The Biocontrol Functions of Bacillus velezensis Strain Bv-25 Against Meloidogyne incognita.

Meloidogyne incognita is obligate parasitic nematode with a wide variety of hosts that causes huge economic losses every year. In an effort to identify novel bacterial biocontrols against M. incognita, the nematicidal activity of Bacillus velezensis strain Bv-25 obtained from cucumber rhizosphere soil was measured. Strain Bv-25 could inhibit the egg hatching of M. incognita and had strong nematicidal activity, with the mortality rate of second-stage M. incognita juveniles (J2s) at 100% within 12 h of exposure to Bv-25 fermentation broth. The M. incognita genes ord-1, mpk-1, and flp-18 were suppressed by Bv-25 fumigation treatment after 48 h. Strain Bv-25 could colonize cucumber roots, with 5.94 × 107 colony-forming units/g attached within 24 h, effectively reducing the infection rate with J2s by 98.6%. The bacteria up-regulated the expression levels of cucumber defense response genes pr1, pr3, and lox1 and induced resistance to M. incognita in split-root trials. Potted trials showed that Bv-25 reduced cucumber root knots by 73.8%. The field experiment demonstrated that disease index was reduced by 61.6%, cucumber height increased by 14.4%, and yield increased by 36.5% in Bv-25-treated plants compared with control. To summarize, B. velezensis strain Bv-25 strain has good potential to control root-knot nematodes both when colonizing the plant roots and through its volatile compounds.

Application of neoadjuvant chemotherapy combined with anlotinib in occult breast cancer: A case report and review of literature.

BACKGROUND: Occult breast cancer (OBC) is a special type of breast cancer presenting as axillary lymph node metastasis with undetectable primary lesions in the breast. Due to its low incidence and unique clinical manifestations, there is a lack of consensus on the diagnosis and treatment of OBC. We report a case of OBC treated with neoadjuvant chemotherapy combined with anlotinib. The treatment was well tolerated, and the patient achieved a pathologic complete response. CASE SUMMARY: A 53-year-old woman presented with a lump in her right axillary area with no primary lesions in the breast. Pathological biopsy confirmed right axillary metastatic carcinoma. Immunohistochemical staining results were positive for progesterone receptor, cytokeratin 7, specific breast markers GATA3 and gross cystic disease fluid protein-15. Tumor cells were negative for estrogen receptor, human epidermal growth factor receptor-2, cytokeratin 5/6, cytokeratin 20, and villin. The patient was diagnosed with OBC, and she underwent neoadjuvant chemotherapy combined with anlotinib. Mastectomy plus axillary lymph node dissection was performed. The patient achieved pathologic complete response with no residual invasive tumor cells in the breast or axillary lymph nodes. Postoperatively, she received adjuvant radiotherapy and endocrine therapy. CONCLUSION: Neoadjuvant chemotherapy and anlotinib had good efficacy and safety in the treatment of OBC and may be a new therapeutic option.

Identification of endophytes with biocontrol potential from Ziziphus jujuba and its inhibition effects on Alternaria alternata, the pathogen of jujube shrunken-fruit disease.

Endophytic strains were isolated from different parts of a healthy "Dongzao" jujube (Ziziphus jujuba Mill. 'Dongzao') to find biocontrol agents against jujube shrunken-fruit disease caused by Alternaria alternata. The strains were screened using A. alternata strain CN193 as the target pathogen. The nutrient competition for all isolates was studied using the dual culture, and their inhibitive capability was tested by measuring the inhibition width of filter paper disks with filtrate. Influence of filtrate from the selected strains with strong inhibition of mycelial growth on spore germination was studied with hanging drop method on concavity slides. Colonization in the jujube leaves was assayed using a rifampicin-resistant mutant of strain St-zn-34 as the screening marker. Strains were identified based on their morphological, physiological, and biochemical characteristics, 16S rDNA sequencing, and phylogenetic analysis. A total of 81 endophytic strains were isolated from the stems, leaves, flowers, and fruits of winter jujube. Among these isolates, 14 strains showed strong antagonism against A. alternata. Further study showed that the filtrate of strains St-zn-9 and St-zn-34 could inhibit the mycelial growth of A. alternata, and the widths of their inhibition zone reached 6.14±0.03 mm and 8.27±0.09 mm, respectively. However, strain St-zn-34 showed stronger inhibition on spore germination than strain St-zn-9. St-zn-34 could significantly reduce the spore germination rate of A. alternata, and the spore did not germinate at all or the germ tube was very short. A rifampicin resistant-derivative of wild-type strain St-zn-34, which was designated as St-zn-34r, was obtained by transferring the strains to media with stepwise-increased rifampicin. Colonization assays indicated that St-zn-34r could colonize in jujube leaves, and the population of St-zn-34r was 1.2×103 CFU/g FW after inoculation for 30 days. Except for its salt tolerance, St-zn-34 was the closest to those of Bacillus subtilis. Thus, the strain was identified as B. subtilis.

Transformation of the endophytic fungus Acremonium implicatum with GFP and evaluation of its biocontrol effect against Meloidogyne incognita.

Acremonium implicatum is an endophytic fungus with biocontrol potential against Meloidogyne incognita based on its opportunistic egg-parasitic, hatching inhibition, and toxic properties. To understand its mode of plant endophytism and opportunistic egg parasitism, GFP-tagged A. implicatum was constructed by PEG-mediated protoplast transformation. By laser scanning confocal microscopy (LSCM), we evaluated the endophytism and opportunistic egg parasitism of a stable gfp transformant (Acr-1). Acr-1 could colonize epidermal tissue, cortical tissue, and xylem of roots and form a mutualistic symbiosis with tomato host plants. LSCM of Acr-1 infecting M. incognita eggs revealed that hyphae penetrated the shell and grew inside eggs to form trophic hyphae. A large number of hyphae enveloped parasitized eggs. In addition, the egg shell integrity was destroyed by fungal penetration. The percentage of egg parasitism was 33.8 %. There were no marked differences between the wild type and mutant in nematode second-stage juvenile mortality and egg hatching and in fungal control efficiency in a pot experiment. In conclusion, gfp-transformation did not change the nematicidal activity of A. implicatum and is a tool to examine the mode of plant endophytism and opportunistic egg parasitism of A. implicatum.

Metagenomic analysis of the pinewood nematode microbiome reveals a symbiotic relationship critical for xenobiotics degradation.

Our recent research revealed that pinewood nematode (PWN) possesses few genes encoding enzymes for degrading α-pinene, which is the main compound in pine resin. In this study, we examined the role of PWN microbiome in xenobiotics detoxification by metagenomic and bacteria culture analyses. Functional annotation of metagenomes illustrated that benzoate degradation and its related metabolisms may provide the main metabolic pathways for xenobiotics detoxification in the microbiome, which is obviously different from that in PWN that uses cytochrome P450 metabolism as the main pathway for detoxification. The metabolic pathway of degrading α-pinene is complete in microbiome, but incomplete in PWN genome. Experimental analysis demonstrated that most of tested cultivable bacteria can not only survive the stress of 0.4% α-pinene, but also utilize α-pinene as carbon source for their growth. Our results indicate that PWN and its microbiome have established a potentially mutualistic symbiotic relationship with complementary pathways in detoxification metabolism.

[Ecological relationships between Bursaphelenchus xylophilus and its companion microorganisms].

Pine wood nematode Bursaphelenchus xylophilus is a notorious invasive species from North America, which can kill a large amount of pine trees and causes economic losses and ecosystem destruction. There is a close relationship and ecological interaction between B. xylophilus and its companion microorganisms. This paper listed the species of companion microorganisms, reviewed their important ecological roles in the propagation and pathogenicity of the nematode, and discussed the pine wilt disease from the viewpoint of microecosystem. The companion fungi can supply food for B. xylophilus, hold the cycle of second infection of the nematode, increase the proportions of dauer juveniles, and benefit the infection and distribution of B. xylophilus. The companion bacteria can enhance the pathogenicity of B. xylophilus, promote the propagation of the nematode, benefit the pinene degradation, and thereby, promote the adaptability of the nematode.