The root system architecture of wheat establishing in soil is associated with varying elongation rates of seminal roots: quantification using 4D magnetic resonance imaging.

Seedling establishment is the first stage of crop productivity, and root phenotypes at seed emergence are critical to a successful start of shoot growth as well as for water and nutrient uptake. In this study, we investigate seedling establishment in winter wheat utilizing a newly developed workflow based on magnetic resonance imaging (MRI). Using the eight parents of the MAGIC (multi-parent advanced generation inter-cross) population we analysed the 4D root architecture of 288 individual seedlings grown in natural soils with plant neighbors over 3 d of development. Time of root and shoot emergence, total length, angle, and depth of the axile roots varied significantly among these genotypes. The temporal data resolved rates of elongation of primary roots and first and second seminal root pairs. Genotypes with slowly elongating primary roots had rapidly elongating first and second seminal root pairs and vice versa, resulting in variation in root system architecture mediated not only by root angle but also by initiation and relative elongation of axile roots. We demonstrated that our novel MRI workflow with a unique planting design and automated measurements allowed medium throughput phenotyping of wheat roots in 4D and could give new insights into regulation of root system architecture.

A Mobile NMR Sensor and Relaxometric Method to Non-destructively Monitor Water and Dry Matter Content in Plants.

Water content (WC) and dry matter content (DMC) are some of the most basic parameters to describe plant growth and yield, but are exceptionally difficult to measure non-invasively. Nuclear Magnetic Resonance (NMR) relaxometry may fill this methodological gap. It allows non-invasive detection of protons in liquids and solids, and on the basis of these measures, can be used to quantify liquid and dry matter contents of seeds and plants. Unfortunately, most existing NMR relaxometers are large, unwieldy and not suitable to measure intact plants or to be used under field conditions. In addition, currently the appropriate NMR relaxometric methods are poorly suited for non-expert use. We here present a novel approach to overcome these drawbacks. We demonstrate that a basic NMR relaxometer with the capability to accept intact plants, in combination with straightforward NMR and data processing methods, can be used as an NMR plant sensor to continuously, quantitatively and non-invasively monitor changes in WC and DMC. This can be done in vivo, in situ, and with high temporal resolution. The method is validated by showing that measured liquid and solid proton densities accurately reflect WC and DMC of reference samples. The NMR plant sensor is demonstrated in an experimental context by monitoring WC of rice leaves under osmotic stress, and by measuring the dynamics of water and dry matter accumulation during seed filling in a developing wheat ear. It is further demonstrated how the method can be used to estimate leaf water potential on the basis of changes in leaf water content.

An integrated magnetic resonance plant imager for mobile use in greenhouse and field.

In this contribution we demonstrate a mobile, integrated MR plant imager that can be handled by one single person and used in the field. Key to the construction of it was a small and lightweight gradient amplifier, specifically tailored to our combination of magnet, gradient coils and the requirements of the desired pulse sequences. To allow imaging of branches and stems, an open C-shaped permanent magnet was used. In the design of the magnet, pole gap width, low weight and robustness were prioritized over homogeneity and field strength. To overcome the adverse effects of short T2*, multi-spin echo imaging was employed, using short echo times and high spectral widths. To achieve microscopic resolution under these constraints requires fast switching field gradients, driven by strong and fast gradient amplifiers. While small-scale spectrometers and RF amplifiers are readily available, appropriate small-scale gradient amplifiers or designs thereof currently are not. We thus constructed a small, 3-channel gradient amplifier on the basis of a conventional current-controlled AB amplifier design, using cheap and well-known parts. The finished device weighs 5 kg and is capable of delivering 40 A gradient pulses of >6 ms in duration. With all components built onto an aluminum hand trolley, the imaging setup weighs 45 kg and is small enough to fit into a car. We demonstrate the mobility and utility of the device imaging quantitative water content and T2, first of an apple tree in an orchard; second, of a beech tree during spring leaf flushing in a greenhouse. The latter experiment ran for a continuous period of 62 days, acquiring more than 6000 images.

Spatio-Temporal Variation in Water Uptake in Seminal and Nodal Root Systems of Barley Plants Grown in Soil.

The spatial and temporal dynamics of root water uptake in nodal and seminal roots are poorly understood, especially in relation to root system development and aging. Here we non-destructively quantify 1) root water uptake and 2) root length of nodal and seminal roots of barley in three dimensions during 43 days of growth. We developed a concentric split root system to hydraulically and physically isolate the seminal and nodal root systems. Using magnetic resonance imaging (MRI), roots were visualized, root length was determined, and soil water depletion in both compartments was measured. From 19 days after germination and onwards, the nodal root system had greater water uptake compared to the seminal root system due to both greater root length and greater root conductivity. At 29 days after germination onwards, the average age of the seminal and nodal root systems was similar and no differences were observed in water uptake per root length between seminal and nodal root systems, indicating the importance of embryonic root systems for seedling establishment and nodal root systems in more mature plants. Since nodal roots perform the majority of water uptake at 29 days after germination and onwards, nodal root phenes merit consideration as a selection target to improve water capture in barley and possibly other crops.

Spatially Resolved Root Water Uptake Determination Using a Precise Soil Water Sensor.

To answer long-standing questions about how plants use and regulate water, an affordable, noninvasive way to determine local root water uptake (RWU) is required. Here, we present a sensor, the soil water profiler (SWaP), which can determine local soil water content (θ) with a precision of 6.10-5 cm3 ⋅ cm-3, an accuracy of 0.002 cm3 ⋅ cm-3, a temporal resolution of 24 min, and a one-dimensional spatial resolution of 1 cm. The sensor comprises two copper sheets, integrated into a sleeve and connected to a coil, which form a resonant circuit. A vector network analyzer, inductively coupled to the resonant circuit, measures the resonance frequency, against which θ was calibrated. The sensors were integrated into a positioning system, which measures θ along the depth of cylindrical tubes. When combined with modulating light (4-h period) and resultant modulating plant transpiration, the SWaP enables quantification of the component of RWU distribution that varies proportionally with total plant water uptake, and distinguishes it from soil water redistribution via soil pores and roots. Additionally, as a young, growing maize (Zea mays) plant progressively tapped its soil environment dry, we observed clear changes in plant-driven RWU and soil water redistribution profiles. Our SWaP setup can measure the RWU and redistribution of sandy-soil water content with unprecedented precision. The SWaP is therefore a promising device offering new insights into soil-plant hydrology, with applications for functional root phenotyping in nonsaline, temperature-controlled conditions, at low cost.

5-Carbonyl-1,3-oxazine-2,4-diones from N-Cyanosulfoximines and Meldrum's Acid Derivatives.

At elevated temperatures, N-cyanosulfoximines react with Meldrum's acid derivatives to give sulfoximines with N-bound 5-carbonyl-1,3-oxazine-2,4-dione groups. A representative product was characterized by single-crystal X-ray structure analysis. The product formation involves an unexpected molecular reorientation requiring several sequential bond-forming and -cleaving processes.

Precise Volumetric Measurements of Any Shaped Objects with a Novel Acoustic Volumeter.

We introduce a novel technique to measure volumes of any shaped objects based on acoustic components. The focus is on small objects with rough surfaces, such as plant seeds. The method allows measurement of object volumes more than 1000 times smaller than the volume of the sensor chamber with both high precision and high accuracy. The method is fast, noninvasive, and easy to produce and use. The measurement principle is supported by theory, describing the behavior of the measured data for objects of known volumes in a range of 1 to 800 µL. In addition to single-frequency, we present frequency-dependent measurements that provide supplementary information about pores on the surface of a measured object, such as the total volume of pores and, in the case of cylindrical pores, their average radius-to-length ratio. We demonstrate the usefulness of the method for seed phenotyping by measuring the volume of irregularly shaped seeds and showing the ability to "look" under the husk and inside pores, which allows us to assess the true density of seeds.

phenoSeeder - A Robot System for Automated Handling and Phenotyping of Individual Seeds.

The enormous diversity of seed traits is an intriguing feature and critical for the overwhelming success of higher plants. In particular, seed mass is generally regarded to be key for seedling development but is mostly approximated by using scanning methods delivering only two-dimensional data, often termed seed size. However, three-dimensional traits, such as the volume or mass of single seeds, are very rarely determined in routine measurements. Here, we introduce a device named phenoSeeder, which enables the handling and phenotyping of individual seeds of very different sizes. The system consists of a pick-and-place robot and a modular setup of sensors that can be versatilely extended. Basic biometric traits detected for individual seeds are two-dimensional data from projections, three-dimensional data from volumetric measures, and mass, from which seed density is also calculated. Each seed is tracked by an identifier and, after phenotyping, can be planted, sorted, or individually stored for further evaluation or processing (e.g. in routine seed-to-plant tracking pipelines). By investigating seeds of Arabidopsis (Arabidopsis thaliana), rapeseed (Brassica napus), and barley (Hordeum vulgare), we observed that, even for apparently round-shaped seeds of rapeseed, correlations between the projected area and the mass of seeds were much weaker than between volume and mass. This indicates that simple projections may not deliver good proxies for seed mass. Although throughput is limited, we expect that automated seed phenotyping on a single-seed basis can contribute valuable information for applications in a wide range of wild or crop species, including seed classification, seed sorting, and assessment of seed quality.

Quantitative 3D Analysis of Plant Roots Growing in Soil Using Magnetic Resonance Imaging.

Precise measurements of root system architecture traits are an important requirement for plant phenotyping. Most of the current methods for analyzing root growth require either artificial growing conditions (e.g. hydroponics), are severely restricted in the fraction of roots detectable (e.g. rhizotrons), or are destructive (e.g. soil coring). On the other hand, modalities such as magnetic resonance imaging (MRI) are noninvasive and allow high-quality three-dimensional imaging of roots in soil. Here, we present a plant root imaging and analysis pipeline using MRI together with an advanced image visualization and analysis software toolbox named NMRooting. Pots up to 117 mm in diameter and 800 mm in height can be measured with the 4.7 T MRI instrument used here. For 1.5 l pots (81 mm diameter, 300 mm high), a fully automated system was developed enabling measurement of up to 18 pots per day. The most important root traits that can be nondestructively monitored over time are root mass, length, diameter, tip number, and growth angles (in two-dimensional polar coordinates) and spatial distribution. Various validation measurements for these traits were performed, showing that roots down to a diameter range between 200 µm and 300 µm can be quantitatively measured. Root fresh weight correlates linearly with root mass determined by MRI. We demonstrate the capabilities of MRI and the dedicated imaging pipeline in experimental series performed on soil-grown maize (Zea mays) and barley (Hordeum vulgare) plants.