Root rot disease caused by Fusarium oxysporum is a devastating disease of Salvia miltiorrhiza and dramatically affected the production and quality of Sa. miltiorrhiza. Besides the agricultural and chemical control, biocontrol agents can be utilized as an additional solution. In the present study, an actinomycete that highly inhibited F. oxysporum was isolated from rhizosphere soil and identified as based on morphological and molecular characteristics. Greenhouse assay proved that the strain had significant biological control effect against Sa. miltiorrhiza root rot disease and growth-promoting properties on Sa. miltiorrhiza seedlings. To elucidate the biocontrol and plant growth-promoting properties of St-220, we employed an analysis combining genome mining and metabolites detection. Our analyses based on genome sequence and bioassays revealed that the inhibitory activity of St-220 against F. oxysporum was associated with the production of enzymes targeting fungal cell wall and metabolites with antifungal activities. Strain St-220 possesses phosphate solubilization activity, nitrogen fixation activity, siderophore and indole-3-acetic acid production activity in vitro, which may promote the growth of Sa. miltiorrhiza seedlings. These results suggest that St. albidoflavus St-220 is a promising biocontrol agent and also a biofertilizer that could be used in the production of Sa. miltiorrhiza.
Callicarpa formosana is a species of beauty-berry with large medicinal value belonging to the family Verbenaceae. In this study, the complete chloroplast genome of C. formosana was sequenced using Illumina Hiseq X Ten platform. The chloroplast genome was 1,54,210 bp in length, containing two short inverted repeat (IRa and IRb) regions of 25,701 bp, which was separated by a large single copy (LSC) region of 84,938 bp and a small single copy (SSC) region of 17,870 bp. The GC content of the whole chloroplast genome was 38.1%. The chloroplast DNA of C. formosana comprised 113 genes, including 79 protein-coding genes, 4 ribosomal RNA genes, and 30 transfer RNA genes. Phylogenetic analysis indicated that the genus Callicarpa L. was located in the basal position within the family Verbenaceae. The chloroplast genome (cpDNA) of C. formosana was closely related to Callicarpa nudiflora.
The genus Coptis belongs to the Ranunculaceae family, containing 15 recognized species highly diverse in morphology. It is a conspicuous taxon with special evolutionary position, distribution pattern and medicinal value, which makes it to be of great research and conservation significance. In order to better understand the evolutionary dynamics of Coptis and promote more practical conservation measures, we performed plastome sequencing and used the sequencing data in combination with worldwide occurrence data of Coptis to estimate genetic diversity and divergence times, rebuild biogeographic history and predict its potential suitable distribution area. The average nucleotide diversity of Coptis was 0.0067 and the hotspot regions with the highest hypermutation levels were located in the ycf1 gene. Coptis is most likely to have originated in North America and Japanese archipelago and has a typical Eastern Asian and North American disjunct distribution pattern, while the species diversity center is located in Mid-West China and Japan. The crown age of the genus is estimated at around 8.49 Mya. The most suitable climatic conditions for Coptis were as follows: precipitation of driest quarter > 25.5 mm, annual precipitation > 844.9 mm and annual mean temperature -3.1 to 19 °C. The global and China suitable area shows an upward trend in the future when emission of greenhouse gases is well controlled, but the area, especially in China, decreases significantly without greenhouse gas policy interventions. The results of this study provide a comprehensive insight into the Coptis evolutionary dynamics and will facilitate future conservation efforts.
Quantitative acetyl-proteomics, a newly identified post-translational modification, is known to regulate transcriptional activity in different organisms. Neurospora crassa is a model ascomycete fungus maintained for biochemistry and molecular biology research; however, extensive studies of the functions of its acylation proteins have yet to be performed. In this study, using LC-MS/MS qualitative proteomics strategies, we identified 1909 modification sites on 940 proteins in N. crassa and analysed the functions of these proteins using GO enrichment, KEGG pathway, and subcellular location experiments. We classified the acetylation protein involvement in diverse pathways, and protein-protein interaction (PPI) network analysis further demonstrated that these proteins participate in diverse biological processes. In summary, our study comprehensively profiles the crosstalk of modified sites, and PPI among these proteins may form a complex network with both similar and distinct regulatory mechanisms, providing improved understanding of their biological functions in N. crassa. K E Y W O R D S acetylated proteins, Neurospora crassa, protein-protein interaction, quantitative acetylproteomics Neurospora crassa is an ascomycete fungus that has been used extensively in modern genetics, biochemistry and molecular biology research, maintaining a central role as a model organism for more than 80 years [1, 2]. Neurospora was first discovered as a contaminant in bakeries, and approximately 80 years later it was developed as an experimental organism in the 1920s [3-6]. In the latter 50 years, Neurospora has been used as a model fungus and has contributed to our fundamental understanding of genomic defines systems, such as DNA repair, DNA methylation, post-transcriptional gene silencing, mitochondrial protein import and circadian rhythms [7]. Moreover, Neurospora has also been used as a model system to study cellular differentiation and development, as well as other aspects of eukaryotic biology [2]. Subsequently, a high-quality draft sequence of the N. crassa genome has aided in efforts to investigative the molecular biology and genetics of filamentous fungi [8]. Studies of the genes regulating various aspects of Neurospora biology include the cAMP-dependent and heterotrimeric G-protein pathways [9, 10], mitogen-activated protein kinase pathway [11-13], light photobiology and circadian rhythmicity [14-17], secondary metabolism [18-20], cell wall structure and synthesis [21] and so on. With respect to N. crassa DNA, approximately 1.5% of the cytosines are methylated, facilitating genetic studies [22]. However, to
Lithocarpus litseifolius (Hance) Chun has been used as tea for 1500 years in China since the Southern and Northern Dynasties (AD 423). In mid-May 2021, a leaf spot disease of L. litseifolius (Hance) Chun was observed in Dexing County in Jiangxi Province, China (28°56'54.1"N 117°45'08.0"E). The disease incidence was estimated to be above 30%. The symptom of natural damage to plant leaves is early necrotic lesions, with a grayish-white center and a brown halo (<5mm) around it. Later, the lesion expanded to the edge of the leaf, with a yellowish-brown center and brown surrounding. Samples were collected from the field, and small tissues around the lesion were cut off and sterilized in 75% ethanol for 3 min, then washed with sterile water 3 times. The sterilized tissues were placed on PDA plates and incubated at 28°C in darkness. After 3 days of incubation, the hyphal tips from the edges of growing colonies were transferred to fresh PDA plates for further purification. Finally, five isolates showing the same morphology were obtained, and one pure single-colony fungal strain JM7 was selected for further analysis. After 7 days of incubation on PDA at 28°C, the fungal colonies showed white aerial mycelium and gelatinous orange conidial masses. The conidia were unicellular, smooth-walled, hyaline, cylindrical with obtuse to rounded ends, and their size was from 8.44 to 15.48 µm (mean = 10.33 µm, n = 60) × 2.95 to 3.42 µm (mean = 3.14 µm, n = 60). These morphological characteristics correspond to those of Colletotrichum nymphaeae. For molecular identification, the internal transcribed spacer, partial actin, β-tubulin 2, glyceraldehyde-3-phosphate dehydrogenase, chitin synthase, and histone H3 regions of JM7 were amplified by PCR using primer pairs ITS1/ITS4, ACT-512F/ACT-783R, T1/Bt2b, GDF/GDR, CHS-79F/CHS-354R and CYLH3F/CYLH3R, respectively (Weir et al. 2012;Crous et al. 2004). The sequences of the amplified fragments were deposited in GenBank (MZ496772, MZ561462-MZ561466), and showed 98 to 100% identities with the corresponding sequences of C. nymphaeae. A phylogenetic tree based on the above six genes of type specimens or ex-type of Colletotrichum (Damm et al. 2012) was generated using the maximum likelihood method by MEGA X, in which the strain JM7 and other C. nymphaeae strains clustered in the same branch. Thus, the strain JM7 was identified as C. nymphaeae based on its morphological and molecular characteristics. Pathogenicity tests were conducted on the leaf of L. litseifolius (Hance) Chun. Six leaflets were superficially disinfected with 75% ethanol and wound-inoculated with 6-mm plugs of PDA medium containing C. nymphaeae acervuli and conidia, while the control treatments received only PDA discs. The inoculated leaves were wrapped with a plastic bag containing moistened cotton and incubated in a greenhouse at 26°C 60% humidity with a photoperiod of 12 h. Symptoms similar to the original observations were observed on leaflets inoculated with C. nymphaeae after 5 days. No symptoms were observed in the control treatments. The pathogen was re-isolated from symptomatic tissues, and was identified as C. nymphaeae, which full-filled the Koch’s postulates. To our knowledge, this is the first report of C. nymphaeae infection on L. litseifolius (Hance) Chun in China. These findings will help develop better preventive measures in accordance with the emergence of this new pathogen.
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