction of lignin, but this was compensated for by a 15% increase in cellulose. As a result, the total lignin–cellulose mass remained essentially unchanged. Leaf, root, and stem growth were substantially enhanced, and structural integrity was maintained both at the cellular and whole-plant levels in the transgenic lines. Our results indicate that lignin and cellulose deposition could be regulated in a compensatory fashion, which may contribute to metabolic flexibility and a growth advantage to sustain the long-term structural integrity of woody perennials.

Biotechnology on genetic control of lignin biosynthesis in trees -Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees-

The current carbon supply status of temperate forest trees was assessed by analysing the seasonal variation of nonstructural carbohydrate (NSC) concentrations in leaves, branch wood and stem sapwood of 10 tree species (six deciduous broad-leafed, one deciduous conifer and three evergreen conifer trees) in a temperate forest that is approximately 100 years old. In addition, all woody tissue was analysed for lipids (acylglycerols). The major NSC fractions were starch, sucrose, glucose and fructose, with other carbohydrates (e.g. raffinose and stachyose) and sugar alcohols (cyclitols and sorbitol) playing only a minor quantitative role. The radial distribution of NSC within entire stem cores, assessed here for the first time in a direct interspecific comparison, revealed large differences in the size of the active sapwood fraction among the species, reflecting the specific wood anatomy (ring-porous versus diffuse-porous xylem). The mean minimum NSC concentrations in branch wood during the growing season was 55% of maximum, and even high NSC concentrations were maintained during times of extensive fruit production in masting Fagus sylvestris . The NSC in stem sapwood varied very little throughout the season (cross species mean never below 67% of maximum), and the small reductions observed were not significant for any of the investigated species. Although some species contained substantial quantities of lipids in woody tissues (‘fat trees’; Tilia , Pinus , Picea , Larix ), the lipid pools did not vary significantly across the growing season in any species. On average, the carbon stores of deciduous trees would permit to replace the whole leave canopy four times. These data imply that there is not a lot of leeway for a further stimulation of growth by ongoing atmospheric CO 2 enrichment. The classical view that deciduous trees rely more on C-reserves than evergreen trees, seems unwarranted or has lost its justification due to the greater than 30% increase in atmospheric CO 2 concentrations over the last 150 years.

https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.0016-8025.2003.01032.x

Non‐structural carbon compounds in temperate forest trees

Nonstructural carbon (NSC) provides the carbon and energy for plant growth and survival. In woody plants, fundamental questions about NSC remain unresolved: Is NSC storage an active or passive process? Do older NSC reserves remain accessible to the plant? How is NSC depletion related to mortality risk? Herein we review conceptual and mathematical models of NSC dynamics, recent observations and experiments at the organismal scale, and advances in plant physiology that have provided a better understanding of the dynamics of woody plant NSC. Plants preferentially use new carbon but can access decade-old carbon when the plant is stressed or physically damaged. In addition to serving as a carbon and energy source, NSC plays important roles in phloem transport, osmoregulation, and cold tolerance, but how plants regulate these competing roles and NSC depletion remains elusive. Moving forward requires greater synthesis of models and data and integration across scales from -omics to ecology.

/pdf/nonstructural-carbon-in-woody-plants-hgp08zhlmn.pdf

Nonstructural carbon in woody plants.

Plants store large amounts of non-structural carbohydrates (NSC). While multiple functions of NSC have long been recognized, the interpretation of NSC seasonal dynamics is often based on the idea that stored NSC is a reservoir of carbon that fluctuates depending on the balance between supply via photosynthesis and demand for growth and respiration (the source–sink dynamics concept). Consequently, relatively high NSC concentrations in some plants have been interpreted to reflect excess supply relative to demand. An alternative view, however, is that NSC accumulation reflects the relatively high NSC levels required for plant survival; an important issue that remains highly controversial. Here, we assembled a new global database to examine broad patterns of seasonal NSC variation across organs (leaves, stems, and belowground), plant functional types (coniferous, drought-deciduous angiosperms, winter deciduous angiosperms, evergreen angiosperms, and herbaceous) and biomes (boreal, temperate, Mediterranean, and tropical). We compiled data from 121 studies, including seasonal measurements for 177 species under natural conditions. Our results showed that, on average, NSC account for ~10% of dry plant biomass and are highest in leaves and lowest in stems, whereas belowground organs show intermediate concentrations. Total NSC, starch, and soluble sugars (SS) varied seasonally, with a strong depletion of starch during the growing season and a general increase during winter months, particularly in boreal and temperate biomes. Across functional types, NSC concentrations were highest and most variable in herbaceous species and in conifer needles. Conifers showed the lowest stem and belowground NSC concentrations. Minimum NSC values were relatively high (46% of seasonal maximums on average for total NSC) and, in contrast to average values, were similar among biomes and functional types. Overall, although starch depletion was relatively common, seasonal depletion of total NSC or SS was rare. These results are consistent with a dual view of NSC function: whereas starch acts mostly as a reservoir for future use, soluble sugars perform immediate functions (e.g., osmoregulation) and are kept above some critical threshold. If confirmed, this dual function of NSC will have important implications for the way we understand and model plant carbon allocation and survival under stress.

/pdf/dynamics-of-non-structural-carbohydrates-in-terrestrial-27a42hy9ms.pdf

Dynamics of non-structural carbohydrates in terrestrial plants: a global synthesis

Low temperature driven carbon shortage is often assumed to explain slow growth and treeline formation at high elevations. To test this hypothesis, we analysed mobile carbon pools in Pinus cembra across the treeline ecotone in the Swiss Alps. Concentrations of non-structural carbohydrates (NSC) in needles, branches, stems and roots, as well as lipids (acylglycerols) in all woody tissues were measured throughout the growing season. Starch was the most prominent non-structural carbon compound in needles, whereas lipids represented 50–75% of the mobile carbon compounds in wood. The relative seasonal variation of the lipid fraction was very small, but due to the high absolute amount of lipids, the annual variability of carbon in lipids exceeded that of NSC in woody tissues. Mobile carbon compounds were highly abundant throughout the year and were never significantly depleted. Across a 110 m altitudinal transect from timberline to the uppermost site of tree existence, NSC and lipid concentrations generally increased. This trend became even more pronounced when the increasing structural density of tissues at higher elevations was accounted for. An estimation of the whole tree mobile carbon concentration (fraction of mobile carbon compounds within the whole tree biomass) also revealed an increasing trend of NSC and lipid pools with elevation. We therefore conclude that carbon limitation is unlikely to be responsible for reduced tree growth at the alpine treeline studied. Increased concentrations of NSC and lipids at the upper tree limit rather suggest that sink activity is limited. Hence, treeline formation is most likely the result of a direct thermal restriction of tissue formation (investment in structures) under otherwise sufficient carbon assimilation during the growing season.

Altitudinal increase of mobile carbon pools in Pinus cembra suggests sink limitation of growth at the Swiss treeline

Non-structural carbohydrates in silver birch (Betula pendula Roth) wood were analysed in a 7-year-old clone and in five mature stems. The analysis was conducted to obtain more detailed information on seasonal fluctuation of these components and of the tree-to-tree variation and within stem variation. The sugars were analysed by GLC-MS. The smallest total soluble sugar amounts (consisting of sucrose, fructose, glucose, raffinose and myo-inositol) in young trees were measured during mid-summer (ca. 0.3%) and the largest while in dormancy (ca. 1.6% on wood dry weight basis). Raffinose was detected in autumn as a minor component. The proportion of monosaccharides and the amount of myo-inositol were largest during growth. Compared to other studies silver birch showed more evident seasonal fluctuation in soluble sugars than evergreen tree species. The sugar amount in mature stems was approximately at the same level as in young trees that had the same felling time. Tree-to-tree variation in the non-structural carbohydrates in the mature wood was fairly large. However, the amount of total soluble sugars, sucrose and glucose showed significant variation within the stem. The amount of these sugars was largest in samples that were taken close to the cambium. Starch was also detected close to pith. According to the heartwood definition and starch measurement results in this paper, it could be stated that silver birch does not form heartwood.

Variation of non-structural carbohydrates in silver birch (Betula pendula Roth) wood

Predicting wood and tracheid properties of Norway spruce

Variations in the concentration and composition of triacylglycerols, free fatty acids and phospholipids were analyzed in Scots pine (Pinus sylvestris L.) trees at five sites. Disks were taken at breast height or at a height of 4 m from the stems of 81 trees differing in diameter and growth rate. The mean concentration of triacylglycerols in sapwood was 26 mg g(-1) dry mass; however, variation among trees was large (16-51 mg g(dm)(-1)). The concentration of triacylglycerols was slightly larger at 4 m height in the stem than at breast height. Concentrations of triacylglycerols did not differ between the sapwood of young and small-diameter stems (DBH 36 cm). Concentrations of free fatty acids were negligible in the outer sapwood, but ranged between 5 and 18 mg g(dm)(-1) in the heartwood. The most abundant fatty acids of triacylglycerols were oleic (18:1), linoleic (18:2omega6, 18:2Delta5,9), linolenic (pinolenic, 18:3Delta5,9,12 and 18:3omega3) and eicosatrienoic acid (20:3Delta5,11,14 and 20:3omega6). The concentration of linoleic acid comprised 39-46% of the triacylglycerol fatty acids and the concentration was higher in the slow-growing stem from northern Finland than in the stems from southern Finland. Major phospholipids were detected only in sapwood, and only traces of lipid phosphorus were detected in heartwood.

Neutral lipids and phospholipids in Scots pine (Pinus sylvestris) sapwood and heartwood

Three-year old Betula pendula Roth clones were grown at two nutrient levels in a field experiment to investigate the responses and recovery in growth and wood properties to a range of defoliation levels (0100%). No general threshold value of defoliation level for negative effects in growth was found, since the sensitivity of saplings to defoliation varied according to plant traits studied. However, responses were related to defoliation intensity. Saplings compensated for 25% defoliation in terms of height growth and number of current branches and were able to tolerate 50% defoliation without effects on diameter growth 1 year after the defoliation. Nutrient availability was significant only in determining how total biomass responded to defoliation. Fertilized saplings were able to tolerate 25% defoliation without reduction in total biomass, but nonfertilized saplings were not. The interaction between defoliation and fertilization disappeared in the second growing season after the defoliation. Saplings wer...

Effects of defoliation on growth, biomass allocation, and wood properties of Betula pendula clones grown at different nutrient levels

The effects of Agrobacterium pRiA4 rol and aux genes, controlled by their endogenous promoters, on tree growth and wood anatomy and chemistry were studied in 5- and 7-year-old silver birch (Betula pendula Roth) plants. Southern hybridization confirmed the following rol and aux gene combinations: control plants (no genes transferred); plants with rolC and rolD genes; plants with rolA, rolB, rolC and rolD genes; and plants with rolA, rolB, rolC, rolD, aux1 and aux2 genes. Transgene mRNA was most abundant in phloem/cambium samples and in the developing xylem, whereas no expression was detected in leaves. Plants with rolC and rolD genes or with all the rol genes were significantly shorter and had smaller leaves and a more bushy growth habit than control plants or plants with both aux and rol genes. Morphological observations and wood chemistry analyses revealed that plants with rol genes produced less xylem and broke bud later than control plants or plants with both aux and rol genes. Tension wood was detected in both control and transgenic plants irrespective of their gene combination, probably as a result of greenhouse cultivation. Xylem fibers were shorter in transgenic plants than in control plants, and plants with all the rol genes were characterized by shorter vessels compared with the control plants and a smaller proportional area of vessels compared with the other groups. In addition, silver birch plants with all the rol genes had approximately a 3.3% lower concentration of total acid soluble carbohydrates than control plants. We conclude that the rolC and rolD genes induced the typical "rol-phenotype," and that this was emphasized by concomitant expression of the rolA and rolB genes and alleviated by the presence of aux1 and aux2 genes. We observed consistent phenotypic effects of rol and aux genes on the morphology, anatomy and cell wall chemistry of the plants.

/pdf/silver-birch-betula-pendula-plants-with-aux-and-rol-genes-23b31ten3c.pdf

Riikka Piispanen

Papers

Variation of non-structural carbohydrates in silver birch (Betula pendula Roth) wood

Predicting wood and tracheid properties of Norway spruce

Neutral lipids and phospholipids in Scots pine (Pinus sylvestris) sapwood and heartwood

Effects of defoliation on growth, biomass allocation, and wood properties of Betula pendula clones grown at different nutrient levels

Silver birch (Betula pendula) plants with aux and rol genes show consistent changes in morphology, xylem structure and chemistry.