Rhizosphere and endophytic fungal communities are considered critically important for plant health and soil fertility. In response to continuous cropping, Panax notoginseng becomes vulnerable to attack by fungal pathogens. In the present study, culture-independent Illumina MiSeq was used to investigate the rhizospheric and root endophytic fungi in response to continuous Panax notoginseng cropping practices. The results demonstrated that fungal diversity is increased inside the roots and in rhizospheric. Ascomycota, Zygomycota, Basidiomycota and Chytridiomycota were the dominant phyla detected during the continuous cropping of Panax notoginseng. The fungal diversity in the rhizospheric soil and roots of root-rot P. notoginseng plants are less than that of healthy plants in the same cultivating year, thus showing that root-rot disease also affects the community structure and diversity of rhizospheric and root endophytic fungi. Similarities in the major fungal components show that endophytic fungal communities are similar to rhizospheric soil fungal community based on a specialized subset of organisms. Canonical correspondence analysis on the fungal communities in root-rot rhizospheric from both healthy plants and rotation soils reveals that the soil pH and organic matter have the greatest impact upon the microbial community composition during continuous cropping, whereas soil nutrition status does not significantly affect the fungal community composition in response to continuous cropping practices. In addition, the results suggest that the unclassified genera Leotiomycetes, Cylindrocarpon, Fusarium and Mycocentrospora are shown as the potential pathogens which are responsible for the obstacles in continuous cropping of P. notoginseng. Further exploration of these potential pathogens might be useful for the biological control of continuous cropping of P. notoginseng.

Rhizobacteria and endophytic bacteria play important roles in protecting host plants from infection by phytopathogens, which cause soil-borne diseases and severely impair plant health. Panax notoginseng is negatively affected by continuous cropping and becomes vulnerable to attack by microbial pathogens. In the present study, culture-independent Illumina MiSeq was used to investigate root-endophytic and rhizospheric bacteria in response to continuous cropping of P. notoginseng. Numbers of rhizospheric bacteria decreased with continuous P. notoginseng cropping, while the effects of continuous cropping on endophytic bacteria were not statistically significant. Bacterial diversity was higher in healthy P. notoginseng rhizospheric soils and roots than in those of diseased P. notoginseng. The most dominant phyla detected during continuous cropping were Proteobacteria, Cyanobacteria, Actinobacteria, and Acidobacteria. The genera Pseudomonas, Rhodoplanes, Candidatus Solibacter, and Streptomyces were dominant in P. notoginseng rhizospheric soils and roots. Erwinia, Stenotrophomonas, Pseudomonas, and Sphingobium were specifically detected in relatively high proportions among root-rot rhizospheric bacteria and endogenous root bacteria in plants under continuous cropping, suggesting that they may be the pathogens responsible for the negative effects of continuous cropping on P. notoginseng. Based on canonical correspondence analysis of the bacterial communities that were identified from healthy plants and fallow soils, total phosphorus (P), pH, and organic soil matter exerted the greatest impacts upon bacterial community composition during continuous cropping. In general, continuous cropping practices for P. notoginseng and severe root-rot disease notably affected the community structure and the diversity of rhizospheric and root endophytic bacteria. Our study elucidated the ecological significance of microbial communities in healthy plant maintenance, and our results may inform biological control strategies during the continuous cropping of P. notoginseng.

Large-scale intensive cultivation has made continuous cropping soil sickness more serious for Panax notoginseng in Yunnan. Autotoxic substances can promote the occurrence of continuous cropping soil sickness. Phenolic acids exert a strong autotoxic effect on P. notoginseng. Based on UPLC-MS/MS, the levels of six phenolic acids with the strongest autotoxicity of P. notoginseng rhizospheric soil were tested. Based on Illumina MiSeq high-throughput sequencing technology, the variation in the microbial diversity in the rhizospheric soil was used as an index to explore the interactions between phenolic acids and the soil microorganisms of the P. notoginseng rhizosphere. (1) Continuous P. notoginseng cropping significantly changed the microbial community structure. Continuous cropping increased bacterial Chao1 index and Shannon index and decreased fungal Shannon index. After P. notoginseng disease, bacterial Shannon index reduced and fungal Chao1 index decreased. (2) Phenolic acid significantly changed the bacterial community structure. VA significantly reduced the bacterial Shannon index. Exogenous p-HA, FA, SA, and VA significantly increased the fungal Chao1 index and p-HA showed the most significant effect. Para-HA affected bacterial specificity, and VA affected fungal specificity. (3) VA was positively correlated with most fungi and bacteria. Para-HA was positively correlated with Lelliottia and Flavobacterium. Para-HA was also positively correlated with plant pathogens (Fusarium and Ilyonectria). Para-HA and VA were able to promote the growth of primary pathogenic bacteria. Thus, p-HA and VA are the main phenolic acid-autotoxin substances in P. notoginseng under continuous cropping. (4) A correlation analysis of soil environmental factors associated with fungal and bacterial communities showed that AK, TN, OM, and HN were most strongly correlated with soil microorganisms. (5) The microorganisms in the rhizosphere of 3-year-old soil planted with P. notoginseng exhibited obvious effects on the degradation of the four phenolic acids. The effect of soil microorganisms on phenolic acids was first-order kinetic degradation with a high degradation rate and a half-life of less than 4.5 h. The results showed that phenolic acids could promote the growth of pathogenic bacteria. And the interaction between rhizospheric soil microorganisms and phenolic acids was the main cause of the disturbance of P. notoginseng rhizosphere microflora.

A class 1 non-symbiotic hemoglobin family gene, SoHb, was isolated from spinach. qRT-PCR showed that SoHb was induced by excess nitrate, polyethylene glycol, NaCl, H2O2, and salicylic acid. Besides, SoHb was strongly induced by application of nitric oxide (NO) donor, while was suppressed by NO scavenger, nitrate reductase inhibitor, and nitric oxide synthase inhibitor. Overexpression of SoHb in Arabidopsis resulted in decreased NO level and sensitivity to nitrate stress, as shown by reduced root length, fresh weight, the maximum photosystem II quantum ratio of variable to maximum fluorescence (Fv/Fm), and higher malondialdehyde contents. The activities and gene transcription of superoxide dioxidase, and catalase decreased under nitrate stress. Expression levels of RD22, RD29A, DREB2A, and P5CS1 decreased after nitrate treatment in SoHb-overexpressing plants, while increased in the WT plants. Moreover, SoHb-overexpressing plants showed decreased tolerance to NaCl and osmotic stress. In addition, the SoHb-overexpression lines showed earlier flower by regulating the expression of SOC, GI and FLC genes. Our results indicated that the decreasing NO content in Arabidopsis by overexpressing SoHb might be responsible for lowered tolerance to nitrate and other abiotic stresses.

Schematic of the synthetic route to fluorescent Pn-CDs and Pn N-CDs and their applications in the bioimaging and detection of Cr6+.