Exogenous inoculation of endophytic bacterium Bacillus cereus suppresses clubroot (Plasmodiophora brassicae) occurrence in pak choi (Brassica campestris sp. chinensis L.).

MAIN CONCLUSION: The presence of Bacillus cereus plays a key role in clubroot suppression and improves plant biomass in pak choi. B. cereus is reported for the first time as a novel biocontrol agent against clubroot. Plasmodiophora brassicae Woronin causes a devastating infectious disease known as clubroot that is damaging to cruciferous vegetables. This study aimed to isolate beneficial bacteria from the rhizosphere soil of pak choi (Brassica campestris sp. chinensis) and to evaluate the ability of the isolate to reduce the severity of clubroot. Strains obtained from the rhizosphere of symptomless pak choi were first selected on the basis of their germination inhibition rate and effects on the viability of P. brassicae resting spores. Eight bacterial isolates had inhibitory effects against the resting spores of clubroot causing pathogen. However, MZ-12 showed the highest inhibitory effect at 73.4%. Inoculation with MZ-12 enhanced the plant biomass relative to plants grown without MZ-12 as well as P. brassicae infected plants. Furthermore, enhanced antioxidant enzymatic activities were observed in clubroot-infected plants during bacterial association. Co-inoculation of the plant with both P. brassicae and MZ-12 resulted in a 64% reduction of gall formation in comparison to plants inoculated with P. brassicae only. Three applications of MZ-12 to plants infected with P. brassicae at 7, 14 and 21 days after seeding (DAS) were more effective than one application and repressed root hair infection. According to 16S rDNA sequence analysis, strain MZ-12 was identified as had a 100% sequence similarity with type strain Bacillus cereus. The findings of the present study will facilitate further investigation into biological mechanisms of cruciferous clubroot control.

Involvement of IDA-HAE Module in Natural Development of Tomato Flower Abscission.

The unwanted detachment of organs such as flowers, leaves, and fruits from the main body of a plant (abscission) has significant effects on agricultural practice. Both timely and precise regulation of organ abscission from a plant is crucial as it influences the agricultural yield. The tomato (Solanum lycopersicum) has become a model system for research on organ abscission. Here, we characterized four tomato natural abscission variants named jointless (j), functionally impaired jointless (fij), functionally impaired jointless like (fij like), and normal joint (NJ), based on their cellular features within the flower abscission zones (AZ). Using eight INFLORESCENCE DEFICIENT IN ABSCISSION (SlIDA) genes and eight HAESA genes (SlHAE) identified in the genome sequence of tomato, we analyzed the pattern of gene expression during flower abscission. The AZ-specific expression for three tomato abscission polygalacturonases (SlTAPGs) in the development of flower AZ, and the progression of abscission validated our natural abscission system. Compared to that of j, fij, and fij like variants, the AZ-specific expression for SlIDA, SlIDL2, SlIDL3, SlIDL4, and SlIDL5 in the NJ largely corelated and increased with the process of abscission. Of eight SlHAE genes examined, the expression for SlHSL6 and SlHSL7 were found to be AZ-specific and increased as abscission progressed in the NJ variant. Unlike the result of gene expression obtained from natural abscission system, an in silico analysis of transcriptional binding sites uncovered that SlIDA genes (SlIDA, SlIDL6, and SlIDL7) are predominantly under the control of environmental stress, while most of the SlHSL genes are affiliated with the broader context in developmental processes and stress responses. Our result presents the potential bimodal transcriptional regulation of the tomato IDA-HAE module associated with flower abscission in tomatoes.

Controllable synthesis and stabilization of Tamarix aphylla-mediated copper oxide nanoparticles for the management of Fusarium wilt on musk melon.

Excessive use of pesticides and mineral fertilizers poses a serious threat to ecoenvironment sustainability and human health. Nano pesticides or Nano fungicides have attained great attention in the field of agriculture due to their unique characteristics, by improving crop growth with enhancing pathogenesis-related defense system. However, there is a need to develop a sustainable mechanism for the synthesis of fungicides which replace the chemical pesticides to avoid their hazardous impact. Here in, Tamarix aphylla mediated CuO-Nanoparticles (NPs) were synthesized, characterized and their activity was evaluated under in-vitro and in-vivo conditions. The structural and elemental analysis of NPs were carried out by using X-ray powder diffraction (XRD), Fourier transformed infrared spectroscopy (FTIR), UV-visible spectrophotometer, Scanning electron microscope (SEM) and Transmission electron microscope (TEM). In the greenhouse, at an optimum concentration of 50 mg/L reduced disease severity very effectively and enhanced plant growth. Application of NPs also assisted in the induction of systemic response of defense-related genes in melon. Under In vitro condition at 100 mg/L significantly reduced mycelial growth (84.5%) by directly acting on the pathogenic cell wall. Our work confirmed that dosedependent concentration of T. aphylla extract based biological CuO-NPs enhance plant growth and help to effectively resist against F. oxysporum infection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03189-0.

Reprisal of Schima superba to Mn stress and exploration of its defense mechanism through transcriptomic analysis.

One of the most diverse protein families, ATP-binding cassette (ABC) transporters, play a role in disease resistance, heavy metal tolerance, and food absorption.Differentially expressed genes contribute in the investigation of plant defense mechanisms under varying stress conditions. To elucidate the molecular mechanisms involved in Mn metal stress, we performed a transcriptomic analysis to explore the differential gene expression in Schima superba with the comparison of control. A total of 79.84 G clean data was generated and 6558 DEGs were identified in response to Mn metal stress. Differentially expressed genes were found to be involved in defense, signaling pathways, oxidative burst, transcription factors and stress responses. Genes important in metal transport were more expressive in Mn stress than control plants. The investigation of cis-acting regions in the ABC family indicated that these genes might be targeted by a large variety of trans-acting elements to control a variety of stress circumstances. Moreover, genes involved in defense responses, the mitogen-activated protein kinase (MAPK) signaling and signal transduction in S. superba were highly induced in Mn stress. Twenty ABC transporters were variably expressed on 1st, 5th, and 10th day of Mn treatment, according to the qRT PCR data. Inclusively, our findings provide an indispensable foundation for an advanced understanding of the metal resistance mechanisms. Our study will enrich the sequence information of S. superba in a public database and would provide a new understanding of the molecular mechanisms of heavy metal tolerance and detoxification.

PacBio Single-Molecule Long-Read Sequencing Reveals Genes Tolerating Manganese Stress in Schima superba Saplings.

Schima superba (Theaceae) is a subtropical evergreen tree and is used widely for forest firebreaks and gardening. It is a plant that tolerates salt and typically accumulates elevated amounts of manganese in the leaves. With large ecological amplitude, this tree species grows quickly. Due to its substantial biomass, it has a great potential for soil remediation. To evaluate the thorough framework of the mRNA, we employed PacBio sequencing technology for the first time to generate S. Superba transcriptome. In this analysis, overall, 511,759 full length non-chimeric reads were acquired, and 163,834 high-quality full-length reads were obtained. Overall, 93,362 open reading frames were obtained, of which 78,255 were complete. In gene annotation analyses, the Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Genes (COG), Gene Ontology (GO), and Non-Redundant (Nr) databases were allocated 91,082, 71,839, 38,914, and 38,376 transcripts, respectively. To identify long non-coding RNAs (lncRNAs), we utilized four computational methods associated with protein families (Pfam), Cooperative Data Classification (CPC), Coding Assessing Potential Tool (CPAT), and Coding Non-Coding Index (CNCI) databases and observed 8,551, 9,174, 20,720, and 18,669 lncRNAs, respectively. Moreover, nine genes were randomly selected for the expression analysis, which showed the highest expression of Gene 6 (Na_Ca_ex gene), and CAX (CAX-interacting protein 4) was higher in manganese (Mn)-treated group. This work provided significant number of full-length transcripts and refined the annotation of the reference genome, which will ease advanced genetic analyses of S. superba.

Cd-tolerant SY-2 strain of Stenotrophomonas maltophilia: a potential PGPR, isolated from the Nanjing mining area in China.

Microbial and plant assisted bioremediation is an emerging way for the remediation of soils polluted with heavy metals. To screen the cadmium tolerant bacteria, soil samples were collected from Nanjing mining area, China. The average cadmium content of the mine soil reached 45.71 mg/kg, which was indicating serious pollution and potential ecological risk. From the mine soil, six cadmium tolerant plant growth-promoting rhizobacteria (PGPR) were isolated. The isolated bacterial strain "SY-2" showed maximum cadmium tolerance and it was selected for further experimentation. This strain was identified as Stenotrophomonas maltophilia by 16S rRNA gene sequencing (GenBank accession number MG597057). SY-2 was found to tolerate maximum cadmium at 1.0 mM concentration. This strain also exhibited good adsorption capacity (up to 35.7%) of heavy metal at 0.5 mM concentration. The results of this study exhibited organic phosphorus solubilization (37.08 mg/L) and IAA biosynthesis (15.11 mg/L) ability of isolated S. maltophilia. Scanning electron microscopy (SEM) revealed cell shrinkage and the cell wall of S. maltophilia was very rough. Moreover, the energy dispersive X-ray (EDX) analysis endorsed the adsorption of Cd ions on the surface of biomass. FT-IR study described the presence of functional groups and the nature of chemical bonds, before and after cadmium stress. At 0.25 mM cadmium concentration, S. maltophilia treated seeds of Capsicum annuum L. developed 1.46 times longer roots than untreated seeds. The results of this study helped us to conclude that SY-2 strain of S. maltophilia possesses significant metal tolerance and bioremediation potential against cadmium. In the future, this strain can be used as a microbial remediation agent to detoxify heavy metals in contaminated soils. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-020-02524-7s) contains supplementary material, which is available to authorized users.

A calmodulin-like CmCML13 from Cucumis melo improved transgenic Arabidopsis salt tolerance through reduced shoot's Na+, and also improved drought resistance.

The calmodulin-like proteins (CMLs) are a large family involved in plant biological processes. A calmodulin-like gene CmCML13 (GenBank accession number: MT340534) from melon (Cucumis melo L.) was isolated and functionally analyzed. CmCML13 was predicted to possess 3 EF-hands in which only the first EF-hand could bind with Ca2+. Subcellular localization assay revealed that CmCML13 was localized in nucleus, cell membrane, vacuolar membrane and cytoplasmic strand. The transcript level of CmCML13 was temporally and spatially regulated under salt stress. Constitutive expression of CmCML13 in the Arabidopsis thaliana enhanced salt tolerance at seeds germination. CmCML13 improved the transgenic Arabidopsis plants salt tolerance by significantly reducing Na+ content of shoots, which was unrelated to HKT1-involving pathway. Moreover, overexpressing of CmCML13 in Arabidopsis showed stronger drought tolerance. This study demonstrates that the CmCML13 is an important multifunctional protein associated with salt and drought stress, which may play a key role in stress signaling pathway.

Evaluation of Metal Tolerance of Fungal Strains Isolated from Contaminated Mining Soil of Nanjing, China.

Rapidly increasing industry has resulted in greater discharge of hazardous chemicals in the soil. In the current study, soil samples were collected from Nanjing mine (32°09'19.29″ N 118°56'57.04″ E) and subjected to heavy metal analysis and microbe isolation. A total of 460 fungi were isolated, and five of these were yeast strains. Most of the strains exhibited tolerance to one metal. Five multimetal tolerant strains were selected and identified as Aspergillus sclerotiorum, Aspergillus aculeatus, Komagataella phaffii, Trichoderma harzianum, and Aspergillus niger. Isolated strains were grown in high concentrations of cadmium (Cd), chromium (Cr) and lead (Pb), for induced-tolerance training. The tolerance index (TI) revealed the highest Cd tolerance of novel K. phaffii strain at 5500 ppm (TI: 0.2). K. phaffii also displayed resistance at 4000 ppm against Cr (TI: 0.32) and Pb (TI: 0.32). In contrast, tolerance training for A. niger was not that successful. K. phaffii also displayed the highest bioaccumulation capacity for Cd (25.23 mg/g), Cu (21.63 mg/g), and Pb (20.63 mg/g) at 200 ppm. Scanning electron microscopy (SEM) explored the morphological changes in the mycelia of stressed fungi. Results of this study describe this delicate approach to be species and metal dependent and suggest a potential utilization of this fungal strain for the bioremediation of contaminated soils.