Efficient In Vitro Regeneration System and Comparative Transcriptome Analysis Offer Insight into the Early Development Characteristics of Explants from Cotyledon with Partial Petiole in Small-Fruited Pepper (Capsicum annuum).

In our research, we utilized six small-fruited pepper germplasms as materials, selected cotyledons with the petiole and hypocotyls as explants, and conducted in vitro regeneration studies. Our outcomes specify that the most suitable explant is cotyledon with the petiole, and the suitable genotype is HNUCA341. The optimal medium for inducing and elongating adventitious buds for this genotype is Murashige and Skoog medium (MS) + 9.12 µM Zeatin (ZT) + 0.57 µM 3-Indoleacetic acid (IAA), with a bud induction rate of 44.4%. The best rooting induction medium is MS + 1.14 µM IAA, with a rooting rate of 86.7%. Research on the addition of exogenous hormones has revealed that the induction speed of buds in small-fruited pepper (HNUCA341) in the combination of ZT and IAA hormones (abbreviated as ZI) is quicker, and the induction effect is better. The histological observations indicate that ZI treatment accelerates the initiation of explant division and differentiation, causing a shorter duration of vascular-bundle tissue production. The plant hormone signaling pathway was significantly enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, including ARR9 (LOC107843874, LOC107843885), ARR4 (LOC107848380, LOC107862455), AHK4 (LOC107870540), AHP1 (LOC107839518), LAX2 (LOC107846008), SAUR36 (LOC107852624), IAA8 (LOC107841020), IAA16 (LOC107839415), PYL4 (LOC107843441), and PYL6 (LOC107871127); these significantly enriched genes may be associated with in vitro regeneration. In addition, the carbon metabolism pathway and plant mitogen-activated protein kinase (MAPK) signaling pathway are also significantly enriched in KEGG. The results of the Gene Ontology (GO) analysis revealed that differentially expressed genes related to carbon metabolism and fixation, photosynthesis and MAPK signaling pathways were upregulated under ZI treatment. It was found that they might be associated with enhanced regeneration in vitro. Furthermore, we also screened out differentially expressed transcription factors, primarily from the MYB, bHLH, AP2/ERF, and NAC families. Overall, our work accumulated important data for the in-depth analysis of the molecular mechanism of in vitro regeneration of pepper, and provides valuable germplasm for establishing an efficient stable pepper genetic-transformation system based on tissue culture.

Intensive masculinization caused by chronic heat stress in juvenile Cynoglossus semilaevis: Growth performance, gonadal histology and gene responses.

The sea temperature has been observed to chronically increase during the past decades, leaving unpredictable influences to the marine biological resources. Thus, it is of vital significance to study the biological responses of ocean inhabited organisms with the artificially stimulated heat stress environment. Cynoglossus semilaevis provides us with an ideal model to study the influence of chronic heat stress on the sexual differentiation in marine teleosts for its genetic sex determination (GSD) + environmental effected (EE) sex determination system. In this study, the comparative experiment was conducted employing heated seawater (HT group) and ambient seawater (CT group) to cultivate juvenile C. semilaevis respectively. Significant differences were exhibited in growth performance and a delayed germ cell development effect was found in pseudomales formed under chronic heat stress. Using transcriptome analysis, the transcription profile of 55 days post fertilization (dpf) and 100 dpf juveniles' gonads were studied. A total of 47 libraries were constructed with an average mapping rate of 94.63% after assembling. GO and KEGG enrichment were proceeded using DEGs screened out between (1) pseudomale gonads at 55 dpf and 100 dpf in HT and CT group (2) pseudomale and female gonads at 55 dpf and 100 dpf in HT and CT group. Terms and pathways involved in steroid stimulation, reproduction ability, germ cell proliferation et al. were shed light on. The expression pattern of 29 DEGs including amh, hsp90b1, pgr et al. were also provided to supplement the results of functional enrichment. Weighted gene co-expression networks analysis (WGCNA) was constructed and hspb8-like, histone H2A.V were exhibited to play vital roles in the heat-induced masculinization. Our findings facilitate the understanding for transcriptional variations in intensive masculinization cause by chronic heat stress of C. semilaevis and provide referable study of the influences on the teleosts in elevated sea temperature.

Histological and Gene Expression Analyses in Banana Reveals the Pathogenic Differences between Races 1 and 4 of Banana Fusarium Wilt Pathogen.

Fusarium oxysporum f. sp. cubense, the causative agent of Panama disease, is classified into three races: Foc1, Foc2, and Foc4. However, the histological characteristics, the accumulation of fusaric acid (FA), and resistant gene expression in banana infected with different races remain unclear. In this study, we compared the infection processes, FA contents, and gene expression levels in a Cavendish banana cultivar (Musa AAA Brazilian) inoculated with Foc1 and Foc4. Results showed that Foc4 can rapidly extend from the roots to the leaves, whereas Foc1 expands slowly from the roots to the rhizomes but cannot expand further upward. In addition, the colonization of plants by Foc4 was significantly higher compared with Foc1, as was the content of FA in those infected plant tissues. We observed that a large amount of starch granules was produced in the rhizomes and the number of starch granules was significantly higher after infection with Foc1 than after infection with Foc4. We further found that starch has an important inhibitory effect on the phytotoxicity induced by FA, thus leading to more resistance to the pathogens in the plants with high amounts of starch accumulation than in those with a low amount of starch accumulation. Moreover, the expression levels of 10 defense-related genes were analyzed and the results showed that the induction levels of those genes were higher after infection with Foc1 than after infection with Foc4. These results suggest that the observed differences in the invasion of host tissues and FA accumulation, and the number of starch granules and expression of defense-related genes, may contribute to a difference in virulence between the two races and the resulting difference in host resistance response, respectively.

Virus infection reduces shoot proliferation of in vitro stock cultures and ability of cryopreserved shoot tips to regenerate into normal shoots in 'Gala' apple (Malus × domestica).

Plant cryopreservation has provide secure back-ups of germplasm collections of vegetatively propagated crops. Often, recovery levels vary among laboratories when the same cryogenic procedures are used for the same genotypes. The present study investigated the effects of Apple stem grooving virus (ASGV) on shoot proliferation of in vitro stock cultures and recovery of cryopreserved shoot tips of 'Gala' apple. Results showed that virus infection reduced shoot proliferation of in vitro stock cultures and cell ability to regenerate normal shoots in cryopreserved shoot tips. Virus infection increased total soluble protein, total soluble sugar and free proline levels and altered endogenous levels of indoleacetic acid (IAA) and zeatin riboside (ZR), but induced severe cell membrane damage and caused alternation in mitochondria shape of the in vitro stock shoots. The altered levels of IAA and ZR were most likely to be responsible for the reduced shoot proliferation of in vitro stock culture. Cell damage and alternations in mitochondria shape in ASGV-infected shoot tips were most likely responsible for the reduced cell ability to regenerate normal shoots following cryopreservation. To the best of our knowledge, this is the first study on effects of virus infection on recovery of cryopreserved shoot tips. Results reported here emphasize that healthy in vitro stock cultures should be used for cryopreservation.

Study on the effects of microencapsulated Lactobacillus delbrueckii on the mouse intestinal flora.

OBJECTIVE: To evaluate the protective effects of microencapsulation on Lactobacillus delbrueckii by random, parallel experimental design. MATERIALS AND METHODS: Lincomycin hydrochloride-induced intestinal malfunction mouse model was successfully established; then the L. delbrueckii microcapsule was given to the mouse. The clinical behaviour, number of intestinal flora, mucous IgA content in small intestine, IgG and IL-2 level in peripheral blood were monitored. The histological sections were also prepared. RESULTS: The L. delbrueckii microcapsule could have more probiotic effects as indicated by higher bifidobacterium number in cecal contents. The sIgA content in microcapsule treated group was significantly higher than that in non-encapsulated L. delbrueckii treated group (p < 0.05). Intestine pathological damage of the L. delbrueckii microcapsule-treated group showed obvious restoration. CONCLUSION: The L. delbrueckii microcapsules could relieve the intestinal tissue pathological damage and play an important role in curing antibiotic-induced intestinal flora dysfunction.

Combined effect of polystyrene nanoplastic and di-n-butyl phthalate on testicular health of male Swiss albino mice: analysis of sperm-related parameters and potential toxic effects.

Plastics, especially polystyrene nanoplastic particles (PSNPs), are known for their durability and absorption properties, allowing them to interact with environmental pollutants such as di-n-butyl phthalate (DBP). Previous research has highlighted the potential of these particles as carriers for various pollutants, emphasizing the need to understand their environmental impact comprehensively. This study focuses on the subchronic exposure of male Swiss albino mice to PSNP and DBP, aiming to investigate their reproductive toxicity between these pollutants in mammalian models. The primary objective of this study is to examine the reproductive toxicity resulting from simultaneous exposure to PSNP and DBP in male Swiss albino mice. The study aims to analyze sperm parameters, measure antioxidant enzyme activity, and conduct histopathological and morphometric examinations of the testis. By investigating the individual and combined effects of PSNP and DBP, the study seeks to gain insights into their impact on the reproductive profile of male mice, emphasizing potential synergistic interactions between these environmental pollutants. Male Swiss albino mice were subjected to subchronic exposure (60 days) of PSNP (0.2 mg/m, 50 nm size) and DBP (900 mg/kg bw), both individually and in combination. Various parameters, including sperm parameters, antioxidant enzyme activity, histopathological changes, and morphometric characteristics of the testis, were evaluated. The Johnsen scoring system and histomorphometric parameters were employed for a comprehensive assessment of spermatogenesis and testicular structure. The study revealed non-lethal effects within the tested doses of PSNP and DBP alone and in combination, showing reductions in body weight gain and testis weight compared to the control. Individual exposures and the combination group exhibited adverse effects on sperm parameters, with the combination exposure demonstrating more severe outcomes. Structural abnormalities, including vascular congestion, Leydig cell hyperplasia, and the extensive congestion in tunica albuginea along with both ST and Leydig cell damage, were observed in the testis, underscoring the reproductive toxicity potential of PSNP and DBP. The Johnsen scoring system and histomorphometric parameters confirmed these findings, providing interconnected results aligning with observed structural abnormalities. The study concludes that simultaneous exposure to PSNP and DBP induces reproductive toxicity in male Swiss albino mice. The combination of these environmental pollutants leads to more severe disruptions in sperm parameters, testicular structure, and antioxidant defense mechanisms compared to individual exposures. The findings emphasize the importance of understanding the interactive mechanisms between different environmental pollutants and their collective impact on male reproductive health. The use of the Johnsen scoring system and histomorphometric parameters provides a comprehensive evaluation of spermatogenesis and testicular structure, contributing valuable insights to the field of environmental toxicology.

Micro-Computed Tomography Analysis and Histological Observation of the Screw-Bone Interface of Novel Porous Scaffold Core Pedicle Screws and Hollow Lateral Hole Pedicle Screws: A Comparative Study in Bama Pigs.

OBJECTIVE: Screw loosening is a common complication of pedicle screw internal fixation surgery. This study aimed to investigate whether the application of a porous scaffold structure can increase the contact area between screws and bone tissue by comparing the bone ingrowth and screw-bone interface of porous scaffold core pedicle screws (PSCPSs) and hollow lateral hole pedicle screws (HLHPSs) in the lumbar spine of Bama pigs. METHODS: Sixteen pedicle screws of both types were implanted into the bilateral pedicles of the L1-4 vertebrae of 2 Bama pigs. All Bama pigs were sacrificed and the lumbar spine was freed into individual vertebrae at 16 weeks postoperatively. After the vertebrae were made into screw-centered specimens, micro-computed tomography analysis and histological observation were performed to assess the screw-bone interface and bone growth around and within the screws. RESULTS: We found that the bone condition around PSCPSs and HLHPSs did not show significant differences on micro-computed tomography three-dimensional reconstruction images. CT transverse views showed different bone growth inside the 2 screws. In PSCPSs, bone tissue was seen to fill the internal pores and was evenly distributed around each strut. Inside HLHPSs, bone growth was confined to 1 side of the screw and did not fill the entire cavity. Osteometric analysis showed that bone volume fraction and trabecular number, the parameters representing bone mass, were higher in PSCPSs than in HLHPSs. These differences were not statistically significant (P > 0.05). Histological observations visualized that the osseointegration within PSCPSs was superior to that of HLHPSs, and the tight integration of bone tissue with the porous scaffold resulted in a larger screw-bone integration area in PSCPSs than in HLHPSs. CONCLUSIONS: Compared with HLHPSs, PSCPSs possessing a porous scaffold core could promote bone ingrowth and osseointegration, resulting in an effective enhancement of the combined area of the screw-bone interface.

Effect of Growth Stages on Anthocyanins and Polyphenols in the Root System of Sweet Potato.

The storage roots of purple-fleshed sweet potato contain a variety of anthocyanins and polyphenols. Little is known about changes in the total content and composition of anthocyanins and polyphenols in the early growth stages of the root system. In this study, we investigated the changes in anthocyanins and polyphenols in the root system of purple-fleshed sweet potato cultivars at 15, 30, 45, and 60 days after transplant (DAT). Unexpectedly, the highest percentage of acylated anthocyanins in three purple-fleshed cultivars among all growth stages was at 15 DAT. On the other hand, the total polyphenol content in the early growth stages of the root system increased rapidly toward 45 DAT, just before the beginning of storage root enlargement, and then decreased rapidly as the storage roots began to enlarge. These data indicate that the early growth stage of the root system is a critical time. This timing may present a strategy to maximize the accumulation of polyphenols with high antioxidant activity, as well as acylated anthocyanins, to protect against abiotic and biotic stresses.

Survey anatomy and histological observation of the nasal cavity of Tupaia belangeri chinensis (Tupaiidae, Scandentia, Mammalia).

This study aimed to provide researchers with an atlas of the survey anatomy, histology, and imaging of the nasal cavity of Tupaia belangeri chinensis. Seven T. b. chinensis adult males were euthanized and scanned using micro-computed tomography (CT). The nose was separated, and tissue sections were made on the coronal and axial planes to observe the survey anatomy and histological and imaging characteristics of the nose. T. b. chinensis contains one maxilloturbinal and three ethmoturbinals, one nasoturbinal, one interturbinal, two frontoturbinals, and one lamina semicircularis in the unilateral nasal cavity. Other identified structures were the ostiomeatal complex, vomeronasal organ, superior nasal vault, maxillary sinus, and frontal recess. The drainage pathways of the sinuses and nasal airflow in T. b. chinensis were confirmed. The vault epithelium consisted of the squamous epithelium, respiratory epithelium, transitional epithelium, and olfactory epithelium. Micro-CT confirmed our findings of the coronal tissue sections. The nasal cavity anatomy of T. b. chinensis is similar to that of some strepsirrhine primates. However, the airflow and olfactory function are quite different from that of humans. Our gross and histological atlas of the nasal septum, turbinals, maxillary sinus, and frontal recess provides a reference for researchers to use T. b. chinensis for nasal cavity functional research.

Transcriptomic Study of Spermatogenesis in the Testis of Hu Sheep and Tibetan Sheep.

Numerous genes involved in male reproduction regulate testis development and spermatogenesis. In this study, the testis tissue transcriptome was used to identify candidate genes and key pathways associated with fecundity in sheep. Histological analysis of testis tissue using hematoxylin-eosin (HE) routine staining was performed for two sheep breeds. Overall, 466 differentially expressed genes (DEGs) were identified between Hu sheep (HS) and Tibetan sheep (TS) through RNA sequencing technology (RNA-Seq), including 226 upregulated and 240 downregulated genes. Functional analysis showed that several terms and pathways, such as "protein digestion and absorption", "cAMP signaling pathway", "focal adhesion", and "p53 signaling pathway" were closely related to testis development and spermatogenesis. Several genes (including COL1A1, COL1A2, COL3A1, SOX9, BCL2, HDC, and GGT5) were significantly enriched in these terms and pathways and might affect the reproduction of sheep by regulating the migration of spermatogenic cells, apoptosis of spermatogenic cells, and secretion of sterol hormones via testicular interstitial cells. Our results provide a theoretical basis for better understanding the molecular mechanisms of reproduction in sheep.

Comparative Transcriptome Analyses Characterize Expression Signatures Among Males, Females, Neo-Males, and Gynogenetic Females in the Yellow Drum (Nibea albiflora).

The yellow drum (Nibea albiflora) is one of the most important marine economic fish in China, and its sexually dimorphic growth makes it preferable for mono-sex culture. Although gynogenesis and neo-male induction techniques have been established, the molecular pathways and regulatory mechanisms of sex determination and maintenance in gynogenetic females and neo-males remains far from fully understood. In this study, the gene expression profiles were investigated in the gonads and brains of wild-type male, wild-type female, neo-male, and gynogenetic female yellow drum using comparative transcriptome analyses. Generally, a total of 52,999 novel transcripts were obtained in RNA-seq, of which 45,651 were isoforms of known protein-coding genes, 1,358 novel protein-coding genes, and 5,990 long non-coding RNAs. We found that the differences between wild-type males and neo-males and between wild-type females and gynogenetic females were relatively small at both the histological and transcriptomic levels, indicating that artificial gynogenesis or hormonal sex reversal may have minimal effects on normal female or male life function, respectively. In the brain, pathways such as "Oocyte meiosis", "Cell cycle", and "Riboflavin metabolism" were found to be significantly enriched. In the gonads, pathways such as "Prolactin signaling pathway", "PPAR signaling pathway", "Cholesterol metabolism", and "Jak-STAT signaling pathway" were found to play important roles in maintaining the regular proliferation and differentiation of females and males in yellow drum. In particular, we found that zp4 might be an effective molecular marker to differentiate between gynogenetic and normal females owing to its unique expression pattern. The results of this study may help to elucidate the molecular mechanisms involved in sex maintenance in the gonads and brain and provide basic data for genetic breeding of the yellow drum.

Key infection stages defending heat stress in high-temperature-resistant Blumeria graminis f. sp. tritici isolates.

With the increase of temperature in the winter wheat-growing regions in China, the high-temperature-resistant Blumeria graminis f. sp. tritici (Bgt) isolates developed in the fields. To clarify the key infection stages and the roles of heat shock protein (HSP) genes of high-temperature-resistant Bgt isolates defending high temperature, 3 high-temperature-resistant and 3 sensitive Bgt isolates were selected from 55 isolates after determination of temperature sensitivity. And then they were used to investigate the infection stages and the expression levels of HSP genes, including Bgthsp60, Bgthsp70, Bgthsp90, and Bgthsp104, at 18°C and 25°C. The formation frequency of abnormal appressoria and inhibition rate of haustoria formation of high-temperature-resistant isolates at 25°C were lower than those of high-temperature-sensitive isolates, while major axis of microcolonies of high-temperature-resistant isolates was higher than those of high-temperature-sensitive isolates at 25°C. The results indicated that haustoria formation and hyphal expansion were the key infection stages of defense against heat stress in high-temperature-resistant isolates. Further analyses of HSP genes found the expression levels of Bgthsp60 and Bgthsp70c were upregulated at 24 and 72 h post-inoculation in high-temperature-resistant isolates, while no significant difference was observed for Bgthsp90 and Bgthsp104 genes. Taken together, the basis of high-temperature-resistant Bgt isolates is associated with induced expression of Bgthsp60 and Bgthsp70c response to heat stress in haustoria formation and hyphal expansion stages.

Histological and proteome analyses of Microbacterium foliorum-mediated decrease in arsenic toxicity in Melastoma malabathricum.

Arsenic (As) is an increasing threat across the globe, widely known as a non-threshold carcinogen, and it is reaching harmful values in several areas of the world. In this study, the effect of plant growth promoting bacteria (Microbacterium foliorum) on inorganic arsenic (Arsenate) phytoremediation by Melastoma malabathricum plants was investigated through histological analysis and proteome profiling of the M. malabathricum plants. Two-dimensional gel electrophoresis and transmission electron microscopy were used to conduct the proteome and histological analysis. When arsenic-treated cells were compared to untreated cells, substantial changes were found (1) severely altered the morphology of the cells, intensely disturbed; (2) the cell wall was thicker; (3) drastically changed the cytoplasm, the cells were polygonal in shape, different in size (scattered), and relatively dense. Compared to the control group, the ultra-structure of the root cells of the control group revealed intact cytoplasm, vacuole, and cell wall under exposure to As + bacteria that had a minor effect on the cell form. To further understand As + bacteria interaction, proteome profiling of the root cell was analyzed. The As-induced oxidative stress enrichment was confirmed by the up-regulation of tubulin, nucleoside diphosphate kinase, and major allergen during As + bacteria exposure It was observed that the profusion of proteins involved in defence, protein biogenesis, signaling, photosynthesis, nucleoside and energy metabolism was greater in As + bacteria as compared to the rooting out of As only. Overall, it can be obviously seen that the current study demonstrates the effectiveness of phytoremediation by M. foliorum on proteins involved and responsive pathways in dealing with As toxicity in M. malabathricum plant.

Identification and Characterization of Dimorphic Expression of Sex-Related Genes in Rock Bream, a Fish With Multiple Sex Chromosomes.

The rock bream (Oplegnathus fasciatus) is a typical fish with a unique multiple sex chromosome system. In this study, we investigated the gene expression profiling in the gonads and brains of both males and females using RNA-Seq to identify sex-related genes and pathways. In accordance with the dimorphic expression profiles, combined with Gene ontology and KEGG enrichment analyses, a number of potential genes and pathways associated with sex determination were obtained from transcriptional analysis, especially some sex-biased genes and pathways. Next, we selected 18 candidate genes and analyzed their expression in different tissues and developmental stages. We found that the expression levels of Amh, Dmrt1, Sox9, Dmrtb1, and Nanos2 were significantly higher in the testis than those in the ovary or other tissues, whereas the expression levels of ZP4, Bouncer, RNF208, FoxH1, and TOB were significantly higher in the ovary than those in the testis. Furthermore, the expression levels of these genes in different developmental stages of gonads also showed sexually dimorphic patterns, suggesting that they might play important roles during gonadal development. These genes are useful markers for investigating sex determination and differentiation in rock bream. The findings of this study can provide insights into the molecular mechanisms of sex determination and differentiation in fish with multiple sex chromosome systems.

Histological Evaluation of Porous Additive-Manufacturing Titanium Artificial Bone in Rat Calvarial Bone Defects.

Jaw reconstruction using an additive-manufacturing titanium artificial bone (AMTAB) has recently attracted considerable attention. The synthesis of a titanium artificial bone is based on three-dimensional computed tomography images acquired before surgery. A histological evaluation of porous AMTAB (pAMTAB) embedded in rat calvarial bone defects was conducted. This study examined three groups: rats implanted with mixed-acid and heat-treated pAMTAB, rats implanted with untreated pAMTAB, and rats with no implant. In both pAMTAB groups, bone defects were created in rat calvarial bones using a 5-mm trephine bar, followed by pAMTAB implantation. The pAMTAB was fixed to the defect using the fitting force of the surrounding bones. The rats were sacrificed at 4, 8, and 16 weeks after implantation, and the skull was dissected. Undecalcified ground slides were prepared and stained with Villanueva Goldner. Compared with the no implant control group, both pAMTAB groups exhibited new bone formation inside the defect, with greater bone formation in the mixed-acid and heat-treated pAMTAB group than in the untreated pAMTAB group, but the difference was not significant. These data suggest that pAMTAB induces bone formation after implantation in bone defects. Bone formation appears to be enhanced by prior mixed-acid and heat-treated pAMTAB.

Reduced cadmium accumulation in tobacco by sodium chloride priming.

Cadmium (Cd) pollution threatens agricultural security worldwide. This study tested the efficacy of priming chemicals to decrease Cd uptake by tobacco plants (Nicotiana tabacum). After initial screening from nine different chemicals (NaCl, Cd(CH3COO)2, Cd(NO3)2, CdCl2, KHNO3, polyethylene glycol 6000 (PEG-6000), indole-3-acetic acid (IAA), ß-aminobutyric acid (BABA), and glutathione (GSH)), NaCl and PEG-6000 were further investigated because of their low risks to plant growth and efficiency to Cd reduction. Priming procedures (concentrations) were optimized for both chemicals and the best one (100 mM NaCl) was used to test both soil and hydroponic media. The results showed 31.3% lower Cd concentrations in shoots after priming with 100 mM NaCl. Phenotype parameters of the plants were also measured and showed no significant impacts of the priming procedures on the shoot biomass and the uptakes of nitrogen (N), phosphorus (P), and potassium (K), nor the photosynthetic capacity (net photosynthesis rate (Pn) and chlorophyll concentration (SPAD)). Histological observations of the roots showed a significant increase of the stele diameter after NaCl priming and a subsequent negative correlation between shoot Cd concentration and stele diameter was found after NaCl priming at different levels. This study confirmed 100 mM NaCl as an efficient priming treatment to decrease Cd uptake and the coarsening of the root stele was identified as a potential explanation for the observed decrease of Cd in tobacco shoots.

Analysis of testis metabolome and transcriptome from the oriental river prawn (Macrobrachium nipponense) in response to different temperatures and illumination times.

A better understanding of the mechanisms underlying the male sexual differentiation of Macrobrachium nipponense is urgently needed in order to maintain sustainable development of the M. nipponense industry. Environmental factors, especially temperature and illumination, have dramatic effects on gonadal development. The aim of the present study was to identify key genes and metabolites involved in the male sexual differentiation and development of M. nipponense through integrated metabolomics and transcriptome analyses of the testis in response to different temperatures and illumination times. A total of 268 differentially abundant metabolites and 11,832 differentially expressed genes (DEGs) were identified. According to integrated metabolomics and transcriptome analyses, glycerophospholipid and sphingolipid metabolism was predicted to have dramatic effects on the male sexual differentiation and development of M. nipponense. According to the KEGG enrichment analysis of DEGs, oxidative phosphorylation, glycolysis/gluconeogenesis, the HIF-1 signaling pathway, the citrate cycle, steroid hormone synthesis, and the spliceosome complex were predicted to promote male differentiation and development by providing adenosine triphosphate, promoting the synthesis of steroid hormones, and providing correct gene products. Quantitative polymerase chain reaction analysis and in situ hybridization showed that the SDHB, PDE1, HSDL1, CYP81F2, SRSF, and SNRNP40 genes were differentially expressed, suggesting roles in the male sexual differentiation and development of M. nipponense. Strong candidate sex-related metabolic pathways and genes in M. nipponense were identified by integrated metabolomics and transcriptome analyses of the testis in response to different temperatures and illumination times, as confirmed by PCR analysis and in situ hybridization.

Effects of ammonia nitrogen on gill mitochondria in clam Ruditapes philippinarum.

Ammonia nitrogen exposure has been found to significantly increase the early apoptosis rates of gill cells, affect the contents of ATP and disturb expressions of calcium-related genes in clam Ruditapes philippinarum. Mitochondria are the centers for energy production, initiation of apoptosis and calcium signal regulation. It is hypothesized that gill mitochondrion is a target organelle for the ammonia nitrogen. Thus, ATP metabolism together with ATP-consuming functions would be interfered by ammonia exposure. In the present study, mitochondrial transmembrane potential (MTP), ATPase activities, gill functions in clearance and respiration, and histological changes were detected to characterize the effects of ammonia to the gill mitochondria in clam R. philippinarum. Results indicated that ammonia exposure led to significant decreases in MTP, Ca2+-ATPase activity and clearance rates. However, different concentrations of ammonia nitrogen induced different variations on H+, K+-ATPase activity and respiration rates. Histological observation revealed that subacute exposure of ammonia damaged the microstructure of gill tissues. Therefore, ammonia exposure dramatically damaged the normal structure and function of mitochondria, resulting in irreversible damage in energy formation and supply. In addition, it affected Ca2+ and K+ metabolism and inhibited food intake and respiration in clam R. philippinarum.

Comparison of Cell and Nuclear Size Difference between Diploid and Induced Triploid in Marine Medaka, Oryzias dancena.

The influence of triploidization on cell and nucleus size characteristics of the same tissues of erythrocyte, retina, kidney, hepatocyte and midgut epithelium in marine medaka, Oryzias dancena has been determined histologically. Induced triploid fish are produced by cold shock treatments. Likewise, the size of horizontal cell nucleus in inner nuclear layer of retina, ganglion cell nucleus in ganglion cell layer of retina, proximal tubule cell of kidney, hepatocytes and nuclear height of midgut epithelium all appear to be significantly larger than diploid (p<0.05). On the other hand, retina thickness is larger in diploid than induced triploid (p<0.05). Induced triploid shows low density of cell number. Results of this study suggest that same characteristics in the induced triploid exhibiting larger cells and nucleus sizes with fewer number of cells than the diploid can be useful criteria for the distinction between diploid and induced triploid, and also the ploidy level in marine medaka.

Recovery patterns, histological observations and genetic integrity in Malus shoot tips cryopreserved using droplet-vitrification and encapsulation-dehydration procedures.

A droplet-vitrification procedure is described for cryopreservation of Malus shoot tips. Survival patterns, recovery types, histological observations, and genetic integrity were compared for Malus shoot tips cryopreserved using this droplet-vitrification procedure and an encapsulation-dehydration procedure that was previously reported by us. In both procedures, three types of shoot tip recovery were observed following cryopreservation: callus formation without shoot regrowth, leaf formation without shoot regrowth, and shoot regrowth. Three categories of histological observations were also identified in cross-sections of shoot tips recovered after cryopreservation using the two cryogenic procedures. In category 1, almost all of the cells (94-95%) in the apical dome (AD) were damaged or killed and only some cells (30-32%) in the leaf primordia (LPs) survived. In category 2, only a few cells (18-20%) in the AD and some cells (30-31%) in the LPs survived. In category 3, majority of the cells (60-62%) in the AD and some cells (30-33%) in the LPs survived. These data suggest that shoot regrowth is correlated to the presence of a majority of surviving cells in the AD after liquid nitrogen exposure. No polymorphic bands were detected by inter-simple sequence repeats or by random amplified polymorphic DNA assessments, and ploidy levels analyzed by flow cytometry were unchanged when plants recovered after cryoexposure were compared to controls. The droplet-vitrification procedure appears to be robust since seven genotypes representing four Malus species and one hybrid recovered shoots following cryopreservation. Mean shoot regrowth levels of these seven genotypes were 48% in the droplet-vitrification method, which were lower than those (61%) in the encapsulation-dehydration procedure reported in our previous study, suggesting the latter may be preferred for routine cryobanking applications for Malus shoot tips.