Soybean Seedling Root Segmentation Using Improved U-Net Network.

Soybean seedling root morphology is important to genetic breeding. Root segmentation is a key technique for identifying root morphological characteristics. This paper proposed a semantic segmentation model of soybean seedling root images based on an improved U-Net network to address the problems of the over-segmentation phenomenon, unsmooth root edges and root disconnection, which are easily caused by background interference such as water stains and noise, as well as inconspicuous contrast in soybean seedling images. Soybean seedling root images in the hydroponic environment were collected for annotation and augmentation. A double attention mechanism was introduced in the downsampling process, and an Attention Gate mechanism was added in the skip connection part to enhance the weight of the root region and suppress the interference of background and noise. Then, the model prediction process was visually interpreted using feature maps and class activation mapping maps. The remaining background noise was removed by connected component analysis. The experimental results showed that the Accuracy, Precision, Recall, F1-Score and Intersection over Union of the model were 0.9962, 0.9883, 0.9794, 0.9837 and 0.9683, respectively. The processing time of an individual image was 0.153 s. A segmentation experiment on soybean root images was performed in the soil-culturing environment. The results showed that this proposed model could extract more complete detail information and had strong generalization ability. It can achieve accurate root segmentation in soybean seedlings and provide a theoretical basis and technical support for the quantitative evaluation of the root morphological characteristics in soybean seedlings.

Mass Biosynthesis of Coumestrol Derivatives and Their Isomers via Soybean Adventitious Root Cultivation in Bioreactors.

Coumestrol (CMS) derivatives are unique compounds, which function as phytoalexins; they are derived from soybean roots, following abiotic and biotic stresses. As a phytoalexin, CMS forms a defense system that enables plants to maintain their viability. However, it is still challenging to achieve the mass production of phytoalexins, which exhibit pharmacological values, via plant breeding. Here, the synthesis of CMS derivatives from the seedling, plant, and adventitious root (AR) of Glycine max were investigated under artificial light, as well as via a chemical elicitor treatment. In the presence of constant light, as well as under treatment with methyl jasmonate, the CMS monoglucoside (coumestrin; CMSN) and malonyl CMSN (M-CMSN) contents of the AR culture (4 weeks) increased drastically. The two CMS derivatives, CMSN and M-CMSN, were obtained as a mixture of isomers, which were identified via nuclear magnetic resonance analysis. These derivatives were also observed in a soybean plant that was grown on artificial soil (AS; 5 weeks) and a Petri dish (9 days) although in considerably lesser quantities than those observed in the AR culture. Compared with the two other media (AS and the Petri dish), the AR culture achieved the superior synthesis of CMSN and M-CMSN within a relatively short cultivation period (<1 month) in laboratory-scale (3 L) and pilot-scale (1,000 L) bioreactors. The isoflavone content of AR under the constant light conditions was three-fold that under dark conditions. Significant quantities of malonyl daidzin and malonyl genistin were produced in the root of AS and the seedling of Petri dish, respectively. Flavonol glycosides were not produced in the AR culture under the dark and light conditions, as well as in AS under the dark condition. However, significant contents of kaempferol glycosides were produced in the leaves of AS and seedling of Petri dish, following the light treatment. Thus, we proposed that the established soybean AR-cultivation approach represented a better method for biosynthesizing phytoalexins, such as the CMS derivatives, as plant-derived functional materials.

Reduction of Pythium Damping-Off in Soybean by Biocontrol Seed Treatment.

Pythium spp. is one of the major groups of pathogens that cause seedling diseases on soybean, leading to both preemergence and postemergence damping-off and root rot. More than 100 species have been identified within this genus, with Pythium irregulare, P. sylvaticum, P. ultimum var ultimum, and P. torulosum being particularly important for soybean production given their aggressiveness, prevalence, and abundance in production fields. This study investigated the antagonistic activity of potential biological control agents (BCAs) native to the U.S. Midwest against Pythium spp. First, in vitro screening identified BCAs that inhibit P. ultimum var. ultimum growth. Scanning electron microscopy demonstrated evidence of mycoparasitism of all potential biocontrol isolates against P. ultimum var. ultimum and P. torulosum, with the formation of appressorium-like structures, short hyphal branches around host hyphae, hook-shaped structures, coiling, and parallel growth of the mycoparasite along the host hyphae. Based on these promising results, selected BCAs were tested under field conditions against six different Pythium spp. Trichoderma afroharzianum 26 used alone and a mix of T. hamatum 16 + T. afroharzianum 19 used as seed treatments protected soybean seedlings from Pythium spp. infection, as BCA-treated plots had on average 15 to 20% greater plant stand and vigor than control plots. Our results also indicate that some of these potential BCAs could be added with a fungicide seed treatment with minimum inhibition occurring, depending on the fungicide active ingredient. This research highlights the need to develop tools incorporating biological control as a facet of soybean seedling disease management programs. The harnessing of native BCAs could be integrated with other management strategies to provide efficient control of seedling diseases.

Development of a novel bio-organic fertilizer for the removal of atrazine in soil.

The accumulation of atrazine in farmland is prone to cause phytotoxicity to kinds of sensitive crops, such as soybean. In addition, some kinds of agricultural solid wastes have long been considered as the important non-point pollution source. The aim of this experiment was to investigate the feasibility of removing atrazine from soil and alleviating the stress of atrazine on the growth of soybean by application a novel bio-organic fertilizer developed by agricultural solid wastes, such as cow manure organic fertilizer, biochar and poly-(γ-glutamic acid), as well as an atrazine-degrading strain Arthrobacter sp. DNS10. Sixteen potential bio-organic fertilizer formulations were designed by D-optimal mixture design of Design Expert software and atrazine-removal ability was selected to single out the optimal formulation. As a result, the optimal formulation of bio-organic fertilizer (named as DNBF10) was produced by the cow manure organic fertilizer 76.20%, biochar 4.46%, poly-(γ-glutamic acid) 8.63% (m/m) and the number of Arthrobacter sp. DNS10 with 0.91 × 108 CFU/g. The atrazine removal percentage of DNBF10 for the atrazine in soil with the initial atrazine concentration 15.26 ±â€¯0.49 mg/kg was 95.05% after 10 days' application with DNBF10 at the adding dosage of 5 mg/kg (relative to the dry weight of the soil). Furthermore, pot experiment results suggest that the growth of soybean seedlings in the soil (initial atrazine was 8.14 ±â€¯0.16 mg/kg) that adding both of DNBF10 (25%) and chemical fertilizer (75%) were better than those of the treatment only adding chemical fertilizer (100%) under the same nutrient addition level. All the results indicate that the application of DNBF10 was a new alternative to reuse the typical agricultural solid wastes, as well as to reduce the harm caused by residual atrazine to soybean.

Combined acid rain and lanthanum pollution and its potential ecological risk for nitrogen assimilation in soybean seedling roots.

Rare earth elements (REEs) are used in various fields, resulting in their accumulation in the environment. This accumulation has affected the survival and distribution of crops in various ways. Acid rain is a serious global environmental problem. The combined effects on crops from these two types of pollution have been reported, but the effects on crop root nitrogen assimilation are rarely known. To explore the impact of combined contamination from these two pollutants on crop nitrogen assimilation, the soybean seedlings were treated with simulated environmental pollution from acid rain and a representative rare earth ion, lanthanum ion (La3+), then the indexes related to plant nitrogen assimilation process in roots were determined. The results showed that combined treatment with pH 4.5 acid rain and 0.08 mM La3+ promoted nitrogen assimilation synergistically, while the other combined treatments all showed inhibitory effects. Moreover, acid rain aggravated the inhibitory effect of 1.20 or 0.40 mM La3+ on nitrogen assimilation in soybean seedling roots. Thus, the effects of acid rain and La3+ on crops depended on the combination levels of acid rain intensity and La3+ concentration. Acid rain increases the bioavailability of La3+, and the combined effects of these two pollutants were more serious than that of either pollutant alone. These results provide new evidence in favor of limiting overuse of REEs in agriculture. This work also provides a new framework for ecological risk assessment of combined acid rain and REEs pollution on soybean crops.

Stomatal and non-stomatal factors regulated the photosynthesis of soybean seedlings in the present of exogenous bisphenol A.

Bisphenol A (BPA) is an emerging environmental endocrine disruptor that has toxic effects on plants growth. Photosynthesis supplies the substances and energy required for plant growth, and regulated by stomatal and non-stomatal factors. Therefore, in this study, to reveal how BPA affects photosynthesis in soybean seedlings (Glycine max L.) from the perspective of stomatal and non-stomatal factors, the stomatal factors (stomatal conductance and behaviours) and non-stomatal factors (Hill reaction, apparent quantum efficiency, Rubisco activity, carboxylation efficiency, the maximum Rubisco carboxylation velocity, ribulose-1,5-bisphospate regeneration capacities mediated by maximum electron transport rates, and triose phosphate utilization rate) were investigated using a portable photosynthesis system. Moreover, the pollution of BPA in the environment was simulated. The results indicate that low-dose BPA enhanced net photosynthetic rate (Pn) primarily by promoting stomatal factors, resulting in increased relative growth rates and accelerated soybean seedling growth. High-dose BPA decreases the Pn by simultaneously inhibiting stomatal and non-stomatal factors, and this inhibition decreases the relative growth rates further reducing soybean seedling growth. Following the withdrawal of BPA, all of the indices were restored to varying degrees. In conclusion, low-dose BPA increased the Pn by promoting stomatal factors while high-dose BPA decreased the Pn by simultaneously inhibiting stomatal and non-stomatal factors. These findings provide a model (or, hypothesis) for the effects of BPA on plant photosynthesis.

Effects of bisphenol A on key enzymes in cellular respiration of soybean seedling roots.

The environmental endocrine disrupter bisphenol A (BPA) is ubiquitous in the environment, with potential toxic effects on plants. Previous studies have found a significant effect of BPA on levels of mineral nutrients in plant roots, but the underlying mechanism remains unknown. To determine how BPA influences root mineral nutrients, the effects of BPA (1.5 mg L(-1) , 3.0 mg L(-1) , 6.0 mg L(-1) , 12.0 mg L(-1) , 24.0 mg L(-1) , 48.0 mg L(-1) , and 96.0 mg L(-1) ) on activities of critical respiratory enzymes (hexokinase, phosphofructokinase, pyruvate kinase, isocitrate dehydrogenase, and cytochrome c oxidase) were investigated in soybean seedling roots. After BPA exposure for 7 d, the low concentrations of BPA increased the activities of critical respiratory enzymes in roots, whereas opposite effects were observed in roots exposed to high concentrations of BPA, and the inhibitory effect was greater for higher BPA concentrations. In addition, evident morphological anomalies and decreases in root lengths and volumes were induced by high concentrations of BPA. Following withdrawal of BPA exposure after 7 d, the activities of respiratory enzymes and visible signs of toxicity recovered, and the extent of recovery depended on the type of enzyme and the BPA concentration. Furthermore, correlation analysis showed that the disturbance by BPA to activities of respiratory enzymes, which led to interference in the energy metabolism in roots, might be an effect mechanism of BPA on mineral element accumulation in plant roots.

Reduced growth of soybean seedlings after exposure to weak microwave radiation from GSM 900 mobile phone and base station.

The aim of this work was to study possible effects of environmental radiation pollution on plants. The association between cellular telephone (short duration, higher amplitude) and base station (long duration, very low amplitude) radiation exposure and the growth rate of soybean (Glycine max) seedlings was investigated. Soybean seedlings, pre-grown for 4 days, were exposed in a gigahertz transverse electromagnetic cell for 2 h to global system for mobile communication (GSM) mobile phone pulsed radiation or continuous wave (CW) radiation at 900 MHz with amplitudes of 5.7 and 41 V m(-1) , and outgrowth was studied one week after exposure. The exposure to higher amplitude (41 V m(-1)) GSM radiation resulted in diminished outgrowth of the epicotyl. The exposure to lower amplitude (5.7 V m(-1)) GSM radiation did not influence outgrowth of epicotyl, hypocotyls, or roots. The exposure to higher amplitude CW radiation resulted in reduced outgrowth of the roots whereas lower CW exposure resulted in a reduced outgrowth of the hypocotyl. Soybean seedlings were also exposed for 5 days to an extremely low level of radiation (GSM 900 MHz, 0.56 V m(-1)) and outgrowth was studied 2 days later. Growth of epicotyl and hypocotyl was found to be reduced, whereas the outgrowth of roots was stimulated. Our findings indicate that the observed effects were significantly dependent on field strength as well as amplitude modulation of the applied field.

Effects of bisphenol A on mineral nutrition in soybean seedling roots.

Bisphenol A (BPA) is a ubiquitous chemical in the environment and potentially harmful to plants. However, relevant studies of the effects of BPA on plants are relatively scarce. In the present work, the effects of BPA on the biomass (fresh wt and dry wt), absorptive function (activity and absorptive area), and mineral element levels in soybean (Glycine max L.) seedling roots treated with 1.5 mg L(-1) , 3.0 mg L(-1) , 6.0 mg L(-1) , 12.0 mg L(-1) , 24.0 mg L(-1) , 48.0 mg L(-1) , and 96.0 mg L(-1) BPA were investigated. Treatment with 1.5 mg L(-1) BPA increased the levels of nitrate and other mineral elements (P, K, Mg, Mn, Zn, and Mo) in the roots, whereas treatments with BPA at higher concentrations decreased the levels of these elements in the roots. All treatments with BPA caused increases in the levels of ammonium, Ca, Fe, and Cu in the roots. Moreover, treatment with 1.5 mg L(-1) BPA increased the fresh weight, dry weight, activity, and absorptive area of the roots, whereas treatments with BPA at higher concentrations decreased these indices in a dose-dependent manner. Furthermore, correlation analysis data showed that BPA affected the levels of mineral elements and absorptive function of soybean seedling roots, which may be the physiological basis of BPA action on plants.

Effect of lanthanum(III) on the production of ethylene and reactive oxygen species in soybean seedlings exposed to the enhanced ultraviolet-B radiation.

The enhanced ultraviolet-B (UV-B) radiation caused by ozone depletion may exert deleterious effects on plants. Therefore, studies on the effect of UV-B radiation on plants, as well as studies on the methods for alleviating the deleterious effects by chemical control, are of great significance. In this study, after soybean (Glycine max) seedlings were exposed to UV-B radiation (10.2 and 13.8kJ m(-2)day(-1)) for 5 days and the followed 6 days of restoration, respectively, the effects of 20mg L(-1) lanthanum (III) [La(III)] on leaf phenotype, photosynthetic rate, and production of ethylene and reactive oxygen species (ROS) were investigated. The results indicated that the exposure to 10.2 and 13.8kJ m(-2)day(-1) UV-B radiation could cause injury to the leaf phenotype, and lead to the decrease in the content of chlorophyll and the net photosynthetic rate, and the increase in the contents of ROS, ethylene and 1-aminocyclopropanecarboxylic acid, and 1-aminocyclopropanecarboxylic acid synthase activity in soybean seedlings. Following the withdrawal of the enhanced UV-B radiation, the above mentioned parameters gradually recovered, and the recovery of soybean seedlings exposed to 10.2kJ m(-2)day(-1) UV-B radiation was faster than those in soybean seedlings exposed to 13.8kJ m(-2)day(-1) UV-B radiation. The leaf injury and the changes in the above indices that were induced by the enhanced UV-B radiation, especially at 10.2kJ m(-2)day(-1), were alleviated after the pretreatment of soybean seedlings with 20mg L(-1) La(III). The results of the correlation analysis demonstrated that the injury to the leaf phenotype and the decrease in the photosynthetic rate of soybean seedlings were correlated with the increase in the ROS content that was induced by ethylene in soybean seedlings. The pretreatment with 20mg L(-1) La(III) alleviated the injury caused by the enhanced UV-B radiation through the regulation of the ROS production.