3D reconstruction and dynamic modeling of root architecture in situ and its application to crop phosphorus research

Root architecture plays important roles in plant water and nutrient acquisition. However, accurate modeling of the root system that provides a realistic representation of roots in the soil is limited by a lack of appropriate tools for the non‐destructive and precise measurement of the root system architecture in situ. Here we describe a root growth system in which the roots grow in a solid gel matrix that was used to reconstruct 3D root architecture in situ and dynamically simulate its changes under various nutrient conditions with a high degree of precision. A 3D laser scanner combined with a transparent gel‐based growth system was used to capture 3D images of roots. The root system skeleton was extracted using a skeleton extraction method based on the Hough transformation, and mesh modeling using Ball‐B spline was employed. We successfully used this system to reconstruct rice and soybean root architectures and determine their changes under various phosphorus (P) supply conditions. Our results showed that the 3D root architecture parameters that were dynamically calculated based on the skeletonization and simulation of root systems were significantly correlated with the biomass and P content of rice and soybean based on both the simulation system and previous reports. Therefore, this approach provides a novel technique for the study of crop root growth and its adaptive changes to various environmental conditions.     

Mykoplasmaähnliche Organismen in Früchten proliferationskranker Apfelbäume Kurze Mitteilung

The potential of an in vitro technique to study root‐knot nematode infection on banana roots was investigated. Regenerated banana plants were placed horizontally on Gamborg B5 (GB5)‐medium and incubated under a light‐dark regime of 16h‐8h. Temperature fluctuated between 24 and 33 °C. Banana roots were inoculated with Meloidogyne incognita race 1 coming from roots of a transgenic tomato (Lycopersicon esculentum cv. Moneymaker) grown on GB5‐medium at 28 °C in complete darkness. Root‐knots appeared on primary and secondary banana roots two to seven days after nematode inoculation. After 28 days, egg masses protruded through the cortex and two days later juveniles hatched and reinfected banana roots. This method holds promise for dynamic studies of banana root infection with root‐knot nematodes.     

Shoot phytochrome B modulates reactive oxygen species homeostasis in roots via abscisic acid signaling in Arabidopsis

Underground roots normally reside in darkness. However, they are often exposed to ambient light that penetrates through cracks in the soil layers which can occur due to wind, heavy rain or temperature extremes. In response to light exposure, roots produce reactive oxygen species (ROS) which promote root growth. It is known that ROS‐induced growth promotion facilitates rapid escape of the roots from non‐natural light. Meanwhile, long‐term exposure of the roots to light elicits a ROS burst, which causes oxidative damage to cellular components, necessitating that cellular levels of ROS should be tightly regulated in the roots. Here we demonstrate that the red/far‐red light photoreceptor phytochrome B (phyB) stimulates the biosynthesis of abscisic acid (ABA) in the shoots, and notably the shoot‐derived ABA signals induce a peroxidase‐mediated ROS detoxification reaction in the roots. Accordingly, while ROS accumulate in the roots of the phyb mutant that exhibits reduced primary root growth in the light, such an accumulation of ROS did not occur in the dark‐grown phyb roots that exhibited normal growth. These observations indicate that mobile shoot‐to‐root ABA signaling links shoot phyB‐mediated light perception with root ROS homeostasis to help roots adapt to unfavorable light exposure. We propose that ABA‐mediated shoot‐to‐root phyB signaling contributes to the synchronization of shoot and root growth for optimal propagation and performance in plants.     

Identification of genes regulated by dark adaptation and far‐red light illumination in roots of Arabidopsis thaliana*

Roots in the soil are illuminated by far‐red (FR) light passed through plant tissues in the daytime, and are in complete darkness at night. To evaluate whether gene expression of roots is affected by a dark‐FR light cycle, gene expression profiles were analysed for dark‐adapted versus light‐grown plants and for FR light‐illuminated versus dark‐adapted plants using the RIKEN Arabidopsis full‐length cDNA microarray (containing approximately 7000 independent, full‐length cDNA groups). Among candidate dark‐ and FR‐regulated genes, several were further analysed. Eleven dark‐inducible and five dark‐repressed genes were characterized. Almost all the dark‐inducible and –repressed genes were oppositely regulated by FR light illumination. The functions of dark‐ and FR‐responsive genes and the significance of FR light‐regulated gene expression in roots under ground are discussed.     

Blue‐light‐induced PIN3 polarization for root negative phototropic response in Arabidopsis

Root negative phototropism is an important response in plants. Although blue light is known to mediate this response, the cellular and molecular mechanisms underlying root negative phototropism remain unclear. Here, we report that the auxin efflux carrier PIN‐FORMED (PIN) 3 is involved in asymmetric auxin distribution and root negative phototropism. Unilateral blue‐light illumination polarized PIN3 to the outer lateral membrane of columella cells at the illuminated root side, and increased auxin activity at the illuminated side of roots, where auxin promotes growth and causes roots bending away from the light source. Furthermore, root negative phototropic response and blue‐light‐induced PIN3 polarization were modulated by a brefeldin A‐sensitive, GNOM‐dependent, trafficking pathway and by phot1‐regulated PINOID (PID)/PROTEIN PHOSPHATASE 2A (PP2A) activity. Our results indicate that blue‐light‐induced PIN3 polarization is needed for asymmetric auxin distribution during root negative phototropic response.     

The Role of Phytophthora cryptogea and Waterlogging in Root Rot Kiwifruit

A 48 h or 96 h period of waterlogging induced a partial rotting of the root system of 18 month old potted kiwifruit cuttings. However, root regeneration compensated for the loss of damaged roots and the plants remained healthy. Inoculation with Phytophthora cryptogea under low soil moisture conditions caused a small amount of root rotting (11 %) and regeneration of the roots occurred. Root rot was significantly increased when inoculations were combined with extended periods of waterlogging. Root rot incidence was highest (89 %) in inoculated plants exposed to periods of 96 h waterlogging. Regeneration did not occur with any of these plants and consequently, the shoot systems were severely damaged.     

Non-photosynthetic effects of red and far-red light on root-nodule formation by leguminous plants

Summary Nodulation of pea and broad bean plants grown in the light was found to be reduced when the roots were exposed to far-red light for 5–15 minutes daily during 5 consecutive days following inoculation with nodule bacteria. Similar results were obtained following a single exposure to far-red light during a period of 15 minutes at the 3rd or 4th day after inoculation. When the roots were exposed to far-red light either before inoculation or during the first two days afterwards there were either no effects or only slight effects on nodulation The inhibitory effect of far-red light on nodulation was partly reduced by subsequent exposure to red light, provided that the same part of the plant was exposed to both red and far-red light,viz either the root or the shoot. When different parts of the plant were exposed to red and far-red light respectively, there was no interaction between the two kinds of light on nodulation. Plants whose roots were exposed to far-red light did not subsequently show stem elongation.Nodules were found to develop on the roots of pea plants grown in the dark, provided that the plants were kept at or below 22°C. At 25°C nodulation was almost absent. Nodulation was decreased by addition of kinetin and IAA. In contrast to plants grown in the light pea plants grown in the dark, inoculated with either an effective or ineffective strain of Rhizobium, developed equal numbers of nodules. Exposure to red light slightly increased the percentage of nodulated plants but decreased the number of nodules per plant. Exposure to far-red light slightly decreased both the percentage of nodulated plants and the number of nodules per plant. The effect of far-red light was counteracted by red light andvice versa.     

In Vitro Haustorium Development in Roots and Root Cultures of the Hemiparasitic Plant Triphysaria Versicolor

Parasitic plants in the Orobanchaceae invade host plant roots through root organs called haustoria. Parasite roots initiate haustorium development when exposed to specific secondary metabolites that are released into the rhizosphere by host plant roots. While molecular approaches are increasingly being taken to understand the genetic mechanism underlying these events, a limitation has been the lack of a transformation system for parasitic plants. Since the haustorium development occurs in roots of Orobanchaceae, root cultures may be suitable material for transient or stable transformation experiments. To this end, root cultures were obtained from explants, and subsequently calluses, from the hemiparasitic plant Triphysaria versicolor. The cultured roots retained their competence to form haustoria when exposed to host roots, host root exudates, or purified haustorium-inducing factors. The root culture haustoria invaded host roots and initiated a vascular continuity between the parasite and host roots. The ontogeny of haustoria development on root cultures was indistinguishable from that on seedlings roots. Root cultures should provide useful material for molecular studies of haustorium development.     

Light-Dark Changes in Proline Content of Barley Leaves under Salt Stress

Proline accumulation in leaves of barley (Hordeum vulgare L. cv. Alfa) seedlings treated with 150 mM NaCl was promoted in the light and suppressed in the dark. The light/dark changes of proline content was enhanced with each 12 h light/12 h dark cycle and the proline content increased steadily. Root and shoot concentrations of Na⁺ and Cl^– in salt treated plants increased about 10 to 25 times as compared to the control. The content of these ions and the content of malondialdehyde were higher in the shoot of seedlings exposed to salt stress for 4 d in the light in comparison with the seedlings exposed to NaCl for 4 d in darkness. Light stimulated both ions and proline accumulation in the leaves and has no effect in the roots. Oxygen uptake was higher in the seedlings kept 4 d in the light which have higher endogenous free proline content. Chlorophyll fluorescence measurements showed that the photochemical activity of PS 2 slightly decreased as a result of salt stress and was not influenced by light regimes during plant growth.     

Arbuscular mycorrhizal fungal-induced alteration to root architecture in strawberry and induced resistance to the root pathogenPhytophthora fragariae

Three strawberry cultivars Elsanta, Cambridge Favourite and Rhapsody were inoculated with eitherGlomus fasciculatum orGlomus etunicatum and their growth compared with non-inoculated plants. The roots of all inoculated plants were 55 to 70% colonised after 98 days. Increases in both root and shoot dry weights were measured. Root architecture was also determined and increases in branching were evident in AMF colonised root systems. The remaining plants were then inoculated with the root pathogenPhytophthora fragariae and allowed to grow for a further 58 days before harvest. In two of the cultivars, Cambridge Favourite and Elsanta, AMF reduced root necrosis by approximately 60 and 30% respectively. Only in the least susceptible cultivar, Rhapsody, was no reduction measured in AMF colonised plants. There were differences in the control conferred by the two arbuscular mycorrhizal fungi and this suggests there may be practical benefits of inoculation. Relationships between the presence of roots of different orders, on inoculation with the pathogen, and subsequent necrosis provided a mechanism for identifying root-architecture driven alteration to susceptibility. Root system necrosis was positively correlated with the proportion of the root system made up of higher order roots (3° to 4°) in non-colonised plants and negatively correlated in AMF colonised plants. These data suggest that root-architecture changes are not important per se but factors expressed concurrently may be.     

Application of heat stress in situ demonstrates a protective role of irradiation on photosynthetic performance in alpine plants

The impact of sublethal heat on photosynthetic performance, photosynthetic pigments and free radical scavenging activity was examined in three high mountain species, Rhododendron ferrugineum, Senecio incanus and Ranunculus glacialis using controlled in situ applications of heat stress, both in darkness and under natural solar irradiation. Heat treatments applied in the dark reversibly reduced photosynthetic performance and the maximum quantum efficiency of photosystem II (F_v/F_m), which remained impeded for several days when plants were exposed to natural light conditions subsequently to the heat treatment. In contrast, plants exposed to heat stress under natural irradiation were able to tolerate and recover from heat stress more readily. The critical temperature threshold for chlorophyll fluorescence was higher under illumination (T_c^′) than in the dark (T_c). Heat stress caused a significant de‐epoxidation of the xanthophyll cycle pigments both in the light and in the dark conditions. Total free radical scavenging activity was highest when heat stress was applied in the dark. This study demonstrates that, in the European Alps, heat waves can temporarily have a negative impact on photosynthesis and, importantly, that results obtained from experiments performed in darkness and/or on detached plant material may not reliably predict the impact of heat stress under field conditions.     

Investigations into rhizosphere microflora

Summary Effect of light and dark treatments on rhizosphere and rhizoplane microflora of two crops,viz Triticum aestivum. Linn. andHordeum vulgare Linn. has been investigated. Higher microbial population was recorded from the rhizosphere of plants exposed to continuous light while continuous dark set harboured lesser microflora. The microflora in 12 hours alternate light and dark set exhibited an intermediate condition. Amino acids (both qualitatively and quantitatively) in root extracts were the least in continuous dark and the maximum in continuous light set. The variation in root leakage, physiological and morphological condition of the plants was thought to be the cause for change in microbial activity. A part of the Ph. D. thesis approved by University of Gorakhpur, Gorakhpur, India (1969).     

Regulation of root anaerobiosis and carbon translocation by light and root aeration in Isoetes alpinus

The activity of alcohol dehydrogenase (ADH) was measured in corms and roots of the submerged freshwater macrophyte Isoetes alpinus Kirk. growing in situ, and related to its capacity for internal oxygen transport and to carbohydrate translocation. ADH activity was present in roots but not corms at uniform activity (0.15–0.35 × 10^?6 mol g^?1 fresh weight s^?1) over the entire plant depth range (3–7 m depth), and was intermediate to that developed in excised roots after 1‐week exposure to either dissolved oxygen at air‐saturation or to anoxia. Responses of photosynthesis and root oxygen release to light intensity confirmed that shoot‐to‐root oxygen transport saturated at similar light intensities to photosynthetic oxygen evolution, but was positive in the dark and at irradiances below the compensation point for photosynthesis, due to contributions to transport by oxygen diffusion from the external medium. Transport of ¹⁴C‐labelled photo‐assimilates to roots nevertheless ceased when intact plants were exposed to a combination of leaf darkness and root external anoxia, even when high ¹⁴C concentrations were present in shoots, but remained high when the roots were provided with external oxygen. The lack of any control over permeability of the root surface to gases in this species suggested that ADH activity and reduced translocation is most likely caused by development of hypoxic tissues in the apical tissue. These results suggest that reductions in ambient light intensity may have indirect effects on I. alpinus viability by increasing the degree of root hypoxia and impairing carbon partitioning.     

The signal transducer NPH3 integrates the phototropin1 photosensor with PIN2-based polar auxin transport in Arabidopsis root phototropism

Under blue light (BL) illumination, Arabidopsis thaliana roots grow away from the light source, showing a negative phototropic response. However, the mechanism of root phototropism is still unclear. Using a noninvasive microelectrode system, we showed that the BL sensor phototropin1 (phot1), the signal transducer NONPHOTOTROPIC HYPOCOTYL3 (NPH3), and the auxin efflux transporter PIN2 were essential for BL-induced auxin flux in the root apex transition zone. We also found that PIN2-green fluorescent protein (GFP) localized to vacuole-like compartments (VLCs) in dark-grown root epidermal and cortical cells, and phot1/NPH3 mediated a BL-initiated pathway that caused PIN2 redistribution to the plasma membrane. When dark-grown roots were exposed to brefeldin A (BFA), PIN2-GFP remained in VLCs in darkness, and BL caused PIN2-GFP disappearance from VLCs and induced PIN2-GFP-FM4-64 colocalization within enlarged compartments. In the nph3 mutant, both dark and BL BFA treatments caused the disappearance of PIN2-GFP from VLCs. However, in the phot1 mutant, PIN2-GFP remained within VLCs under both dark and BL BFA treatments, suggesting that phot1 and NPH3 play different roles in PIN2 localization. In conclusion, BL-induced root phototropism is based on the phot1/NPH3 signaling pathway, which stimulates the shootward auxin flux by modifying the subcellular targeting of PIN2 in the root apex transition zone.     

Effect of belowground herbivory on parasitoid associative learning of plant odours

Root herbivores can influence both the performance and the behaviour of parasitoids of aboveground insect herbivores through changes in aboveground plant quality and in the composition of the plant's odour blend. Here we show that root herbivory by Delia radicum larvae did not influence the innate preferences for plant odours of the two closely related parasitoid species Cotesia glomerata and C. rubecula, but did affect their learned preferences, and did so in an opposite direction. While C. glomerata learned to prefer the odour of plants with intact roots, C. rubecula learned to prefer the odour of root‐infested plants. The learned preference of C. glomerata for the odour of plants with intact roots matches our previously published result of its better performance when developing in P. brassicae hosts feeding on this plant type. In contrast, the relatively stronger learned preference of C. rubecula for the odour of root‐infested plants cannot be merely explained by its performance, as the results of our present study indicate that D. radicum root herbivory did not influence the performance of C. rubecula nor of its host P. rapae. Our results stress the importance of assessing the influence of root herbivores on both innate and learned responses of parasitoids to plant odours.     

Salt-induced plasticity of root hair development is caused by ion disequilibrium in Arabidopsis thaliana

Root hair development is controlled by environmental signals. Studies on root hair plasticity in Arabidopsis thaliana have mainly focused on phosphate and iron deficiency. Root hair growth and development and their physiological role in response to salt stress are largely unknown. Here, we show that root epidermal cell types and root hair development are highly regulated by salt stress. Root hair length and density decreased significantly in a dose-dependent manner on both primary roots and junction sites between roots and shoots. The root hair growth and development were sensitive to inhibition by ions but not to osmotic stress. High salinity also alters anatomical structure of roots, leading to a decrease in cell number in N positions and enlargement of the cells. Moreover, analysis of the salt overly sensitive mutants indicated that salt-induced root hair response is caused by ion disequilibrium and appears to be an adaptive mechanism that reduces excessive ion uptake. Finally, we show that genes WER, GL3, EGL3, CPC, and GL2 might be involved in cell specification of root epidermis in stressed plants. Taken together, data suggests that salt-induced root hair plasticity represents a coordinated strategy for early stress avoidance and tolerance as well as a morphological sign of stress adaptation.     

Systemic signalling in photosynthetic induction of Rumex K‐1 (Rumex patientia × Rumex tianschaious) leaves

The rapid induction of photosynthesis is critical for plants under light‐fleck environment. Most previous studies about photosynthetic induction focused upon single leaf, but they did not consider the systemic integrity of plant. Here, we verified whether systemic signalling is involved in photosynthetic induction. Rumex K‐1 (Rumex patientia × Rumex tianschaious) plants were grown under light‐fleck condition. After whole night dark adaptation, different numbers of leaves (system leaf or SL) were pre‐illuminated with light, and then the photosynthetic induction of other leaves (target leaf or TL) was investigated. This study showed that the pre‐illumination of SL promoted photosynthetic induction in TL. This promotion was independent of the number of SL, the light intensity on SL and the distance between SL and TL, indicating that this systemic signalling is non‐dose‐dependent. More interestingly, the photosynthetic induction was promoted by only the pre‐illumination of morphological upper leaf rather than the pre‐illumination of morphological lower leaf, indicating that the transfer of this signal is directional. The results showed that the transfer of this systemic signalling depends upon the phloem. This systemic signalling helps plants to use light energy more efficiently under light flecks.     

Effect of Photoperiod, Auxins and Gibberellic Acid on Rooting of Stem Cuttings of Bryophyllum tubiflorum

Photoperiod controls the initiation and development of roots on cuttings of Bryophyllum tubiflorum. Root initiation occurred when either the mother plant or the cuttings were exposed subsequently to SD conditions. Cuttings from LD plants exposed to LD did not root at all even after 4 weeks showing that short days are necessary for rooting of cuttings of this plant. The short day requirement can be substituted by IAA or IBA as roots were initiated in cuttings from LD plants when treated with the auxins under LD conditions. GA stimulates rooting of cuttings under SD conditions but does not replace the SD requirement. It failed to induce rooting under LD conditions.