Impact of freeze-thaw-induced pit aspiration on stem water transport in the subalpine conifer Abies veitchii.

During winter, subalpine conifers experience frequent freeze-thaw cycles in stem xylem that may cause embolism and pit aspiration due to increased water volume during the sap to ice transition. This study examined the occurrence and ecological impacts of a combination of freeze-thaw-induced pit aspiration and embolism triggered by natural and artificial stem freezing. In subalpine Veitch's fir (Abies veitchii) trees, the fraction of closed pits and embolized tracheids as well as conductivity losses were measured to examine pit aspiration and its effects. When trees incurred mild drought stress in February and early March, 70%-80% of stem conductivity was lost. Cryo-scanning electron microscopy indicated <20% embolized tracheids but ∼90% closed pits. Severe drought stress in late March caused 96% ± 1.2% (mean ± standard error) loss of stem conductivity, while the fraction of embolized tracheids increased to 64% ± 6.6%, and aspirated pit fraction decreased to 23% ± 5.6%. Experimental freeze-thaw cycles also increased pit aspiration from 7.1% ± 0.89% to 49% ± 10%, and the fraction of closed pits was positively correlated to the percent loss of stem hydraulic conductivity. The results indicated that freezing-induced pit aspiration is an important factor for stem xylem dysfunction under mild drought, and upon severe drought in winter; stem water transport is predominantly inhibited by xylem embolism.

Pit aspiration causes an apparent loss of xylem hydraulic conductivity in a subalpine fir (Abies mariesii Mast.) overwintering at the alpine timberline.

Conifers growing at the alpine timberline are exposed to combinatorial stresses that induce embolism in xylem during winter. We collected branches of Abies mariesii Mast. at the timberline on Mt Norikura of central Japan to evaluate the seasonal changes in the loss of xylem hydraulic conductivity (percent loss of hydraulic conductivity; PLC). Concurrently, we evaluated the distribution of embolized tracheids in native samples via cryo-scanning electron microscopic (cryo-SEM) observation in comparison with the drought-induced embolism samples used for the vulnerability curve. The twigs collected in late winter showed 100 PLC at a water potential of ~-3 MPa, and air-filled tracheids were observed sporadically in the cryo-SEM images. The cryo-SEM images also showed that nearly all pits of the samples from the timberline were aspirated in the xylem with 100 PLC. Conversely, in drought-induced samples used for vulnerability analysis, lower frequency of aspirated pits was observed at similar water potentials and all tracheids in the earlywood of xylem with 100 PLC were filled with air. We hypothesized that pit aspiration is the primary cause of the pronounced winter xylem conductivity loss at the timberline when water potential is near, but still above, the drought-induced vulnerability threshold. Pit aspiration before water loss may be an adaptation to severe winter conditions at timberlines to prevent widespread xylem embolism. The possible causes of pit aspiration are discussed in relation to complex stresses under harsh winter conditions at timberlines.

Hydraulic architecture with high root-resistance fraction contributes to efficient carbon gain of plants in temperate habitats.

PREMISE: The hydraulic architecture in the leaves, stems and roots of plants constrains water transport and carbon gain through stomatal limitation to CO2 absorption. Because roots are the main bottleneck in water transport for a range of plant species, we assessed the ecophysiological mechanism and importance of a high fraction of root hydraulic resistance in woody and herbaceous species. METHODS: Biomass partitioning and hydraulic conductance of leaves, stems, and roots of Japanese knotweed (Fallopia japonica, a perennial herb) and Japanese zelkova (Zelkova serrata, a deciduous tall tree) were measured. Theoretical analyses were used to examine whether the measured hydraulic architecture and biomass partitioning maximized the plant photosynthetic rate (the product of leaf area and photosynthetic rate per leaf area). RESULTS: Root hydraulic resistance accounted for 83% and 68% of the total plant resistance for Japanese knotweed and Japanese zelkova, respectively. Comparisons of hydraulic and biomass partitioning revealed that high root-resistance fractions were attributable to low biomass partitioning to root organs rather than high mass-specific root conductance. The measured partitioning of hydraulic resistance closely corresponded to the predicted optimal partitioning, maximizing the plant photosynthetic rate for the two species. The high fraction of root resistance was predicted to be optimal with variations in air humidity and soil water potential. CONCLUSIONS: These results suggest that the hydraulic architecture of plants growing in mesic and fertile habitats not only results in high root resistance due to small biomass partitioning to root organs, but contributes to efficient carbon gain.

Genetic Dissection of Growth and Eco-Physiological Traits Associated with Altitudinal Adaptation in Sakhalin Fir (Abies sachalinensis) Based on QTL Mapping.

(1) Background: The genetic basis of local adaptation in conifers remains poorly understood because of limited research evidence and the lack of suitable genetic materials. Sakhalin fir (Abies sachalinensis) is an ideal organism for elucidating the genetic basis of local adaptation because its altitudinal adaptation has been demonstrated, and suitable materials for its linkage mapping are available. (2) Method: We constructed P336 and P236 linkage maps based on 486 and 516 single nucleotide polymorphisms, respectively, that were derived from double digest restriction site-associated DNA sequences. We measured the growth and eco-physiological traits associated with morphology, phenology, and photosynthesis, which are considered important drivers of altitudinal adaptation. (3) Results: The quantitative trait loci (QTLs) for growth traits, phenology, needle morphology, and photosynthetic traits were subsequently detected. Similar to previous studies on conifers, most traits were controlled by multiple QTLs with small or moderate effects. Notably, we detected that one QTL for the crown area might be a type-A response regulator, a nuclear protein responsible for the cytokinin-induced shoot elongation. (4) Conclusion: The QTLs detected in this study include potentially important genomic regions linked to altitudinal adaptation in Sakhalin fir.

Imperforate tracheary elements and vessels alleviate xylem tension under severe dehydration: insights from water release curves for excised twigs of three tree species.

PREMISE: Water stored in the xylem of woody plants is important for supporting the transpiration stream under prolonged drought, yet the source of stored water within the xylem during drought remains unclear. Insights into xylem water utilization during drought will uncover the adaptation strategies of the test species to stress. METHODS: To fill the existing knowledge gap, we excised twigs of Abies firma (Japanese fir, conifer), Cercidiphyllum japonicum (katsura tree, diffuse-porous) and Quercus serrata (konara oak, ring-porous) to quantify interspecific variation of water transfer in xylem corresponding with increasing cumulative water release (CWR) using micro x-ray computed tomography and cryo-SEM. RESULTS: For all species studied, the main components of water storage within the operating range of water potential were not living cells but cavitation release and capillaries. Abies firma maintained water in the earlywood-like cells, for possible maintenance of the transpiration stream. Cercidiphyllum japonicum maintained water in its vessels over 200 kg m-3 of CWR, while Q. serrata lost most of its water in vessels with increasing CWR up to 100 kg m-3 . Cercidiphyllum japonicum exhibited a higher water storage capacity than Q. serrata. Under high CWR, narrow conduits stored xylem water in C. japonicum and imperforate tracheary elements in Q. serrata. CONCLUSIONS: Among the species examined, increasing CWR appears to indicate differential utilization of stored water in relation to variation of xylem structure, thereby providing insight into the interspecific responses of tree species to drought.

Repair of severe winter xylem embolism supports summer water transport and carbon gain in flagged crowns of the subalpine conifer Abies veitchii.

Xylem embolism induced by winter drought is a serious dysfunction in evergreen conifers growing at wind-exposed sites in the mountains. Some coniferous species can recover from winter embolism. The aim of this study was to determine whether wind direction influences embolism formation and/or repair dynamics on short windward and long leeward branches of asymmetrical `flagged' crowns. We analyzed the effect of branch orientation on percentage loss of xylem conductive area (PLC), leaf functional traits and the xylem:leaf area ratio for subalpine, wind-exposed flagged-crown Abies veitchii trees in the northern Yatsugatake Mountains of central Japan. In late winter, the shoot water potential was below -2.5 MPa, and the PLC exceeded 80% in 2-year-old branches, independent of branch orientation within a flagged crown. Both of these parameters almost fully recovered by summer. At branch internodes 4 years of age and older, seasonal changes in PLC were not found in either windward or leeward branches, but the PLC was higher in less leafy windward branches. The leaf nitrogen content and water-use efficiency of mature leaves were comparable between windward branches and leafy leeward branches. The ratio of xylem conductive area to total leaf area was the same for windward and leeward branches. These results indicate that the repair of winter xylem embolism allows leaf physiological functions to be maintained under sufficient leaf water supply, even on winter-wind-exposed branches. This permits substantial photosynthetic carbon gain during the following growing season on both windward and leeward branches. Thus, xylem recovery from winter embolism is a key trait for the survival of harsh winters and to support productivity on the individual level in flagged-crown A. veitchii trees.

Leaf structural and hydraulic adjustment with respect to air humidity and canopy position in silver birch (Betula pendula).

Climate change scenarios predict an increase in air temperature and precipitation in northern temperate regions of Europe by the end of the century. Increasing atmospheric humidity inevitably resulting from more frequent rainfall events reduces water flux through vegetation, influencing plants' structure and functioning. We investigated the extent to which artificially elevated air humidity affects the anatomical structure of the vascular system and hydraulic conductance of leaves in Betula pendula. A field experiment was carried out at the Free Air Humidity Manipulation (FAHM) site with a mean increase in relative air humidity (RH) by 7% over the ambient level across the growing period. Leaf hydraulic properties were determined with a high-pressure flow meter; changes in leaf anatomical structure were studied by means of conventional light microscopy and digital image processing techniques. Leaf development under elevated RH reduced leaf-blade hydraulic conductance and petiole conductivity and had a weak effect on leaf vascular traits (vessel diameters decreased), but had no significant influence on stomatal traits or tissue proportions in laminae. Both hydraulic traits and relevant anatomical characteristics demonstrated pronounced trends with respect to leaf location in the canopy-they increased from crown base to top. Stomatal traits were positively correlated with several petiole and leaf midrib vascular traits. The reduction in leaf hydraulic conductance in response to increasing air humidity is primarily attributable to reduced vessel size, while higher hydraulic efficiency of upper-crown foliage is associated with vertical trends in the size of vascular bundles, vessel number and vein density. Although we observed co-ordinated adjustment of vascular and hydraulic traits, the reduced leaf hydraulic efficiency could lead to an imbalance between hydraulic supply and transpiration demand under the extreme environmental conditions likely to become more frequent in light of global climate change.

Bundle sheath lignification mediates the linkage of leaf hydraulics and venation.

The lignification of the leaf vein bundle sheath (BS) has been observed in many species and would reduce conductance from xylem to mesophyll. We hypothesized that lignification of the BS in lower-order veins would provide benefits for water delivery through the vein hierarchy but that the lignification of higher-order veins would limit transport capacity from xylem to mesophyll and leaf hydraulic conductance (Kleaf ). We further hypothesized that BS lignification would mediate the relationship of Kleaf to vein length per area. We analysed the dependence of Kleaf , and its light response, on the lignification of the BS across vein orders for 11 angiosperm tree species. Eight of 11 species had lignin deposits in the BS of the midrib, and two species additionally only in their secondary veins, and for six species up to their minor veins. Species with lignification of minor veins had a lower hydraulic conductance of xylem and outside-xylem pathways and lower Kleaf . Kleaf could be strongly predicted by vein length per area and highest lignified vein order (R2  = .69). The light-response of Kleaf was statistically independent of BS lignification. The lignification of the BS is an important determinant of species variation in leaf and thus whole plant water transport.

Intra-specific trends of lumen and wall resistivities of vessels within the stem xylem vary among three woody plants.

Water flow through xylem vessels encounters hydraulic resistance when passing through the vessel lumen and end wall. Comparative studies have reported that lumen and end wall resistivities co-limit water flow through stem xylem in several angiosperm woody species that have vessels of different average diameter and length. This study examined the intra-specific relationship between the lumen and end wall resistivities (Rlumen and Rwall) for vessels within the stem xylem using three deciduous angiosperm woody species found in temperate forest. Morus australis Poir. and Acer rufinerve Siebold et Zucc. are early- and late-successional species, and Vitis coignetiae Pulliat ex Planch is a woody liana. According to the Hagen-Poiseuille equation, Rlumen is proportional to the fourth power of vessel diameter (D), whereas vessel length (L) and inter-vessel pit area (Apit) determine Rwall. To estimate Rlumen and Rwall, the scaling relationships between the L and D and between Apit and D were measured. The scaling exponents between L and D were 1.47, 3.19 and 2.86 for A. rufinerve, M. australis and V. coignetiae, respectively, whereas those between Apit and D were 0.242, 2.11 and 2.68, respectively. Unlike the inter-specific relationships, the wall resistivity fraction (Rwall/(Rlumen + Rwall)) within xylem changed depending on D. In M. australis and V. coignetiae, this fraction decreased with increasing D, while in A. rufinerve, it increased with D. Vessels with a high wall resistivity fraction have high Rwall and total resistivity but are expected to have low susceptibility to xylem cavitation due to a small cumulative Apit. In contrast, vessels with a low wall resistivity fraction have low Rwall and total resistivity but high susceptibility to xylem cavitation. Because the wall resistivity fraction varies with D, the stem xylem contains vessels with different hydraulic efficiencies and safety to xylem cavitation. These features produce differences in the hydraulic properties of plants with different life forms.

Structures of Bordered Pits Potentially Contributing to Isolation of a Refilled Vessel from Negative Xylem Pressure in Stems of Morus australis Poir.: Testing of the Pit Membrane Osmosis and Pit Valve Hypotheses.

Two hypotheses have been proposed to explain the mechanism preventing the refilling vessel water from being drained to the neighboring functional vessels under negative pressure. The pit membrane osmosis hypothesis proposes that the xylem parenchyma cells release polysaccharides that are impermeable to the intervessel pit membranes into the refilling vessel; this osmotically counteracts the negative pressure, thereby allowing the vessel to refill. The pit valve hypothesis proposes that gas trapped within intervessel bordered pits isolates the refilling vessel water from the surrounding functional vessels. Here, using the single-vessel method, we assessed these hypotheses in shoots of mulberry (Morus australis Poir.). First, we confirmed the occurrence of xylem refilling under negative pressure in the potted mulberry saplings. To examine the pit membrane osmosis hypothesis, we estimated the semi-permeability of pit membranes for molecules of various sizes and found that the pit membranes were not semi-permeable to polyethylene glycol of molecular mass <20,000. For the pit valve hypothesis, we formed pit valves in the intervessel pits in the short stem segments and measured the maximum liquid pressure up to which gases in bordered pits were retained. The threshold pressure ranged from 0.025 to 0.10 MPa. These values matched the theoretical value calculated from the geometry of the pit chamber (0.0692-0.101 MPa). Our results suggest that gas in the pits is retained by surface tension, even under substantial positive pressure to resolve gases in the refilling vessel, whereas the molecule size required for the pit membrane osmosis mechanism in mulberry would be unrealistically large.