Molecular Cloning, Expression Analysis, and Functional Analysis of Nine IbSWEETs in Ipomoea batatas (L.) Lam.

Sugar Will Eventually be Exported Transporter (SWEET) genes play an important regulatory role in plants' growth and development, stress response, and sugar metabolism, but there are few reports on the role of SWEET proteins in sweet potato. In this study, nine IbSWEET genes were obtained via PCR amplification from the cDNA of sweet potato. Phylogenetic analysis showed that nine IbSWEETs separately belong to four clades (Clade I~IV) and contain two MtN3/saliva domains or PQ-loop superfamily and six~seven transmembrane domains. Protein interaction prediction showed that seven SWEETs interact with other proteins, and SWEETs interact with each other (SWEET1 and SWEET12; SWEET2 and SWEET17) to form heterodimers. qRT-PCR analysis showed that IbSWEETs were tissue-specific, and IbSWEET1b was highly expressed during root growth and development. In addition to high expression in leaves, IbSWEET15 was also highly expressed during root expansion, and IbSWEET7, 10a, 10b, and 12 showed higher expression in the leaves. The expression of SWEETs showed a significant positive/negative correlation with the content of soluble sugar and starch in storage roots. Under abiotic stress treatment, IbSWEET7 showed a strong response to PEG treatment, while IbSWEET10a, 10b, and 12 responded significantly to 4 °C treatment and, also, at 1 h after ABA, to NaCl treatment. A yeast mutant complementation assay showed that IbSWEET7 had fructose, mannose, and glucose transport activity; IbSWEET15 had glucose transport activity and weaker sucrose transport activity; and all nine IbSWEETs could transport 2-deoxyglucose. These results provide a basis for further elucidating the functions of SWEET genes and promoting molecular breeding in sweet potato.

Transcriptomic analysis of tuberous root in two sweet potato varieties reveals the important genes and regulatory pathways in tuberous root development.

BACKGROUND: Tuberous root formation and development is a complex process in sweet potato, which is regulated by multiple genes and environmental factors. However, the regulatory mechanism of tuberous root development is unclear. RESULTS: In this study, the transcriptome of fibrous roots (R0) and tuberous roots in three developmental stages (Rl, R2, R3) were analyzed in two sweet potato varieties, GJS-8 and XGH. A total of 22,914 and 24,446 differentially expressed genes (DEGs) were identified in GJS-8 and XGH respectively, 15,920 differential genes were shared by GJS-8 and XGH. KEGG pathway enrichment analysis showed that the DEGs shared by GJS-8 and XGH were mainly involved in "plant hormone signal transduction" "starch and sucrose metabolism" and "MAPK signal transduction". Trihelix transcription factor (Tai6.25300) was found to be closely related to tuberous root enlargement by the comprehensive analysis of these DEGs and weighted gene co-expression network analysis (WGCNA). CONCLUSION: A hypothetical model of genetic regulatory network for tuberous root development of sweet potato is proposed, which emphasizes that some specific signal transduction pathways like "plant hormone signal transduction" "Ca2+signal" "MAPK signal transduction" and metabolic processes including "starch and sucrose metabolism" and "cell cycle and cell wall metabolism" are related to tuberous root development in sweet potato. These results provide new insights into the molecular mechanism of tuberous root development in sweet potato.

Proper Glyphosate Application at Post-anthesis Lowers Grain Moisture Content at Harvest and Reallocates Non-structural Carbohydrates in Maize.

Glyphosate (GP)-based herbicides have been widely applied to crops for weed control and pre-harvest desiccation. The objective of this research was to evaluate the effects of pre-harvest GP application on maize or how it physiologically alters this crop. Here, we applied four GP treatment (Control, GP150, GP200, and GP250) on maize lines of Z58 and PH6WC belonging to different maturity groups at grain-filling stages form DAP30 to DAP45. GP application significantly decreased the grain moisture content at harvest by 22-35% for Z58 and by 15-41% for PH6WC. However, the responses of grain weight to glyphosate vary with inbred lines and application time. A high concentration of glyphosate (GP250) reduced the grain weight of Z58 and low concentrations (GP150 and GP200) did not affect, while the grain weight of PH6WC significantly decreased under glyphosate treatment. In summary, our results revealed that timely and appropriate GP application lowers grain moisture content without causing seed yield and quality loss. GP application adversely affected photosynthesis by promoting maturation and leaf senescence. Meanwhile, it also enhanced non-structural carbohydrate (soluble sugars and starch) remobilization from the vegetative organs to the grains. Hence, GP treatment coordinates plant senescence and assimilate remobilization. RNA sequencing revealed that glyphosate regulated the transcript levels of sugar signaling-related genes and induced assimilate repartitioning in grains. This work indicates the practical significance of GP application for maize seed production and harvest, which highlights the contributions of source-sink communication to maize yield in response to external stress or pre-harvest desiccant application.

Thermo/pH dual-stimuli-responsive drug delivery for chemo-/photothermal therapy monitored by cell imaging.

A thermo/pH dual-stimuli-responsive drug delivery system (DDS) based on polymer coated mesoporous silica nanostructures (MSNs) is developed for facilitating chemotherapy and photothermal therapy. Thermo/pH-responsive polymer, poly((N-isopropylacrylamide, NIPAM)-co-methacrylic acid, MA), is grafted onto MSNs by in situ polymerization, followed by loading a chemotherapeutic drug (doxorubicin hydrochloride, DOX) and a near-infrared-absorbing phototherapeutic agent (indocyanine green, ICG) to construct the intelligent drug delivery system, shortly as DOX-ICG-MSN@p(NIPAM-co-MA). At NIR irradiation, the photothermal conversion capability of ICG raises the temperature of the DDS and opens the gatekeeper by shrinkage of the copolymer p(NIPAM-co-MA), which triggers controlled release of DOX at an elevated temperature. On the other hand, drug release is also realized at pH 5.3, a characteristic pH value in cancer cell microenvironment, at which it not only causes the shrinkage of the pH-sensitive polymeric moiety of methacrylic acid in MSN@p(NIPAM-co-MA) but also deteriorates electrostatic interaction of DOX molecules in the mesoporous channel by protonation of silanols. In addition, ICG further ensures photothermal therapy (PTT) and photodynamic therapy (PDT). The cytotoxicity assay of HeLa cells shows obvious synergistic effect by demonstrating that the combined use of DOX and ICG is more effective in killing HeLa cells than free DOX and ICG. The endocytosis of the drug is monitored by cell imaging.

Redescription of 13 holotypes of Rheocricotopus Brundin, 1956 (Diptera: Chironomidae) from the Sino-Indian Region.

Thirteen holotypes of the orthoclad genus Rheocricotopus from Sino-Indian Region: R. (Psilocricotopus) hidakadeeus Sasa & Suzuki, R. (P.) isigadeeus Sasa & Suzuki, R. (P.) kurocedeus Sasa, R. (P.) tokarakeleus Sasa & Suzuki, R. (P.) tobatervicesimus Kikuchi & Sasa, R. (Rheocricotopus) inaquereus Sasa, Kitami & Suzuki, R. (R.) inaxeyeus Sasa, Kitami & Suzuki, R. (R.) shoufukusecundus Sasa, R. (R.) tamahumeralis Sasa, R. (R.) tatequintus Sasa, R. (R.) tedorisecundus Sasa, R. (R.) togapeniculus Sasa & Okazawa and R. (R.) yakulemeus Sasa & Suzuki are re-examined and illustrated, Some additional descriptions, corrections and a key of these thirteen holotypes are given.

Trace metals in the surface sediments of the intertidal Jiaozhou Bay, China: Sources and contamination assessment.

The major (Al) and trace metal (Cu, Pb, Zn, Cr, Cd, and As) concentrations in 29 surface sediment samples from the intertidal Jiaozhou Bay (JZB) are evaluated to assess the contamination level. The results show that the overall sediment quality in the area has been obviously impacted by trace metal contamination. The geoaccumulation index and the enrichment factor values indicate that no Cr or Cu contamination has occurred on the whole, only a few stations have been polluted by As, and some areas have been polluted by Cd, Pb, and Zn. Principal component analysis suggests that the Cu, Pb, Zn, and Cd are derived from anthropogenic inputs and that Cr, As, Cu, and Zn are influenced by natural weathering processes. Cu and Zn may originate from both natural and anthropogenic sources. The contamination in the northeastern JZB is higher than that in other areas of the bay.

Stimulation of expression of the intestinal glutamine transporter ATB0 in tumor-bearing rats.

BACKGROUND: Glutamine supplementation ameliorates host catabolic response in tumor bearing states. The purpose of this in vivo study was to investigate intestinal glutamine transport and expression of glutamine transporter ATB(0) in methyl-cholanthrene (MCA)-sarcoma bearing rats. METHODS: Fisher-344 rats underwent subcutaneous flank implantation of MCA-sarcoma cells (saline as control) and were pair-fed an equal quantity of chow as controls, to account for tumor-induced anorexia, until tumors reached 10 or 20% body weight. Intestinal mucosal brush border membrane [3H]-Glutamine transport was measured. Glutamine transporter ATB(0) mRNA and protein levels were measured by real-time PCR and western blot techniques, respectively. RESULTS: Glutamine transport activity across the intestinal brush border membrane (BBM) was 3.7-fold higher in tumor-bearing rats (TBR) than in controls (TBR 153 +/- 22.6 vs. Control 41.9 +/- 9.7 pmol/mg protein/10s, P < .01). Transporter ATB(0) mRNA levels were 1.4-fold higher in tumor-bearing rats (Relative value TBR .61 +/- .12 vs. Control .43 +/- .1, P < .05). A 1.4-fold increase in transporter ATB(0) protein levels was observed in the tumor-bearing rats (Relative value TBR .52 +/- .07 vs. Control .37 +/- .04, P < .05). Circulating aortic plasma glutamine levels were 1.3-fold higher in tumor bearing rats ([Glutamine] = .63 +/- .02 Control vs. [Glutamine] = .74 +/- .01 mmol/l TBR, P < .0001). Portal venous plasma glutamine levels were also higher in tumor bearing rats ([Glutamine] = .47 +/- .01 Control vs. [Glutamine] = .60 +/- .02 mmol/l TBR, P < .0001). CONCLUSION: Intestinal brush border membrane glutamine transport activity, transporter ATB(0) mRNA and protein levels are up-regulate in tumor-bearing rats.

Regulation of amino acid arginine transport by lipopolysaccharide and nitric oxide in intestinal epithelial IEC-6 cells.

As a precursor for nitric oxide (NO) synthesis and an immune-enhancing nutrient, amino acid L-arginine plays a critical role in maintaining intestine mucosal integrity and immune functions in sepsis. However, the relationship between intestinal arginine transport and NO synthesis in sepsis remains unclear. In the present study, we investigated the effects of lipopolysaccharide (LPS) and NO on the arginine transport in cultured rat intestinal epithelial IEC-6 cell. Near-confluent IEC-6 cells were incubated with LPS (0-50 microg/ml) in serum-free Dulbecco's modified Eagles's medium, in the presence and absence of the NO donor sodium nitroprusside (SNP, 0-500 micromol/L) and the inducible nitric oxide synthase (iNOS) inhibitor N-omega-nitro-L-arginine (NNA, 0-1000 micromol/L) for various periods of time (0-48 hours). Arginine transport activity, arginine transporter CAT1 mRNA and protein levels were measured with transport assay, Northern blot analysis, and Western blot analysis, respectively. LPS increased arginine transport activity in a time- and dose-dependent fashion. Prolonged incubation of LPS (24 hours, 25 microg/ml) resulted in a 3-fold increase of arginine transport activity (control: 28 +/- 5; LPS: 92 +/- 20 pmol/mg/min, P < 0.05), with the System y(+) as the predominant arginine transport system, and a 2-fold increase of System y(+)CAT1 mRNA and transporter protein levels (P < 0.05). LPS increased the arginine transport System y(+) maximal velocity (V(max), control: 1484 +/- 180; LPS: 2800 +/- 230 pmol/mg/min, P < 0.05) without affecting the transport affinity (K(m), control: 76 +/- 8; LPS: 84 +/- 14 micromol/L, p = NS). The LPS-induced arginine transport activity was blocked by sodium nitroprusside (SNP) (control: 25 +/- 6; LPS: 97 +/- 26 *; SNP: 22 +/- 0.4(+); LPS+SNP: 33 +/- 10.3(+) pmole/mg/min, *P < 0.01 and (+)p = NS, compared with control). In contrary, the LPS-induced arginine transport activity was further augmented by NNA (control: 18 +/- 3.2; LPS: 59 +/- 2.7 *; NNA: 26.3 +/- 5.8; LPS + NNA: 127 +/- 18(+) pmol/mg/min; *P < 0.01 compared with control and (+)P < 0.01 compared with control or LPS). LPS-stimulates arginine transport activity in IEC-6 cells via a mechanism that involves increase of transport System y(+) mRNA levels and transporter protein levels. The LPS-stimulated arginine transport activity is regulated by the availability of nitric oxide.