Asymmetric wall ingrowth deposition in Arabidopsis phloem parenchyma transfer cells is tightly associated with sieve elements

In Arabidopsis, polarized deposition of wall ingrowths in phloem parenchyma (PP) transfer cells (TCs) occurs adjacent to cells of the sieve element/companion cell (SE/CC) complex. However, the spatial relationships between these different cell types in minor veins, where phloem loading occurs, are poorly understood. PP TC development and wall ingrowth localization were compared to other phloem cells in leaves of Col-0 and the transgenic lines AtSUC2::AtSTP9-GFP and AtSWEET11::AtSWEET11-GFP that identify CCs and PP respectively. The development of PP TCs in minor veins, indicated by deposition of wall ingrowths, proceeded basipetally in leaves. However, not all PP develop ingrowths and higher levels of wall ingrowth deposition occur in abaxial- compared to adaxial-positioned PP TCs. Furthermore, the deposition of wall ingrowths was exclusively initiated on and preferentially covered the PP TC/SE interface, rather than the PP TC/CC interface, and only occurred in PP cells that were adjacent to SEs. Collectively, these results demonstrate the dominant impact of SEs on wall ingrowth deposition in PP TCs and suggest the existence of two sub-types of PP cells in leaf minor veins. Compared to PP cells, PP TCs showed more abundant accumulation of AtSWEET11-GFP, indicating functional differences in phloem loading between PP and PP TCs.

Loading regulation prevents phloem failure during drought and widens the range of phloem and stomatal traits

Plant carbon transport is controlled by a multitude of parameters both internal and external to the sugar transporting phloem tissue. Sucrose transporter kinetics, conduit hydraulic resistance, and xylem water stress are all hypothesized to impact the amount of carbon delivered to sink tissues. However, the most important traits determining carbon export under drought are not well understood, especially for species with active molecular regulation of sucrose transport. This in turn limits our ability to assess species’ resistances to phloem dysfunction under drought. Here, we use an integrated xylem-phloem-stomatal model to calculate leaf water potential from soil dryness, which is then used to determine gas exchange and phloem pressure gradients. We quantitatively compare the impacts of phloem loading kinetics, including feedbacks between loading and phloem pressure, phloem conduit resistances, and stomatal responses to water stress, on the total carbon export to sinks during drought. Regulating sucrose transporter kinetics which downregulates loading at high phloem pressures prevented runaway viscosity in the phloem sap and was the most important determinant of export rates under drought. In contrast to previous models, we found this feedback mechanism decoupled stomatal traits from phloem export efficiency during drought and increased the operational range of phloem hydraulic resistances.

Identification and functional analysis of SWEET gene family in Averrhoa carambola L. fruits during ripening

Sugar Will Eventually be Exported Transporters (SWEETs), a type of sugar efflux transporters, have been extensively researched upon due to their role in phloem loading for distant sugar transport, fruit development, and stress regulation, etc. Several plant species are known to possess the SWEET genes; however, little is known about their presence in Averrhoa Carambola L. (Oxalidaceae), an evergreen fruit crop (star fruit) in tropical and subtropical regions of Southeast Asia. In this study, we established an Averrhoa Carambola L. unigenes library from fruits of ‘XianMiyangtao’ (XM) by RNA sequencing (RNA-seq). A total of 99,319 unigenes, each longer than 200 bp with a total length was 72.00 Mb, were identified. A total of 51,642 unigenes (52.00%) were annotated. Additionally, 10 AcSWEET genes from the Averrhoa Carambola L. unigenes library were identified and classified, followed by a comprehensive analysis of their structures and conserved motif compositions, and evolutionary relationships. Moreover, the expression patterns of AcSWEETs in ‘XM’ cultivars during fruit ripening were confirmed using quantitative real-time PCR (qRT-PCR), combined with the soluble sugar and titratable acids content during ripening, showed that AcSWEET2a/2b and AcSWEET16b might participate in sugar transport during fruit ripening. This work presents a general profile of the AcSWEET gene family in Averrhoa Carambola L., which can be used to perform further studies on elucidating the functional roles of AcSWEET genes.

Coupling kinetic models and advection-diffusion equations. 2. Sensitivity analysis of an advection-diffusion-reaction model

The accompanying paper (Uys et al., in silico Plants, XXXX) presented a core model of sucrose accumulation within the advection-diffusion-reaction framework, which is able to capture the spatio-temporal evolution of the system from a set of initial conditions. This paper presents a sensitivity analysis of this model. Because this is a non-steady-state model based on partial differential equations, we performed the sensitivity analysis using two approaches from engineering. The Morris method is based on a one-at-a-time design, perturbing parameters individually and calculating the influence on model output in terms of elementary effects. FAST is a global sensitivity analysis method, where all parameters are perturbed simultaneously, oscillating at different frequencies, enabling the calculation of the contribution of each parameter through Fourier analysis. Overall, both methods gave similar results. Perturbations in reactions tended to have a large influence on their own rate, as well as on directly connected metabolites. Sensitivities varied both with the time of the simulation and the position along the sugarcane stalk. Our results suggest that vacuolar sucrose concentrations are most sensitive to vacuolar invertase in the centre of the stalk, but that phloem unloading and vacuolar sucrose uptake also contribute, especially towards the stalk edges. Sucrose in the phloem was most sensitive to phloem loading at the nodes, but most sensitive to phloem unloading in the middle of the internodes. Sink concentrations of sucrose in the symplast were most sensitive to phloem unloading in the middle of the internodes, but at the nodes cytosolic invertase had the greatest effect.

SWEET Transporters and the Potential Functions of These Sequences in Tea (Camellia sinensis)

Tea (Camellia sinensis) is an important economic beverage crop. Its flowers and leaves could be used as healthcare tea for its medicinal value. SWEET proteins were recently identified in plants as sugar transporters, which participate in diverse physiological processes, including pathogen nutrition, seed filling, nectar secretion, and phloem loading. Although SWEET genes have been characterized and identified in model plants, such as Arabidopsis thaliana and Oryza sativa, there is very little knowledge of these genes in C. sinensis. In this study, 28 CsSWEETs were identified in C. sinensis and further phylogenetically divided into four subfamilies with A. thaliana. These identified CsSWEETs contained seven transmembrane helixes (TMHs) which were generated by an ancestral three-TMH unit with an internal duplication experience. Microsynteny analysis revealed that the large-scale duplication events were the main driving forces for members from CsSWEET family expansion in C. sinensis. The expression profiles of the 28 CsSWEETs revealed that some genes were highly expressed in reproductive tissues. Among them, CsSWEET1a might play crucial roles in the efflux of sucrose, and CsSWEET17b could control fructose content as a hexose transporter in C. sinensis. Remarkably, CsSWEET12 and CsSWEET17c were specifically expressed in flowers, indicating that these two genes might be involved in sugar transport during flower development. The expression patterns of all CsSWEETs were differentially regulated under cold and drought treatments. This work provided a systematic understanding of the members from the CsSWEET gene family, which would be helpful for further functional studies of CsSWEETs in C. sinensis.

Drought Intensity-Responsive Salicylic Acid and Abscisic Acid Crosstalk with the Sugar Signaling and Metabolic Pathway in Brassica napus

The aim of this study was to characterize hormonal crosstalk with the sugar signaling and metabolic pathway based on a time course analysis of drought intensity. Drought intensity-responsive changes in the assimilation of newly fixed carbon (C) into soluble sugar, the content of sugar and starch, and expression of genes involved in carbohydrate metabolism were interpreted as being linked to endogenous abscisic acid (ABA) and salicylic acid (SA) levels and their signaling genes. The ABA and SA levels in the drought-stressed leaves increased together during the early drought period (days 0–6), and additional ABA accumulation occurred with depressed SA during the late period (days 6–14). Although drought treatment decreased the assimilation of newly fixed C into soluble sugar, representing a 59.9%, 33.1%, and 62.9% reduction in 13C-glucose, 13C-fructose, and 13C-sucrose on day 14, respectively, the drought-responsive accumulation of soluble sugars was significant. During the early period, the drought-responsive accumulation of hexose and sucrose was concurrent with the upregulated expression of hexokinase 1 (HXK1), which, in turn, occurred parallel to the upregulation of ABA synthesis gene 9-sis-epoxycarotenoid dioxygenase (NCED3) and SA-related genes (isochorismate synthase 1 (ICS1) and non-expressor of pathogenesis-related gene (NPR1)). During the late period, hexose accumulation, sucrose phloem loading, and starch degradation were dominant, with a highly enhanced expression of the starch degradation-related genes β-amylase 1 (BAM1) and α-amylase 3 (AMY3), which were concomitant with the parallel enhancement of sucrose non-fermenting−1 (Snf1)-related protein kinase 2 (SnRK2).2 and ABA-responsive element binding 2 (AREB2) expression in an ABA-dependent manner. These results indicate that the drought-responsive accumulation of sugars (especially SA-mediated sucrose accumulation) is part of the acclamatory process during the early period. Conversely, ABA-responsive hexose accumulation and sucrose phloem loading represent severe drought symptoms during the late drought period.