Christopher M. Cohu

May photoinhibition be a consequence, rather than a cause, of limited plant productivity?

Photoinhibition in leaves in response to high and/or excess light, consisting of a decrease in photosynthesis and/or photosynthetic efficiency, is frequently equated to photodamage and often invoked as being responsible for decreased plant growth and productivity. However, a review of the literature reveals that photoinhibited leaves characterized for foliar carbohydrate levels were invariably found to possess high levels of sugars and starch. We propose that photoinhibition should be placed in the context of whole-plant source–sink regulation of photosynthesis. Photoinhibition may represent downregulation of the photosynthetic apparatus in response to excess light when (1) more sugar is produced in leaves than can be utilized by the rest of the plant and/or (2) more light energy is harvested than can be utilized by the chloroplast for the fixation of carbon dioxide into sugars.

Foliar phloem infrastructure in support of photosynthesis

Acclimatory adjustments of foliar minor loading veins in response to growth at different temperatures and light intensities are evaluated. These adjustments are related to their role in providing infrastructure for the export of photosynthetic products as a prerequisite for full acclimation of photosynthesis to the respective environmental conditions. Among winter-active apoplastic loaders, higher photosynthesis rates were associated with greater numbers of sieve elements per minor vein as well as an increased apparent total membrane area of cells involved in phloem loading (greater numbers of cells and/or greater cell wall invaginations). Among summer-active apoplastic loaders, higher photosynthesis rates were associated with increased vein density and, possibly, a greater number of sieve elements and companion cells per minor vein. Among symplastic loaders, minor loading vein architecture (number per vein and arrangement of cells) was apparently constrained, but higher photosynthesis rates were associated with higher foliar vein densities and larger intermediary cells (presumably providing a greater volume for enzymes involved in active raffinose sugar synthesis). Winter-active apoplastic loaders thus apparently place emphasis on adjustments of cell membrane area (presumably available for transport proteins active in loading of minor veins), while symplastic loaders apparently place emphasis on increasing the volume of cells in which their active loading step takes place. Presumably to accommodate a greater flux of photosynthate through the foliar veins, winter-active apoplastic loaders also have a higher number of sieve elements per minor loading vein, whereas symplastic loaders and summer-active apoplastic loaders have a higher total number of veins per leaf area. These latter adjustments in the vasculature (during leaf development) may also apply to the xylem (via greater numbers of tracheids per vein and/or greater vein density per leaf area) serving to increase water flux to mesophyll tissues in support of high rates of transpiration typically associated with high rates of photosynthesis.

Association between photosynthesis and contrasting features of minor veins in leaves of summer annuals loading phloem via symplastic versus apoplastic routes

Foliar vascular anatomy and photosynthesis were evaluated for a number of summer annual species that either load sugars into the phloem via a symplastic route (Cucumis sativus L. cv. Straight Eight; Cucurbita pepo L. cv. Italian Zucchini Romanesco; Citrullus lanatus L. cv. Faerie Hybrid; Cucurbita pepo L. cv. Autumn Gold) or an apoplastic route (Nicotiana tabacum L.; Solanum lycopersicum L. cv. Brandywine; Gossypium hirsutum L.; Helianthus annuus L. cv. Soraya), as well as winter annual apoplastic loaders (Spinacia oleracea L. cv. Giant Nobel; Arabidopsis thaliana (L.) Heynhold Col-0, Swedish and Italian ecotypes). For all summer annuals, minor vein cross-sectional xylem area and tracheid number as well as the ratio of phloem loading cells to phloem sieve elements, each when normalized for foliar vein density (VD), was correlated with photosynthesis. These links presumably reflect (1) the xylems role in providing water to meet foliar transpirational demand supporting photosynthesis and (2) the importance of the driving force of phloem loading as well as the cross-sectional area for phloem sap flux to match foliar photosynthate production. While photosynthesis correlated with the product of VD and cross-sectional phloem cell area among symplastic loaders, photosynthesis correlated with the product of VD and phloem cell number per vein among summer annual apoplastic loaders. Phloem cell size has thus apparently been a target of selection among symplastic loaders (where loading depends on enzyme concentration within loading cells) versus phloem cell number among apoplastic loaders (where loading depends on membrane transporter numbers).

Low temperature acclimation of photosynthetic capacity and leaf morphology in the context of phloem loading type

Carbon export from leaf mesophyll to sugar-transporting phloem occurs via either an apoplastic (across the cell membrane) or symplastic (through plasmodesmatal cell wall openings) pathway. Herbaceous apoplastic loaders generally exhibit an up-regulation of photosynthetic capacity in response to growth at lower temperature. However, acclimation of photosynthesis to temperature by symplastically loading species, whose geographic distribution is particularly strong in tropical and subtropical areas, has not been characterized. Photosynthetic and leaf anatomical acclimation to lower temperature was explored in two symplastic (Verbascum phoeniceum, Cucurbita pepo) and two apoplastic (Helianthus annuus, Spinacia oleracea) loaders, representing summer- and winter-active life histories for each loading type. Regardless of phloem loading type, the two summer-active species, C. pepo and H. annuus, exhibited neither foliar anatomical nor photosynthetic acclimation when grown under low temperature compared to moderate temperature. In contrast, and again irrespective of phloem loading type, the two winter-active mesophytes, V. phoeniceum and S. oleracea, exhibited both a greater number of palisade cell layers (and thus thicker leaves) and significantly higher maximal capacities of photosynthetic electron transport, as well as, in the case of V. phoeniceum, a greater foliar vein density in response to cool temperatures compared to growth at moderate temperature. It is therefore noteworthy that symplastic phloem loading per se does not prevent acclimation of intrinsic photosynthetic capacity to cooler growth temperatures. Given the vagaries of weather and climate, understanding the basis of plant acclimation to, and tolerance of, low temperature is critical to maintaining and increasing plant productivity for food, fuel, and fiber to meet the growing demands of a burgeoning human population.

Leaf anatomical and photosynthetic acclimation to cool temperature and high light in two winter versus two summer annuals

Acclimation of foliar features to cool temperature and high light was characterized in winter (Spinacia oleracea L. cv. Giant Nobel; Arabidopsis thaliana (L.) Heynhold Col-0 and ecotypes from Sweden and Italy) versus summer (Helianthus annuus L. cv. Soraya; Cucurbita pepo L. cv. Italian Zucchini Romanesco) annuals. Significant relationships existed among leaf dry mass per area, photosynthesis, leaf thickness and palisade mesophyll thickness. While the acclimatory response of the summer annuals to cool temperature and/or high light levels was limited, the winter annuals increased the number of palisade cell layers, ranging from two layers under moderate light and warm temperature to between four and five layers under cool temperature and high light. A significant relationship was also found between palisade tissue thickness and either cross-sectional area or number of phloem cells (each normalized by vein density) in minor veins among all four species and growth regimes. The two winter annuals, but not the summer annuals, thus exhibited acclimatory adjustments of minor vein phloem to cool temperature and/or high light, with more numerous and larger phloem cells and a higher maximal photosynthesis rate. The upregulation of photosynthesis in winter annuals in response to low growth temperature may thus depend on not only (1) a greater volume of photosynthesizing palisade tissue but also (2) leaf veins containing additional phloem cells and presumably capable of exporting a greater volume of sugars from the leaves to the rest of the plant.

Association between minor loading vein architecture and light- and CO2-saturated rates of photosynthetic oxygen evolution among Arabidopsis thaliana ecotypes from different latitudes

Through microscopic analysis of veins and assessment of light- and CO2-saturated rates of photosynthetic oxygen evolution, we investigated the relationship between minor loading vein anatomy and photosynthesis of mature leaves in three ecotypes of Arabidopsis thaliana grown under four different combinations of temperature and photon flux density (PFD). All three ecotypes exhibited greater numbers and cross-sectional area of phloem cells as well as higher photosynthesis rates in response to higher PFD and especially lower temperature. The Swedish ecotype exhibited the strongest response to these conditions, the Italian ecotype the weakest response, and the Col-0 ecotype exhibited an intermediate response. Among all three ecotypes, strong linear relationships were found between light- and CO2-saturated rates of photosynthetic oxygen evolution and the number and area of either sieve elements or of companion and phloem parenchyma cells in foliar minor loading veins, with the Swedish ecotype showing the highest number of cells in minor loading veins (and largest minor veins) coupled with unprecedented high rates of photosynthesis. Linear, albeit less significant, relationships were also observed between number and cross-sectional area of tracheids per minor loading vein versus light- and CO2-saturated rates of photosynthetic oxygen evolution. We suggest that sugar distribution infrastructure in the phloem is co-regulated with other features that set the upper limit for photosynthesis. The apparent genetic differences among Arabidopsis ecotypes should allow for future identification of the gene(s) involved in augmenting sugar-loading and -transporting phloem cells and maximal rates of photosynthesis.

Minor loading vein acclimation for three Arabidopsis thaliana ecotypes in response to growth under different temperature and light regimes

In light of the important role of foliar phloem as the nexus between energy acquisition through photosynthesis and distribution of the products of photosynthesis to the rest of the plant, as well as communication between the whole plant and its leaves, we examined whether foliar minor loading veins in three Arabidopsis thaliana ecotypes undergo acclimation to the growth environment. As a winter annual exhibiting higher rates of photosynthesis in response to cooler vs. warmer temperatures, this species might be expected to adjust the structure of its phloem to accommodate greater fluxes of sugars in response to growth at low temperature. Minor (fourth- and third-order) veins had 14 or fewer sieve elements and phloem tissue comprised 50% or more of the cross-sectional area. The number of phloem cells per minor loading vein was greater in leaves grown under cool temperature and high light vs. warm temperature and moderate light. This effect was greatest in an ecotype from Sweden, in which growth under cool temperature and high light resulted in minor veins with an even greater emphasis on phloem (50% more phloem cells with more than 100% greater cross-sectional area of phloem) compared to growth under warm temperature and moderate light. Likewise, the number of sieve elements per minor vein increased linearly with growth temperature under moderate light, almost doubling over a 27°C temperature range (21°C leaf temperature range) in the Swedish ecotype. Increased emphasis on cells involved in sugar loading and transport may be critical for maintaining sugar export from leaves of an overwintering annual such as A. thaliana, and particularly for the ecotype from the northern-most population experiencing the lowest temperatures.

Associations between the acclimation of phloem-cell wall ingrowths in minor veins and maximal photosynthesis rate

The companion cells (CCs) and/or phloem parenchyma cells (PCs) in foliar minor veins of some species exhibit invaginations that are amplified when plants develop in high light (HL) compared to low light (LL). Leaves of plants that develop under HL also exhibit greater maximal rates of photosynthesis compared to those that develop under LL, suggesting that the increased membrane area of CCs and PCs of HL-acclimated leaves may provide for greater levels of transport proteins facilitating enhanced sugar export. Furthermore, the degree of wall invagination in PCs (Arabidopsis thaliana) or CCs (pea) of fully expanded LL-acclimated leaves increased to the same level as that present in HL-acclimated leaves 7 days following transfer to HL, and maximal photosynthesis rates of transferred leaves of both species likewise increased to the same level as in HL-acclimated leaves. In contrast, transfer of Senecio vulgaris from LL to HL resulted in increased wall invagination in CCs, but not PCs, and such leaves furthermore exhibited only partial upregulation of photosynthetic capacity following LL to HL transfer. Moreover, a significant linear relationship existed between the level of cell wall ingrowths and maximal photosynthesis rates across all three species and growth light regimes. A positive linear relationship between these two parameters was also present for two ecotypes (Sweden, Italy) of the winter annual A. thaliana in response to growth at different temperatures, with significantly greater levels of PC wall ingrowths and higher rates of photosynthesis in leaves that developed at cooler versus warmer temperatures. Treatment of LL-acclimated plants with the stress hormone methyl jasmonate also resulted in increased levels of wall ingrowths in PCs of A. thaliana and S. vulgaris but not in CCs of pea and S. vulgaris. The possible role of PC wall ingrowths in sugar export versus as physical barriers to the movement of pathogens warrants further attention.

Growth temperature impact on leaf form and function in Arabidopsis thaliana ecotypes from northern and southern Europe.

The plasticity of leaf form and function in European lines of Arabidopsis thaliana was evaluated in ecotypes from Sweden and Italy grown under contrasting (cool versus hot) temperature regimes. Although both ecotypes exhibited acclimatory adjustments, the Swedish ecotype exhibited more pronounced responses to the two contrasting temperature regimes in several characterized features. These responses included thicker leaves with higher capacities for photosynthesis, likely facilitated by a greater number of phloem cells per minor vein for the active loading and export of sugars, when grown under cool temperature as opposed to leaves with a higher vein density and a greater number of tracheary elements per minor vein, likely facilitating higher rates of transpirational water loss (and thus evaporative cooling), when grown under hot temperature with high water availability. In addition, only the Swedish ecotype exhibited reduced rosette growth and greater levels of foliar tocopherols under the hot growth temperature. These responses, and the greater responsiveness of the Swedish ecotype compared with the Italian ecotype, are discussed in the context of redox signalling networks and transcription factors, and the greater range of environmental conditions experienced by the Swedish versus the Italian ecotype during the growing season in their native habitats.

Habitat Temperature and Precipitation of Arabidopsis thaliana Ecotypes Determine the Response of Foliar Vasculature, Photosynthesis, and Transpiration to Growth Temperature

Acclimatory adjustments of foliar vascular architecture, photosynthetic capacity, and transpiration rate in Arabidopsis thaliana ecotypes (Italian, Polish [Col-0], Swedish) were characterized in the context of habitat of origin. Temperatures of the habitat of origin decreased linearly with increasing habitat latitude, but habitat precipitation was greatest in Italy, lowest in Poland, and intermediate in Sweden. Plants of the three ecotypes raised under three different growth temperature regimes (low, moderate, and high) exhibited highest photosynthetic capacities, greatest leaf thickness, highest chlorophyll a/b ratio and levels of β-carotene, and greatest levels of wall ingrowths in phloem transfer cells, and, in the Col-0 and Swedish ecotypes, of phloem per minor vein in plants grown at the low temperature. In contrast, vein density and minor vein tracheary to sieve element ratio increased with increasing growth temperature – most strongly in Col-0 and least strongly in the Italian ecotype – and transpirational water loss correlated with vein density and number of tracheary elements per minor vein. Plotting of these vascular features as functions of climatic conditions in the habitat of origin suggested that temperatures during the evolutionary history of the ecotypes determined acclimatory responses of the foliar phloem and photosynthesis to temperature in this winter annual that upregulates photosynthesis in response to lower temperature, whereas the precipitation experienced during the evolutionary history of the ecotypes determined adjustment of foliar vein density, xylem, and transpiration to temperature. In particular, whereas photosynthetic capacity, leaf thickness, and foliar minor vein phloem features increased linearly with increasing latitude and decreasing temperature of the habitats of origin in response to experimental growth at low temperature, transpiration rate, foliar vein density, and minor vein tracheary element numbers and cross-sectional areas increased linearly with decreasing precipitation level in the habitats of origin in response to experimental growth at high temperature. This represents a situation where temperature acclimation of the apparent capacity for water flux through the xylem and transpiration rate in a winter annual responded differently from that of photosynthetic capacity, in contrast to previous reports of strong relationships between hydraulic conductance and photosynthesis in other studies.