[Effect of Exogenous Chitosan on Physiological Properties, Antioxidant Activity, and Cadmium Uptake of Wheat (Triticum aestivum L.) Seedlings Under Cadmium Stress].

This research aimed to clarify the effects of exogenously applied chitosan on the physiological characteristics, antioxidant activities, and Cd accumulation of wheat (Triticum aestivum L.) seedlings under cadmium (Cd) stress and to identify the key indicators based on the partial least squares model. The wheat variety studied was Bainong207 (BN207), and Cd-stress was achieved by growing seedlings in a hydroponic culture experiment with 10 and 25 µmol·L-1 Cd2+ added to the culture solution. It was found that both Cd-stress at 10 and 25 µmol·L-1 significantly inhibited the chlorophyll content, photosynthesis, and biomass accumulation of wheat seedlings. Seedling roots became shorter and thicker, and the lateral roots decreased under Cd-stress. The Cd-stress also increased H2O2 and MDA accumulation and the degree of cell membrane lipid peroxidation and affected the activities of antioxidant enzymes such as superoxide dismutase (SOD) and peroxidase (POD). Under Cd stress, exogenous chitosan decreased the Cd content in the aboveground and underground parts of wheat by 13.22 %-21.63 % and 7.92 %-28.32 % and reduced Cd accumulation in the aboveground and underground parts by 5.37 %-6.71 % and 1.91 %-4.09 %, respectively. Whereas exogenous chitosan application significantly reduced the content of H2O2 in roots and aboveground parts of wheat by 38.21 %-47.46 % and 45.81 %-55.73 % and MDA content by 37.65 %-48.12 % and 29.87 %-32.51 %, it increased the activities of SOD and POD in roots by 2.78 %-5.61 % and 13.81 %-18.33 %, respectively. In summary, exogenous chitosan can improve the photosynthetic characteristics and antioxidant enzyme activities of wheat seedlings under Cd stress, reduce the content and accumulation of Cd in the root and aboveground parts of wheat, and alleviate the damage of lipid peroxidation to the cell membrane. All of these results provide the basal data for the application of exogenous chitosan to alleviate Cd toxicity to wheat seedlings.

[Mitigative Effect of Rare Earth Element Cerium on the Growth of Zinc-stressed Wheat （Triticum aestivum L.）Seedlings].

This research aimed to clarify the mitigative effect of exogenously applied rare earth element cerium （Ce） on the growth， zinc （Zn） accumulation， and physiological characteristics of wheat （Triticum aestivum L.） seedlings under Zn stress. The wheat variety studied was Bainong307 （BN307）， and Zn stress was achieved by growing seedlings in a hydroponic culture experiment with 500 µmol·L-1 Zn2 + added to the culture solution. It was found that Zn stress at 500 µmol·L-1 significantly inhibited the chlorophyll content， photosynthesis， and biomass accumulation of wheat seedlings. Seedling roots became shorter and thicker， and the lateral roots decreased under Zn stress. The Zn stress also increased MDA accumulation and the degree of cell membrane lipid peroxidation and reduced soluble protein contents and the activities of antioxidant enzymes such as superoxide dismutase （SOD）， catalase （CAT）， and ascorbate peroxidase （APX）. On the contrary， exogenous Ce decreased the adsorption and transport of Zn by the root system and alleviated the damage of Zn stress to wheat seedlings. Specifically， the increase in chlorophyll content （chlorophyll a， chlorophyll b， and total chlorophyll） and photosynthetic parameters， the enhancement of antioxidant enzymes activities and soluble protein levels， and the reduction in MDA content and the damage of lipid peroxidation to the cell membrane were all driven by exogenous Ce， which ultimately led to the increase in dry matter biomass of the root system and shoot. In summary， these results provide basic data for the application of exogenous Ce to alleviate Zn toxicity to plants.

[Effects of Exogenous Zinc on Growth and Root Architecture Classification of Maize Seedlings Under Cadmium Stress].

To explore the effects of different concentrations of zinc （Zn） on the growth and root architecture classification of maize seedlings under cadmium （Cd） stress， a hydroponic experiment was conducted to study the effects of different concentrations of Zn （0， 10， 25， 50， 100， 200， and 400 µmol·L-1） on the growth， root architecture and classification characteristics， Cd content， root Cd uptake capacity， and photosynthetic system of maize seedlings under Cd stress （50 µmol·L-1） by using Zhengdan 958 as the experimental material. Principal component analysis and the membership function method were used for comprehensive evaluation. The results showed that the 50 µmol·L-1 Cd stress had a significant toxic effect on maize seedlings， which significantly reduced chlorophyll content and photosynthetic parameters. The main root length， plant height， biomass， root forks， and root tips， including the root length and root surface area of the grade Ⅰ-Ⅲ diameter range and the root volume of the grade Ⅰ-Ⅱ diameter range， decreased significantly， which hindered the normal growth and development of maize seedlings. Compared with that under no Zn application， 100 µmol·L-1 and 200 µmol·L-1 Zn application reduced the uptake of Cd by maize seedlings， significantly reduced the Cd content in shoots and roots and the Cd uptake efficiency. The toxic effect on maize seedlings was alleviated， and the fresh weight， dry weight， tolerance index， and root forks of shoots and roots were significantly increased. The photosynthesis of maize seedlings was significantly enhanced， and the photosynthetic rate and the total chlorophyll content was significantly increased. The RL， SA， and RV in the Ⅰ-Ⅱ diameter range reached the maximum at 100 µmol·L-1 Zn， and the RL， SA， and RV in the Ⅲ diameter range reached the maximum at 200 µmol·L-1 Zn， which were significantly higher than those without Zn treatment. The comprehensive evaluation of the growth tolerance of maize seedlings showed that 100 µmol·L-1 and 200 µmol·L-1 Zn had better effects on alleviating Cd toxicity. Comprehensive analysis showed that the application of appropriate concentration of Zn could reduce the Cd content in maize seedlings， the Cd uptake capacity， and Cd uptake efficiency of roots； increase the biomass accumulation of maize seedlings； reduce the effect of Cd toxicity on root architecture； reduce the effect on the light and system； and improve the tolerance of maize seedlings to Cd.

Antibiotics and antibiotic resistance genes in the water sources of the Wuhan stretch of the Yangtze River: Occurrence, distribution, and ecological risks.

Given the ubiquitous detection of antibiotics and antibiotic resistance genes (ARGs) in waterbodies worldwide and increasing public attention to water resource safety, this study investigated the presence of antibiotics and ARGs in the water sources of the Wuhan stretch of the Yangtze River (YR) as well as potential ecological risks. In this study, 15 antibiotics and 10 ARGs in a source of drinking water were analyzed using solid-phase extraction-ultra performance liquid chromatography-mass spectrometry technology (SPE-UPLC-MS/MS) and real-time fluorescence quantitative polymerase chain reaction (qPCR). Fourteen antibiotics were detected in the samples from 18 water sources, with the highest concentration detected for tetracycline, reaching up to 1708.33 ng/L. The detection rates of norfloxacin, enrofloxacin, ofloxacin, tetracycline, and roxithromycin were 100%. The concentrations of antibiotics were highest in She Shui, followed by the Wuhan stretch of the lower reaches of the YR, whereas the lowest concentrations were found in the Wuhan stretch of the upper reaches of the YR which were approximately equal to those in the Han River (HR). Ofloxacin and roxithromycin presented a substantial threat to aquatic organisms with high sensitivity at the majority of the sampling sites. The overall abundance of ARGs was notably greater in the lower reaches of the YR compared with the upper reaches and the HR. The highest absolute abundance was observed for sulfa ARGs. Integron intl1 strongly correlated with sul1, sul2, ermB, and qnrS, and antibiotics, strongly correlated with multiple ARGs, suggesting that antibiotics and ARGs are present in water sources in Wuhan and may present a plausible hazard to both human and ecological well-being. Hence, regulating the spread and dissemination of antibiotics and ARGs in the environment is imperative. The findings of this research offer significant insights into the stewardship and safeguarding of aquatic reserves in the Wuhan stretch of the YR.

Interactive effect of silicon and zinc on cadmium toxicity alleviation in wheat plants.

Silicon (Si) and Zinc (Zn) have been frequently used to alleviate cadmium (Cd) toxicity, which are feasible strategies for crop safety production. However, the mechanisms underlying the interaction of Si and Zn on alleviating Cd toxicity are not well understood. A hydroponic system was adopted to evaluate morphological, physiological-biochemical responses, and related gene expression of wheat seedlings to Si (1 mM) and Zn (50 µM) addition under Cd stress (10 µM). Cd induced obvious inhibition of wheat growth by disturbing photosynthesis and chlorophyll synthesis, provoking generation of reactive oxygen species (ROS) and interfering ion homeostasis. Cd concentration was decreased by 68.3%, 43.1% and 73.3% in shoot, and 78.9%, 44.1% and 85.8% in root by Si, Zn, and combination of Si with Zn, relative to Cd only, respectively. Si and Zn effectively ameliorated Cd toxicity and enhanced wheat growth; but single Si or combination of Si with Zn had more efficient ability on alleviating Cd stress than only Zn, indicating Si and Zn have synergistic effect on Cd toxicity; Interaction of them alleviated oxidative stress by reducing ROS content, improving AsA-GSH cycle and antioxidant enzymes activities, and regulating Cd into vacuole through PC-Cd complexes transported by HMA3 transporter. Our results suggest that fertilizers including Si and Zn should be made to reduce Cd content, which will beneficial for food production and safety.

Integrated physio-biochemical and transcriptomic analysis revealed mechanism underlying of Si-mediated alleviation to cadmium toxicity in wheat.

Cadmium (Cd) contamination has resulted in serious reduction of crop yields. Silicon (Si), as a beneficial element, regulates plant growth to heavy metal toxicity mainly through reducing metal uptake and protecting plants from oxidative injury. However, the molecular mechanism underlying Si-mediated Cd toxicity in wheat has not been well understood. This study aimed to reveal the beneficial role of Si (1 mM) in alleviating Cd-induced toxicity in wheat (Triticum aestivum) seedlings. The results showed that exogenous supply of Si decreased Cd concentration by 67.45% (root) and 70.34% (shoot), and maintained ionic homeostasis through the function of important transporters, such as Lsi, ZIP, Nramp5 and HIPP. Si ameliorated Cd-induced photosynthetic performance inhibition through up-regulating photosynthesis-related genes and light harvesting-related genes. Si minimized Cd-induced oxidative stress by decreasing MDA contents by 46.62% (leaf) and 75.09% (root), and helped re-establish redox homeostasis by regulating antioxidant enzymes activities, AsA-GSH cycle and expression of relevant genes through signal transduction pathway. The results revealed molecular mechanism of Si-mediated wheat tolerance to Cd toxicity. Si fertilizer is suggested to be applied in Cd contaminated soil for food safety production as a beneficial and eco-friendly element.

Wheat PHT1;9 acts as one candidate arsenate absorption transporter for phytoremediation.

Arsenate (AsV) is one of the most common forms of arsenic (As) in environment and plant high-affinity phosphate transporters (PHT1s) are the primary plant AsV transporters. However, few PHT1s involved in AsV absorption have been identified in crops. In our previous study, TaPHT1;3, TaPHT1;6 and TaPHT1;9 were identified to function in phosphate absorption. Here, their AsV absorption capacities were evaluated using several experiments. Ectopic expression in yeast mutants indicated that TaPHT1;9 had the highest AsV absorption rates, followed by TaPHT1;6, while not for TaPHT1;3. Under AsV stress, further, BSMV-VIGS-mediated TaPHT1;9-silencing wheat plants exhibited higher AsV tolerance and lower As concentrations than TaPHT1;6-silenced plants, whereas TaPHT1;3-silencing plants had similar phenotype and AsV concentrations to control. These suggested that TaPHT1;9 and TaPHT1;6 possessed AsV absorption capacity with the former showing higher activities. Under hydroponic condition, furthermore, CRISPR-edited TaPHT1;9 wheat mutants showed the enhanced tolerance to AsV with decreased As distributions and concentrations, whereas TaPHT1;9 ectopic expression transgenic rice plants had the opposite results. Also, under AsV-contaminated soil condition, TaPHT1;9 transgenic rice plants exhibited depressed AsV tolerance with increased As concentrations in roots, straws and grains. Moreover, Pi addition alleviated the AsV toxicity. These suggested that TaPHT1;9 should be a candidate target gene for AsV phytoremediation.

Abscisic acid alleviates mercury toxicity in wheat (Triticum aestivum L.) by promoting cell wall formation.

Mercury (Hg) is a heavy metal (HM) that affects crop growth and productivity. In a previous study, we found that application of exogenous abscisic acid (ABA) alleviated growth inhibition in Hg-stressed wheat seedlings. However, the physiological and molecular mechanisms underlying ABA-mediated Hg detoxification remained unclear. In this study, Hg exposure reduced the plant fresh and dry weights and root numbers. Exogenous ABA treatment significantly resumed the plant growth, increased the plant height and weight, and enriched the roots numbers and biomass. The application of ABA enhanced Hg absorption and raised the Hg levels in the roots. In addition, exogenous ABA decreased Hg-induced oxidative damage and significantly brought down the activities of antioxidant enzymes, such as SOD, POD and CAT. Global gene expression patterns in the roots and leaves exposed to HgCl2 and ABA treatments were examined via RNA-Seq. The data showed that genes related to ABA-mediated Hg detoxification were enriched in functions related to cell wall formation. Weighted gene co-expression network analysis (WGCNA) further indicated that the genes implicated in Hg detoxification were related to cell wall synthesis. Under Hg stress, ABA significantly induced expression of the genes encoding cell wall synthesis enzymes, regulated the activity of hydrolase, and increased the concentrations of cellulose and hemicellulose, hence promoting cell wall synthesis. Taken together, these results suggest that exogenous ABA could alleviate Hg toxicity in wheat by promoting cell wall formation and suppressing translocation of Hg from roots to shoots.

Genome-Wide Identification and Analysis of FKBP Gene Family in Wheat (Triticum asetivum).

FK506-binding protein (FKBP) genes have been found to play vital roles in plant development and abiotic stress responses. However, limited information is available about this gene family in wheat (Triticum aestivum L.). In this study, a total of 64 FKBP genes were identified in wheat via a genome-wide analysis involving a homologous search of the latest wheat genome data, which was unevenly distributed in 21 chromosomes, encoded 152 to 649 amino acids with molecular weights ranging from 16 kDa to 72 kDa, and was localized in the chloroplast, cytoplasm, nucleus, mitochondria, peroxisome and endoplasmic reticulum. Based on sequence alignment and phylogenetic analysis, 64 TaFKBPs were divided into four different groups or subfamilies, providing evidence of an evolutionary relationship with Aegilops tauschii, Brachypodium distachyon, Triticum dicoccoides, Arabidopsis thaliana and Oryza sativa. Hormone-related, abiotic stress-related and development-related cis-elements were preferentially presented in promoters of TaFKBPs. The expression levels of TaFKBP genes were investigated using transcriptome data from the WheatExp database, which exhibited tissue-specific expression patterns. Moreover, TaFKBPs responded to drought and heat stress, and nine of them were randomly selected for validation by qRT-PCR. Yeast cells expressing TaFKBP19-2B-2 or TaFKBP18-6B showed increased influence on drought stress, indicating their negative roles in drought tolerance. Collectively, our results provide valuable information about the FKBP gene family in wheat and contribute to further characterization of FKBPs during plant development and abiotic stress responses, especially in drought stress.

Sulfur enhances cadmium bioaccumulation in Cichorium intybus by altering soil properties, heavy metal availability and microbial community in contaminated alkaline soil.

Cadmium (Cd) contamination seriously threatens the soil health and food safety. Combination of amendment and accumulator plant is a green and effective technique to improve phytoremediation of Cd-contaminated alkaline soil. In this study, a potting experiment was conducted to investigate the effect of sulfur on Cd phytoextraction by Cichorium intybus (chicory). Soil chemical and microbial properties were determined to reveal the mechanism of sulfur-assisting Cd phytoremediation by chicory. Soil pH decreased from 7.77 to the lowest 7.30 with sulfur addition (0.6, 0.9 and 1.2 g kg-1, LS, MS and HS treatment); Electric conductivity, sulfate anion and available cadmium concentration increased gradually with increasing sulfur doses. Cd concentration of shoot and root significantly increased from 1.47 to 4.43 mg kg-1, 6.15 to 20.16 mg kg-1 by sulfur treatment relative to CK, which were attributed to increased available Cd concentration induced by decreased pH. Sulfur treatments significantly increased the Cd bioconcentration factor by 64.1%, 118.6%, 201.0% for shoot, 76.3%, 145.6% and 227.7% for root under LS, MS and HS relative to CK treatment, respectively (P < 0.05). However, only MS treatment significantly improved the Cd removal efficiency by 82.9% in comparison of CK treatment (P < 0.05). Microbial community diversity measured by 16SrRNA showed that Thiobacillus and Actinobacteria were the key and dominant strains of soil microbial communities after sulfur addition, which played a pivotal role in the process of sulfur oxidation involved in decrease of soil pH and the transformation of Cd forms. Correlation analysis and path analysis by structural equation model indicated that soil sulfate anion and Thiobacillus directly affected Cd removal efficiency by chicory in Cd-contaminated alkaline soil. This suggests that combination of sulfur and chicory may provide a way to promote Cd bioaccumulation for phytoremediation of Cd-contaminated alkaline soil.

WRKY74 regulates cadmium tolerance through glutathione-dependent pathway in wheat.

Cadmium (Cd) is a toxic heavy metal to plants and human health. Ascorbate (ASA)-glutathione (GSH) synthesis pathway plays key roles in Cd detoxification, while its molecular regulatory mechanism remains largely unknown, especially in wheat. Here, we found a WRKY transcription factor-TaWRKY74, and its function in wheat Cd stress is not clear in previous studies. The expression levels of TaWRKY74 were significantly induced by Cd stress. Compared to control, the activities of GST, GR, or APX were significantly increased by 1.55-, 1.43-, or 1.75-fold and 1.63-, 2.65-, or 2.30-fold in shoots and roots of transiently TaWRKY74-silenced wheat plants under Cd stress. Similarly, the contents of hydrogen peroxide (H2O2), malondialdehyde (MDA), GSH, or Cd were also significantly increased by 2.39- or 1.25-fold, 1.54- or 1.20-fold, and 1.34- or 5.94-fold in shoots or roots in transiently TaWRKY74-silenced wheat plants, while ASA content was decreased by 47.4 or 43.3% in shoots, 10.7 or 6.5% in roots in these silenced wheat plants, respectively. Moreover, the expression levels of GSH, GPX, GR, DHAR, MDHAR, and APX genes, which are involved in ASA-GSH synthesis, were separately induced by 2.42-, 2.16-, 3.28-, 2.08-, 1.92-, and 2.23-fold in shoots, or by 10.69-, 3.33-, 3.26-, 1.81-, 16.53-, and 3.57-fold in roots of the BSMV-VIGS-TaWRKY74-inoculated wheat plants, respectively. However, the expression levels of TaNramp1, TaNramp5, TaHMA2, TaHMA3, TaLCT1, and TaIRT1 metal transporters genes were decreased by 21.2-76.3% (56.6%, 59.2%, 76.3%, 53.6%, 35.8%, and 21.2%) in roots of the BSMV-VIGS-TaWRKY74-inoculated wheat plants. Taken together, our results suggested that TaWRKY74 alleviated Cd toxicity in wheat by affecting the expression of ASA-GSH synthesis genes and suppressing the expression of Cd transporter genes, and further affecting Cd uptake and translocation in wheat plants.

Exogenous melatonin mitigates cadmium toxicity through ascorbic acid and glutathione pathway in wheat.

Cadmium (Cd) is a dispensable element that can be absorbed by crops, posing a threat to human health through the food chains. Melatonin (MT), as a plant growth regulator, has been used to alleviate Cd toxicity in many plant species; however, the underlying molecular mechanisms responsible for Cd toxicity in wheat are still poorly understood. In this study, the suitable exogenous MT concentration (50 µM) was screened to mitigate Cd toxicity of wheat plants by increasing the plant height, root length, fresh or dry weight and chlorophyll content, or decreasing the malondialdehyde (MDA) content. In addition, MT application significantly increased ascorbic acid (ASA) and glutathione (GSH) content by reducing ROS production, especially in roots, further decreasing Cd content in fraction of organelles. Moreover, the expression levels of ASA-GSH synthesis genes, APX, GR, and GST were significantly increased by 171.5%, 465.2%, and 256.8% in roots, respectively, whereas GSH, DHAR, or MDHAR were significantly decreased by 48.5%, 54.3%, or 60.0% in roots under MT + Cd stress. However, the expression levels of Cd-induced metal transporter genes TaNramp1, TaNramp5, TaHMA2, TaHMA3, and TaLCT1 were significantly decreased by 53.7%, 50.1%, 86.5%, 87.2%, and 94.5% in roots under MT + Cd stress compared with alone Cd treatment, respectively. In conclusion, our results suggesting that MT alleviate Cd toxicity in wheat by enhancing ASA-GSH metabolism, suppressing Cd transporter gene expression, and regulating Cd uptake and translocation in wheat plants.

Hormetic effects of zinc on growth and antioxidant defense system of wheat plants.

Although hormesis induced by heavy metals is a well-known phenomenon, the involved biological mechanisms are not fully understood. Zinc (Zn) is an essential micronutrient for wheat, an important crop contributing to food security as a main staple food; however, excessive Zn is detrimental to the growth of wheat. The aim of this study was to evaluate morphological and physiological responses of two wheat varieties exposed to a broad range of Zn concentrations (0-1000 µM) for 14 days. Hormesis was induced by Zn in both wheat varieties. Treatment with 10-100 µM Zn promoted biomass accumulation by enhancing the photosynthetic ability, the chlorophyll content and the activities of antioxidant enzymes. Increased root/shoot ratio suggested that shoot growth was severely inhibited when Zn concentration exceeded 300 µM by reducing photosynthetic ability and the content of photosynthetic pigments. Excessive Zn accumulation (Zn treatment of 300-1000 µM) in leaf and root induced membrane injuries through lipid peroxidation as malondialdehyde (MDA) content increased with increasing Zn concentration. The results show that MDA content was higher than other treatments by 16.1-151.1% and 15.0-88.3% (XN979) and 36.8-235.7% and 20.6-83.8% (BN207) in the leaves and roots under 1000 µM Zn treatment. To defend against Zn toxicity, ascorbate (AsA), glutathione (GSH), non-protein thiols (NPT) and phytochelatin (PC) content of both wheat varieties (except leaf GSH content of BN207) was increased, while, the activities of superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, and the content of soluble protein decreased by 300-1000 µM Zn. The results showed that AsA-GSH cycle and NPT and PC content of wheat seedlings play important roles in defending against Zn toxicity. This study contributes new insights into the physiological mechanisms underlying the hormetic response of wheat to Zn, which could be beneficial for optimizing plant health in changing environments and improving risk assessments.

Exogenous ascorbic acid application alleviates cadmium toxicity in seedlings of two wheat (Triticum aestivum L.) varieties by reducing cadmium uptake and enhancing antioxidative capacity.

The aggravation of soil cadmium (Cd) pollution is a serious threat to human food health and safety. To reduce Cd uptake and alleviate Cd toxicity in staple food of wheat, a completely random experiment was performed to investigate the effect of exogenous ascorbic acid (AsA) on Cd toxicity in two wheat varieties (L979 and H27). In this study, the treatments with combinations of Cd (0, 5, and 10 µmol L-1) and AsA (0, 50, and 200 µmol L-1) were applied in a hydroponic system. Toxicity induced by Cd inhibited biomass accumulation; decreased wheat growth, photosynthesis, and chlorophyll content; increased lipid peroxidation; and reduced activity of superoxide dismutase (SOD), but stimulated catalase (CAT) and peroxidase (POD). The addition of AsA significantly improved the growth status by increasing the wheat biomass, chlorophyll content, photosynthetic rate, protein concentrations, and antioxidant enzyme activity. Besides, AsA significantly decreased Cd concentration of shoot and root by 14.1-53.9% and 20.8-59.5% in L979 and 23.7-58.8% and 22.1-58.1% in H27 under Cd5, and 23.7-53.6% and 16.6-57.1% in L979 and 21.5-51.6% and 15.3-54.0% in H27 under Cd10, respectively. Malondialdehyde (MDA) accumulation was decreased remarkably with the addition of AsA by 31.2-32.9% in L979 and 27.1-45.2% in H27 under Cd10, respectively. Overall, exogenous application of AsA alleviated the Cd toxicity in wheat plants by improving the wheat growth, soluble protein content, photosynthesis, and antioxidant defense systems, and decreasing MDA accumulation.

Total thyroidectomy for giant nodular goiter guided by pre-operative 3D computed tomography reconstruction and 3D printing: A case report.

BACKGROUND: Nodular goiter is a common clinical problem, and thyroidectomy is preferred in patients with obstructive symptoms. Thyroidectomy is a complex procedure with some common complications. Three-dimensional (3D) computed tomography (CT) reconstruction and 3D printing provide visualized 3D anatomical structure, posing an enormously valuable potential in precise surgery with optimal efficacy and minimum complications. Here, we aimed to perform a precise thyroidectomy guided by this technology. METHODS: The patient was an 80-year-old woman with 10 years of goiter, 1 year of labored dyspnea, and a history of thyroid surgery 62 years ago. In addition to ultrasonography examination, CT images were obtained to construct the 3D model to identify the 3D relationship between the lesion and adjacent structures, and a 3D model of the trachea was created and printed using a 3D printer. RESULTS: The 3D model clearly presented the diffuse enlargement of the two lobes and isthmus and the compression of the goiter. Under the 3D guidance, the operative resection specimen of the right lobe and isthmus was 12 ´ 7 ´ 5 cm, whereas the left lobe specimen was 12 ´ 9 ´ 6 cm. Nodular goiter and lymphocytic thyroiditis were confirmed by postoperative histopathology. There were no complications after total thyroidectomy except for non-permanent hypocalcemia and hypoparathyroidism. CONCLUSION: Our results proved that total thyroidectomy for giant goiter is challenging, and 3D image-guided thyroidectomy facilitates precise and safe resection with fewer complications. 3D CT reconstruction and 3D printing can provide anatomical details and may be considered in thyroidectomy planning for patients with giant goiter.

TaWRKY74 participates copper tolerance through regulation of TaGST1 expression and GSH content in wheat.

Glutathione S-transferase (GST) is the key enzyme in glutathione (GSH) synthesis, and plays a crucial role in copper (Cu) detoxification. Nonetheless, its regulatory mechanisms remain largely unclear. In this study, we identified a Cu-induced glutathione S-transferase 1 (TaGST1) gene in wheat. Yeast one-hybrid (Y1H) screened out TaWRKY74, which was one member from the WRKY transcription factor family. The bindings between TaGST1 promoter and TaWRKY74 were further verified by using another Y1H and luciferase assays. Expression of TaWRKY74 was induced more than 30-folds by Cu stress. Functions of TaWRKY74 were tested by using transiently silence methods. In transiently TaWRKY74-silenced wheat plants, TaWRKY74 and TaGST1 expression, GST activity, and GSH content was significantly inhibited by 25.68%, 19.88%, 27.66%, and 12.68% in shoots, and 53.81%, 52.11%, 23.47%, and 17.11% in roots, respectively. However, contents of hydrogen peroxide, malondialdehyde, or Cu were significantly increased by 2.58%, 12.45%, or 37.74% in shoots, and 25.24%, 53.84%, and 103.99% in roots, respectively. Notably, exogenous application of GSH reversed the adverse effects of transiently TaWRKY74-silenced wheat plants during Cu stress. Taken together, our results suggesting that TaWRKY74 regulated TaGST1 expression and affected GSH accumulation under Cu stress, and could be useful to ameliorate Cu toxicity for crop food safety.

TaPHT1;9-4B and its transcriptional regulator TaMYB4-7D contribute to phosphate uptake and plant growth in bread wheat.

Efficient phosphate (Pi) uptake and utilisation are essential for promoting crop yield. However, the underlying molecular mechanism is still poorly understood in complex crop species such as hexaploid wheat. Here we report that TaPHT1;9-4B and its transcriptional regulator TaMYB4-7D function in Pi acquisition, translocation and plant growth in bread wheat. TaPHT1;9-4B, a high-affinity Pi transporter highly upregulated in roots by Pi deficiency, was identified using quantitative proteomics. Disruption of TaPHT1;9-4B function by BSMV-VIGS or CRISPR editing impaired wheat tolerance to Pi deprivation, whereas transgenic expression of TaPHT1;9-4B in rice improved Pi uptake and plant growth. Using yeast-one-hybrid assay, we isolated TaMYB4-7D, a R2R3 MYB transcription factor that could activate TaPHT1;9-4B expression by binding to its promoter. Silencing TaMYB4-7D decreased TaPHT1;9-4B expression, Pi uptake and plant growth. Four promoter haplotypes were identified for TaPHT1;9-4B, with Hap3 showing significant positive associations with TaPHT1;9-4B transcript level, growth performance and phosphorus (P) content in wheat plants. A functional marker was therefore developed for tagging Hap3. Collectively, our data shed new light on the molecular mechanism controlling Pi acquisition and utilisation in bread wheat. TaPHT1;9-4B and TaMYB4-7D may aid further research towards the development of P efficient crop cultivars.

Exogenous Glutathione Alleviates Cadmium Toxicity in Wheat by Influencing the Absorption and Translocation of Cadmium.

Cadmium (Cd), a toxic heavy metal, is harmful to plants and human health. Glutathione (GSH) could alleviate Cd toxicity of plant species, whereas its mechanism responsible for wheat remains poorly understood. Here, we found that exogenous GSH application significantly increased the fresh and dry weight, root elongation, chlorophyll contents, while decreased the contents of malondialdehyde (MDA) and GSH, and translocation factor of Cd compared with Cd treatment. Moreover, GSH application significantly increased activities of antioxidant enzymes and expression of related genes, which involved in GSH synthesis, especially in roots. In addition, we found that GSH application suppressed Cd-induced expression of metal transporter genes TaNramp1, TaNramp5, TaHMA2, TaHMA3, TaLCT1 and TaIRT2 in roots. Taken together, our results suggested that GSH could alleviate Cd toxicity in wheat by increasing GSH synthesis gene expression or suppressing Cd transporter genes expression, and further affecting Cd uptake and translocation in wheat plants.

Melatonin promotes potassium deficiency tolerance by regulating HAK1 transporter and its upstream transcription factor NAC71 in wheat.

Melatonin (MT) is involved in various physiological processes and stress responses in animals and plants. However, little is known about the molecular mechanisms by which MT regulates potassium deficiency (DK) tolerance in crops. In this study, an appropriate concentration (50 µmol/L) was found to enhance the tolerance of wheat plants against DK. RNA-seq analysis showed that a total of 6253 and 5873 differentially expressed genes (DEGs) were separately identified in root and leaf tissues of the DK + MT-treated wheat plants. They functionally involved biological processes of secondary metabolite, signal transduction, and transport or catabolism. Of these, an upregulated high-affinity K transporter 1 (TaHAK1) gene was next characterized. TaHAK1 overexpression markedly enhanced the K absorption, while its transient silencing exhibited the opposite effect, suggesting its important role in MT-mediated DK tolerance. Moreover, yeast one-hybrid (Y1H) was used to screen the upstream regulators of TaHAK1 gene and the transcription factor TaNAC71 was identified. The binding between TaNAC71 and TaHAK1 promoter was evidenced by using Y1H, LUC, and EMSA assays. Transient overexpression of TaNAC71 in wheat protoplasts activated the TaHAK1 expression, whereas its transient silencing inhibited the TaHAK1 expression and aggravated the sensitivity to DK. Exogenous MT application greatly upregulated the expression of TaHAK1 in both transient overexpression and silencing systems. Our findings revealed some molecular mechanisms underlying MT-mediated DK tolerance and helped broaden its practical application in agriculture.

Functional characterization and regulatory mechanism of wheat CPK34 kinase in response to drought stress.

BACKGROUND: Drought is one of the most adverse environmental factors limiting crop productions and it is important to identify key genetic determinants for food safety. Calcium-dependent protein kinases (CPKs) are known to be involved in plant growth, development, and environmental stresses. However, biological functions and regulatory mechanisms of many plant CPKs have not been explored. In our previous study, abundance of the wheat CPK34 (TaCPK34) protein was remarkably upregulated in wheat plants suffering from drought stress, inferring that it could be involved in this stress. Therefore, here we further detected its function and mechanism in response to drought stress. RESULTS: Transcripts of the TaCPK34 gene were significantly induced after PEG-stimulated water deficiency (20% PEG6000) or 100 µM abscisic acid (ABA) treatments. The TaCPK34 gene was transiently silenced in wheat genome by using barley stripe mosaic virus-induced silencing (BSMV-VIGS) method. After 14 days of drought stress, the transiently TaCPK34-silenced wheat seedlings showed more sensitivity compared with control, and the plant biomasses and relative water contents significantly decreased, whereas soluble sugar and MDA contents increased. The iTRAQ-based quantitative proteomics was employed to measure the protein expression profiles in leaves of the transiently TaCPK34-silenced wheat plants after drought stress. There were 6103 proteins identified, of these, 51 proteins exhibited significantly altered abundance, they were involved in diverse function. And sequence analysis on the promoters of genes, which encoded the above identified proteins, indicated that some promoters harbored some ABA-responsive elements. We determined the interactions between TaCPK34 and three identified proteins by using bimolecular fluorescent complementation (BiFC) method and our data indicated that TaCPK34directly interacted with the glutathione S-transferase 1 and prx113, respectively. CONCLUSIONS: Our study suggested that the TaCPK34 gene played positive roles in wheat response to drought stress through directly or indirectly regulating the expression of ABA-dependent manner genes, which were encoding identified proteins from iTRAQ-based quantitative proteomics. And it could be used as one potential gene to develop crop cultivars with improved drought tolerance.