Abstract
Measurements of vein density and foliar minor vein phloem cell numbers, minor vein phloem cell sizes, and transfer cell wall ingrowths provide quantitative proxies for the leaf’s capacities to load and export photosynthates. While overall infrastructural capacity for sugar loading and sugar export correlated positively and closely with photosynthetic capacity, the specific targets of the adjustment of minor vein organization varied with phloem-loading mechanism, plant life-cycle characteristics, and environmental growth conditions. Among apoplastic loaders, for which sugar loading into the phloem depends on cell membrane-spanning transport proteins, variation in minor vein density, phloem cell number, and level of cell wall ingrowth (when present) were consistently associated with photosynthetic capacity. Among active symplastic loaders, for which sugar loading into the phloem depends on cytosolic enzymes, variation in vein density and phloem cell size were consistently associated with photosynthetic capacity. All of these anatomical features were also subject to acclimatory adjustment depending on species and environmental conditions, with increased levels of these features supporting higher rates of photosynthesis. We present a procedure for the preparation of leaf tissue for minor vein analysis, using both light and transmission electron microscopy, that facilitates quantification of not only phloem features but also xylem features that provide proxies for foliar water import capacity.
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Abbreviations
- CA:
-
Cross-sectional area of cells per minor vein
- CC:
-
Companion cell
- CN:
-
Cell number per minor vein
- EM:
-
Electron microscopy
- PC:
-
Phloem parenchyma cell
- SE:
-
Sieve element
- TE:
-
Tracheary element
- VD:
-
Minor vein density
- VP:
-
Vascular parenchyma
- XP:
-
Xylem parenchyma
References
Amiard V, Mueh KE, Demmig-Adams B, Ebbert V, Turgeon R, Adams WW III (2005) Anatomical and photosynthetic acclimation to the light environment in species with differing mechanisms of phloem loading. Proc Natl Acad Sci U S A 102:12968–12973
Brodribb TJ, Feild TS, Jordan GJ (2007) Leaf maximum photosynthetic rate and venation are linked by hydraulics. Plant Physiol 144:1890–1898
Adams WW III, Cohu CM, Muller O, Demmig-Adams B (2013) Foliar phloem infrastructure in support of photosynthesis. Front Plant Sci 4:194
Adams WW III, Stewart JJ, Polutchko SK, Demmig-Adams B (2018) Leaf vasculature and the upper limit of photosynthesis. In: Adams WW III, Terashima I (eds) The leaf: a platform for performing photosynthesis. Advances in photosynthesis and respiration, vol 44, pp 27–54. Springer, Dordrecht
Cohu CM, Muller O, Stewart JJ, Demmig-Adams B, Adams WW III (2013) Association between minor loading vein architecture and light- and CO2-saturated rates of photosynthetic oxygen evolution among Arabidopsis thaliana ecotypes from different latitudes. Front Plant Sci 4:264
Caringella MA, Bongers FJ, Sack L (2015) Leaf hydraulic conductance varies with vein anatomy across Arabidopsis thaliana wild-type and leaf vein mutants. Plant Cell Environ 38:2735–2746
Scoffoni C, Chatelet DS, Pasquet-kok J, Rawls M, Donoghue MJ, Edwards EJ, Sack L (2016) Hydraulic basis for the evolution of photosynthetic productivity. Nat Plant 2:16072
McAdam SAM, Eléouët MP, Best M et al (2017) Linking auxin with photosynthetic rate via leaf venation. Plant Physiol 175:351–360
Polutchko SK, Stewart JJ, Demmig-Adams B, Adams WW III (2018) Evaluating the link between photosynthetic capacity and leaf vascular organization with principal component analysis. Photosynthetica 56:392–403
Adams WW III, Cohu CM, Amiard V, Demmig-Adams B (2014) Associations between phloem-cell wall ingrowths in minor veins and maximal photosynthesis rate. Front Plant Sci 5:24
Adams WW III, Stewart JJ, Cohu CM, Muller O, Demmig-Adams B (2016) Habitat temperature and precipitation of Arabidopsis thaliana ecotypes determine the response of foliar vasculature, photosynthesis, and transpiration to growth temperature. Front Plant Sci 7:1026
Muller O, Cohu CM, Stewart JJ, Protheroe JA, Demmig-Adams B, Adams WW III (2014a) Association between photosynthesis and contrasting features of minor veins in leaves of summer annuals loading phloem via symplastic versus apoplastic routes. Physiol Plant 152:174–183
Muller O, Stewart JJ, Cohu CM, Polutchko SK, Demmig-Adams B, Adams WW III (2014b) Leaf architectural, vascular, and photosynthetic acclimation to temperature in two biennials. Physiol Plant 152:763–772
Stewart JJ, Demmig-Adams B, Cohu CM, Wenzl CA, Muller O, Adams WW III (2016) Growth temperature impact on leaf form and function in Arabidopsis thaliana ecotypes from northern and southern Europe. Plant Cell Environ 39:1549–1558
Stewart JJ, Polutchko SK, Adams WW III, Cohu CM, Wenzl CA, Demmig-Adams B (2017a) Light, temperature and tocopherol status influence foliar vascular anatomy and leaf function in Arabidopsis thaliana. Physiol Plant 160:98–110
Stewart JJ, Polutchko SK, Adams WW III, Demmig-Adams B (2017b) Acclimation of Swedish and Italian ecotypes of Arabidopsis thaliana to light intensity. Photosynth Res 134:215–229
Stewart JJ, Adams WW III, Cohu CM, Demmig-Adams B (2018a) Tocopherols modulate leaf vein arrangement and composition without impacting photosynthesis. Photosynthetica 56:382–391
Stewart JJ, Polutchko SK, Demmig-Adams B, Adams WW III (2018b) Arabidopsis thaliana Ei-5: altered minor vein architecture compensates for low vein density and supports maintenance of photosynthetic capacity. Front Plant Sci 9:693
Demmig-Adams B, Stewart JJ, Adams WW III (2017) Environmental regulation of intrinsic photosynthetic capacity: an integrated view. Curr Opin Plant Biol 37:34–41
Cohu CM, Muller O, Demmig-Adams B, Adams WW III (2014) Leaf anatomical and photosynthetic acclimation to cool temperature and high light in two winter versus two summer annuals. Physiol Plant 152:164–173
Dumlao MR, Darehshouri A, Cohu CM, Muller O, Mathias J, Adams WW III, Demmig-Adams B (2012) Low temperature acclimation of photosynthetic capacity and leaf morphology in the context of phloem loading type. Photosynth Res 113:181–189
Cohu CM, Muller O, Demmig-Adams B, Adams WW III (2013b) Minor loading vein acclimation for three Arabidopsis thaliana ecotypes in response to growth under different temperature and light regimes. Front Plant Sci 4:240
Amiard V, Demmig-Adams B, Mueh KE, Turgeon R, Combs AF, Adams WW III (2007) Role of light and jasmonic acid signaling in regulating foliar phloem cell wall ingrowth development. New Phytol 173:722–731
Adams WW III, Watson AM, Mueh KE, Amiard V, Turgeon R, Ebbert V, Logan BA, Combs AF, Demmig-Adams B (2007) Photosynthetic acclimation in the context of structural constraints to carbon export from leaves. Photosynth Res 94:455–466
Fu Q, Chen L, Guo Y, Turgeon R (2011) Phloem loading strategies and water relations in trees and herbaceous plants. Plant Physiol 157:1518–1527
Demmig-Adams B, Stewart JJ, Adams WW III (2018) Optimization of photosynthetic productivity in contrasting environments by regulons controlling plant form and function. Int J Mol Sci 19:872
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675
Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43
Winey M, Meehl JB, O’Toole ET, Giddings TH Jr (2014) Conventional transmission electron microscopy. Mol Biol Cell 25:319–323
Sack L, Scoffoni C (2013) Leaf venation: structure, function, development, evolution, ecology, and applications in the past, present and future. New Phytol 198:983–1000
Sack L, Caringella M, Scoffoni C, Mason C, Rawls M, Markesteijn L, Poorter L (2014) Leaf vein length per unit area is not intrinsically dependent on image magnification: avoiding measurement artifacts for accuracy and precision. Plant Physiol 166:829–838
Zhang C, Turgeon R (2018) Mechanisms of phloem loading. Curr Opin Plant Biol 43:71–75
Acknowledgments
We thank Dr. Thomas Giddings, Dr. Véronique Amiard, and Prof. Robert Turgeon for instruction, advice, and valuable discussion. This work was supported by the National Science Foundation (Award Numbers IOS-0841546, DEB-0122236, and IBN-0235351) and the University of Colorado at Boulder.
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Stewart, J.J., Muller, O., Cohu, C.M., Demmig-Adams, B., Adams, W.W. (2019). Quantification of Leaf Phloem Anatomical Features with Microscopy. In: Liesche, J. (eds) Phloem. Methods in Molecular Biology, vol 2014. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9562-2_5
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DOI: https://doi.org/10.1007/978-1-4939-9562-2_5
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