Molecular Soil Biology 2025, Vol.16, No.1, 45-54 http://bioscipublisher.com/index.php/msb 49 It is not easy to control environmental conditions. Temperature, humidity and light must be adjusted in place. A slight fluctuation may affect the activity of microorganisms. Sometimes when the environment changes, the beneficial bacteria may be reduced, but the harmful bacteria may become active (Wang et al., 2020). Microbial inoculation should be precise. For example, the inoculation location, dosage and time should be well controlled, otherwise beneficial bacteria are easy to be "squeezed out" by other bacteria in the soil (Wang et al., 2022). 5.2 Successfully inoculated microorganisms and their growth promoting effects In the existing studies, some microorganisms have been successfully applied in Anoectochilus roxburghii tissue culture seedlings. For example, Bacillus subtilis can effectively inhibit Fusarium, a common pathogenic fungus, so as to make Anoectochilus roxburghii grow healthier (Dong et al., 2018). The method of inoculation can be seed wrapping, rhizosphere injection or soil root irrigation to directly send the bacteria around the plant roots (Wang et al., 2022). In terms of effect, obvious changes can usually be seen after inoculation. Beneficial bacteria can not only improve the nutrient utilization rate, but also improve the soil structure, such as making the soil more loose and better water retention. Mortierella alpina was found to improve the availability of nitrogen and phosphorus in soil, and these two elements are very important for Anoectochilus roxburghii (Wang et al., 2022). Such microorganisms can also secrete plant hormones and other active substances to directly stimulate the growth of roots and stems (Hakim et al., 2022). 5.3 How the culture environment affects microbial activity and plant growth The culture environment has a great influence on the growth of microorganisms and Anoectochilus roxburghii. If the temperature is too high or too low, and the humidity is insufficient, some microorganisms will lose activity, or even die. After microbial inactivation, rhizosphere ecology will also change, beneficial bacteria will decrease, and bacteria will easily spread (Wang et al., 2020). Light is also a key factor. Plants grow by light cooperation. The more sufficient the light, the higher the photosynthetic efficiency of plants and the more energy they produce. These energies are not only supplied to themselves, but also released in the form of root exudates, further affecting the species and activities of root microorganisms (Sun et al., 2023) (Figure 2). Light intensity may also affect the synthesis of secondary metabolites such as ginsenosides in Anoectochilus roxburghii, which are the source of medicinal value of Anoectochilus roxburghii (Sun et al., 2022). Figure 2 Relative abundances of fungal phyla (A) and genera (B) in the rhizosphere soil of ginseng under different root exudate concentrations (n = 3) (Adopted from Sun et al., 2023) Image caption: Root exudate concentrations: CK, 0 mg·g−1; T1: 0.3 mg·g−1; T2: 1.5 mg·g−1; T3: 3mg·g−1; T4: 6mg·g−1;T5,15 mg·g−1 (Adopted from Sun et al., 2023)
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