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We determined the phosphate (P) uptake, substrate P concentration, phytase activity, acid phosphatase activity and expression of three predicted secreted purple acid phosphatase genes (PAPs) from trifoliate orange (Poncirus trifoliata L. Raf) through Glomus versiforme and phytin treatments to evaluate phytate-phosphorus utilization promoted by arbuscular mycorrhizal fungi (AMF, G. versiforme). The results showed that under inorganic phosphate (Pi) deficiency, trifoliate orange seedlings utilized phytin and G. versiforme colonization promoted trifoliate orange phytin utilization by increasing phytase activity and acid phosphatase activity in the roots and the substrate. Although the expression of the three secreted PtPAPs in the seedlings was significantly upregulated in the roots and the leaves after 2 months of -P solution application, the genes exhibited different expression patterns. In general, PtPAP1 was regulated more strongly in the roots than in the leaves, which was upregulated by G. versiforme colonization but downregulated by phytin treatment in the roots. PtPAP2 was activated by Pi deficiency in the leaves but was almost constitutive expressed in the roots. PtPAP3, which might encode a phytase, was also regulated more strongly in the roots than in the leaves and reduced by phytin treatment, but unaffected by G. versiforme colonization. The expression patterns of the PtPAP genes in response to the AMF and organic phosphate (Po) treatments suggested that the three PtPAPs had different functions in the Po utilization of trifoliate orange. Although the phytase activity and acid phosphatase activity in the roots and the substrate were not consistent with PtPAPs expression, AMF promoted the activities of the two enzymes in the substrate, which was at least partly ascribed to the regulation of PtPAPs expression.
This study focused on the microbiota and chemical compounds of the fermented Pinelliae Rhizoma produced in Longchang (LC), Zizhong (ZZ) and Xindu (XD), in Sichuan Province (China). High-throughput sequencing was used to analyze the microbiota. GC-MS and LC-MS were used to detect the compounds produced during the three different Pinelliae Rhizoma fermentation processes. The bacteria and fungi of the three fermented Pinelliae Rhizoma differed substantially, with the bacterial content mainly composed of the Bacillus genus, while the common fungi were only included in four OTUs, which belong to three species of Eurotiomycetes and Aspergillus cibarius. 51 volatile compounds were detected; they varied between LC, XD, and ZZ fermented Pinelliae Rhizoma. C10 and C15 terpenes were most frequently detected, and only curcumene and β-bisabolene were detected in the three fermented Pinelliae Rhizoma. 65 non-volatile compounds were detected by LC-MS, most were of C16, C18, C20, C21 and C22 structures. Cluster analysis showed more similarity between LC and XD fermented Pinelliae Rhizoma with regards to volatile compound content, but more similarity between the XD and ZZ fermented Pinelliae Rhizoma for non-volatiles. Moreover, no correlation between geographical distance and microflora or compounds of fermented Pinelliae Rhizoma was observed. These results showed that hundreds of compounds are produced by the natural mixed fermentation of Pinelliae Rhizoma, and may mostly relate to the microorganisms of five species.
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