Phloem Transport Research Articles

Phloem Delivery of Fludioxonil by Plant Amino Acid Transporter-Mediated Polysuccinimide Nanocarriers for Controlling Fusarium Wilt in Banana.

Fusarium wilt disease poses a serious threat to the global production of bananas. The targeted delivery of fungicides to banana phloem tissues may offer new hope for controlling this hard-to-treat vascular disease. In this study, fludioxonil (FLU)-loaded glycine methyl ester-conjugated polysuccinimide nanoparticles (PGA) were prepared with a loading efficiency (LE) of 27.9%. The obtained nanoparticles (FLU@PGA) exhibited pH-sensitive controlled release, specifically under an alkaline pH in plant phloem. In vivo experiments in potted bananas demonstrated that FLU@PGA can achieve the downward delivery of FLU to banana rhizomes and roots after foliar application, reducing disease severity by 50.4%. The phloem transport studies showed that the phloem loading of FLU@PGA was involved in an active transport mechanism at the organ level (castor bean seedlings). The observation of fluorescein-5-isothiocyanate cadaverine-labeled PGA nanocarriers showed that they could be absorbed by mesophyll cells and loaded into vascular tissues through the symplastic pathway. Furthermore, the interaction of FLU@PGA with the plant amino acid transporter AtLHT1 was observed to enhance transmembrane uptake at the cellular level (Xenopus oocytes). These results suggested that the phloem-targeted delivery of fungicide by transporter-mediated nanocarriers could be a promising new strategy for the management of Fusarium wilt in bananas.

Recent Advances in Carbon and Nitrogen Metabolism in C3 Plants.

C and N are the most important essential elements constituting organic compounds in plants. The shoots and roots depend on each other by exchanging C and N through the xylem and phloem transport systems. Complex mechanisms regulate C and N metabolism to optimize plant growth, agricultural crop production, and maintenance of the agroecosystem. In this paper, we cover the recent advances in understanding C and N metabolism, regulation, and transport in plants, as well as their underlying molecular mechanisms. Special emphasis is given to the mechanisms of starch metabolism in plastids and the changes in responses to environmental stress that were previously overlooked, since these changes provide an essential store of C that fuels plant metabolism and growth. We present general insights into the system biology approaches that have expanded our understanding of core biological questions related to C and N metabolism. Finally, this review synthesizes recent advances in our understanding of the trade-off concept that links C and N status to the plant’s response to microorganisms.

Stage-specific events in tomato graft formation and the regulatory effects of auxin and cytokinin

Grafting is widely used worldwide because of its obvious advantages, especially in solanaceous vegetable crops. However, the molecular mechanisms underlying graft formation are unknown. In this study, internode tissues from above and below the graft junction were harvested, and we performed weighted gene co-expression network analysis (WGCNA) to describe the temporal and spatial transcriptional dynamics that occur during graft formation in tomato. The wounding stress response involved in JA, ETH, and oxylipins mainly occurred at 1 h after grafting (HAG). From 3 to 12 HAG, the biological processes of snRNA and snoRNA modification and the gibberellin-mediated signaling pathway functioned both above and below the graft junction. However, auxin transport and signaling, DNA replication, and xylem and phloem pattern formation were restricted to the scion, whereas the cytokinin-activated signaling pathway and the cellular response to sucrose starvation was restricted to the rootstock. At 24−72 HAG, cell division occurred above the graft junction, and photosynthesis-related pathways were activated below the graft junction. The levels of auxin and cytokinin reached their maxima above and below the graft junction at 12 HAG, respectively. Exogenous application of certain concentrations of IAA and 6-BA will promote xylem and phloem transport capacity. The current work has analyzed the stage-specific events and hub genes during the developmental progression of tomato grafting. We found that auxin and cytokinin levels respond to grafting, above and below the graft junction, respectively, to promote the formation of xylem and phloem patterning. In addition, the accumulation of auxin above the graft junction induced cells to prepare for mitosis and promoted the formation of callus. In short, our work provides an important reference for theoretical research and production application of tomato grafting in the future.

Metabolic profiling reveals local and systemic responses of kiwifruit to Pseudomonas syringae pv. actinidiae.

Pseudomonas syringae pv. actinidiae (Psa), a bacterial pathogen, causes bacterial canker disease in kiwifruit. To elucidate the local and systemic influences of Psa infection on kiwifruit, comprehensive analyses were conducted by combining metabolomic and physiological approach under Psa‐infected treatment and mock‐inoculated control in leaves, stems, and bleeding saps. Our results show that Psa infection stimulated kiwifruit metabolic reprogramming. Levels of many sugars, fumarate, and malic acid were decreased in Psa‐infected leaves and stems, accompanied by the increased level of amino acids (AAs), which implies the anaplerotic reaction to replenish the TCA cycle generating energy and intermediates for defense‐related metabolic pathways, such as phenylpropanoid metabolism. The inconsistent results were observed in bleeding saps, which may be attributed to the induced phloem transport of carbon (C) out of leaves and such a transport benefits bacterium movement. Arg, Gln, and pyroglutamic acid systematically were accumulated in long‐distance leaves, which probably confers to systemic acquired resistance (SAR) and Psa inoculation accelerated the nitrogen (N) cycling in kiwifruit. Moreover, Psa infection specifically affected the content of phenolic compounds and lignin. Phenolic compounds were negatively and lignin was positively related to kiwifruit Psa resistance, respectively. Our results first reveal that Psa enhances infection by manipulating C/N metabolism and sweet immunity, and that host lignin synthesis is a major physical barrier for restricting bacterial infection. This study provides an insight into the complex remodeling of plant metabolic response to Psa stress.

A Proposed Drought Response Equation Added to the Münch-Horwitz Theory of Phloem Transport.

Theoretical and experimental evidence for an effect of sieve tube turgor pressure on the mechanisms of phloem unloading near the root tips during moderate levels of drought stress is reviewed. An additional, simplified equation is proposed relating decreased turgor pressure to decreased rate kinetics of membrane bound transporters. The effect of such a mechanism would be to decrease phloem transport speed, but increase concentration and pressure, and thus prevent or delay negative pressure in the phloem. Experimental evidence shows this mechanism precedes and exceeds a reduction in stomatal conductance.

Metabolic Responses to Waterlogging Differ between Roots and Shoots and Reflect Phloem Transport Alteration in Medicago truncatula.

Root oxygen deficiency that is induced by flooding (waterlogging) is a common situation in many agricultural areas, causing considerable loss in yield and productivity. Physiological and metabolic acclimation to hypoxia has mostly been studied on roots or whole seedlings under full submergence. The metabolic difference between shoots and roots during waterlogging, and how roots and shoots communicate in such a situation is much less known. In particular, the metabolic acclimation in shoots and how this, in turn, impacts on roots metabolism is not well documented. Here, we monitored changes in the metabolome of roots and shoots of barrel clover (Medicago truncatula), growth, and gas-exchange, and analyzed phloem sap exudate composition. Roots exhibited a typical response to hypoxia, such as γ-aminobutyrate and alanine accumulation, as well as a strong decline in raffinose, sucrose, hexoses, and pentoses. Leaves exhibited a strong increase in starch, sugars, sugar derivatives, and phenolics (tyrosine, tryptophan, phenylalanine, benzoate, ferulate), suggesting an inhibition of sugar export and their alternative utilization by aromatic compounds production via pentose phosphates and phosphoenolpyruvate. Accordingly, there was an enrichment in sugars and a decline in organic acids in phloem sap exudates under waterlogging. Mass-balance calculations further suggest an increased imbalance between loading by shoots and unloading by roots under waterlogging. Taken as a whole, our results are consistent with the inhibition of sugar import by waterlogged roots, leading to an increase in phloem sugar pool, which, in turn, exert negative feedback on sugar metabolism and utilization in shoots.

The interplay of phloem-mobile signals in plant development and stress response.

Plants integrate a variety of biotic and abiotic factors for optimal growth in their given environment. While some of these responses are local, others occur distally. Hence, communication of signals perceived in one organ to a second, distal part of the plant and the coordinated developmental response require an intricate signaling system. To do so, plants developed a bipartite vascular system that mediates the uptake of water, minerals, and nutrients from the soil; transports high-energy compounds and building blocks; and traffics essential developmental and stress signals. One component of the plant vasculature is the phloem. The development of highly sensitive mass spectrometry and molecular methods in the last decades has enabled us to explore the full complexity of the phloem content. As a result, our view of the phloem has evolved from a simple transport path of photoassimilates to a major highway for pathogens, hormones and developmental signals. Understanding phloem transport is essential to comprehend the coordination of environmental inputs with plant development and, thus, ensure food security. This review discusses recent developments in its role in long-distance signaling and highlights the role of some of the signaling molecules. What emerges is an image of signaling paths that do not just involve single molecules but rather, quite frequently an interplay of several distinct molecular classes, many of which appear to be transported and acting in concert.

Sparse dynamic tomography: a shearlet-based approach for iodine perfusion in plant stems

In this paper we propose a motion-aware variational approach to reconstruct moving objects from sparse dynamic data. The motivation of this work stems from x-ray imaging of plants perfused with a liquid contrast agent, aimed at increasing the contrast of the images and studying the phloem transport in plants over time. The key idea of our approach is to deploy 3D shearlets as a space-temporal prior, treating time as the third dimension. The rationale behind this model is that a continuous evolution of a cartoon-like object is well suited for the use of 3D shearlets. We provide a basic mathematical analysis of the variational model for the image reconstruction. The numerical minimization is carried out with primal-dual scheme coupled with an automated choice of the regularization parameter. We test our model on different measurement setups: a simulated phantom especially designed to resemble a plant stem, with spreading points to simulate a spreading contrast agent; a measured agarose gel phantom to demonstrate iodide diffusion and geometry prior to imaging living sample; a measured living tree grown in vitro and perfused with a liquid sugar–iodine-mix. The results, compared against a 2D static model, show that our approach provides reconstructions that capture well the time dynamic of the contrast agent onset and are encouraging to develop microCT as a tool to study phloem transport using iodine tracer.

The Chromosome-Scale Genome of Melon Dissects Genetic Architecture of Important Agronomic Traits.

SummaryComparative and evolutionary genomics analyses are the powerful tools to provide mechanistic insights into important agronomic traits. Here, we completed a chromosome-scale assembly of the “neglected” but vital melon subspecies Cucumis melo ssp. agrestis using single-molecule real-time sequencing, Hi-C, and an ultra-dense genetic map. Comparative genomics analyses identified two targeted genes, UDP-sugar pyrophosphorylase and α-galactosidase, that were selected during evolution for specific phloem transport of oligosaccharides in Cucurbitaceae. Association analysis of transcriptome and the DNA methylation patterns revealed the epigenetic regulation of sucrose accumulation in developing fruits. We constructed the melon recombinant inbred lines to uncover Alkaline/Neutral Invertase (CINV), Sucrose-Phosphatase 2 (SPP2), α-galactosidase, and β-galactosidase loci related to sucrose accumulation and an LRR receptor-like serine/threonine-protein kinase associated with gummy stem blight resistance. This study provides essential genomic resources enabling functional genomics studies and the genomics-informed breeding pipelines for improving the fruit quality and disease resistance traits.

Aspartate Residues in a Forisome-Forming SEO Protein Are Critical for Protein Body Assembly and Ca2+ Responsiveness.

Forisomes are protein bodies known exclusively from sieve elements of legumes. Forisomes contribute to the regulation of phloem transport due to their unique Ca2+-controlled, reversible swelling. The assembly of forisomes from sieve element occlusion (SEO) protein monomers in developing sieve elements and the mechanism(s) of Ca2+-dependent forisome contractility are poorly understood because the amino acid sequences of SEO proteins lack conventional protein-protein interaction and Ca2+-binding motifs. We selected amino acids potentially responsible for forisome-specific functions by analyzing SEO protein sequences in comparison to those of the widely distributed SEO-related (SEOR), or SEOR proteins. SEOR proteins resemble SEO proteins closely but lack any Ca2+ responsiveness. We exchanged identified candidate residues by directed mutagenesis of the Medicago truncatula SEO1 gene, expressed the mutated genes in yeast (Saccharomyces cerevisiae) and studied the structural and functional phenotypes of the forisome-like bodies that formed in the transgenic cells. We identified three aspartate residues critical for Ca2+ responsiveness and two more that were required for forisome-like bodies to assemble. The phenotypes observed further suggested that Ca2+-controlled and pH-inducible swelling effects in forisome-like bodies proceeded by different yet interacting mechanisms. Finally, we observed a previously unknown surface striation in native forisomes and in recombinant forisome-like bodies that could serve as an indicator of successful forisome assembly. To conclude, this study defines a promising path to the elucidation of the so-far elusive molecular mechanisms of forisome assembly and Ca2+-dependent contractility.

Precision Delivery of Multiscale Payloads to Tissue-Specific Targets in Plants.

The precise deployment of functional payloads to plant tissues is a new approach to help advance the fundamental understanding of plant biology and accelerate plant engineering. Here, the design of a silk‐based biomaterial is reported to fabricate a microneedle‐like device, dubbed “phytoinjector,” capable of delivering a variety of payloads ranging from small molecules to large proteins into specific loci of various plant tissues. It is shown that phytoinjector can be used to deliver payloads into plant vasculature to study material transport in xylem and phloem and to perform complex biochemical reactions in situ. In another application, it is demonstrated Agrobacterium‐mediated gene transfer to shoot apical meristem (SAM) and leaves at various stages of growth. Tuning of the material composition enables the fabrication of another device, dubbed “phytosampler,” which is used to precisely sample plant sap. The design of plant‐specific biomaterials to fabricate devices for drug delivery in planta opens new avenues to enhance plant resistance to biotic and abiotic stresses, provides new tools for diagnostics, and enables new opportunities in plant engineering.

Role of ureides in source-to-sink transport of photoassimilates in non-fixing soybean.

Nitrogen (N)-fixing soybean plants use the ureides allantoin and allantoic acid as major long-distance transport forms of N, but in non-fixing, non-nodulated plants amino acids mainly serve in source-to-sink N allocation. However, some ureides are still synthesized in roots of non-fixing soybean, and our study addresses the role of ureide transport processes in those plants. In previous work, legume ureide permeases (UPSs) were identified that are involved in cellular import of allantoin and allantoic acid. Here, UPS1 from common bean was expressed in the soybean phloem, which resulted in enhanced source-to-sink transport of ureides in the transgenic plants. This was accompanied by increased ureide synthesis and elevated allantoin and allantoic acid root-to-sink transport. Interestingly, amino acid assimilation, xylem transport, and phloem partitioning to sinks were also strongly up-regulated. In addition, photosynthesis and sucrose phloem transport were improved in the transgenic plants. These combined changes in source physiology and assimilate partitioning resulted in increased vegetative growth and improved seed numbers. Overall, the results support that ureide transport processes in non-fixing plants affect source N and carbon acquisition and assimilation as well as source-to-sink translocation of N and carbon assimilates with consequences for plant growth and seed development.

Studi Keragaman Struktur Morfologi dan Anatomi Petiole (Tangkai Daun) Dari Berbagai Kultivar Pisang Kepok (Musa Paradisiaca L.)

Kepok bananas are one type of banana that is found and used in Indonesia. These bananas contain high nutritional value so they are suitable for use as food ingredients. This study aims to determine the anatomical structure and morphology of petiole from various banana kepok cultivars. These cultivars are: kepok abu, batu, kapas, kuning, Libanon, and Manado. However, Kepok Libanon cannot be found in the previous collection area. The research was conducted in December 2018 to January 2019. The taking of plant samples using the method of exploration and observation of anatomical structures using fresh preparations across the incision and the results were analyzed descriptively. The observation of the structure of morphology of petiole is the largest petiole diameter, namely kepok kapas while the smallest is kepok batu. Kepok batu is the only one that has a canal leaf shape overlaps while kepok abu, kapas, kuning, and manado have the leaf shape of a curved canal inside. The fifth color of the kepok banana cultivar is the same which is green. Kepok abu have blackish brown spots while kepok batu, kapas, kuning, and manado have brown spots. The anatomical structure of the petiole includes the xylem diameter, transport tissue, support tissue, basic tissue, and protector tissue. Kepok kapas has the largest xylem diameter while kepok kuning has the smallest xylem diameter. Five kepok banana cultivars, has xylem and phloem transport tissue were clearly seen by the sclerenchyma sheath. Epidermal, parenchymal, and sclerenchymal tissue were clearly seen in the five cultivars.

Autumn flooding disrupts seasonal nitrogen storage and impacts spring growth in Quercus texana seedlings

Autumn flooding reduced nitrogen uptake and fine root mass. Leaf nitrogen resorption increased, but overall autumn nitrogen accumulation was negligible in flooded seedlings. Subsequently, spring survival and growth were decreased. Since nitrogen (N) is often limiting in terrestrial ecosystems, N is conserved in trees by resorption before leaf senescence. Bottomland forests are prone to flooding, which could reduce N uptake, and may decrease phloem transport, which is essential for N resorption. Therefore, we hypothesized that autumn flooding may diminish both N uptake and N resorption in trees, and this would reduce spring growth. Two-year-old Quercus texana seedlings either had no flood or had their complete root system flooded during dormancy induction, the period in autumn when trees prepare metabolically and physiologically for winter dormancy. We measured seedling growth and nutrient contents before flooding and after leaf fall and determined impacts of autumn flooding on growth during the subsequent spring. Autumn flooding resulted in a small increase in N resorption from leaves. In non-flooded seedlings, much more N accumulated in stems and roots than the amount resorbed from leaves, suggesting that there was substantial N uptake during the autumn dormancy induction period. However, flooding severely reduced accumulation of N in roots and stems during autumn, probably by directly reducing uptake and by increasing fine root mortality. Winter survival was reduced 50% by autumn flooding. Autumn-flooded seedlings that survived winter had greater new root growth in spring than non-flooded seedlings, but substantially decreased stem diameter growth. Our results indicate that Q. texana seedlings which are flooded during dormancy induction may be less competitive the following spring due to fine root mortality and reduced nutrient storage, which negatively impact spring growth.

Effect of introducing amino acids into phenazine-1-carboxylic acid on phloem mobility

To develop new phenazine carboxylic acid derivatives with better phloem mobility, five novel 7-amino acid substituted phenazine-1-carboxylic acids were synthesised by introducing amino acids into PCA at the 7-position. The phloem mobility experiments in Ricinus communis seedlings showed that retaining the carboxyl group of PCA and conjugating amino acids to its phenazine ring can also endow PCA with phloem mobility. Comparing our previous research, we found the amino acids substituted at 7-position on phenazine ring of PCA could clearly enhance the phloem mobility of PCA than that of amino acids conjugated with carboxyl group. Especially, the phloem transport concentration of the compound 7-L-isoleucine substituted PCA (7d) was 21 times higher than PCA-L-isoleucine conjugate (8d). These data suggest that the introduction of amino acids at different structural sites on the phenazine ring could effectively enhance the phloem mobility of PCA and it is worth a further study.

Woody tissue photosynthesis reduces stem CO2 efflux by half and remains unaffected by drought stress in young Populus tremula trees.

A substantial portion of locally respired CO2 in stems can be assimilated by chloroplast-containing tissues. Woody tissue photosynthesis (Pwt ) therefore plays a major role in the stem carbon balance. To study the impact of Pwt on stem carbon cycling along a gradient of water availability, stem CO2 efflux (EA ), xylem CO2 concentration ([CO2 ]), and xylem water potential (Ψxylem ) were measured in 4-year-old Populus tremula L. trees exposed to drought stress and different regimes of light exclusion of woody tissues. Under well-watered conditions, local Pwt decreased EA up to 30%. Axial CO2 diffusion (Dax ) induced by distant Pwt caused an additional decrease in EA of up to 25% and limited xylem [CO2 ] build-up. Under drought stress, absolute decreases in EA driven by Pwt remained stable, denoting that Pwt was not affected by drought. At the end of the dry period, when transpiration was low, local Pwt and Dax offset 20% and 10% of stem respiration on a daily basis, respectively. These results highlight (a) the importance of Pwt for an adequate interpretation of EA measurements and (b) homeostatic Pwt along a drought stress gradient, which might play a crucial role to fuel stem metabolism when leaf carbon uptake and phloem transport are limited.

Defenses against Virus and Vector: A Phloem-Biological Perspective on RTM- and SLI1-Mediated Resistance to Potyviruses and Aphids.

Combining plant resistance against virus and vector presents an attractive approach to reduce virus transmission and virus proliferation in crops. Restricted Tobacco-etch virus Movement (RTM) genes confer resistance to potyviruses by limiting their long-distance transport. Recently, a close homologue of one of the RTM genes, SLI1, has been discovered but this gene instead confers resistance to Myzus persicae aphids, a vector of potyviruses. The functional connection between resistance to potyviruses and aphids, raises the question whether plants have a basic defense system in the phloem against biotic intruders. This paper provides an overview on restricted potyvirus phloem transport and restricted aphid phloem feeding and their possible interplay, followed by a discussion on various ways in which viruses and aphids gain access to the phloem sap. From a phloem-biological perspective, hypotheses are proposed on the underlying mechanisms of RTM- and SLI1-mediated resistance, and their possible efficacy to defend against systemic viruses and phloem-feeding vectors.

Detection and in vitro studies of Cucurbita maxima phloem serpin-1 RNA-binding properties

Apart from being a conduit for photoassimilate transport in plants, the phloem serves as a pathway for transport of proteins and RNAs from sites of their synthesis to distant plant parts. As demonstrated for mRNAs and small RNAs such as miRNA and siRNA, their phloem transport is largely involved in responses to environmental cues including stresses and pathogen attacks. RNA molecules are believed to be transported in the phloem in the form of complexes with RNA-binding proteins; however, proteins forming such complexes are generally poorly studied. Here, we demonstrate that the Cucurbita maxima phloem serpin-1 (CmPS1), which has been previously described as a functional protease inhibitor capable of long-distance transport via the phloem, is able to bind RNA in vitro. Among different RNAs tested, CmPS1 exhibits a preference for imperfect RNA duplexes and the highest affinity to tRNA. A characteristic complex formed by CmPS1 with tRNA is not observed upon CmPS1 binding to tRNA-like structures of plant viruses. Mutational analysis demonstrates that the CmPS1 N-terminal region is not involved in RNA binding. Since antithrombin-III, the human protease inhibitor of serpin family most closely sequence-related to CmPS1, is found to be unable to bind RNA, one can suggest that, in its evolution, CmPS1 has gained the RNA binding capability as an additional function likely relevant to its specific activities in the plant phloem.

The morphological characteristics and classification of vascular parenchyma cells in bamboo, Phyllostachys edulis (Carr.) J. Houz

Abstract Vascular parenchyma cells (VPCs) provide a critical metabolic and energetic link for xylem transport of water and mineral nutrients and phloem transport of photoassimilates. Understanding the morphology of these cells is required to understand their function. This study describes the morphology and classification of VPCs of moso bamboo by light microscopy (LM) and scanning electron microscopy (SEM). The key results revealed that vascular parenchyma could be divided into three categories according to cell geometric morphology: cells with two transverse end walls, cells with one oblique end and one transverse end walls, and cells with two oblique end walls. Additionally, there were two types of thickening patterns of the secondary wall, uniform and reticulate thickening, and both diffuse pitting and opposite-alternate pitting were observed. The average length, width, lumen diameter, double wall thickness, and area of the VPCs were 139.0 μm, 17.3 μm, 10.4 μm, 6.9 μm, and 51.1 μm2, respectively. Most VPCs were slender and thin-walled, and growth of the VPCs was not correlated in either the length or the width directions.

Potassium in Root Growth and Development.

Potassium is an essential macronutrient that has been partly overshadowed in root science by nitrogen and phosphorus. The current boom in potassium-related studies coincides with an emerging awareness of its importance in plant growth, metabolic functions, stress tolerance, and efficient agriculture. In this review, we summarized recent progress in understanding the role of K+ in root growth, development of root system architecture, cellular functions, and specific plant responses to K+ shortage. K+ transport is crucial for its physiological role. A wide range of K+ transport proteins has developed during evolution and acquired specific functions in plants. There is evidence linking K+ transport with cell expansion, membrane trafficking, auxin homeostasis, cell signaling, and phloem transport. This places K+ among important general regulatory factors of root growth. K+ is a rather mobile element in soil, so the absence of systemic and localized root growth response has been accepted. However, recent research confirms both systemic and localized growth response in Arabidopsis thaliana and highlights K+ uptake as a crucial mechanism for plant stress response. K+-related regulatory mechanisms, K+ transporters, K+ acquisition efficiency, and phenotyping for selection of K+ efficient plants/cultivars are highlighted in this review.