Pathological findings of mammary gland carcinoma and hemangiosarcoma cases in two African wild dogs (Lycaon pictus).

There are few studies on diseases affecting endangered African wild dogs. We report our findings on malignant tumors in two African wild dogs. Case 1 was a 6-year-old intact female diagnosed with inflammatory mammary carcinoma with pulmonary metastasis. Case 2 was an 11-year-old male diagnosed with primary hemangiosarcoma of the left atrial coronary sulcus with metastasis to multiple organs. Additionally, the tumor had grown through the cardiac wall, causing cardiac tamponade. The identification of disease incidence trends provides important information which will allow for the early detection and treatment of malignant tumors, and aid in the conservation of this species.

Fibrinogen Aα-chain amyloidosis outbreaks in Japanese squirrels (Sciurus lis): a potential disease model.

Fibrinogen Aα-chain amyloidosis is a hereditary systemic amyloidosis characterized by glomerular amyloid depositions, which are derived from the fibrinogen Aα-chain variant in humans. Despite its unique pathology, the pathogenic mechanisms of this disease are only partially understood. This is in part because comparative pathological studies on fibrinogen Aα-chain amyloidosis are currently unavailable as there is a lack of reported cases in animals other than humans. In this study, mass spectrometry-based proteomic analyses of Japanese squirrels (Sciurus lis) that died in five Japanese zoos showed that they developed glomerular-associated fibrinogen Aα-chain amyloidosis with an extremely high incidence rate (29/38 cases, 76.3%). The condition was found to be age-dependent in the Japanese squirrels, with 89% of individuals over 4 years of age affected. Mass spectrometry revealed that the C-terminal region of the fibrinogen Aα-chain was involved in amyloidogenesis in Japanese squirrels as well as humans. No gene variations were identified between amyloid-positive and amyloid-negative squirrels, which contrasted with the available data for humans. The results indicate that fibrinogen Aα-chain amyloidosis is a senile amyloidosis in Japanese squirrels. The results have also provided comparative pathological support that the amyloidogenic C-terminal region of the fibrinogen Aα-chain is involved in the characteristic glomerular pathology, regardless of the animal species. This study elucidates the potential causes of death in Japanese squirrels and will contribute to future comparative pathological studies of fibrinogen Aα-chain amyloidosis. © 2023 The Pathological Society of Great Britain and Ireland.

Neoplasia in captive Tsushima leopard cats (Prionailurus bengalensis euptilurus).

The Tsushima leopard cat (Prionailurus bengalensis euptilurus) is a subspecies of the mainland leopard cat that lives on the small island of Tsushima, Japan. Captive breeding has been attempted in zoos in Japan because only approximately 100 animals remain in the wild and the Tsushima leopard cat is an endangered species. There are very few reports on diseases, including tumours, of this species. We analysed the deaths of 58 Tsushima leopard cats and confirmed that nine had neoplastic disease. The average age at death of the animals with neoplasia was 14 years and tumours were the primary cause of death in all animals. Eight of the nine cases involved primary tumours of the pancreas, liver, gallbladder, tongue and salivary glands, suggesting that Tsushima leopard cats may have a predilection for digestive system tumours. This is the first report of neoplastic disease in the Tsushima leopard cat.

Dietary Weizmannia coagulans Strain SANK70258 Ameliorates Coccidial Symptoms and Improves Intestinal Barrier Functions of Broilers by Modulating the Intestinal Immunity and the Gut Microbiota.

To determine the mechanisms by which Weizmannia coagulans SANK70258 (WC) supplementation improved growth performance and coccidial symptoms, we assessed the gene expressions and the microbiota compositions in the small intestinal tissues and digestas of coccidium-infected broilers previously given WC or lasalocid-A sodium (AM). WC supplementation significantly upregulated the gene expressions related to intestinal immunity and barrier functions, such as IL17A, IL17F, IL10, cathelicidin-2 and pIgR. Body weights, and Claudin-1 and IL10 expressions were positively correlated (r = 0.41, p < 0.05 and r = 0.37, p = 0.06, respectively), whereas lesion scores of the small intestine and IL17A expression were negatively correlated (r = −0.33, p = 0.09). The microbiota analysis detected that genus Alistipes was more abundant in WC-supplemented broilers than in control, and positively correlated with body weights and Claudin-1 expression (r = 0.61, p < 0.05 and r = 0.51, p < 0.05, respectively). Intriguingly, genus Enterococcus was most abundant in WC-supplemented broilers and positively correlated with IL17A expression (r = 0.49, p < 0.05). Interestingly, Escherichia-Shigella was significantly more abundant in the small intestinal digestas of AM-administered broilers than in those of control. To summarize, WC supplementation modulated and immunostimulated the microbiotas of broilers, specifically genera Alistipes and Enterococcus, which led to the improvement of weight gain and coccidial symptoms, without disrupting the intestinal microbiota compositions, as AM did.

Impacts of Subchronic and Mild Social Defeat Stress on Plasma Putrefactive Metabolites and Cardiovascular Structure in Male Mice.

Psychosocial stress precipitates mental illnesses, such as depression, and increases the risk of other health problems, including cardiovascular diseases. In this study, we observed the effects of psychosocial stress on the histopathological features of systemic organs and tissues in a mouse psychosocial stress model, namely the subchronic and mild social defeat stress (sCSDS) model. There were several pathological findings in the tissues of both sCSDS and control mice. Mild fibrosis of the heart was observed in sCSDS mice but not in control mice. Extramedullary hematopoiesis in the spleen and hemorrhage in the lungs were observed in both the control and sCSDS mice. Focal necrosis of the liver was seen only in control mice. Furthermore, putrefactive substances in the blood plasma were analyzed because these metabolites originating from intestinal fermentation might be linked to heart fibrosis. Among them, plasma p-cresyl glucuronide and p-cresyl sulfate concentrations significantly increased owing to subchronic social defeat stress, which might influence cardiac fibrosis in sCSDS mice. In conclusion, several pathological features such as increased cardiac fibrosis and elevated plasma putrefactive substances were found in sCSDS mice. Thus, sCSDS mice are a potential model for elucidating the pathophysiology of psychosocial stress and heart failure.

Salivary microRNA and Metabolic Profiles in a Mouse Model of Subchronic and Mild Social Defeat Stress.

Identification of early biomarkers of stress is important for preventing mood and anxiety disorders. Saliva is an easy-to-collect and non-invasive diagnostic target. The aim of this study was to characterize the changes in salivary whole microRNAs (miRNAs) and metabolites in mice subjected to subchronic and mild social defeat stress (sCSDS). In this study, we identified seven upregulated and one downregulated miRNAs/PIWI-interacting RNA (piRNA) in the saliva of sCSDS mice. One of them, miR-208b-3p, which is reported as a reliable marker for myocardial infarction, was upregulated in the saliva of sCSDS mice. Histological analysis showed frequent myocardial interstitial fibrosis in the heart of such mice. In addition, gene ontology and pathway analyses suggested that the pathways related to energy metabolism, such as the oxidative phosphorylation and the pentose phosphate pathway, were significantly related to the miRNAs affected by sCSDS in saliva. In contrast, salivary metabolites were not significantly changed in the sCSDS mice, which is consistent with our previous metabolomic study on the plasma of sCSDS mice. Taken in the light of previous studies, the present study provides novel potential stress biomarkers for future diagnosis using saliva.

The Effect of Supplementation with Weizmannia coagulans Strain SANK70258 to Coccidia-Infected Broilers Is Similar to That of a Coccidiostat Administration.

To determine whether it could also improve the production performance of Eimeria-infected broilers, Weizmannia coagulans strain SANK70258 (WC) supplementation was compared with coccidiostat lasalocid-A sodium (AM) administration. First, to determine the optimum WC dose, newly hatched broiler chick groups (n = 10) were untreated or consecutively given WC (0.005%, 0.01%, 0.03%, and 0.1%) and AM until slaughter (31 days of age). At day 21, all chicks were infected with coccidia. From the economical and practical viewpoints, 0.03% WC supplementation was the best dose. Second, newly hatched broiler chick groups (n = 10) were untreated or given 0.03% WC and AM. Each group was run in triplicate. At day 21, two chicks/pen with the farthest body weights as per the group's mean body weight were spared, and the remaining inoculated with coccidia. At days 42 and 49, the WC and AM groups had significantly greater body weights and daily weight gains. Intestinal lesion scores were lower in 29-day-old AM and WC. Oocyst numbers were lower in 29- and 49-day-old AM and WC, but only 29- and 49-day-old AM had higher Escherichia coli levels. To conclude, although WC and AM induced similar growth performance in coccidium-infected chicks, unlike AM, the E. coli levels did not increase with WC.

Detection and characterization of Hyaloklossia kasumiensis (Apicomplexa: Sarcocystidae) from Nagoya Daruma pond frog, Pelophylax porosus brevipodus.

The Nagoya Daruma pond frog (Pelophylax porosus brevipodus) is an endangered anuran species endemic to the semi-aquatic environment of Japan. On June 11, 2021, four of the six Nagoya Daruma pond frogs captured from the wild and kept in an aquarium died in rapid succession. Pathological and/or parasitological examinations of two of the four carcasses revealed protozoan infection in their kidneys. The pathogen identified as Hyaloklossia kasumiensis (Apicomplexa: Sarcocystidae: Hyaloklossiinae) based on their morphological and molecular characterization. This is a new host record of H. kasumiensis. Mitochondrial cox1 sequence analysis suggest that the present species detected from Nagoya Daruma pond frog belongs to the same lineage as those found in the Tokyo Daruma pond frog (P. p. porosus) from Ibaraki Prefecture, Japan. This particular lineage may be prevalent among Pelophylax porosus ssp. distributed in Japan.

High-fat diet-induced nonalcoholic steatohepatitis is accelerated by low carnitine and impaired glucose tolerance in novel murine models.

Carnitine deficiency and impaired glucose tolerance (IGT) exacerbate liver steatosis. Given the current lack of ideal murine nonalcoholic steatohepatitis (NASH) models, we investigated new NASH models using jvs/+ mice with low carnitine and wild-type mice with low-dose alloxan-induced IGT. The jvs/+ and wild-type mice were divided into jvs/+ mice fed a high-fat diet (HFD) from 3 weeks of age (HF hetero group), wild-type mice with low-dose alloxan treatment fed HFD (AL + HF wild group), wild-type mice fed HFD (HF wild group), and two types of mice fed a normal diet-jvs/+ and wild-type (intact group). All mice were sacrificed at 20 or 40 weeks of age. All male HFD-fed mice showed obesity, IGT, high blood insulin levels, homeostatic model assessment of insulin resistance (HOMA-IR), high liver enzyme levels, and high cholesterol levels. The degree of IGT was the worst in the AL + HF wild group, and blood insulin levels and HOMA-IR score were remarkably increased from 20 to 40 weeks of age. Almost all HFD-fed mice showed steatosis, fibrosis, and lobular inflammation in the centrilobular zone. These changes were accompanied by hepatocyte ballooning and were enhanced at 40 weeks of age. Furthermore, the incidence rate of nodular hyperplasia and adenoma in both the HF hetero and AL + HF wild groups was nearly 30%. We successfully established two novel murine models of NASH using male jvs/+ mice with low carnitine and male wild-type mice with IGT that eventually developed obesity, fatty liver, insulin resistance, liver fibrosis, and tumorigenesis. These results suggest that low carnitine levels and early-stage induction of IGT are important factors in the progression of NASH to tumorigenesis, similar to human NASH.

Temporal Evolution of Microscopic Structure and Functionality during Crystallization of Amorphous Indium-Based Oxide Films.

Understanding the crystallization mechanism of amorphous metal-oxide thin films remains of importance to avoid the deterioration of multifunctional flexible electronics. We derived the crystallization mechanism of indium-based functional amorphous oxide films by using in situ X-ray diffraction (XRD) and transmission electron microscopy (TEM) measurements. Crystallization begins with surface nucleation, especially at low annealing temperatures, and proceeds simultaneous nucleation and growth in the bulk. Three-dimensional crystal growth in the film was observed when the crystallite size was sufficiently smaller than the film thickness. When the growing crystallites reached the film surface, the crystallization was dominated by two- or lower-dimensional growth. Such crystallization can be explained within the framework of the modified Avrami theory and can be varied for tailoring the electrical properties of the amorphous In2O3 film. After tailoring the film crystallinity and crystallite size, the carrier mobility was improved to >100 cm2/V·s in 30 min. Our results show that a carrier mobility of >90 cm2/V·s can be implemented for the In2O3 film with a crystallinity of >40% and a crystallite size of >70 nm by an optimized annealing process. The incorporation of Ga element into amorphous In2O3 films obviously increases the activation energy of nucleation and migration. In contrast, Sn dopants can promote the crystal growth. This is attributed to two kinds of migration mechanisms during the annealing in air, one of which is the dominant migration mechanism of oxygen interstitials in crystallized indium-tin oxide (ITO) films and the other dominated by oxygen vacancies in In2O3 and IGO films. Combining the modified Avrami theory with TEM observations, we predicted the structural evolution kinetics for indium-based amorphous oxide films and gained new insights for understanding the temporal structure-functionality relationship during crystallization.

Theileria infection with severe anemia and unhealed fracture in a sika deer Cervus nippon aplodontus (Cervidae: Cetartiodactyla).

An adult female sika deer (Cervus nippon aplodontus) inhabiting Nara Park, Nara, Japan, had broken bone injuries from a car accident. During its treatment, we found that the sika deer had severe anemia and the fracture remained unhealed throughout. Peripheral blood smear revealed piroplasms in the erythrocytes, which were identified as merozoites of undescribed Theileria species, widely found in sika deer in Japan. This is the report of a clinical case of Theileria infection, accompanied by severe anemia in a sika deer.

Shoot base responds to root-applied glutathione and functions as a critical region to inhibit cadmium translocation from the roots to shoots in oilseed rape (Brassica napus).

Glutathione (GSH) is a tripeptide involved in controlling heavy metal movement in plants. Our previous study showed that GSH, when site-specifically applied to plant roots, inhibits Cd translocation from the roots to shoots in hydroponically cultured oilseed rape (Brassica napus) plants. A factor that led to this inhibitory effect was the activation of Cd efflux from root cells. To further investigate the molecular mechanism triggered by root-applied GSH, Cd movement was non-invasively monitored using a positron-emitting tracer imaging system. The Cd absorption and efflux process in the roots were visualized successfully. The effects of GSH on Cd efflux from root cells were estimated by analyzing imaging data. Reanalysis of image data suggested that GSH applied to roots, at the shoot base, activated Cd return. Cutting the shoot base significantly inhibited Cd efflux from root cells. These experimental results demonstrate that the shoot base plays an important role in distributing Cd throughout the plant body. Furthermore, microarray analysis revealed that about 400 genes in the roots responded to root-applied GSH. Among these, there were genes for transporter proteins related to heavy metal movement in plants and proteins involved in the structure modification of cell walls.

Eosinophilic Leukaemia and Systemic Mycobacterium marinum Infection in an African Pygmy Hedgehog (Atelerix albiventris).

A two-year-old male African pygmy hedgehog (Atelerix albiventris) presented with lethargy and anorexia. Ultrasonographic and radiographic examinations revealed splenomegaly and pneumonia, respectively. Prominent leucocytosis, consisting mainly of large atypical eosinophils, was observed in a peripheral blood smear. Necropsy revealed a black scab on the left hindlimb, which was swollen, an enlarged left inguinal lymph node, firm oedematous lungs, splenomegaly and multiple nodules in the lung, spleen and liver. Histopathologically, infiltration of numerous eosinophils was seen in lung, spleen, liver, kidney, lymph node and adrenal medulla tissues. Necrotizing granulomatous lesions, with intralesional acid-fast bacilli, were found in the lung, spleen, liver, left inguinal lymph node and left hindlimb. Mycobacterium marinum was identified by broad-range polymerase chain reaction targeting of the bacterial 16S rRNA gene. To the best of our knowledge, this is only the second reported case of systemic M. marinum infection in any mammalian species.

Consecutive intra-gingival injections of lipopolysaccharide and butyric acid to mice induce abnormal behavior and changes in cytokine concentrations.

BACKGROUND: Periodontopathic bacteria such as Porphyromonas gingivalis produce several metabolites, including lipopolysaccharide (LPS) and n-butyric acid (BA). Past work suggested that periodontal infection may cause cognitive impairment in mice. AIMS: To elucidate the mechanisms by which metabolites such as LPS and BA, resulting from Porphyromonas gingivalis activity, induce immunological and physiological abnormalities in mice. METHODS: In the present work, 28 male ICR mice were placed in an open-field arena and the total distance (cm/600 s) they covered was recorded. Based on their moving distances, mice were divided into 4 groups (n = 7) and injected the following substances into their gingival tissues for 32 consecutive days: saline (C), 5 mmol/L of BA (B), 1 µg/mouse of LPS (L), and BA-LPS (BL) solutions. Distances covered by mice were also measured on days 14 and 21, with their habituation scores considered as "(moving distance on day 14 or 21)/(moving distance on day 0)". Afterwards, mice were dissected, and hippocampal gene expression and the concentrations of short-chain fatty acids, neurotransmitters and cytokines in their blood plasma and brains were analyzed. In addition, mouse brain and liver tissues were fixed and visually assessed for histopathological abnormalities. RESULTS: Group BL had significantly higher habituation scores than C and B on day 14. LPS induced higher habituation scores on day 21. LPS induced significant decreases in the mRNA levels of interleukin (IL)-6 and brain-derived neurotrophic factors, and an increase in neurotrophic tyrosine kinase receptor type 2. In both plasma and brain, LPS induced a significant acetate increase. Moreover, LPS significantly increased acetylcholine in brain. In plasma alone, LPS and BA significantly decreased monocyte chemoattractant protein 1 (MCP-1). However, while LPS significantly decreased tyrosine, BA significantly increased it. Lastly, LPS significantly decreased IL-6 and tumor necrosis factor in plasma. No histopathological abnormalities were detected in liver or brain tissues of mice. CONCLUSION: We showed that injections of LPS and/or BA induced mice to move seemingly tireless and that both LPS and BA injections strongly induced a reduction of MCP-1 in blood plasma. We concluded that LPS and BA may have been crucial to induce and/or aggravate abnormal behavior in mice.

Distinct deposition of ester-linked ferulic and p-coumaric acids to the cell wall of developing sorghum internodes.

Sorghum is important as a cereal crop, and also as livestock feed and a renewable energy crop because it produces a large amount of biomass. In grass plants like sorghum, hydroxycinnamates such as ferulic acids (FA) and p-coumaric acids (pCA) are characteristically ester-linked to the cell wall, and are believed to affect cell wall digestibility. Genetic manipulation of the esterification of FA and pCA to the cell wall appears to be one of the solutions to increase the digestibility of the cell wall so as to utilize sorghum biomass effectively. In this study, we measured esterified FA and pCA in each stage of internode elongation and determined the accumulation pattern of each hydroxycinnamate. The results revealed that FA were mainly accumulated during the cell elongation stage, and that pCA accumulation was increased after the cell elongation stage. Furthermore, 6 of the 12 sorghum BAHD acyltransferase genes were significantly expressed in the elongating internodes, suggesting that these genes might be involved in the feruloylation and/or p-coumaroylation of the cell wall in sorghum internodes.

Mutation of rice bc1 gene affects internode elongation and induces delayed cell wall deposition in developing internodes.

A rice COBRA-like gene, BRITTLE CULM1 (BC1) has been shown to be involved in assembling cell wall components and cellulose crystallinity, which determines mechanical strength in above ground organs. However, the detailed roles of BC1 in rice development are poorly understood. In this study, we found that, unlike the known brittle culm mutants, the internode length of the bc1 mutant was ~1.27 times longer than that of wild type in rice. In order to analyze the effects of bc1 mutation on internode development, we compared the deposition of cell wall components among each developmental stage of the elongating second internodes from wild type, Kinmaze, and the bc1 mutant. In wild type, histochemical observations of lignin revealed that lignin deposition was gradually increased after the cell elongation stage of the internodes. Cellulose and p-coumaric acid (pCA) content also gradually increased along with the progress of the developmental stage. The ferulic acid (FA) content rapidly increased in the cell elongation stage and decreased at the late secondary cell wall formation stage. In the bc1 mutant, the contents of cell wall components were lower than those of wild type from the cell elongation stage, in which the BC1 started to express at this stage in wild type. In the bc1 mutant, the deposition patterns of cell wall components, especially phenolic components including lignin, pCA, and FA, were delayed compared with those of wild type. These results suggest that the BC1 gene plays a role in synthesizing appropriate cell walls at each stage in the developing internode.

Effects of enhancing endogenous and exogenous glutathione in roots on cadmium movement in Arabidopsis thaliana.

Glutathione (GSH) is a thiol-containing compound involved in many aspects of plant metabolism. In the present study, we investigated how enhancing endogenous and exogenous GSH affects cadmium (Cd) movement and distribution in Arabidopsis plants cultured hydroponically. Transgenic Arabidopsis plants with a strong ability to synthesize GSH in roots were generated　by transforming the gene encoding the bifunctional γ-glutamylcysteine synthetase-glutathione synthetase enzyme from Streptococcus thermophiles (StGCS-GS). Enhancing endogenous and exogenous GSH decreased the Cd translocation ratio in different ways. Only exogenous GSH significantly inhibited Cd translocation from roots to shoots in wild-type and transgenic Arabidopsis plants. Our study demonstrated that GSH mainly functions outside root cells to inhibit Cd translocation from roots to shoots.

Spontaneous abdominal hemangiosarcoma in a ferret.

Herein, we describe the case of a 6-year-old female ferret that died within a few days of the onset of anorexia and reduced spontaneous locomotor activity. Necropsy revealed a dark red abdominal mass of unknown origin between the right lobes of the pancreas and the proximal jejunum, with massive blood retention in the peritoneal cavity. Histopathologically, spindle-shaped or sometimes polygonal tumor cells were proliferating with irregularly shaped vascular spaces containing blood components and surrounding-tissue infiltration. In some areas, tumor cells formed distinctly dilated blood vessel-like structures. Immunohistochemically, most of the tumor cells were strongly positive for CD31, but factor VIII-related antigen immunoreactivity was confined to the area with dilated blood vessel-like structures. Based on these findings, the tumor was diagnosed as an abdominal hemangiosarcoma. Abdominal hemangiosarcoma excluding cases of the liver and spleen are rare in ferrets.

Phloem-specific overexpression of AtOPT6 in Arabidopsis enhances Zn transport into shoots.

The Arabidopsis oligopeptide transporter AtOPT6 is membrane transport protein that mediated transport of glutathione in both the reduced (GSH) and oxidized (GSSG) forms. In this study, the role of AtOPT6 in glutathione distribution throughout the plant was investigated. We found that transgenic Arabidopsis overexpressing AtOPT6 under the control of a phloem-specific promoter of sucrose-proton symporter 2 (pSUC2), remarkably increased AtOPT6 transcript levels, ranging from 30- to 40-fold in shoots and 6- to 10-fold in roots, relative to the wild type. AtOPT6-overexpressing lines could elevate the foliar glutathione content; however, glutathione content in the phloem did not change. We observed that the ratio of shoot glutathione content to total glutathione content increased in AtOPT6-overexpressing lines, but not in transgenic Arabidopsis with elevated foliar GSH synthesis. These results indicate the possibility that loading and unloading of glutathione in phloem tissues are enhanced in AtOPT6-overexpressing lines under the control of pSUC2. The results of heavy metal analysis revealed that transgenic Arabidopsis overexpressing AtOPT6 under the control of pSUC2 could promote the transport of Zn into shoots as effectively as transgenic Arabidopsis with elevated foliar GSH synthesis, or wild-type plants with exogenous foliar application of GSH.

Elevated glutathione synthesis in leaves contributes to zinc transport from roots to shoots in Arabidopsis.

Glutathione (GSH) is a vital compound involved in several plant metabolic pathways. Our previous study indicated that foliar GSH application can increase zinc (Zn) levels in leafy vegetables. The objective of this study was to determine the mode of action of GSH as it relates to Zn transport from roots to shoots. Two types of transgenic Arabidopsis plants with genes for GSH synthesis, including StGCS-GS or AtGSH1 driven by the leaf-specific promoter of chlorophyll a/b-binding protein (pCab3) gene were generated. Both types of transgenic Arabidopsis plants showed significant increases in shoot GSH concentrations compared to the wild type (WT). Monitoring 65Zn movement by positron-emitting tracer imaging system (PETIS) analysis indicated that the 65Zn amount in the shoots of both types of transgenic Arabidopsis plants were higher than that in the WT. GSH concentration in phloem sap was increased significantly in WT with foliar applications of 10 mM GSH (WT-GSH), but not in transgenic Arabidopsis with elevated foliar GSH synthesis. Both types of transgenic Arabidopsis with elevated foliar GSH synthesis and WT-GSH exhibited increased shoot Zn concentrations and Zn translocation ratios. These results suggest that enhancement of endogenous foliar GSH synthesis and exogenous foliar GSH application affect root-to-shoot transport of Zn.