Analysis of carotenoid profile changes and carotenogenic genes transcript levels in Rhodosporidium toruloides mutants from an optimized Agrobacterium tumefaciens-mediated transformation method.

Rhodosporidium toruloides has been reported as a potential biotechnological microorganism to produce carotenoids. The most commonly used molecular and genetic manipulation methods based on Agrobacterium-mediated transformation (ATMT). However, this method was of relatively lower transformation efficiency. In this study, we optimized the ATMT method for R. toruloides on account of the promoter on T-DNA, the ratio of A. tumefaciens to R. toruloides NP11, acetosyringone concentration, cocultivation temperature and time, and a transformation efficiency of 2,369 cells per 105 recipient cells was obtained and was 24 times as that of the previous report. With this optimized method, four redder mutants and four yellower mutants were selected out with torularhodin and ß-carotene production preference, respectively. The highest torularhodin production was 1,638.15 µg/g dry cell weight in A1-13. The yellower mutants were found to divert the metabolic flux from torularhodin and torulene to γ-carotene and ß-carotene, and the proportion of γ-carotene and ß-carotene were all over 92%. TAIL-PCR was carried out to found T-DNA insertion in these mutants, and insertion hotspot was found. RT-qPCR results showed that CTA1 genes in these mutants were closely related to the synthesis of total carotenoids, especially torularhodin, and was a potenial metabolic engineering site in the future.

[The clinical significance of leucocytosis and increase in interleukin content in acute mountain sickness complicated with multiple organ dysfunction syndrome].

OBJECTIVE: To study the role of white blood cell (WBC) and interleukins (ILs) in acute mountain sickness (AMS) complicated with multiple organ dysfunction syndrome (MODS). METHODS: Three thousand one hundred and eighty-four patients suffering from severe AMC in the past 50 years were surveyed with questionnaire. Correlation analysis was done to explore the relationship between differential count of WBC and elevation of ILs contents and MODS. RESULTS: There was no difference in differential count of WBC between essential high altitude pulmonary edema (HAPE) and secondary HAPE. No difference was also found between simple HAPE and high altitude cerebral edema (HACE). However, obvious difference in WBC was found between HACE accompanied by HAPE and simple HAPE or simple HACE in the differential counts of WBC, counts of WBC and neutrophil were significantly elevated, while lymphocyte was significantly declined (all P < 0.05). The count of WBC in people suffering from AMS accompanied by MODS was higher than patients only suffering from AMS, lymphocyte was significantly decreased, the difference was obviously (both P < 0.01). The content of tumor necrosis factor-alpha (TNF-alpha), IL-1, IL-2, IL-6 and IL-8 of severe AMS patients were obviously higher than that of normal people, therefore the content of IL-4 in severe AMS patients was obviously lower than that normal people the difference were obvious (all P < 0.01). CONCLUSION: People suffering from AMS also exist systemic inflammatory response syndrome (SIRS). SIRS play an important part in MODS which is a major cause of AMS, and it's a critical factor of high altitude MODS.