Research on the dentification of Panax ginseng and Panax quinquefolium using catalysed hairpin assembly technology.

INTRODUCTION: Panax ginseng and Panax quinquefolium are traditional Chinese herb medicines and similar in morphology and some chemical components but differ in drug properties, so they cannot be mixed. However, the processed products of them are often sold in the form of slices, powder, and capsules, which are difficult to identify by traditional morphological methods. Furthermore, an accurate evaluation of P. ginseng, P. quinquefolium and the processed products have not been conducted. OBJECTIVE: This study aimed to establish a catalysed hairpin assembly (CHA) identification method for authenticating products made from P. ginseng and P. quinquefolium based on single nucleotide polymorphism (SNP) differences. METHOD: By analysing the differences of SNP in internal transcribed spacer 2 (ITS2) in P. ginseng and P. quinquefolium to design CHA-specific hairpins. Establish a sensitive and efficient CHA method that can identify P. ginseng and P. quinquefolium, use the sequencing technology to verify the accuracy of this method in identifying Panax products, and compare this method with high-resolution melting (HRM). RESULTS: The reaction conditions of CHA were as follows: the ratio of forward and reverse primers, 20:1; hairpin concentration, 5 ng/µL. Compared with capillary electrophoresis, this method had good specificity and the limit of detection was 0.5 ng/µL. The result of Panax product identification with CHA method were coincidence with that of the sequencing method; the positive rate of CHA reaction was 100%. CONCLUSION: This research presents an effective identification method for authenticating P. ginseng and P. quinquefolium products, which is helpful to improve the quality of Panax products.

[Functional Genomics Analysis of Nitrogen and Phosphorus Transformation in Maize Rhizosphere Microorganisms].

Fertilizer reduction and efficiency improvement is an important basis for ensuring the safety of the agricultural ecological environment. Microorganisms are the key driving force for regulating the soil nitrogen and phosphorus cycle. Studying the nitrogen and phosphorus transformation function of rhizosphere microorganisms can provide a microbiological regulation approach for further improving the use efficiency of soil nitrogen and phosphorus. Based on the field micro-plot experiments of three typical farmland soils(phaeozem, cambisol, and acrisol), metagenomic sequencing technology was used to study the differences in functional genes and regulatory factors of maize rhizosphere microorganisms during soil nitrogen and phosphorus transformation. The results showed that the functional diversity of maize rhizosphere microorganisms was affected by soil type. The functional diversity of rhizosphere microorganisms in phaeozem and cambisol was mainly affected by water content and nutrient content, and that in acrisol was affected by total phosphorus(TP) and available phosphorus(AP). For soil nitrogen transformation, the gene abundance of related enzymes in the pathway of nitrogen transformation was the highest in the urease gene(ureC) and glucose dehydrogenase gene(gdh), which were 7.25×10-5-12.88×10-5 and 4.47×10-5-7.49×10-5, respectively. The total abundance of assimilatory nitrate reduction functional genes in acrisol was higher than that in phaeozem and cambisol, and the total abundance of functional genes related to other processes was the highest in cambisol. The abundance of functional genes encoding enzymes related to nitrogen metabolism was mainly driven by soil bacterial richness, total potassium(TK), and TP. For soil phosphorus transformation, the number of alkaline phosphatase genes(phoD) catalyzing organic phosphorus mineralization was 1093, and the number of acid phosphatase genes(PHO) was 42. The abundance of phoD was two orders of magnitude higher than that of PHO. In addition, fertilization had no significant effect on the abundance of phoD and PHO in the same soil type. Random forest analysis showed that the abundances of phoD and PHO were significantly affected by soil moisture, organic matter(OM), and total nitrogen(TN), but AP content had the greatest impact on PHO abundance. These results clarified the nitrogen and phosphorus transformation characteristics of maize rhizosphere microorganisms at the functional genomic level and enriched the molecular biological mechanism of the microbial nitrogen and phosphorus transformation function.