RÉSUMÉ
ObjectiveExosomes are microvesicles which could be secreted by all cell types with diameters between 30 and 150 nm. It was widely distributed in body fluids including blood, urine, and breast milk. Exosomes are considered as potential biomarkers and drug carriers by reason of containing nucleic acids, lipids, proteins and other bioactive molecules. Milk-derived exosomes have been widely used as drug delivery carriers to treat targeted diseases with a lower cost, higher biocompatibility and lower immunogenicity. Until now, there is no research about the milk-derived exosomes phosphorylation to reveal the difference of protein phosphorylation in different species of milk. To investigate the pathways and proteins with specific functions, phosphorylated proteomic analysis of milk-derived exosomes from different species is performed, and provide new ideas for exploring diversified treatments of disease. MethodsWhey and exosomes derived from bovine, porcine and caprine milk were performed for proteomics and phosphoproteomics analysis. The relationship between milk exosome proteins from different species and signaling pathways were analyzed using bioinformatics tools. ResultsA total of 4 191 global proteins, 1 640 phosphoproteins and 4 064 phosphosites were identified from 3 species of milk-derived exosomes, and the exosome proteins and phosphoproteins from different species were significantly higher than those of whey. Meanwhile, some special pathways were enriched like Fcγ-mediated phagocytosis from bovine exosomes, pathways related with neural and immune system from caprine exosomes, positive and negative regulation of multiple activities from porcine exosomes. ConclusionIn this study, the proteomic and phosphoproteomic analyses of exosomes and whey from bovine, porcine and caprine milk were carried out to reveal the difference of composition and related signaling pathways of milk exosome from different species. These results provided powerful support for the application of exosomes from different milk sources in the field of disease treatment.
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Objective: To screen and evaluate DNA barcoding of Amomun tsao-ko populations in Yunnan. Methods: ITS, psbA-trnH, matK, rbcL, and ycf1 sequences were screened and evaluated using A. tsao-ko as samples. The samples of A. tsao-ko population were amplified and sequenced. The sequences were spliced with Genestar, and then processed with Mega for data processing. And A. tsao-ko diversity and identification were analyzed and discussed. Results: The length of the amplified fragments of primers ITS5 and ITS4 was approximately 520 bp; The length of the amplified fragments of the primers rbcLa-F and rbcLa-R was approximately 498 bp; The length of the amplified fragments of the primers ycf1-bF and ycf1-bR was approximately 800 bp; The length of the amplified fragments of the primers psbA-trnH-1F and psbA-trnH-1R was approximately 400 bp; The length of the amplified fragments of the primers matK-2F and matK-2R was approximately 470 bp. The success rate of amplification and sequencing was high, and most of the results were available. By analyzing the amplification results of ITS, psbA-trnH, matK and ycf1 sequences of A. tsao-ko, A. tsao-ko and other Amomum genus plants can be clearly distinguished; All samples of the ITS sequence were divided into MG5 white flower A. tsao-ko population and other populations; All samples of the psbA-trnH sequence were divided into MG5 white flower A. tsao-ko population, MG6 yellow flower A. tsao-ko population and other populations; All samples of the matK sequence were divided into MG6 A. tsao-ko population and other populations. The MG5 white flower A. tsao-ko sample failed to be amplified; All samples of the ycf1 sequence were divided into the MG6 yellow flower A. tsao-ko population and other populations, and the MG5 white flower A. tsao-ko population was clustered with the other 22 A. tsao-ko populations; The amplification of rbcL sequence was consistent for all samples. Conclusion: The ITS, matK, psbA-trnH and ycf1 sequences can accurately distinguish A. tsao-ko from other plants of Amomum genus; The sequence site variations were found in matK, psbA-trnH and ycf1 sequences of MG6. This research has contributed to the selection and breeding of A. tsao-ko varieties. ITS and psbA-trnHsequences can distinguish yellow flower and white flower of A. tsao-ko; There is no variation in the rbcL sequence of all samples of white and yellow flowers of A. tsao-ko, and Amomum tsao-ko and other plants of Amomum genus cannot be identified with the rbcL sequence, which can be discarded.
RÉSUMÉ
Objective To evaluate the genetic diversity and phylogenetic relationships of Amomun tsao-ko populations in Yunnan. Methods Seven pairs of microsatellite (SSR) primers were used to analyze 24 A. tsao-ko populations; First, GenALEx was used to calculate genetic diversity parameters, PCoA and AMOVA analysis was carried out; NTsys software was then used to draw population clusters map; And finally, the Structure software was used to calculate the best K value. Results The average of Shannon’s diversity index (H) of the 24 A. tsao-ko populations was 0.49, the average of heterozygosity (He) was 0.32, the genetic differentiation coefficient (Fst) was 0.090, and the gene flow (Nm) was 2.930. Eighty-one percent of the genetic differentiation among the 24 populations of A. tsao-ko existed within the population, and only 19% existed among the populations. The genetic identity (I) of the 23 A. tsao-ko populations of yellow flowers was 0.631 8-0.982 4. The genetic distance (D) was in the range of 0.017 7- 0.459 2, while the consistency degree of the A. tsao-ko population of white flower (MG5) and 23 other yellow flowers was 0.369 7-0.609 0. However, cluster analysis showed that A. tsao-ko population of the white flowers and yellow flowers were clearly separated at the genetic distance of 0.49. Structure clustering showed 209 A. tsao-ko resources can be divided into four groups when K value was 4. Conclusion The genetic diversity of A. tsao-ko populations of yellow flowers of Yunnan is higher on average, and the genetic variation is mainly found in population rather than among populations. According to the genotypes, A. tsao-ko of yellow flower and white flower are clearly divided into two categories, and the genetic distance is very far; and the yellow flower of A. tsao-ko is roughly divided into four groups.