Assessment of Rhizospheric Microorganisms of Transgenic Populus tomentosa with Cowpea Trypsin Inhibitor (CpTI) Gene

To have a preliminary insight into biosafety of genetically transformed hybrid triploid poplars （Populus tomentosa × P bolleana）× P. tomentosa with the cowpea trypsin inhibitor （CpTD gene, two layers of rhizospheric soil （from 0 to 20cm deep and from 20 to 40cm deep, respectively） were collected for microorganism culture, counting assay and PCR analysis to assess the potential impact of transgenic poplars on non-target microorganism population and transgene dispersal. When the same soil layer of suspension stock solution was diluted at both 1：1000 and 1：10000 rates, there were no significant differences in bacterium colony numbers between the inoculation plates of both transgenic and non-transgenic poplars. The uniform results were revealed for both soil layer suspension solutions of identical poplars at both dilution rates except for non-transgenic poplars at 1：10000 dilution rates from the same type of soil. No significant variation in morphology of both Gram-positive and Gram-negative bacteria was observed under the microscope. The potential transgene dispersal from root exudates or fallen leaves to non-target microbes was repudiated by PCR analysis, in which no CpTI gene specific DNA band was amplified for 15 sites of transgenic rhizospheric soil samples. It can be concluded that transgenic poplar with the CpTI gene has no severe impact on rhizospheric microorganisms and is tentatively safe to surrounding soil micro-ecosystem.

Integrated Biological Control of Tobacco Bacterial Wilt (<i>Ralstonia solanacearum</i>) and Its Effect on Rhizosphere Microbial Community

Biological control agents and soil amendments have been applied to control tobacco bacterial wilt, but the mechanism is not well-known. In the present study, a field experiment was performed to investigate the soil physicochemical properties, disease index (DI) and disease incidence of tobacco bacterial wilt, and rhizosphere microbial community. The results show that the control efficacy of single application of antagonistic bacteria and calcium cyanamide was 46.43% and 51.92%, respectively. While the combined control efficacy of antagonistic bacteria and calcium cyanamide was 65.79%. Besides, the combined application of antagonistic bacteria and calcium cyanamide could increase soil pH, total N alkaline N, and exchangeable Ca, which were negatively associated with the microbial diversity, soil-borne pathogenic microorganisms, and incidence of tobacco bacterial wilt. Additionally, the combination of antagonistic bacteria and calcium cyanamide can improve the proportion of some antagonistic microbial species, and these antagonistic microbial species were inversely associated with the DI of tobacco bacterial wilt. In conclusion: The integrated measure may influence soil microbial communities through enhancing soil physicochemical properties and rebuild healthy soil microbial community structure to mitigate tobacco bacterial wilt. The current study presented valuable insights into the mechanisms enhancing soil health in the integrated measure.

Using Phospholipid FattyAcid Technique toAnalysis the Rhizosphere Specific Microbial Community of Seven Hybrid Rice Cultivars

To analyze the intrinsic relationship between rhizosphere microbial community structure and variety of rice, the microbial community structures in rhizosphere of different hybrid rice cultivars were determined with phospholipid fatty acids （PLFA） analysis. Three series of new-breeding hybrid rice cultivars in China were tested in the experiment, Ilyouming 86 （II-32A/Minghui 86）, Ilyouhang 1 （II-32A/Hang 1）, and Ilyouhang 2 （II-32A/Hang 2） with H-32A as female parent, XinyouHK02 （XinA/HK02） and YiyouHK02 （YXA/HK02） with HK02 as male parent, Chuanyou 167 （ChuanxiangA/MR167） and 44you167 （Hunan44A/MR167） with MR167 as male parent. The results showed that the microbial community in rhizosphere of the hybrid rice comprised bacteria, fungi, actinomycetes, and protozoa, according to the 40 PLFA biomarkers detected. Bacteria were more abundant than fungi and actinomycetes in rhizosphere of the hybrid rice tested. Both sulfate-reducing and methane-oxidizing bacteria were found to exist in the hybrid rice rhizosphere. It was also found that the characteristics of PLFA biomarkers had correlation with the biological traits of rice. The cluster analysis suggested that microbial community structure and activity in rhizosphere were associated with genetic background of the rice cultivar.

Rhizosphere microbial markers (micro-markers): A new physical examination indicator for traditional Chinese medicines

Rhizosphere microorganisms,as one of the most important components of the soil microbiota and plant holobiont,play a key role in the medicinal plant-soil ecosystem,which are closely related to the growth,adaptability,nutrient absorption,stress tolerance and pathogen resistance of host plants.In recent years,with the wide application of molecular biology and omics technologies,the outcomes of rhizosphere microorganisms on the health,biomass production and secondary metabolite biosynthesis of medicinal plants have received extensive attention.However,whether or to what extent rhizosphere microorganisms can contribute to the construction of the quality evaluation system of Chinese medicinal materials is still elusive.Based on the significant role of rhizosphere microbes in the survival and quality formation of medicinal plants,this paper proposed a new concept of rhizosphere microbial markers(micro-markers),expounded the relevant research methods and ideas of applying the new concept,highlighted the importance of micro-markers in the quality evaluation and control system of traditional Chinese medicines(TCMs),and introduced the potential value in soil environmental assessment,plant pest control and quality assessment of TCMs.It provides reference for developing ecological planting of TCMs and ensuring the production of high quality TCMs by regulating rhizosphere microbial communities.

Microbial Fertilizer: A Sustainable Strategy for Medicinal Plants Production

Medicinal plants have aroused considerable interest as an alternative to chemical drugs due to the beneficial effects of their active secondary metabolites.However,the extensive use of chemical fertilizers and pesticides in pursuit of yield has caused serious pollution to the environment,which is not conducive to sustainable devel-opment in thefield of medicinal plants.Microbial fertilizers are a type of“green fertilizer”containing specific microorganisms that can improve the soil microbial structure,enhance plant resistance to biological and abiotic stresses,and increase the yield of medicinal plants.The root exudates of medicinal plants attract different micro-organisms to the rhizosphere,which then migrate further to the plant tissues.These microbes can increase the levels of soil nutrients,and improve the physical and chemical properties of soil through nitrogenfixation,and phosphorus and potassium solubilization.In addition,soil microbes can promote the synthesis of hormones that increase plant growth and the accumulation of active compounds,eventually improving the quality of med-icinal plants.In 2022,the total value of the global microbial fertilizer market was$4.6 billion and is estimated to reach$10.36 billion by 2030.In this review,we have summarized the types of microbial fertilizers,the coloniza-tion and migration of microorganisms to plant tissues,and the beneficial effects of microbial fertilizers.In addi-tion,the prospects of developing microbial fertilizers and their application for medicinal plants have also been discussed.It aims to provide a reference for the rational application of microbial fertilizers in thefield of med-icinal plants and the green and sustainable development of medicinal plant resources.

Bacillus and microalgae biofertilizers improved quality and biomass of Salvia miltiorrhiza by altering microbial communities

Objective: Biofertilizers are reliable alternatives to chemical fertilizers due to various advantages. However, the effect of biofertilizers on Salvia miltiorrhiza yield and quality and the possible mechanisms remain little known. Here, an experiment was conducted in S. miltiorrhiza field treated with two kinds of biofertilizers including Bacillus and microalgae.Methods: A field experiment was conducted on S. miltiorrhiza of one year old. The biofertilizers were applied at six treatments:(i) control check, CK;(ii) microalgae, VZ;(iii) Bacillus, TTB;(iv)microalgae + Bacillus(1:1), VTA;(v) microalgae + Bacillus(0.5:1), VTB;(vi) microalgae+Bacillus(1:0.5),VTC. Here, high-throughput sequencing, ICP-MS and UPLC were employed to systematically characterize changes of microbial diversity and structure composition, heavy metals content and bioactive compounds, respectively.Results: Compared to CK, root biomass increased by 29.31%-60.39%(P < 0.001). Meanwhile, bioactive compounds were higher than CK after the application of the biofertilizers, peculiarly in TTB and VTB.However, the content of Pb contents in roots significantly reduced by 46.03% and 37.58% respectively in VTC and TTB(P<0.05). VTA application notably increased the available nitrogen content by 53.03%(P<0.05), indicating the improvement of soil fertility. Significantly, bacterial and fungal Chao I diversity indices showed an increasing trend with biofertilizer application(P < 0.05), and biofertilizer amendment enriched the rhizosphere soil with beneficial microorganisms that have abilities on promoting plant growth(Achromobacter and Penicillium), adsorbing heavy metal(Achromobacter and Beauveria), controlling plant pathogen(Plectosphaerella, Lechevalieria, Sorangium, Phlebiopsis and Beauveria) and promoting the accumulation of metabolites(Beauveria and Phoma).Conclusion: Bacillus and microalgae biofertilizers improved the quality and biomass of S. miltiorrhiza by altering microbial communities in soil.

Biosurfactant-assisted phytoremediation of potentially toxic elements in soil:Green technology for meeting the United Nations Sustainable Development Goals

Biosurfactants are biomolecules produced by microorganisms, low in toxicity, biodegradable, and relatively easy to synthesize using renewable waste substrates. Biosurfactants are of great importance with a wide and versatile range of applications, including the bioremediation of contaminated sites. Plants may accumulate soil potentially toxic elements(PTEs), and the accumulation efficacy may be further enhanced by the biosurfactants produced by rhizospheric microorganisms. Occasionally, the growth of bacteria slows down in adverse conditions, such as highly contaminated soils with PTEs. In this context,the plant's phytoextraction capacity could be improved by the addition of metal-tolerant bacteria that produce biosurfactants. Several sources, categories,and bioavailability of PTEs in soil are reported in this article, with the focus on the cost-effective and sustainable soil remediation technologies, where biosurfactants are used as a remediation method. How rhizobacterial biosurfactants can improve PTE recovery capabilities of plants is discussed, and the molecular mechanisms in bacterial genomes that support the production of important biosurfactants are listed. The status and cost of commercial biosurfactant production in the international market are also presented.

Diclofop-methyl affects microbial rhizosphere community and induces systemic acquired resistance in rice

Diclofop-methyl（DM）,a widely used herbicide in food crops,may partly contaminate the soil surface of natural ecosystems in agricultural area and exert toxic effects at low dose to nontarget plants.Even though rhizosphere microorganisms strongly interact with root cells,little is known regarding their potential modulating effect on herbicide toxicity in plants.Here we exposed rice seedlings（Xiushui 63） to 100 μg/L DM for 2 to 8 days and studied the effects of DM on rice rhizosphere microorganisms,rice systemic acquired resistance（SAR） and rice-microorganisms interactions.The results of metagenomic 16 S rDNA Illumina tags show that DM increases bacterial biomass and affects their community structure in the rice rhizosphere.After DM treatment,the relative abundance of the bacterium genera Massilia and Anderseniella increased the most relative to the control.In parallel,malate and oxalate exudation by rice roots increased,potentially acting as a carbon source for several rhizosphere bacteria.Transcriptomic analyses suggest that DM induced SAR in rice seedlings through the salicylic acid（but not the jasmonic acid） signal pathway.This response to DM stress conferred resistance to infection by a pathogenic bacterium,but was not influenced by the presence of bacteria in the rhizosphere since SAR transcripts did not change significantly in xenic and axenic plant roots exposed to DM.The present study provides new insights on the response of rice and its associated microorganisms to DM stress.