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Hidden Rice Diversity in the Guianas

Traditional crop varieties are an important source of genetic diversity for crop adaptation and modern breeding. Landraces of Asian (Oryza sativa) and African (Oryza glaberrima) rice have been well studied on the continents where they were domesticated. However, their history of cultivation in northern South America is poorly understood. Here we reveal the rice diversity that is maintained by Maroons, descendants of enslaved Africans who fled to the interior forests of the Guianas ca. 300 years ago. We interviewed subsistence farmers who practice shifting cultivation along the Maroni and Lawa rivers that form the natural border between French Guiana and Suriname, and used ethnobotanical and morphological methods to identify around 50 varieties, of which 15 were previously undocumented. The genetic origin of these varieties was explored using the Angiosperms-353 universal probe set. Despite the large distances between sites and relative inaccessibility of the area, phenotypic and genetic diversity did not display any geographic structure, which is consistent with knowledge of seed exchange among members of the same ethnolinguistic group. Although improved US cultivars were introduced in Maroon villages in the 1940s, these have not displaced the traditional landraces, which are cherished for their taste and nutritious qualities and for their importance in Maroon spiritual life. The unique agricultural and ritual practices of Maroons confirm their role as custodians of rice diversity, a role that is currently under threat from external pressures and encroaching globalization. We expect that the rice diversity uncovered in this study represents only a fraction of the total diversity in the Guianas and may constitute a large untapped resource that holds promise for future rice improvement. Further efforts to inventory and preserve these landraces will help to protect a precious cultural heritage and local food security.

plant biology

Flowering behaviour in Arabis alpina ensures the maintenance of a perennating dormant bud bank

Arabis alpina, similar to woody perennials, has a complex architecture with a zone of axillary vegetative branches and a zone of dormant buds that serve as perennating organs. We show that floral development during vernalization is the key for shaping the dormant bud zone by facilitating a synchronized and rapid growth after vernalization and thereby causing an increase in auxin response and transport and endogenous indole-3-acetic acid (IAA) levels in the stem. Floral development during vernalization is associated with the development of axillary buds in subapical nodes. Our transcriptome analysis indicated that these buds are not dormant during vernalization but only attain sustained growth after the return to warm temperatures. Floral and subapical vegetative branches grow after vernalization and inhibit the development of the buds below. Dormancy in these buds is regulated across the A. alpina life cycle by low temperatures and by apical dominance in a BRANCHED 1-dependent mechanism.

plant biology

The effect of drought and intercropping on chicory nutrient uptake from below 2 m studied in a multiple tracer setup

AimsWe tested if chicory acquires nutrients from soil layers down to 3.5 m depth and whether the deep nutrient uptake increases as a result of topsoil drought or topsoil resource competition. We also tested whether application of the trace elements Cs, Li, Rb, Sr, and Se, as tracers result in similar uptake rates. MethodsThe methodological tests were primarily carried out in a pilot experiment where the five tracers were applied to 1 m depth in lucerne and red beet grown in tube rhizotrons. The dynamics of deep nutrient uptake in chicory was studied in large 4 m deep rhizoboxes. A drought was imposed when roots had reached around 2 m depth. ResultsChicory acquired tracers applied to 3.5 m depth, but we found no compensatory tracer uptake with depth during drought. We found some indications of a compensatory tracer uptake from 2.3 and 2.9 m depth in intercropped chicory. Application of equimolar amounts of trace elements resulted in similar excess tracer concentrations within species. ConclusionChicory acquires nutrients from below 3 m but does not increase deep nutrient uptake as a response to limited topsoil nutrient availability.

plant biology

Optical properties of living corals determined with diffuse reflectance spectroscopy

The internal light field and thus light exposure of the photosymbiotic microalgae (Symbiodinium sp.) in corals is strongly modulated by the optical properties of coral tissue and skeleton. While there are numerous studies documenting the light microenvironment in corals, there are only few measurements of the inherent optical properties of corals in the literature, and this has hampered a more quantitative understanding of coral optics. Here we present a study of the optical properties of 26 live coral samples, representative of 11 coral species and spanning a variety of morphotypes. We employed well-established fiber-optic reflectance spectroscopy techniques from biomedical optics using two methods: (1) A source and a detection fiber separated by a variable distance measured the lateral spread of light in corals, dominated by the skeleton; (2) A fiber-optic field radiance probe measured the diffuse reflectance from the coral surface, dominated by the living coral tissue. Analysis based on diffusion theory and Monte Carlo simulation yielded estimates of the bulk scattering and absorption coefficients of the coral tissue and skeleton, in the 750-1030 nm wavelength range. Extrapolating into the spectral region of photosynthetically active radiation (PAR, 400-700 nm) allowed estimation of the optical depth of absorption by the main Symbiodinium photopigment chlorophyll a. Coral tissue scattering was on average ~1.9x stronger than the scattering of the skeleton, consistent with the model that corals trap photons by high scattering to enhance absorption by algal pigments, while the lower scattering of the skeleton allows spread of light to otherwise shaded coral tissue areas.

plant biology

Mechanical stress initiates and sustains the morphogenesis of wavy leaf epidermal cells

Plant cell morphogenesis is governed by the mechanical properties of the cell wall and the resulting cell shape is intimately related to the respective specific function. Pavement cells covering the surface of plant leaves form wavy interlocking patterns in many plants. We use computational mechanics to simulate the morphogenetic process based on experimentally assessed cell shapes, growth dynamics, and cell wall chemistry. The simulations and experimental evidence suggest a multistep process underlying the morphogenesis of pavement cells during tissue differentiation. The mechanical shaping process relies on spatially confined, feedback-augmented stiffening of the cell wall in the periclinal walls, an effect that correlates with experimentally observed deposition patterns of cellulose and de-esterified pectin. We provide evidence for mechanical buckling of the pavement cell walls that can robustly initiate patterns de novo and may precede chemical and geometrical anisotropy.\n\nHighlightsO_LIA multistep mechano-chemical morphogenetic process underlies the wavy pattern of epidermal pavement cells.\nC_LIO_LIMicrotubule polarization is preceded by an event that breaks mechanical isotropy in the cell wall.\nC_LIO_LIMechanical models simulate the formation of wavy cell shapes, predict buckling of the cell walls and spatially confined variations in the mechanical properties of leaf epidermal cells.\nC_LIO_LIStress/strain stiffening following the buckling of the cell walls constitutes a crucial element in a positive feedback loop forming interlocking pavement cells.\nC_LIO_LIPolarization of cortical microtubules, cellulose microfibrils, and de-esterified pectin occur at the necks of wavy pavement cells, matching the in silico prediction of cell wall stiffening.\nC_LI

plant biology

Transcriptional variation in glucosinolate biosynthetic genes and inducible responses to aphid herbivory on field-grown Arabidopsis thaliana

Recently, increasing attempts have been made to understand how plant genes function in natura studies. To determine whether plant defense genes are activated under multiple biotic stimuli, we combined a high-throughput RNA-Seq with insect survey data on 19 accessions of Arabidopsis thaliana growing on the field site of Switzerland. We found that genes with GO annotations "glucosinolate biosynthetic process" and "response to insects" were the most significantly enriched, exhibiting largely variable expression among plant accessions. Nearly half of the total expression variation in glucosinolate biosynthetic genes, AOPs, ESM1, ESP, and TGG1, was explained by among-accession variance. Combined with the field RNA-Seq data, bioassays confirmed that AOP3 was up-regulated in response to the mustard aphid Lipaphis erysimi. In addition, we also found that the expression of a major cis-jasmone activated gene CYP81D11 was positively correlated with the number of the flea beetles Phyllotreta spp. The combined results from RNA-Seq and insect surveys suggested that plants can activate their defenses even when they are exposed to multiple biotic stimuli in natura.

plant biology

Nanotubes effectively deliver siRNA to intact plant cells and protect siRNA against nuclease degradation

Post-transcriptional gene silencing (PTGS) is a powerful tool to understand and control plant metabolic pathways, which is central to plant biotechnology. PTGS is commonly accomplished through delivery of small interfering RNA (siRNA) into cells. While siRNA delivery has been optimized for mammalian systems, it remains a significant challenge for plants due to the plant cell wall. Standard plant siRNA delivery methods (Agrobacterium and viruses) involve coding siRNA into DNA vectors, and are only tractable for certain plant species. Herein, we develop a nanotube-based platform for direct delivery of siRNA, and show high silencing efficiency in intact plant cells. We demonstrate that nanotubes successfully deliver siRNA and silence endogenous genes owing to effective intracellular delivery and nanotube-induced protection of siRNA from nuclease degradation. This study establishes that nanotubes, which are below the size exclusion limit of the plant cell wall, could enable a myriad of plant biotechnology applications that rely on RNA delivery.

plant biology

A framework for predicting soft-fruit yields and phenology using embedded, networked microsensors, coupled weather models and machine-learning techniques

Predicting harvest timing is a key challenge to sustainably develop soft fruit farming and reduce food waste. Soft fruits are perishable, high-value and seasonal, and sales prices are typically time-sensitive. In addition, fruit harvesting is labour-intensive and increasingly expensive making accurate phenological predictions valuable for growers. A novel approach for predicting soft fruit phenology and yields was developed and tested, using strawberries as the model crop. Seedlings were planted in polytunnels, and environmental and yield data were collected throughout the growing season. Over 1.2 million datapoints were collected by networked microsensors which measured spatial and temporal variability in air temperature, relative humidity (RH), soil moisture and photosynthetically active radiation (PAR). Fleeces were added to a subset of the plants to generate additional within-polytunnel variation. Cumulative fruit yields followed logistic growth curves and the coefficients of these curves were dependent on micro-climatic growing conditions. After 10,000 iterations, machine learning revealed that RH was the optimal factor informing the coefficients of these curves, perhaps because it is an integrative metric of air temperature and water status. Trigonometric models transformed weather forecasts, which showed a relatively low agreement with polytunnel air temperature (R2 = 0.6) and RH (R2 = 0.5) measurements, into more accurate polytunnel-specific predictions for temperature and RH (both R2 = 0.8). We present a framework for using machine-learning techniques to calculate curve coefficients and parametrise coupled weather models which can predict fruit yields and timing to a greater degree of accuracy that previously possible. Dataloggers measuring environmental and yield data could infer model parameters using iterative training for novel fruit varieties or crop types growing in different locations without a-priori phenological information. At this stage in the development of artificial intelligence and networked microsensors, this is a step forward in generating bespoke phenological prediction models to inform and support growers.

plant biology

On Pectin Methyl-esterification: Implications for In vitro and In vivo Viscoelasticity

Pectin is a major component of the primary plant cell wall and is important for cell expansion. However, the relationship between its chemistry and mechanical properties is not fully understood, especially in vivo. In this study, a protocol for viscoelastic micro-indentation using atomic force microscopy (AFM) was developed and applied to pectin in vitro and in vivo. After determining that linear viscoelasticity was a suitable theoretical framework for in vitro pectin analyses were conducted with both a standard linear solid and fractional Zener model. These indicated a strong coupling between elastic and viscous properties over a range of degrees of methyl-esterification (DM). Both elasticity and viscosity were found to vary non-linearly with DM which had interesting consequences for pectin gels of mixed DM. In Arabidopsis cell walls, the standard linear solid model was found to be appropriate. In this in vivo composite material a weaker elastic-viscous coupling was exhibited, correlated with DM. The viscoelastic testing in vivo of rapidly elongating cell walls, rich in high DM pectin, displayed a longer viscous time-scale. The implications of the testing method and results are discussed in the context of mechanobiology, mechano-chemistry, and cell growth.

plant biology

Small RNA expression pattern in multiply inbred lines and their hybrids of maize embryo

Heterosis, also known as hybrid vigor or transgression, is the phenomenon wherein an F1 hybrid produced from crossing two cultivars of the same or different species shows superior phenotypes. Heterosis in maize has been found and applied in breeding for more than one hundred years. However, its underlying molecular mechanisms are still poorly understood. To investigate whether small RNAs (sRNAs) participate in the regulation of heterosis, we profiled the sRNA expression patterns in the germ seeds of five inbred lines and theirs three F1 hybrids using high-throughput sequencing technology. The sequencing result show maize sRNAs are enriched in 22-nt length. Nearly 90% of 22-nt small RNA dominated clusters (SRCs) are in repeat regions, which mainly originated from Gypsy and Copia in retrotransposon. About 25% differentially expressed SRCs exist between parents, and hybrid gain almost all differentially expressed 22-nt SRCs. Twenty-four-nt sRNA also enriched in maize, which showed great sequence diversity and overall low expression across the genome. More than half of 24-nt SRCs originate from repeat, and 80% of them come from DNA transposon. Nearly 30% of 24-nt SRCs located in genes or their flanking regions, especially in flanking regions of \"lipid metabolic process\" and \"macromolecule modification\" related genes. Several thousands 24-nt SRCs are paternal or maternal specific expressed, and hybrids gain only half of differentially expressed 24-nt SRCs. Hundreds of 24-nt SRCs show high parent or above high parent expression pattern in different hybrids, and them mainly from Tourist, haT, and CACTA in DNA transposon and enrich nearby \"tRNA aminoacylation for protein translation\" related genes. Also, some 21-nt SRCs show conserved expression pattern in low parent. They were enriched in anti-sense region of some genes, which functions are about oxidative phosphorylation and translation. miRNAs have a global downregulated tendency in hybrids. zma-miR408-5p, zma-miR1432-5p and zma-miR528-5p are significant downregulated in each hybrid, and this phenomenon may cause their target genes more stable and contribute to hybrid vigor. Taken together, our results illustrated that sRNAs may contribute to heterosis at the very early stage of seed germination through repressing of retrotransposon activity, regulation gene activity at gene and genic flanking regions, and promotion some gene expressions by downregulated miRNAs.

plant biology

Synergetic effect of non-complementary 5’ AT-rich sequences on the development of a multiplex TaqMan real-time PCR for specific and robust detection of Clavibacter michiganensis and C. michiganensis subsp. nebraskensis

Clavibacter is an agriculturally important genus comprising a single species, Clavibacter michiganensis, and multiple subspecies, including, C. michiganensis subsp. nebraskensis which causes Gosss wilt/blight of corn and accounts for high yield losses - listed among the five most significant diseases of corn in the United States of America. Our research objective was to develop a robust and rapid multiplex TaqMan real-time PCR (qPCR) to detect C. michiganensis in general and C. michiganensis subsp. nebraskensis with enhanced reliability and accuracy by adding non-complementary AT sequences to the 5 end of the forward and reverse primers. Comparative genomic analyses were performed to identify unique and conserved gene regions for primer and probe design. The unique genomic regions, ABC transporter ATP-binding protein CDS/ABC-transporter permease and MFS transporter were determined for specific detection of C. michiganensis and C. m. subsp. nebraskensis, respectively. The AT-rich sequences at the 5 position of the primers enhanced the reaction efficiency and sensitivity of rapid qPCR cycling; the reliability, accuracy and high efficiency of the developed assay was confirmed after testing with 59 strains from inclusivity and exclusivity panels - no false positives or false negatives were detected. The assays were also validated through naturally and artificially infected corn plant samples; all samples were detected for C. michiganensis and C. m. subsp. nebraskensis with 100% accuracy. The assay with 5 AT-rich sequences detected up to 10- and 100-fg of C. michiganensis and C. michiganensis subsp. nebraskensis genome targets, respectively. No adverse effect was observed when sensitivity assays were spiked with host genomic DNA. Addition of 5 AT rich sequences enhanced the qPCR reaction efficiency from 0.82 (M = -3.83) and 0.91 (M = -3.54) to 1.04 (with optimum slope value; M = -3.23) for both C. michiganensis and C. michiganensis subsp. nebraskensis, respectively; a increase of 10-fold sensitivity was also obtained with C. michiganensis primer set. The methodology proposed here can be used to optimize the reaction efficiency and to harmonize the diagnostic protocols which have prodigious applications in routine diagnostics, biosecurity and microbial forensics.

plant biology

Redundant SCARECROW genes pattern distinct cell layers in roots and leaves of maize

The highly efficient C4 photosynthetic pathway is facilitated by Kranz leaf anatomy. In Kranz leaves, closely spaced veins are encircled by concentric layers of photosynthetic bundle sheath (inner) and mesophyll (outer) cells. Here we demonstrate that in the C4 monocot maize, Kranz patterning is regulated by redundant function of SCARECROW 1 (ZmSCR1) and a previously uncharacterized homeolog ZmSCR1h. ZmSCR1 and ZmSCR1h transcripts accumulate in ground meristem cells of developing leaf primordia and in Zmscr1;Zmscr1h mutant leaves, most veins are separated by one rather than two mesophyll cells; many veins have sclerenchyma above and/or below instead of mesophyll cells; and supernumerary bundle sheath cells develop. The mutant defects are unified by compromised mesophyll cell development. In addition to Kranz defects, Zmscr1;Zmscr1h mutants fail to form an organized endodermal layer in the root. Collectively, these data indicate that ZmSCR1 and ZmSCR1h redundantly regulate cell-type patterning in both leaves and roots of maize. Leaf and root pathways are distinguished, however, by the cell layer in which they operate - mesophyll at a two-cell distance from leaf veins versus endodermis immediately adjacent to root vasculature.\n\nSummary statementTwo duplicated maize SCARECROW genes control the development of the endodermis in roots and the mesophyll in leaves

plant biology

Cryopreservation of Rosa hybrida cv. Helmut Schmidt by PVS2 vitrification method using in vitro fragmented explants (IFEs)

This is the first report on cryopreservation via PVS2 vitrification method on roses using in vitro fragmented explants (IFEs) as the starting material. The aim of this study is to optimize the efficient plant recovery and regeneration system for cryopreservation of Rosa hybrida cv. Helmut Schmidt using IFEs. Some important parameters have been optimized in this study are the effect of ascorbic acid (0.3 mM) examined separately and in combination at all steps in cryopreservation procedure (preculture, loading, unloading and growth recovery), loading type, loading duration, and PVS2 duration. The highest growth recovery of 43.33% was obtained when 3-4 mm size IFEs precultured on 0.25 M sucrose media supplemented with full-strength MS for one (1) day, followed by loading treatment supplemented with 1.5 M glycerol + 0.4 M sucrose + 5% DMSO + 0.3 mM ascorbic acid for 20 minutes, dehydration with PVS2 solution for 30 minutes and then treated with unloading solution supplemented with 1.2 M sucrose + 0.3 mM ascorbic acid for 20 minutes. This finding implies that long-term storage of Rosa Hybrida cv. Helmut Schmidt by PVS2 vitrification method was successful with essential biomolecules.

plant biology

Comprehensive analysis of the internal structure and firmness in American cranberry (Vaccinium macrocarpon Ait.) fruit

Cranberry (Vaccinium macrocarpon Ait.) fruit quality traits encompass many properties. Among these, fruit firmness has become a quality standard due to the recent demand increase for sweetened and dried cranberries (SDC). Traditionally, this trait has been measured by the cranberry industry using compression tests; however, it is poorly understood how fruit firmness is influenced by other characteristics. In this study, we developed a high-throughput computer-vision method to measure the internal structure of cranberry fruit, which may in turn influence cranberry fruit firmness. We measured the internal structure of 16 cranberry cultivars measured over a 40-day period. Internal structure data paired with fruit firmness values at each evaluation period allowed us to explore the correlations between firmness and internal morphological characteristics.

plant biology

Plant-soil feedback contributes to predicting plant invasiveness of 68 alien plant species differing in invasive status

1. Understanding what species characteristics allow some alien plants to become invasive while others fail to is critical to our understanding of community assembly processes. While many characteristics have been shown to predict plant invasiveness, the importance of plant-soil feedbacks (PSFs) in invasions has been difficult to assess since individual studies include only a few species and use disparate methodology.\n\n2. We studied PSFs of 68 invasive and non-invasive alien species in a single two-phase garden experiment, and compared the relative importance of PSF, residence time, phylogenetic novelty and plant traits for plant invasiveness. Additionally, we tested for relationships between PSF, residence time and phylogenetic novelty.\n\n3. PSF for seedling establishment belonged to five best predictors of plant invasiveness, along with specific leaf area, height, seedling growth rate, and residence time. Invasive species had more positive PSF for seedling establishment, but not for biomass, than non-invasive species. Phylogenetically novel species experienced less negative PSF than species with native congeners, suggesting they benefit more from enemy release. PSF of non-invasive species, contrary to that of invasive species, was becoming more negative with increasing residence time.\n\n4. Synthesis. We demonstrated that PSF plays a role in predicting invasiveness that is comparable with other species characteristics that are more commonly studied. PSF should thus receive more attention in studies predicting community structure and in programs assessing the likely invasions of aliens.

plant biology

Arabidopsis phytochelatin synthase 1, but not phytochelatin synthesis, functions in extracellular defense against multiple fungal pathogens

Phytochelatin synthase (PCS) is a key component of heavy metal detoxification in plants. PCS catalyzes both the synthesis of the peptide phytochelatin from glutathione as well as the degradation of glutathione conjugates via peptidase activity. Here, we describe a hitherto uncharacterized role for PCS in disease resistance against plant pathogenic fungi. The pen4 mutant, which is allelic to cadmium insensitive 1 (cad1/pcs1) mutants, was recovered from a screen for Arabidopsis mutants with reduced resistance to the non-adapted barley fungal pathogen, Blumeria graminis f. sp. hordei. PCS1, which is found in the cytoplasm of cells of healthy plants, translocates upon pathogen attack and colocalizes with the PEN2 myrosinase on the surface of immobilized mitochondria. pcs1 and pen2 mutant plants exhibit a similar metabolic defect in the accumulation of pathogen-inducible indole glucosinolate-derived compounds, suggesting that PEN2 and PCS1 act in the same metabolic pathway. The function of PCS1 in this pathway is independent of phytochelatin synthesis and deglycination of glutathione conjugates, as catalytic-site mutants of PCS1 are still functional in indole glucosinolate metabolism. In uncovering a previously unknown function for PCS1, we reveal this enzyme to be a moonlighting protein important for plant responses to both biotic and abiotic stresses.

plant biology

Plant photoreceptors and their signaling components compete for binding to the ubiquitin ligase COP1 using their VP-peptide motifs

Plants sense different parts of the suns light spectrum using specialized photoreceptors, many of which signal through the E3 ubiquitin ligase COP1. Photoreceptor binding modulates COP1s ubiquitin ligase activity towards transcription factors. Here we analyze why many COP1-interacting transcription factors and photoreceptors harbor sequence-divergent Val-Pro (VP) peptide motifs. We demonstrate that VP motifs enable different light signaling components to bind to the WD40 domain of COP1 with various binding affinities. Crystal structures of the VP motifs of the UV-B photoreceptor UVR8 and the transcription factor HY5 in complex with COP1, quantitative binding assays and reverse genetic experiments together suggest that UVR8 and HY5 compete for the COP1 WD40 domain. Photoactivation of UVR8 leads to high-affinity cooperative binding of its VP domain and its photosensing core to COP1, interfering with the binding of COP1 to its substrate HY5. Functional UVR8 - VP motif chimeras suggest that UV-B signaling specificity resides in the UVR8 photoreceptor core, not its VP motif. Crystal structures of different COP1 - VP peptide complexes highlight sequence fingerprints required for COP1 targeting. The functionally distinct blue light receptors CRY1 and CRY2 also compete with downstream transcription factors for COP1 binding using similar VP-peptide motifs. Together, our work reveals that photoreceptors and their components compete for COP1 using a conserved displacement mechanism to control different light signaling cascades in plants.

plant biology

RhizoVision Crown: An Integrated Hardware and Software Platform for Root Crown Phenotyping

Root crown phenotyping measures the top portion of crop root systems and can be used for marker-assisted breeding, genetic mapping, and understanding how roots influence soil resource acquisition. Several imaging protocols and image analysis programs exist, but they are not optimized for high-throughput, repeatable, and robust root crown phenotyping. The RhizoVision Crown platform integrates an imaging unit, image capture software, and image analysis software that are optimized for reliable extraction of measurements from large numbers of root crowns. The hardware platform utilizes a back light and a monochrome machine vision camera to capture root crown silhouettes. RhizoVision Imager and RhizoVision Analyzer are free, open-source software that streamline image capture and image analysis with intuitive graphical user interfaces. RhizoVision Analyzer was physically validated using copper wire and features were extensively validated using 10,464 ground-truth simulated images of dicot and monocot root systems. This platform was then used to phenotype soybean and wheat root crowns. A total of 2,799 soybean (Glycine max) root crowns of 187 lines and 1,753 wheat (Triticum aestivum) root crowns of 186 lines were phenotyped. Principal component analysis indicated similar correlations among features in both species. The maximum heritability was 0.74 in soybean and 0.22 in wheat, indicating differences in species and populations need to be considered. The integrated RhizoVision Crown platform facilitates high-throughput phenotyping of crop root crowns, and sets a standard by which open plant phenotyping platforms can be benchmarked.

plant biology