Analysis of the Differences in Volatile Organic Compounds in Different Rice Varieties Based on GC-IMS Technology Combined with Multivariate Statistical Modelling.

In order to investigate the flavour characteristics of aromatic, glutinous, and nonaromatic rice, gas chromatography-ion mobility spectrometry (GC-IMS) was used to analyse the differences in volatile organic compounds (VOCs) amongst different rice varieties. The results showed that 103 signal peaks were detected in these rice varieties, and 91 volatile flavour substances were identified. Amongst them, 28 aldehydes (28.89~31.17%), 24 alcohols (34.85~40.52%), 14 ketones (12.26~14.74%), 12 esters (2.30~4.15%), 5 acids (7.80~10.85%), 3 furans (0.30~0.68%), 3 terpenes (0.34~0.64%), and 2 species of ethers (0.80~1.78%) were detected. SIMCA14.1 was used to perform principal component analysis (PCA) and orthogonal partial least squares discriminant analysis, and some potential character markers (VIP > 1) were further screened out of the 91 flavour substances identified based on the variable important projections, including ethanol, 1-hexanol, hexanal, heptanal, nonanal, (E)-2-heptenal, octanal, trans-2-octenal, pentanal, acetone, 6-methyl-5-hepten-2-one, ethyl acetate, propyl acetate, acetic acid, and dimethyl sulphide. Based on the established fingerprint information, combined with principal component analysis and orthogonal partial least squares discriminant analysis, different rice varieties were also effectively classified, and the results of this study provide data references for the improvement in aromatic rice varieties.

IbMYB308, a Sweet Potato R2R3-MYB Gene, Improves Salt Stress Tolerance in Transgenic Tobacco.

The MYB (v-myb avian myeloblastosis viral oncogene homolog) transcription factor family plays an important role in plant growth, development, and response to biotic and abiotic stresses. However, the gene functions of MYB transcription factors in sweet potato (Ipomoea batatas (L.) Lam) have not been elucidated. In this study, an MYB transcription factor gene, IbMYB308, was identified and isolated from sweet potato. Multiple sequence alignment showed that IbMYB308 is a typical R2R3-MYB transcription factor. Further, quantitative real-time PCR (qRT-PCR) analysis revealed that IbMYB308 was expressed in root, stem, and, especially, leaf tissues. Moreover, it showed that IbMYB308 had a tissue-specific profile. The experiment also showed that the expression of IbMYB308 was induced by different abiotic stresses (20% PEG-6000, 200 mM NaCl, and 20% H2O2). After a 200 mM NaCl treatment, the expression of several stress-related genes (SOD, POD, APX, and P5CS) was upregulation in transgenic plants, and the CAT activity, POD activity, proline content, and protein content in transgenic tobacco had increased, while MDA content had decreased. In conclusion, this study demonstrated that IbMYB308 could improve salt stress tolerance in transgenic tobacco. These findings lay a foundation for future studies on the R2R3-MYB gene family of sweet potato and suggest that IbMYB308 could potentially be used as an important positive factor in transgenic plant breeding to improve salt stress tolerance in sweet potato plants.

Identification of miRNAs in Response to Sweet Potato Weevil (Cylas formicarius) Infection by sRNA Sequencing.

The sweet potato weevil (Cylas formicarius) is an important pest in the growing and storage of sweet potatoes. It is a common pest in the sweet potato production areas of southern China, causing serious harm to the development of the sweet potato industry. For the existing cultivars in China and abroad, there is no sweet potato variety with complete resistance to the sweet potato weevil. Thus, understanding the regulation mechanisms of sweet potato weevil resistance is the prerequisite for cultivating sweet potato varieties that are resistant to the sweet potato weevil. However, very little progress has been made in this field. In this study, we inoculated adult sweet potato weevils into sweet potato tubers. The infected sweet potato tubers were collected at 0, 24, 48, and 72 h. Then, a miRNA library was constructed for Eshu 6 and Guang 87 sweet potato tubers infected for different lengths of time. A total of 407 known miRNAs and 298 novel miRNAs were identified. A total of 174 differentially expressed miRNAs were screened out from the known miRNAs, and 247 differentially expressed miRNAs were screened out from the new miRNAs. Moreover, the targets of the differentially expressed miRNAs were predicted and their network was further investigated through GO analysis and KEGG analysis using our previous transcriptome data. More importantly, we screened 15 miRNAs and their target genes for qRT-PCR verification to confirm the reliability of the high-throughput sequencing data, which indicated that these miRNAs were detected and most of the expression results were consistent with the sequencing results. These results provide theoretical and data-based resources for the identification of miRNAs in response to sweet potato weevil infection and an analysis of the molecular regulatory mechanisms involved in insect resistance.

A Flavonoid Compound Promotes Neuronal Differentiation of Embryonic Stem Cells via PPAR-ß Modulating Mitochondrial Energy Metabolism.

Relatively little is known regarding mitochondrial metabolism in neuronal differentiation of embryonic stem (ES) cells. By using a small molecule, present research has investigated the pattern of cellular energy metabolism in neural progenitor cells derived from mouse ES cells. Flavonoid compound 4a faithfully facilitated ES cells to differentiate into neurons morphologically and functionally. The expression and localization of peroxisome proliferator-activated receptors (PPARs) were examined in neural progenitor cells. PPAR-ß expression showed robust upregulation compared to solvent control. Treatment with PPAR-ß agonist L165041 alone or together with compound 4a significantly promoted neuronal differentiation, while antagonist GSK0660 blocked the neurogenesis-promoting effect of compound 4a. Consistently, knockdown of PPAR-ß in ES cells abolished compound 4a-induced neuronal differentiation. Interestingly, we found that mitochondrial fusion protein Mfn2 was also abolished by sh-PPAR-ß, resulting in abnormal mitochondrial Ca2+ ([Ca2+]M) transients as well as impaired mitochondrial bioenergetics. In conclusion, we demonstrated that by modulating mitochondrial energy metabolism through Mfn2 and mitochondrial Ca2+, PPAR-ß took an important role in neuronal differentiation induced by flavonoid compound 4a.

[Phenotype-based primary screening for drugs promoting neuronal subtype differentiation in embryonic stem cells with light microscope].

OBJECTIVE: To set up a platform for phenotype-based primary screening of drug candidates promoting neuronal subtype differentiation in embryonic stem cells (ES) with light microscope. METHODS: Hanging drop culture 4-/4+ method was employed to harvest the cells around embryoid body (EB) at differentiation endpoint. Morphological evaluation for neuron-like cells was performed with light microscope. Axons for more than three times of the length of the cell body were considered as neuron-like cells. The compound(s) that promote neuron-like cells was further evaluated. Icariin (ICA, 10(-6)mol/L) and Isobavachin (IBA, 10(-7)mol/L) were selected to screen the differentiation-promoting activity on ES cells. Immunofluorescence staining with specific antibodies (ChAT, GABA) was used to evaluate the neuron subtypes. RESULTS: The cells treated with IBA showed neuron-like phenotype, but the cells treated with ICA did not exhibit the morphological changes. ES cells treated with IBA was further confirmed to be cholinergic and GABAergic neurons. CONCLUSION: Phenotypic screening with light microscope for molecules promoting neuronal differentiation is an effective method with advantages of less labor and material consuming and time saving, and false-positive results derived from immunofluorescence can be avoided. The method confirms that IBA is able to facilitate ES cells differentiating into neuronal cells, including cholinergic neurons and GABAergic neurons.

[Establishment of optimized neuronal differentiation-promoting model derived from P19 embryonic carcinoma cells].

OBJECTIVE: To establish an optimized primary drug screen model of neuronal differentiation using P19 embryonal carcinoma cells. METHODS: The final concentration of retinoid acid (RA), days of suspension culture, manner of adherent culture, suitable cell density and adherent culture medium were tested, respectively. Two stages of neuronal differentiation were examined based on morphological changes and immunocytochemistry analysis of neuronal specific protein ß-tubulin III. RESULTS: On d 8 of differentiation culture, neuron-like cells were observed with final concentration of 1 µmol/L RA. Neuron-like network was formed on d 16 of neuronal differentiation. ß-tubulin III was positively stained on both stages, indicating P19 cells were differentiated into neurons. CONCLUSION: The model using RA to induce P19 embryonic carcinoma cells to differentiate into neuron-like cells has been successfully established, which may provide a rapid, phenotypic cell-based platform for primary screening of neurogenesis-promoting drugs.

Junctophilin 2 knockdown interfere with mitochondrium status in ESC-CMs and cardiogenesis of ES cells.

In the present study, we explored the possible links between Junctophilin 2 (Jp2) and the mitochondrium-sarcoplasmic reticulum (SR) interaction in embryonic stem cell-derived cardiomyocytes (ESC-CMs), as well as the role of Jp2 in cardiogenesis of ES cells. We found that Ca(2+) transient was abnormal and mitochondria were de-energized within siJp2 ESC-CMs. The essential juxtaposition structure of mitochondrium with SR was destroyed accompanied by selectively downregulation of Pgc-1α, Nrf-1, and Mfn-2. Impaired co-localization of the JP2 and sarcomeres (α-Actinin or Troponin-T) appeared in embryoid bodies (EBs) after Jp2 knockdown. Calsequestrin2 and ryanodine receptor 2 within SR were expressed as early as the initiation of differentiation, while triadin and caveolin3 within t-tubules (TTs) did not appear until the terminal, indicating that JP2 probably did not contribute to anchoring the SR to TTs at the early cardiogenesis stage as usual. In addition, Jp2 knockdown selectively decreased gene transcription toward cardiogenesis (Brachyury, Isl1, and Nkx2.5), subsequently weaken EB beating activity by 60%. Taken together, reducing JP2 expression in ESC-CMs resulted in impaired mitochondrial status due to either abnormal cellular Ca(2+) homeostasis or disturbing of juxtaposition. A sensitive time window of JP2 necessary in cardiac differentiation was found at early stage via an extra non-TTs/SR anchor-dependent role.

Response to temperature stress of reactive oxygen species scavenging enzymes in the cross-tolerance of barley seed germination.

A number of studies have shown the existence of cross-tolerance in plants, but the physiological mechanism is poorly understood. In this study, we used the germination of barley seeds as a system to investigate the cross-tolerance of low-temperature pretreatment to high-temperature stress and the possible involvement of reactive oxygen species (ROS) scavenging enzymes in the cross-tolerance. After pretreatment at 0 °C for different periods of time, barley seeds were germinated at 35 °C, and the content of malondialdehyde (MDA) and the activities of ROS scavenging enzymes were measured by a spectrophotometer analysis. The results showed that barley seed germinated very poorly at 35 °C, and this inhibitive effect could be overcome by pretreatment at 0 °C. The MDA content varied, depending on the temperature at which seeds germinated, while barley seeds pretreated at 0 °C did not change the MDA content. Compared with seeds germinated directly at 35 °C, the seeds pretreated first at 0 °C and then germinated at 35 °C had markedly increased activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and glutathione reductase (GR). The SOD and APX activities of seeds germinated at 35 °C after 0 °C-pretreatment were even substantially higher than those at 25 °C, and GR activity was similar to that at 25 °C, at which the highest germination performance of barley seeds was achieved. These results indicate that low-temperature pretreatment can markedly increase the tolerance of barley seed to high temperature during germination, this being related to the increase in ROS scavenging enzyme activity. This may provide a new method for increasing seed germination under stress environments, and may be an excellent model system for the study of cross-tolerance.