StMBF1c positively regulates disease resistance to Ralstonia solanacearum via it's primary and secondary upregulation combining expression of StTPS5 and resistance marker genes in potato.

Multiprotein bridging factor 1 (MBF1) is a transcription coactivator that has a general defense response to pathogens. However, the regulatory mechanisms of MBF1 resistance bacterial wilt remain largely unknown. Here, the role of StMBF1c in potato resistance to Ralstonia solanacearum infection was characterized. qRT-PCR assays indicated that StMBF1c could was elicited by SA, MJ and ABA and the time-course expression pattern of the StMBF1c gene induced by R. solanacearum was found to be twice significant upregulated expression during the early and middle stages of bacterial wilt. Combined with the co-expression analysis of disease-resistant marker genes, gain-of-function and loss-of-function assays demonstrated that StMBF1c was associated with defence priming. Overexpression or silencing the MBF1c could enhance plants resistance or sensitivity to R. solanacearum through inducing or reducing NPR and PR genes related to SA signal pathway. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) experiment results confirmed the interaction of StMBF1c with StTPS5 which played a key role in ABA signal pathway in potato. It is speculated that by combining StTPS5 and resistance marker genes, StMBF1c is activated twice to participate in potato bacterial wilt resistance, in which EPI, PTI involved.

Quantitative analysis of n-3 polyunsaturated fatty acids and their metabolites by chemical isotope labeling coupled with liquid chromatography - mass spectrometry.

n-3 polyunsaturated fatty acids (PUFAs) and their metabolites play the crucial role in a wide range of physiologic and pathologic processes, including cardiovascular, neurodegenerative diseases, and inflammation-associated disorders. However, the quantitative analysis of n-3 PUFAs and their metabolites, oxylipins, is obstructed by high structural similarity, poor ionization efficiency and low abundance. In this study, a sensitive method was developed to quantify 28 n-3 PUFAs/oxylipins using chemical isotope labeling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Standards labeled with cholamine-d9 were used as one-to-one internal standards to achieve accurate quantification. The cholamine-d0-derivatized biological samples were mixed with cholamine d9-labeled standards for LC-MS/MS with multiple reaction monitoring. After cholamine derivatization, both MS sensitivity and chromatographic performance of n-3 PUFAs/oxylipins were substantially improved. Furthermore, the relationship between retention time and substituent position of regioisomers, and their fragmentation patterns were investigated, which may facilitate the identification of unknown oxylipins. Additionally, the developed method was applied to quantify the target n-3 PUFAs/oxylipins in serum and brain tissue from fish oil-supplemented mice, which exhibited its great potential and practicability. Collectively, this sensitive and reliable method may facilitate the elucidation of the roles of n-3 PUFAs/oxylipins in the physiological and pathological processes.

Effect of elemental nano-selenium on semen quality, glutathione peroxidase activity, and testis ultrastructure in male Boer goats.

The objective of this experiment is to study the effects of novel elemental nano-selenium in the diet on testicular ultrastructure, semen quality and GSH-Px activity in male goats. Forty-two 2-month-old bucks were offered a total mixed ration which had been supplemented with nano-Se (0.3mg/kg Se) or unsupplemented (the control group only received 0.06mg/kg Se-background), for a period of 12 weeks (from weaning to sexual maturity). Results showed that the testicular Se level, semen glutathione peroxidase and ATPase activity increased significantly in the nano-Se supplementation group compared with control (P<0.05). The semen quality (volume, density, motility and pH) was not affected by added Se in diets, however, the sperm abnormality rate of control bucks was significantly higher than Se supplemented bucks (P<0.05). The testes of 5 goats in each group were examined by transmission electron microscopy (TEM), and showed that in Se-deficient bucks the membrane was damaged, and showed the occurrence of abnormalities in the mitochondria of the midpiece of spermatozoa. In conclusion, selenium deficiency resulted in abnormal spermatozoal mitochondria, and supplementation with nano-Se enhanced the testis Se content, testicular and semen GSH-Px activity, protected the membrane system integrity and the tight arrayment of the midpiece of the mitochondria. Further studies are required to research the novel elemental nano-Se with characterization of bioavailability and toxicity in small ruminants.