Optimal biochar selection for cadmium pollution remediation in Chinese agricultural soils via optimized machine learning.

Biochar is effective in mitigating heavy metal pollution, and cadmium (Cd) is the primary pollutant in agricultural fields. However, traditional trial-and-error methods for determining the optimal biochar remediation efficiency are time-consuming and inefficient because of the varied soil, biochar, and Cd pollution conditions. This study employed the machine learning method to predict the Cd immobilization efficiency of biochar in soil. The predictive accuracy of the random forest (RF) model was superior to that of the other common linear and nonlinear models. Furthermore, to improve the reliability and accuracy of the RF model, it was optimized by employing a root-mean-squared-error-based trial-and-error approach. With the aid of the optimized model, the empirical categories for soil Cd immobilization efficiency were biochar properties (60.96 %) > experimental conditions (19.6 %) ≈ soil properties (19.44 %). Finally, this study identified the optimal biochar properties for enhancing agricultural soil Cd remediation in different regions of China, which was beneficial for decision-making regarding nationwide agricultural soil remediation using biochar. The immobilization effect of alkaline biochar was pronounced in acidic soils with relatively high organic matter. This study provides insights into the immobilization mechanism and an approach for biochar selection for Cd immobilization in agricultural soil.

Selective and efficient immobilization of cadmium in soil by layered double hydroxides intercalated with the mercaptosuccinic acid.

Cadmium (Cd) contamination in cropland poses a significant threat to the quality of agricultural products, but even though in-situ remediation has been extensively applied, non-selective immobilization remains an issue. In order to develop a material that specifically immobilizes Cd in soil, a layered double hydroxide, intercalated with mercaptosuccinic acid (MSA-CFA), was synthesized through co-precipitation. In this case, the MSA-CFA's maximum adsorption capacity was increased from the 513.8 mg·g-1 for unintercalated hydrotalcite CFA to 692.6 mg·g-1. Besides, MSA-CFA efficiently removed 99.25 % of Cd from soil water-extract solution and immobilized up to 70.03 % of bio-available Cd. However, interestingly, its immobilization effects on beneficial metal elements Fe, Mn and Zn were milder, being equivalent to 2/7, 5/7 and 1/2 that of lime, respectively. Moreover, XRD and XPS techniques revealed isomorphous substitution with calcium and sulfhydryl complexation during the Cd adsorption by MSA-CFA. Compared with CFA, the increased adsorption capacity of MSA-CFA for Cd was due to intercalated MSA acting as a new adsorption site, while the enhanced selectivity was contributed by sulfhydryl's affinity for Cd. Altogether, MSA-CFA showed great promise as a competitive and highly efficient candidate amendment in Cd-contaminated soil remediation.

Optical-Cavity-Incorporated Colorful All-Solid-State Electrochromic Devices for Dual Anti-Counterfeiting.

The fusion of electrochromic technology with optical resonant cavities presents an intriguing innovation in the electrochromic field. However, this fusion is mainly achieved in liquid electrolyte-based or sol-gel electrolyte-based electrochromic devices, but not in all-solid-state electrochromic devices, which have broader industrial applications. Here, a new all-solid-state electrochromic device is demonstrated with a metal-dielectric-metal (MDM) resonant cavity, which can achieve strong thin-film interference effects through resonance, enabling the device to achieve unique structural colors that have rarely appeared in reported all-solid-state electrochromic devices, such as yellow green, purple, and light red. The color gamut of the device can be further expanded due to the adjustable optical constants of the electrochromic layer. What is more, this device exhibits remarkable cycling stability (maintaining 84% modulation capability after 7200 cycles), rapid switching time (coloration in 2.6 s and bleaching in 2.8 s), and excellent optical memory effect (only increasing by 13.8% after almost 36 000 s). In addition, this exquisite structural design has dual-responsive anti-counterfeiting effects based on voltage and angle, further demonstrating the powerful color modulation capability of this device.

Assessment of CuO NPs on soil microbial community structure based on phospholipid fatty acid techniques and phytotoxicity of bok choy seedlings.

In this study, a soil culture and a hydroponic experiment were conducted to assess the toxicology effects of copper oxide nanoparticles (CuO NPs) on soil microbial community structure and the growth of bok choy. Results showed CuO NPs had an inhibitory effect on soil microbial abundance, diversity, and activity, as well as the bok choy seedling growth, whereas CuO NPs at low concentrations did not significantly affect the soil microbial biomass or plant growth. In soil, CuO NPs at high dose (80 mg kg-1) significantly reduced the indexes of Simpson diversity, Shannon-Wiener diversity and Pielou evenness by 3.7%, 4.9% and 4.5%, respectively. In addition, CuO NPs at 20 and 80 mg kg-1 treatment significantly reduced soil enzymes (urease, alkaline phosphatase, dehydrogenase, and catalase) activities by 25.5%-58.9%. Further, CuO NPs at 20 mg L-1 significantly inhibited the growth of plant root by 33.8%, and catalase (CAT) activity by 17.9% in bok choy seedlings. The present study can provide a basis for a comprehensive evaluation of the toxicity effect of CuO NPs on soil microorganisms and phytotoxicity to bok choy seedlings.

Comparative effects of copper nanoparticles and copper oxide nanoparticles on physiological characteristics and mineral element accumulation in Brassica chinensis L.

In this study, the short-term toxicity of Cu NPs, CuO NPs, and CuSO4 on bok choy (Brassica chinensis L.) under hydroponic conditions was evaluated using indicators such as biomass, net photosynthesis rate, root morphology, enzyme activity, and Cu accumulation and subcellular distribution. Results showed that CuO NPs exposure notably increased the biomass, root length, and root tip number by 22.0%, 22.7%, and 82.9%, respectively, whereas Cu NPs and CuSO4 significantly reduced root biomass, net photosynthetic rate (PN), and root length by 31.2% and 44.2%, 24.5% and 32.2%, and 43.4% and 40.6%, respectively. In addition, Cu NPs, CuO NPs and CuSO4 exposure increased the distribution of Cu in soluble component and cell wall. Moreover, short-term exposure to different Cu forms significantly affected mineral element accumulation in bok choy. For instance, Cu NPs exposure reduced the concentrations of Mg, Ca and Mn in edible part by 21.7%, 16.1% and 23.2%, respectively. CuSO4 exposure reduced the concentrations of Mg and Ca in edible part by 12.3% and 50.1%, respectively. CuO NPs caused a significant increase of 30.4% for Ca concentration in root, 34.5% and 34.5% for K and Mn concentration in edible part. Over all, CuO NPs exposure was beneficial for plant growth. These findings help understand the phytotoxic effect of different Cu forms on bok choy, and CuO NPs has the potential to be applied to improve nutrition and prompt growth in edible plants.

Derivation and validation of soil cadmium thresholds for the safe farmland production of vegetables in high geological background area.

Excessive dietary intake of cadmium (Cd) poses toxicity risks to human health, and it is therefore essential to establish accurate and regionally appropriate soil Cd thresholds that ensure the safety of agricultural products grown in different areas. This study investigated the differences in the Cd accumulation in 32 vegetable varieties and found that the Cd content ranged from 0.01 to 0.24 mg·kg-1, and decreased in the order of stem and bulb vegetables > leafy vegetables > solanaceous crops > bean cultivars. A correlation analysis and structural equation model showed that pH, soil organic matter, and the cation exchange capacity had significant effects on Cd accumulation in the vegetables and explained 72.1 % of the variance. In addition, species sensitivity distribution (SSD) curves showed that stem and bulb vegetables were more sensitive to Cd than other types of vegetables. Using the Burr Type III function for curve fitting, we derived Cd thresholds of 6.66, 4.15, and 1.57 mg·kg-1 for vegetable soils. These thresholds will ensure that 20 %, 50 %, and 95 % of these vegetable varieties were risk-free, respectively. The predicted threshold of soil Cd was more than twice that of China's current National Soil Quality Standard (GB 15618-2018) for Cd values. Therefore, soil scenarios and cultivars should be considered comprehensively when determining farmland soil thresholds. The present results provide a new model for setting soil Cd criteria in high geological background areas.

Yellow-Green Luminescence Due to Polarity-Dependent Incorporation of Carbon Impurities in Self-Assembled GaN Microdisk.

Yellow-green luminescence (YGL) competes with near-bandgap emission (NBE) for carrier recombination channels, thereby reducing device efficiency; yet uncovering the origin of YGL remains a major challenge. In this paper, nearly stress-free and low dislocation density self-assembled GaN microdisks were synthesized by Na-flux method. The YGL of GaN microdisks highly depend on their polar facets. Variable accelerating voltage/power CL, variable temperature PL, and Raman spectroscopy are further performed to clarify the origin of polarity dependence of GaN microdisk YGL behavior, which indicates its independence of dislocations, surface effects, stress, crystalline quality, and gallium vacancies. It was found that the incorporation ability of carbon impurities in the polar (0001) facet is greater than that in the semipolar (101̅1) facets, producing higher content of CN or CNON defects, resulting in a more pronounced YGL in the polar (0001) facet of GaN.

Recapitulation of dynamic nanoparticle transport around tumors using a triangular multi-chamber tumor-on-a-chip.

3D tumor models are emerging as valuable tools for drug screening and nanoparticle based personalized cancer treatments. The main challenges in building microfluidic chip-based 3D tumor models currently include the development of bioinks with high bioactivity and the reproduction of the key tumor extracellular matrix (ECM) with heterogeneous tumor microenvironments. In this study, we designed a triangular multi-chamber tumor-on-a-chip (TM-CTC) platform, which consisted of three circular chambers at the vertices of a triangle connected by three rectangular chambers; it significantly improved the culture efficiency of 3D tumor tissues. MCF-7 tumor cells were cultured in a 3D ECM and then dynamically perfused for 7 days of culture to obtain abundant tumor spheroids with uniform size (100 ± 4.1 µm). The biological features of the 3D tumor tissue including epithelial transformation (EMT), hypoxia and proliferation activities were reproduced in the triangular multi-chamber tumor-on-a-chip (TM-CTC) platform. The permeability results of NPs confirmed that the ECM exhibited a significant barrier effect on the transportation of NPs when compared with free drugs, indicating that the ECM barrier should be considered as one of the key factors of drug delivery carrier development. In addition, this TM-CTC model provided a suitable platform for constructing a complex heterogeneous tumor microenvironment with multiple cells (MCF-7, HUVEC and MRC-5) involved, which was beneficial for exploring the dynamic interaction between tumor cells and other cells in the tumor microenvironment. The above results suggest that this TM-CTC model can simulate the dynamic transportation of NPs around 3D tumor tissues, and thus provide a reliable platform for NP evaluation.

Phytotoxicity and Accumulation of Copper-Based Nanoparticles in Brassica under Cadmium Stress.

The widespread use of copper-based nanoparticles expands the possibility that they enter the soil combined with heavy metals, having a toxic effect and posing a threat to the safety of vegetables. In this study, single and combined treatments of 2 mg/L Cd, 20 mg/L Cu NPs and 20 mg/L CuO NPs were added into Hoagland nutrient solution by hydroponics experiments. The experimental results show that copper-based Nanoparticles (NPs) can increase the photosynthetic rate of plants and increase the biomass of Brassica. Cu NPs treatment increased the Superoxide Dismutase (SOD), Peroxidase (POD) and catalase (CAT) activities of Brassica, and both NPs inhibited ascorbate peroxidase (APX) activity. We observed that Cd + Cu NPs exhibited antagonistic effects on Cd accumulation, inhibiting it by 12.6% in leaf and 38.6% in root, while Cd + CuO NPs increased Cd uptake by 73.1% in leaves and 22.5% in roots of Brassica. The Cu content in the shoots was significantly negatively correlated with Cd uptake. The Cd content of each component in plant subcellular is soluble component > cytoplasm > cell wall. Cu NPs + Cd inhibited the uptake of Zn, Ca, Fe, Mg, K and Mn elements, while CuO NPs + Cd promoted the uptake of Mn and Na elements. The results show that copper-based nanoparticles can increase the oxidative damage of plants under cadmium stress and reduce the nutritional value of plants.

Two-step modification (sodium dodecylbenzene sulfonate composites acid-base) of sepiolite (SDBS/ABsep) and its performance for remediation of Cd contaminated water and soil.

To enhance the remediation capability of cadmium (Cd) polluted water and soil, our approach involved two-step modification of sepiolite (Sep) through acid-base compound treatment and sodium dodecylbenzene sulfonate (referring as SDBS/ABsep), and then the batch adsorption and soil culture experiments were conducted to investigate its immobilization potential and mechanisms of Cd. The findings revealed that the SDBS/ABsep had a rougher surface and higher porosity, and the maximum adsorption capacity of Cd2+ onto SDBS/ABsep was 241.39 mg g-1, which was 5.32 times higher than that on Sep. It conformed to the pseudo-second-order kinetic and Redlich-Paterson isotherm models. SDBS/ABsep exhibited a high efficiency for immobilization-induced remediation of Cd polluted soils. Upon the addition of different concentrations of SDBS/ABsep, DTPA-Cd content decreased by 17.41-47.33% compared with the control groups, and the ratio of residual fraction-Cd increased from 4.67% in unamended soil to 14.05% in the presence of 4% SDBS/ABsep. SEM-EDS, TEM, FTIR, XRD, and XPS analyses indicated that the interaction mechanisms between SDBS/ABsep and Cd included the electrostatic force, precipitation, ion exchange, and complexation of sulfonic acid groups. Therefore, SDBS/ABsep can be used as a promising effective passivation agent for remediation of Cd contaminated soil and water.

Selenate regulates the activity of cell wall enzymes to influence cell wall component concentration and thereby affects the uptake and translocation of Cd in the roots of Brassica rapa L.

Selenium (Se) can be used to counteract cadmium (Cd) toxicity in plants. However, mechanisms underlying the alleviation of Cd toxicity by Se have not been completely elucidated, especially those by which Se reduces Cd translocation. A hydroponic experiment was performed to illustrate the regulatory mechanisms of Cd transport by selenate (Se (VI)) in pakchoi (Brassica rapa L., LvYou 102). The results showed that this plant had a high accumulation capacity for Cd, and Se(VI) addition restricted Cd translocation from roots to shoots. Se(VI) exposure stimulated the concentrations of pectins and hemicellulose II but reduced the concentration of hemicellulose I in the roots. In many cases, the enzymes pectin methylesterase, polygalacturonase, and ß-galactosidase were dose-dependently triggered by Se(VI) under Cd exposure, but root calcium concentration was significantly lowered (p < 0.05). Xyloglucan endoglycosidase (hydrolase) was triggered by Se(VI) under 2 mg L-1 Cd exposure and cellulase was generally activated by Se(VI) under Cd stress. The above results suggest that Se(VI) up-regulates pectin methylesterase activity, stimulates synthesis of pectins, and down-regulates root Ca concentration to release free carboxyl groups to combine Cd. In this study, the relationships between enzyme activity (e.g., peroxidase, superoxidase and ß-galactosidase), hydrogen peroxide, cell wall structure strengthening/loosening, and Cd toxicity affected by Se(VI) were also discussed.

Reshaping the murine immunoglobulin heavy chain repertoire with bovine DH genes.

Having a limited number of VH segments, cattle rely on uniquely long DH gene segments to generate CDRH3 length variation (3-70 aa) far greater than that in humans or mice. Bovine antibodies with ultralong CDRH3s (>50 aa) possess unusual structures and abilities to bind to special antigens. In this study, we replaced most murine endogenous DH segments with bovine DH genes, generating a mouse line termed B-DH. The use of bovine DH genes significantly increased the length variation of CDRH3 and consequently the Ig heavy chain repertoire in B-DH mice. However, no ultralong CDRH3 was observed in B-DH mice, suggesting that other factors, in addition to long DH genes, are also involved in the formation of ultralong CDRH3. The B-DH mice mounted a normal humoral immune response to various antigens, although the B-cell developmental paradigm was obviously altered compared with wild-type mice. Additionally, B-DH mice are not predisposed to the generation of autoantibodies despite the interspecies DH gene replacement. The B-DH mice reported in this study provide a unique model to answer basic questions regarding the synergistic evolution of DH and VH genes, VDJ recombination and BCR selection in B-cell development.

In‒situ immobilization remediation, soil aggregate distribution, and microbial community composition in weakly alkaline Cd‒contaminated soils: A field study.

Biochar has advantages of a large specific surface area and micropore structure, which is beneficial for immobilization remediation of heavy metal‒contaminated soils. A field experiment was conducted to investigate the effects of rice husk biochar (BC) (7.5, 15, and 15 t hm-2) on Cd availability in soils and accumulation in maize (Zea mays L), soil aggregate structure, and microbial community abundance. The results show that BC treatment promoted the formation of large aggregates (5-8 and 2-5 mm) and enhanced aggregate stability, whereas it decreased the proportion of ≤0.25 mm soil aggregates. The geometric mean diameter and mean weight diameter under BC‒treated soils increased by 9.9%-40.5% and 3.6%-32.7%, respectively, indicating that the stability of soil aggregates increased. Moreover, BC facilitated the migration of Cd from large particles (>0.5 mm aggregates) to small particles (<0.25 mm aggregates). The application of BC decreased diethylenetriamine pentaacetic acid ‒extractable Cd by 17.6%-32.12% in contrast with the control. The amount of Cd in maize was reduced by 56.7%-81.1% for zhengdan958, 52.4%-85.9% for Sanbei218, and 73.7%-90.4% for Liyu16. When compared with the control groups, BC addition significantly (P < 0.05) increased the number of Ace observed, Shannon diversity indices, and the relative abundances of Proteobacteria, Acidobacteria, and Bacteroidetes. Therefore, rice husk BC exhibited a certain feasibility in immobilizing remediation of weakly alkaline Cd‒contaminated soils.

Cd immobilization and soil quality under Fe-modified biochar in weakly alkaline soil.

Cost-effective and environment-friendly implementation techniques are critical to the success of remediation in large-scale cadmium (Cd) contaminated agricultural soil. Field experiments were conducted to investigate the effect of Fe-modified biochar on Cd bioavailability in soils and uptake by maize (Zea mays L.), soil aggregate distribution and stability, and microbial community composition in weakly alkaline Cd-contaminated soil. Results showed that Fe-modified biochar optimized the structure and stability of soil aggregates. Moreover, the content of soil organic carbon increased by 6.59%-20.36% when compared with the control groups. However, DTPA-Cd concentration under the treatment of Fe-modified biochar was suffered by 37.74%-41.65% reduction in contrast with CK, and the significant decrease (P < 0.05) was obtained at 0.5% Fe-modified biochar. Moreover, sequential extraction procedures showed that the acid soluble and reducible states of Cd was converted into oxidizable and residual form. The addition of Fe-modified biochar inhibited Cd accumulation in maize, being 41.31%-76.64% (Zhengdan 958), 38.19%-70.95% (Liyu 86) and 52.30%-59.95% (Sanbei 218) reduction, respectively, in contrast with CK. The activity of catalase, urease and alkaline phosphatase in soil increased gradually with the addition of Fe-modified biochar. The enhancement in the number of soil bacterial OTUs and the values of Shannon, Chao1, ACE index indicated that Fe-modified biochar promoted the richness and diversity of bacterial communities. Therefore, the improvements of soil environment and biological quality indicated that Fe-modified biochar should be an alternative agent on remediation of Cd-contaminated soils.

Phytoexclusion of heavy metals using low heavy metal accumulating cultivars: A green technology.

Heavy metal (HM) pollution of farmland is a serious problem worldwide and consumption of HM-contaminated food products poses significant public health risks. Phytoexclusion using low HM accumulating cultivars (LACs) is a promising and practical technology to mitigate the risk of HM contamination of agricultural products grown in polluted soils, and does not alter cultivation practices, is easy to apply, and is economical. This review provides an overview of the major scientific advances accomplished in the field of LACs worldwide. The LACs concept and identification criteria are presented, and the known LACs among currently cultivated grain crops and vegetables are re-evaluated. The low HM accumulation by LACs is affected by crop ecophysiological features and soil physicochemical characteristics. Taking low Cd accumulating cultivars as an example, it is known that they can efficiently exclude Cd from entering their edible parts in three ways: 1) decrease in root Cd uptake by reducing organic acids secretion in the rhizosphere and transport protein production; 2) restriction of Cd translocation from roots to shoots via enhanced Cd retention in the cell wall and Cd sequestration in vacuoles; and 3) reduction in Cd translocation from shoots to grains by limiting Cd redirection and remobilization mediated through nodes. We propose an LAC application strategy focused on LACs and optimized to work with other agronomic measures according to the classification of HM risk level for LACs, providing a cost-effective and practical solution for safe utilization of large areas of farmland polluted with low to moderate levels of HMs.

[Combined Effect of Weathered Coal Based Amendments and Soil Water Management on Methylmercury Accumulation in Paddy Soil and Rice Grains].

A pot experiment and field experiment were designed to study the changes in the grain methyl mercury content in paddy soil and rice yield by sowing soil amendments that contained weathered coal, CaCO3, and Na2SeO3 as the main raw materials, combined with water management in a paddy field (80% field capacity after the heading and flowering periods). The results showed that:① In pot experiment, the content of methylmercury in rice rhizosphere soil decreased by 86.6% and the content of methylmercury in the rice grains decreased by 65.2% compared with that of the control. In field experiment, the content of methylmercury in rice rhizosphere soil decreased by 77.4% and the content of methylmercury in rice grains decreased by 60.6% upon adding the amendment+water management compared with that of CK. ② The soil pH increased by more than 0.3 in the pot experiment and 0.2 in the field experiment compared with that of the control. Furthermore, rice yield and plant biomass did not decrease in the two parts of the experiment. It can be inferred that the soil amendment and agronomic regulation measures (water management) used in this study have the advantages of quick effects, convenient use, and remarkable control effects and without secondary pollution. More, they can effectively reduce the risk of rice methylmercury exposure.

Underlying mechanisms responsible for restriction of uptake and translocation of heavy metals (metalloids) by selenium via root application in plants.

Since selenium (Se) was shown to be an essential element for humans in 1957, the biofortification of Se to crops via foliar spraying or soil fertilization has been performed for several decades to satisfy the daily nutritional need of humans. Appropriate doses of Se were found to counteract a number of abiotic and biotic stresses, such as exposure to heavy metals (metalloids) (HMs), via influencing the regulation of antioxidant systems, by stimulation of photosynthesis, by repair of damaged cell structures and functions, by regulating the metabolism of some substances and the rebalancing of essential elements in plant tissues. However, few concerns were paid on why and how Se could reduce the uptake of a variety of HMs. This review will mainly address the migration and transformation of HMs regulated by Se in the soil-plant system in order to present a hypothesis of why and how Se can reduce the uptake of HMs in plants. The following aspects will be examined in greater detail, including 1) how the soil characteristics influences the ability of Se to reduce the bioavailability of HMs in soils and their subsequent uptake by plants, which include soil Se speciation, pH, water regime, competing ions and microbes; 2) how the plant root system influenced by Se affects the uptake or the sequestration of HMs, such as root morphology, root iron plaques and root cell wall; 3) how Se combines with HMs and then sequesters them in plant cells; 4) how Se competes with arsenic (As) and thereby reduces As uptake in plants; 5) how Se regulates the expression of genes encoding functions involved in uptake, translocation and sequestration of HMs by Se in plants.

Performance and mechanisms of immobilization remediation for Cd contaminated water and soil by hydroxy ferric combined acid-base modified sepiolite (HyFe/ABsep).

In order to simultaneously improve the remediation capability of Cd contaminated water and soil, hydroxy iron-ABsep (HyFe/ABsep) was synthesized by a two-step modified (acid-base composite treatment, and hydroxy group was by using NaOH and Fe (NO3)3·9H2O). Results showed that HyFe/ABsep had developed pores and a rougher surface morphology, and the salt-soluble ion content was increased, surface-loaded iron species was mainly composed of FeOOH. Adsorption process of Cd2+ by HyFe/ABsep conformed best to the preudo-second-order model and Redlich-Paterson models, respectively. The behavior over a whole range of adsorption was consistent with chemical adsorption being the rate-controlling step and the theoretical maximum adsorption capacity obtained for the HyFe/ABsep was 220.9 mg·g-1 at 298 K, which was 4.87 times than Sep. HyFe/ABsep also had a more excellent passivation effect on available Cd in soil, being 36.83%-48.46% under the treatments of 0.5%-4% HyFe/ABsep, The structure and morphology of HyFe/ABsep were characterized through SEM-EDS, TEM, FTIR, XRD and XPS indicated that the mainly mechanisms of Cd sorption may include precipitates, ion exchange and complexation of active silanol groups. Therefore, HyFe/ABsep can employ as an effective agent for immobilization remediation of Cd contaminated water and soil.

Genetic removal of the CH1 exon leads to the production of hypofunctional heavy chain-only IgG2a in rats.

Despite great values in many applications, heavy chain-only antibodies (HcAbs) are naturally only produced in camelids and sharks, which are not easy to access and handle. Production of the type of antibodies in small laboratory animals would remarkably facilitate their applications. We previously reported a mouse line in which the CH1 exon of mouse γ1 was deleted that could express heavy chain-only IgG1 antibodies. However, these mice showed an extremely weak IgG1 response to specific antigens when immunized, and we could only achieve single VH domains with low affinity to antigens using these mice. One possibility is that the mouse germline VH repertoire was not sufficient to support the expression of functional heavy chain-only antibodies. In this study, we report the generation of a rat line in which the CH1 exon of the γ2a gene was removed and the γ1 and γ2b genes were silenced. Although the genetically modified rats expressed heavy chain-only IgG2a, they also exhibited a very weak IgG2a response to antigen immunization. Panning of a phage library constructed using IgG2a VH segments amplified from immunized rats identified antigen-specific single VH antibodies, which also exhibited much lower affinity than that of commercial mAbs. Together with our previous report, this study suggests that the simple genetic removal of the CH1 exon does not guarantee the successful expression of functional heavy chain-only antibodies.