In this study, lambda-cyhalothrin (LC) loaded polyurea microcapsules (MCs) with different particle sizes were fabricated.  All of the MCs showed varying degrees of physical collapse, which was more obvious among those with smaller particle sizes.  MCs with particle sizes of 1.38 μm (MC-S), 5.13 μm (MC-M) and 10.05 μm (MC-L) had shell thicknesses of 39.6, 50.3 and 150.1 nm, respectively.  MCs with smaller particles tended to have significantly faster release profiles, and the MC-S group had much higher bioactivity against Agrotis ipsilon and better foliar affinity on the peanut leaves (indicated by rainfastness) than MC-M and MC-L.  All of the MCs exhibited light-enhanced release profiles and had much slower degradation compared with the emulsifiable concentrate (EC) group, among which MC-L had the slowest degradation.  To generate MCs with both favorable quick efficacy and long-lasting efficacy, binary mixtures of MC-S, MC-M and MC-L were produced by mixing them in pairs at ratios of 2:1, 1:1 and 1:2.  The mixture of MC-S:MC-L at 1:2 showed the best comprehensive efficacy in the peanut foliar spray scenario among the nine tested combinations, and its effective duration was three times longer than that of EC.  Overall, the precise combination of MCs with different particle sizes can regulate the efficacy of pesticide control and serve as a strategy for the better utilization of pesticides.

SrUGT76G1, the most well-studied diterpene glycosyltransferase in Stevia rebaudiana, is key to the biosynthesis of economically important steviol glycosides (SGs).  However, the molecular regulatory mechanism of SrUGT76G1 has rarely been explored.  In this study, we identified a MYB transcription factor, SrMYB1, using a yeast one-hybrid screening assay.  SrMYB1 belongs to the typical R2R3-type MYB protein and is specifically localized in the nucleus with strong transactivation activity.  The transcript of SrMYB1 is predominantly accumulated in flowers, but is also present at a lower level in leaves.  Yeast one-hybrid and electrophoretic mobility shift assays verified that SrMYB1 binds directly to the MYB binding sites in the F4-3 fragment (+50–(–141)) of the SrUGT76G1 promoter.  Furthermore, we found that SrMYB1 could significantly repress the expression of SrUGT76G1 in both epidermal cells of tobacco leaves and stevia callus.  Taken together, our results demonstrate that SrMYB1 is an essential upstream regulator of SrUGT76G1 and provide novel insight into the regulatory network for the SGs metabolic pathway in S. rebaudiana.

Perilipin1 (PLIN1) is a major phosphorylated protein that specifically coats the surface of neutral lipid droplets (LDs) in adipocytes and plays a crucial role in regulating the accumulation and hydrolysis of triacylglycerol (TG).  Mammalian studies have shown that Plin1 gene transcription is mainly regulated by peroxisome proliferator-activated receptor-gamma (PPARγ), the master regulator of adipogenesis.  However, the regulatory mechanism of the chicken Plin1 (cPlin1) gene is poorly understood.  The present study aimed to investigate whether Plin1 is regulated by PPARγ in chickens and identify its exact molecular mechanism.  Reporter gene and expression assays showed that PPARγ2, but not PPARγ1, activated (P<0.01) the cPlin1 gene promoter.  An electrophoretic mobility shift assay and mutational analysis revealed that PPARγ2 bound to a special site in the cPlin1 gene promoter to enhance its expression.  In summary, our results show that PPARγ promotes the expression of the cPlin1 gene and that PPARγ2 is the main regulatory isoform.

In agricultural production, temperature and moisture are important factors affecting grain yield and quality.  Although moderate drought at the grain-filling stage can effectively alleviate the damage caused by high temperature, the specific regulatory mechanism driving the effect of moderate drought at the high temperature on starch synthesis is still unclear.  To explore the effects and mechanisms of high temperature and moderate drought on rice starch synthesis at the grain-filling stage, the activities of enzymes and expression levels of the genes involved in starch synthesis under four different treatments involving high temperature and/or water stress (CK, HT, WS, and HT+WS) were investigated in this study.  The starch synthesis of a japonica inbred rice was measured under the four treatments during the grain filling.  The results show that the effects of high temperature and moderate drought on grain filling mainly occur in the inferior grains of rice.  Through the regulation of enzymes involved in starch synthesis and the expression levels of their main genes, the synthesis of rice starch can be affected.  Therefore, the high temperature and moderate drought were antagonistic, and moderate drought can alleviate the damage to grain quality at a high temperature by improving the starch synthesis of inferior grains in japonica rice.  This study provides a basis for stress-resistance cultivation and breeding strategies of rice with high temperature tolerance.

    Pomegranates is abundant in polyphenols and is well-known for its antioxidant activity. Punicalagin (PG) is a major polyphenolic compound in the pomegranate peel. In certain conditions, PG can be hydrolyzed to punicallin (PL) and ellagic acid (EA), and PL can be further hydrolyzed to EA. PG, PL, and EA all play important roles in the antioxidant activity of pomegranate peels. This study was conducted to compare the in vitro antioxidant activity and in vivo anti-oxidative stress effects of PG, PL, and EA. For the in vitro test, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH·) and superoxide anion (O2-.) scavenging capacities, ferric-reducing antioxidant power (FRAP), and lipid peroxidation (LPO) inhibition capacities of PG, PL, and EA were tested. For the in vivo test, oxidatively stressed mice, which were induced by oxidized fish oil, were administrated PG, PL or EA (10 mg kg^–1 d ^–1) for 21 days. The results showed that the in vitro antioxidant activity trends were EA>PG>PL>Trolox in scavenging DPPH?, PG>PL>EA≈Trolox in scavenging O2-. , EA>PG≈PL>Trolox in FRAP, and Trolox>PG>EA>PL in LPO inhibition. In the in vivo test, the EA treatment increased the average daily weight gain and total antioxidant capacity (T-AOC) in the plasma (P<0.05), liver (P<0.05), and intestine (P<0.05) in oxidatively stressed mice. It increased the superoxide dismutase (SOD) activity in the liver (P<0.05) and intestine (P<0.05). It increased the glutathione peroxidase (GSH-Px) activity in the intestine (P<0.05) and the intestinal villus height to crypt depth ratio (P<0.05). EA treatment decreased the malondialdehyde (MDA) content in the plasma (P<0.05), liver (P<0.05), and intestine (P<0.05) and the mRNA expressions of the pro-inflammatory factors, TNF-α (P<0.05), IFN-γ (P<0.05) and IL-6 (P<0.05). PL increased the SOD (P<0.05) and GSH-Px activities (P<0.05) in the intestine and decreased the MDA content (P<0.05) and the mRNA expressions of TNF-α (P<0.05) and IL-6 (P<0.05) in the intestine. PG increased the SOD activity (P<0.05) and GSH-Px activity (P<0.05) in the intestine and decreased the MDA content in the intestine (P<0.05) and IL-6 mRNA expression in the intestine (P<0.05). In summary, EA, PL, and PG all had powerful in vitro antioxidant capacities, and they had different antioxidant advantages in acting against different types of radicals; EA was more effective than PL and PG in protecting against oxidative injury in vivo, especially for intestinal injury. These findings suggest that multiple polyphenol compounds in pomegranate peel may exert superior antioxidant activity than single purified polyphenols; when using pomegranate peels as health-promoting additive in animal feed, raising EA content by methods of hydrolysis or fermentation in advance could achieve better effects.

Seven inorganic amendment materials were added into arsenic (As) contaminated soil at a rate of 0.5% (w/w); the materials used were sepiolite, red mud, iron grit, phosphogypsum, ferrihydrite, iron phosphate, and layered double oxides (LDO). Plant growth trials using rape (edible rape, Brassia campestris L.) as a bio-indicator are commonly used to assess As bioavailability in soils. In this study, B. campestris was grown in a contaminated soil for 50 days. All of the inorganic amendments significantly inhibited the uptake of As by B. campestris. Following soil treatment with the seven aforementioned inorganic ammendments, the As concentrations in the edible parts of B. campestris were reduced by 28.6, 10.5, 8.7, 31.0, 47.4, 25.3, and 28.8%, respectively, as compared with the plants grown in control soil. The most effective amendment was ferrihydrite, which reduced As concentration in B. campestris from 1.84 to 0.97 mg kg–1, compared to control. Furthermore, ferrihydrite-treated soils had a remarkable decrease in both non-specifically sorbed As and available-As by 67 and 20%, respectively, comparing to control. Phosphogypsum was the most cost-effective amendment and it showed excellent performance in reducing the water soluble As in soils by 31% and inhibiting As uptake in B. campestris by 21% comparing to control. Additionally, obvious differences in As transfer rates were observed in the various amendments. The seven amendment materials used in this study all showed potential reduction of As bioavailability and influence on plant growth and other biological processes still need to be further explored in the long term.

Many animal feed grains contain high β-glucan in the cell wall. Pigs do not secret β-glucanase to degrade the β-glucan in their feed. The indigestible β-glucan not only blocks the release of nutrients from the grain cell wall, but also increases the digesta viscosity in the gastrointestinal tract of pigs. Therefore, dietary β-glucan significantly inhibits nutrient digestion and absorption in pigs. Transgenic expression of β-glucanase in the digestive tract of pigs may offer a solution to solve this problem. In the current study, four arti?cial codon-optimized β-glucanases genes was prepared and expressed in porcine cells. Only pBgA and pEgx showed high activity in transfected pig kidney cells. To improve the pH range and pH stability of β-glucanase, the two β-glucanases, pBgA and pEgx, were co-expressed in pig kidney cells and salivary gland cells by Linker A3 or 2A peptide. The resulting dual enzymes of pBgA3pEg and pBg2ApEg showed significantly enlarged pH range and significantly increased pH stability, as compared to parental enzymes. These results provide useful data for future study on increasing the feed digestibility of pigs by transgenic expression of β-glucanase in their salivary glands.

The reverse genetics for classical swine fever virus (CSFV) is currently based on the transfection of in vitro transcribed RNA from a viral genomic cDNA clone, which is inefficient and time-consuming. This study was aimed to develop an improved method for rapid recovery of CSFV directly from cloned cDNA. Full-length genomic cDNA from the CSFV Shimen strain, which was flanked by a T7 promoter, the hepatitis delta virus ribozyme and T7 terminator sequences, was cloned into the lowcopy vector pOK12, producing pOKShimen-RzT . Direct transfection of pOKShimen-RzT into PK/T7 cells, a PK-15- derived cell line stably expressing bacteriophage T7 RNA polymerase, allowed CSFV to be rescued rapidly and efficiently, i.e., at least 12 h faster and 31.6-fold greater viral titer when compared with the in vitro transcription-based rescue system. Furthermore, the progeny virus rescued from PK/T7 cells was indistinguishable, both in vitro and in vivo, from its parent virus and the virus rescued from classical reverse genetics. The reverse genetics based on intracellular transcription is efficient, convenient and cost-effective. The PK/T7 cell line can be used to rescue CSFV directly from cloned cDNA and it can also be used as an intracellular transcription and expression system for studying the structure and function of viral genes.

Plants respond to drought stress with different physical manners, such as morphology and color of leaves. Thus, plants can be considered as a sort of living-sensors for monitoring dynamic of soil water content or the stored water in plant body. Because of difficulty to identify the early wilting symptom of plants from the results in 2D (two-dimension) space, this paper presented a preliminary study with 3D (three-dimension)-based image, in which a laser scanner was used for achieving the morphological information of zucchini (Cucurbita pepo) leaves. Moreover, a leaf wilting index (DLWIF) was defined by fractal dimension. The experiment consisted of phase-1 for observing the temporal variation of DLWIF and phase-2 for the validation of this index. During the experiment, air temperature, luminous intensity, and volumetric soil water contents (VSWC) were simultaneously recorded over time. The results of both phases fitted the bisector (line: 1:1) with R2=0.903 and REMS=0.155. More significantly, the influence of VSWC with three levels (0.22, 0.30, and 0.36 cm3 cm-3) on the response of plant samples to drought stress was observed from separated traces of DLWIF. In brief, two conclusions have been made: (i) the laser scanner is an effective tool for the non-contact detection of morphological wilting of plants, and (ii) defined DLWIF can be a promising indicator for a category of plants like zucchini.

Fertility restoration of cytoplasmic male-sterility in pepper (Capsicum annuum L.) is useful for commercial production ofhybrid seeds. However, the mechanism of fertility restoration has not been determined. We previously constructed acDNA library and identified some genes related to fertility restoration in pepper using suppression subtractive hybridizationtechnology. In this study, the expression patterns of 20 genes were investigated using semi-quantitative RT-PCR. Threegenes expressed only in restorer lines, but not in sterility lines. Four genes expressed only in anther, but not in otherorgans. Among these 7 genes, the clone TG31 was observed to specifically express in anther of restorer lines. The workdescribed here provides a comprehensive overview on the expression pattern of the genes that are induced by restoreralleles in pepper. It will also contribute to the current understanding of molecular networks for the regulation of fertilityrestoration.

Calcium-dependent protein kinases (CDPKs, EC 2.7.1.37) comprise a large family of Ser/Thr kinases in plants and play an important role in plant Ca2+ signal transduction. A full-length CDPK gene, NtCDPK12 (GenBank accession number GQ337420), was isolated from common tobacco (Nicotiana tabacum) leaves by rapid amplification of cDNA ends (RACE). The NtCDPK12 cDNA is 1 816 bp length and contains an open reading frame (ORF) of 1 461 bp encoding 486 amino acids. Sequence alignments indicated that NtCDPK12 contains all conserved regions found in CDPKs and shows a high level of sequence similarity to many other plant CDPKs. The results of real-time quantitative reverse transcription-PCR (qRTPCR) showed that NtCDPK12 was highly expressed in stems and increased in roots treated with high-salt or subjected to drought stress, which indicates that NtCDPK12 was induced by high-salt and drought stresses.

Classical swine fever (CSF) and porcine reproduction and respiratory syndrome (PRRS) are both economically important, highly contagious diseases of swine worldwide. To develop an effective vaccine to control these two diseases, we constructed a recombinant adenovirus rAdV-GP52AE2, using a replication-defective human adenovirus serotype 5 as a delivery vector, to co-express the GP5 protein of highly pathogenic porcine reproduction and respiratory syndrome virus (PRRSV) and the E2 protein of classical swine fever virus (CSFV). Foot-and-mouth disease virus (FMDV) 2A peptide was used as a linker between the GP5 and E2 proteins to allow automatic self-cleavage of the polyprotein. The GP5 and E2 genes were expressed as demonstrated by immunofluorescence assay and Western blotting. Immunization of mice resulted in a CSFV-neutralizing antibody titer of 1:128 and a PRRSV-neutralizing antibody titer of 1:16. The lymphoproliferative responses were detected by Cell Counting Kit-8 assay and the stimulation index of CFSV-specific and PRRSV-specific lymphocytes in the rAdV-GP52AE2 group was significantly higher than that in the negative control group. The results show that rAdV-GP52AE2 can induce both effective humoral and cell-mediated immune responses in mice. The protective efficacy of the recombinant virus against CSF was evaluated in immunized rabbits, which were protected from fever induced by challenge with C-strain. Our study provides supporting evidence for the use of FMDV 2A to develop a bivalent genetically-engineered vaccine.