Phosphate-solubilizing microbes (PSMs) in soil play a crucial role in converting insoluble phosphates into plant-available soluble phosphorus. This paper systematically presents a comprehensive array of qualitative and quantitative techniques to assess the phosphate-decomposing capabilities of microbes. Additionally, it introduces two optimized media, namely improved Monkina medium No. 1 and No. 2, which are particularly suitable for detecting the solubilization abilities of microbes toward insoluble organic phosphates. Talaromyces nanjingensis, a novel fungal species recently isolated from the rhizosphere soil of Pinus massoniana, demonstrates remarkable phosphate-solubilizing abilities. Across multiple temperature gradients (15 °C, 20 °C, 25 °C, 30 °C, and 37 °C), it effectively decomposes both insoluble inorganic and organic phosphates. This is achieved through the secretion of organic acids, including gluconic acid (6.10 g L−1), oxalic acid (0.93 g L−1), and malonic acid (0.17 g L−1), as well as phosphate-solubilizing enzymes. Moreover, under low-, medium-, and high-temperature conditions, T. nanjingensis can decompose insoluble phosphates in three types of soil with varying pH levels, thereby enhancing the overall soil fertility. Genomic analysis of T. nanjingensis has identified approximately 308 genes associated with phosphate decomposition and environmental adaptability, validating its superior capabilities and multi-faceted strategies for phosphate mobilization. These findings underscore the wide applicability of T. nanjingensis in maintaining soil phosphorus homeostasis and optimizing the phosphorus use efficiency, highlighting its promising potential for agricultural and environmental applications.

In this study, a germination test was conducted using the seeds of Tilia miqueliana, a rare tree species in China. The seeds were treated with H2SO4, H2SO4-gibberellic acid (GA3), or H2SO4-magnetically treated water (MTW)GA3 and then incubated in wet sand at a constant temperature (i.e., 5 degrees C, 10 degrees C, 15 degrees C, 20 degrees C, or 25 degrees C) or at alternating temperatures (i.e., 15 degrees C/25 degrees C or 5 degrees C/25 degrees C/5 degrees C). Additionally, the effect of relative humidity (RH) on seed germination was assessed (i.e., at 40 % or 50 % RH). These analyses revealed that the optimal temperature and RH for H2SO4-MTW-GA3 treated T. miqueliana seed germination were 15 degrees C and 50 % RH, respectively. T. miqueliana produces deeply dormant woody seeds. During dormancy release following H2SO4MTW-GA3 treatment, consistent with the observed metabolic enzyme activities, the contents of protein and fat showed a downward-upward-downward trend, whereas the sugar and starch contents showed an upward-downward trend. Hence, the H2SO4-MTW-GA3 treatment enhanced the conversion of glycogen, starch, protein, and fat to nutrients and energy, while also increasing the activities of diverse metabolic enzymes. Furthermore, the abundance of different hormones promoting seed dormancy release peaked at different time points. Specifically, the GA content peaked before the trans-zeatin-riboside (ZR) content, which may be related to the different mechanisms by which various hormones induce dormancy release.

The assessment of seed quality and physiological potential is essential in seed production and crop breeding. In the process of rapid detection of seed viability using tetrazolium (TZ) staining, it is necessary to spend a lot of labour and material resources to explore the pretreatment and staining methods of hard and solid seeds with physical barriers. This study explores the TZ staining methods of six hard seeds (Tilia miqueliana, Tilia henryana, Sassafras tzumu, Prunus subhirtella, Prunus sibirica, and Juglans mandshurica) and summarizes the TZ staining conditions required for hard seeds by combining the difference in fat content between seeds and the kinship between species, thus providing a rapid viability test method for the protection of germplasm resources of endangered plants and the optimization of seed bank construction. The TZ staining of six species of hard seeds requires a staining temperature above 35 degrees C and a TZ solution concentration higher than 1%. Endospermic seeds require shorter staining times than exalbuminous seeds. The higher the fat content of the seeds, the lower the required incubation temperature and TZ concentration for staining, and the longer the staining time. And the closer the relationship between the two species, the more similar their staining conditions become. The TZ staining method of similar species can be predicted according to the genetic distance between the phylogenetic trees, and the viability of new species can be detected quickly.

Soil degradation resulting from human trampling in urban forest parks can negatively impact the taxonomic diversity and function of soil microbial communities. In this study, we established long-term, fixed large plots in Zijin Mountain Urban Forest Park in Nanjing, China, to assess the level of trampling pressure. Soil samples were collected from depths of 0–10 cm, 10–20 cm, and 20–30 cm for light trampling (LD), moderate trampling (MD), severe trampling (SD), extreme trampling (ED), and a no-trampling control (CK). The effects of different trampling pressures on soil were studied, including soil nutrient indices, microbial biomass, and the taxonomic diversity of fungi and bacteria. ANOVA and structural equation modeling (SEM) were employed to investigate the impacts of human trampling on the microbial community structure and function. The results indicated that soil organic carbon, ammonium, and acid phosphatase activity were the primary driving factors of microbial community change. Soil microbial diversity initially increased and then decreased with increasing trampling intensity. The changes in soil microbial function and classification were found to be associated with the intensity of trampling. Moderate trampling could enhance the diversity of the soil microbial community. The succession pattern of the fungi and bacteria communities was distinct, and the composition of the bacteria community remained relatively stable. Trampling impacts vegetation and soil structure, which then affects the structure and function of the microbial community. This study provides an essential foundation for the restoration of compacted soil in urban forest parks through targeted monitoring and management efforts.

Introduction Tilia miqueliana is an endemic species whose population is declining. The permeability barrier and mechanical constraint of the pericarp (seed coat) are important causes of its seed dormancy. Although there has been considerable research on this subject, questions remain regarding how the permeability barrier and mechanical constraint of the seed coat are eliminated during dormancy release and how water enters the seed. Therefore, protecting the species by improving its germination/dormancy breaking in the laboratory is urgent Methods In this study, the changes in the cellular structure, mechanical properties, and components of the Tilia miqueliana seed coat after an H 2 SO 4 -gibberellic acid (GA 3 ) treatment were analyzed during dormancy release. Various analyses (e.g., magnetic resonance imaging, scanning electron microscopy, and paraffin section detection) revealed the water gap and water channel. Results The H 2 SO 4 treatment eliminated the blockage at the micropyle and hilum of the seeds. Water entered the seeds through the water gap (micropyle) rather than through the hilum or seed coat, after which it dispersed along the radicle, hypocotyl, and cotyledon to the endosperm. During the cold stratification period, the cellular structure was damaged and an increasing number of holes appeared on the inner and outer surfaces of the seed coat. Vickers hardness tests showed that GA 3 decreased the seed coat hardness. Additionally, the seed coat lignin and total phenol contents continuously decreased during the cold stratification period. Notably, the Liquid chromatography–mass spectrometry (LC–MS) analysis of the seed coat detected polyethylene glycol (osmoregulator), which may have destabilized the water potential balance inside and outside the seed and increased the water content to levels required for germination, ultimately accelerating seed dormancy release. Discussion This sophisticated and multi-level study reveals how H 2 SO 4 and GA 3 eliminate the permeability barrier and mechanical constraints of the seed coat during dormancy release of Tilia miqueliana seeds. This will be beneficial to artificially assist the natural regeneration and population expansion of Tilia miqueliana .

Persistence in the soil is a function of seed physiology, particularly non-germination and inherent lifespan. However, for seeds with mechanical dormancy, non-germination is also a function of the composition and activity of the soil microbiota. We attempted to screen out microorganisms in the soil that can specifically and rapidly decompose the hard fruit pericarps of Tilia miqueliana Maxim., a unique native tree species in China. Using the classical replica plating method, more than 100 different culturable microorganisms that could rapidly erode the pericarp were collected from the surface of pericarps under different culture conditions. At the same time, we successfully extended the concept of metagenomics and applied it to the identification of mixed artificial cultures. The decomposition process of the pericarps in soil was also simulated artificially. The physical and chemical data suggested a potential mechanism of microbial scarification and cracking in pericarp, whilst the embryos inside the eroded fruits retained good viability. Our discoveries could pave the way for the removal of physical and mechanical obstacles that prevent hard coat seeds from germinating. We anticipate that the use of this technology will improve the germination of other hard coat seeds. More research is needed to investigate the impacts on other seeds. The findings of this research can inform the design of experiments on the seed ecology of persistence.

Introduction Cryo treatment of dry seeds is known to attenuate the structure of fruit and seed coats, but little is known about the microstructural impacts of such treatment. The seeds of Tilia miqueliana are dispersed within a hard pericarp, the manual removal (hulling) of which is time-consuming and inefficient. Rapid hulling technology is urgently needed for sustainable production and convenience of edible nuts. Methods We explored the mechanistic basis of liquid nitrogen (N)-treatment weakening of the pericarp of T. miqueliana fruits using a range of microscopical, biophysical and chemical approaches. Results Liquid N treatment (40 s) resulted in lower pericarp contents of cellulose and hemicellulose, and increased amounts of lignin. Profound changes in cell structure and mechanical properties included the emergence of large holes and gaps between the mesocarp and endocarp cells. Also, the toughness of the pericarp decreased, whilst the hardness and brittleness increased, thereby changing the fracture type from ductile to brittle. Liquid N treatment of dry fruits followed by tapping with a hammer, reduced the number of damaged seeds three-fold and pericarp peeling time four-fold compared with manual hulling, whilst seed viability was not negatively affected. Discussion Comparable findings for the efficient and economical removal of hard covering structures from dispersal units of five more species from three other families following liquid N treatment indicates the potential application of our findings to large-scale production of seeds and seedlings for breeding, forestry and conservation/restoration purposes. Furthermore, it introduces a novel concept for postharvest treatment and pre-treatment of deep processing in nuts.

In May 2019, Aucuba japonica var. variegata, known as an evergreen shrub, was found to have multiple leaf spots in Nanjing City, Jiangsu Province, China. A fungus was isolated from the margins of the lesions and one isolate JS-RL3 was selected for identification and pathogenicity tests. The morphological characteristics of the fungus were similar to Botrytis cinerea. Phylogenetic analysis using concatenated sequences of ITS and G3PDH also showed that isolate JS-RL3 clustered with strains of B. cinerea, a relationship supported by a 100% bootstrap value. This report is the first of B. cinerea causing leaf spots on A. japonica var. variegata.

Rhizosphere fungi have the beneficial functions of promoting plant growth and protecting plants from pests and pathogens. In our preliminary study, rhizosphere fungus JP-NJ4 was obtained from the soil rhizosphere of Pinus massoniana and selected for further analyses to confirm its functions of phosphate solubilization and plant growth promotion. In order to comprehensively investigate the function of this strain, it is necessary to ascertain its taxonomic position. With the help of genealogical concordance phylogenetic species recognition (GCPSR) using five genes/regions (ITS, BenA, CaM, RPB1, and RPB2) as well as macro-morphological and micro-morphological characters, we accurately determined the classification status of strain JP-NJ4. The concatenated phylogenies of five (or four) gene regions and single gene phylogenetic trees (ITS, BenA, CaM, RPB1, and RPB2 genes) all show that strain JP-NJ4 clustered together with Talaromyces brevis and Talaromyces liani, but differ markedly in the genetic distance (in BenA gene) from type strain and multiple collections of T. brevis and T. liani. The morphology of JP-NJ4 largely matches the characteristics of genes Talaromyces, and the rich and specific morphological information provided by its colonies was different from that of T. brevis and T. liani. In addition, strain JP-NJ4 could produce reduced conidiophores consisting of solitary phialides. From molecular and phenotypic data, strain JP-NJ4 was identified as a putative novel Talaromyces fungal species, designated T. nanjingensis.

Liriodendron chinense × tulipifera black spot is a newly discovered disease that causes yellowing and early shedding of leaves, affecting the growth of Liriodendron trees, and significantly reducing their ornamental value as a garden species. The pathogen responsible for this disease, and how it can be prevented and controlled, are not clear. In this study, the occurrence of this disease was first investigated according to Koch’s postulates, and the primary pathogens causing Liriodendron black spot were determined to be Colletotrichum gloeosporioides and Alternaria alternata. Biocontrol strains antagonistic to these two pathogens were then screened from the leaf microorganisms of L. chinense × tulipifera, and a preliminary investigation of the biological control of Liriodendron black spot was performed. Through the screening of antagonistic microorganisms on the leaf surface of L. chinense × tulipifera, the strain Trichoderma koningiopsis T2, which displayed strong antagonism against C. gloeosporioides and A. alternata, was obtained. The T2 strain could inhibit the growth of the two pathogens via three mechanisms: hyperparasitism, volatile and nonvolatile metabolite production, and environmental acidification. The biocontrol experiments in the greenhouse and field showed that initial spraying with a T. koningiopsis T2 spore suspension followed by the two pathogens resulted in the lowest disease incidence. These results confirmed the black spot pathogens of L. chinense × tulipifera, clarified the antagonistic mechanism of T. koningiopsis T2 against the two pathogens, and provided a theoretical basis and technical support for the biological control of the disease.

Plant growth-promoting rhizobacteria can potentially be used as an alternative strategy to control plant disease. In this study, strain ST-TJ4 isolated from the rhizosphere soil of a healthy poplar was found to have strong antifungal activity against 11 phytopathogenic fungi in agriculture and forestry. Strain ST-TJ4 was identified as Pseudomonas chlororaphis subsp. aurantiaca based on 16S rDNA sequences. The bacterium can produce siderophores, cellulase, and protease, and has genes involved in the synthesis of phenazine, 1-phenazinecarboxylic acid, pyrrolnitrin, and hydrogen cyanide. Moreover, the volatile compounds released by strain ST-TJ4 can inhibit the mycelial growth of plant pathogenic fungi more than diffusible substances can. Based on volatile compound profiles of strain ST-TJ4 obtained from headspace collection and GC-MS/MS analysis, 1-undecene was identified. In summary, the results suggested that P. chlororaphis subsp. aurantiaca ST-TJ4 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Plant growth-promoting rhizobacteria (PGPR) can potentially be used as an alternative strategy to control plant diseases. In this study, strain ST–TJ4 isolated from the rhizosphere soil of a healthy poplar was found to have a strong antifungal activity against 11 phytopathogenic fungi in agriculture and forestry. Strain ST–TJ4 was identified as Pseudomonas sp. based on 16S rRNA-encoding gene sequences. The bacterium can produce siderophores, cellulase, and protease, and has genes involved in the synthesis of phenazine, 1–phenazinecarboxylic acid, pyrrolnitrin, and hydrogen cyanide. Additionally, the volatile compounds released by strain ST–TJ4 can inhibit the mycelial growth of plant pathogenic fungi more than diffusible substances can. Based on volatile compound profiles of strain ST–TJ4 obtained from headspace collection and GC–MS/MS analysis, 1-undecene was identified. In summary, the results suggested that Pseudomonas sp. ST–TJ4 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Microbes in soil can degrade insoluble inorganic and organic phosphorus, which are components of the soil phosphorus cycle and play an important role in plant growth. Pinus massoniana is a pioneer tree species used for afforestation in southern China and grows in poor, acidic soil. A shortage of available phosphorus in soil limits the growth of P. massoniana. To alleviate this situation, it is necessary to improve soil fertility. A fungal strain (JP-NJ2) with the ability to solubilize phosphate was isolated from the P. massoniana rhizosphere. The ability of JP-NJ2 to solubilize inorganic and organic phosphorus and promote the growth of P. massoniana was evaluated. It showed that JP-NJ2 could grow in NBRIP inorganic phosphate (AlPO4, FePO4·4H2O, and Ca3[PO4]2) fermentation broths, with the highest phosphorus concentration (1.93 mg/ml) and phosphate-solubilizing rate (43.7%) for AlPO4 and in Monkina organic phosphate fermentation broth with a phosphorus concentration of 0.153 mg/ml. The phosphate-solubilizing capability in inorganic and organic fermentation broths was negatively correlated with pH. JP-NJ2-produced acids at a total concentration of 4.7 g/l, which included gluconic (2.3 g/l), oxalic (1.1 g/l), lactic (0.7 g/l) and malonic (0.5 g/l) acids. It prioritized extracellular acidic phosphatase and combined with phytase to solubilize organic phosphates. The fungal suspension and extracellular metabolites from phosphate-solubilizing fungi promoted the shoot length of P. massoniana seedlings by 97.7% and 59.5%, respectively, while increasing the root crown diameter by 46.8% and 27.7%. JP-NJ2 was identified as Penicillium guanacastense based on its morphology and phylogenetic analyses of five genes/regions (ITS, ben A, cmd, cox1 and tef). This is the first report on P. guanacastense isolated from pine tree rhizosphere soil in China and its high phosphate-solubilizing capability, which promoted the growth of P. massoniana. P. guanacastense JP-NJ2 has potential use as a biological fertilizer in forestry and farming.