Multiomics Reveals Mechanisms of Alternaria oxytropis Inhibiting Pathogenic Fungi in Oxytropis ochrocephala.

Endophytic fungi can benefit the host plant and increase the plant resistance. Now, there is no in-depth study of how Alternaria oxytropis (A. oxytropis) is enhancing the ability of inhibiting pathogenic fungi in Oxytropis ochrocephala (O. ochrocephala). In this study, the fungal community and metabolites associated with endophyte-infected (EI) and endophyte-free (EF) O. ochrocephala were compared by multiomics. The fungal community indicated that there was more A. oxytropis, less phylum Ascomycota, and less genera Leptosphaeria, Colletotrichum, and Comoclathris in the EI group. As metabolic biomarkers, the levels of swainsonine and apigenin-7-O-glucoside-4-O-rutinoside were significantly increased in the EI group. Through in vitro validation experiments, swainsonine and apigenin-7-O-glucoside-4-O-rutinoside can dramatically suppress the growth of pathogenic fungi Leptosphaeria sclerotioides and Colletotrichum americae-borealis by increasing the level of oxidative stress. This work suggested that O. ochrocephala containing A. oxytropis could increase the resistance to fungal diseases by markedly enhancing the content of metabolites inhibiting pathogenic fungi.

High Diversity of Epicoccum Associated with Leaf Spot on Italian Ryegrass in Southwestern China: Three New Species and Six New Records.

Italian ryegrass is widely cultivated for the productions of forage, hay and silage, due to its high nutritional value and good palatability. Leaf spots caused by fungi pose a serious threat to forage crops. In order to expand knowledge of fungi causing leaf spot in ryegrass (Lolium multiforum) in Sichuan, Yunnan, Chongqing and Guizhou of southwestern China, a comprehensive survey was undertaken from 2015 to 2022. Survey discovered that Epicoccum leaf spot (ELS) was a common and widespread disease, more serious at the late stage of growth (after late May); symptomatic leaf samples collected from the four different provinces were analyzed, and a total of 202 Epicoccum isolates were obtained; based on both multilocus phylogeny (ITS, LSU, TUB2, and RPB2) and morphology, 10 Epicoccum species were finally identified, including three novel species (E. endololii sp. nov., E. lolii sp. nov. and E. loliicola sp. nov.), six new host records (E. draconis, E. endophyticum, E. oryzae, E. plurivorum, E. thailandicum and E. tobaicum), and an unknown species (Epicoccum sp.1). Pathogenicity tests showed that E. endophyticum, E. endololii and Epicoccum sp.1 were non-pathogenic to Italian ryegrass, which were confirmed as endophytes in this study; other six species could infect Italian ryegrass and cause leaf lesions to different degrees, of which E. draconis was more aggressive (P ≤ 0.05). Coupling with the isolation rates and geographical distributions of these species, E. plurivorum was the predominant pathogen in Yunnan while E. oryzae and E. tobaicum in other three provinces. This work provides an initial understanding of the taxonomies, virulence and distributions of Epicoccum species associated with ELS of southwestern China, and lays a solid foundation for the diagnosis in the field, and scientific control of ELS on Italian ryegrass.

First Report of Bipolaris zeae Causing Leaf Spot on Setaria viridis in China.

Green bristlegrass (Setaria viridis) is an annual dominant herb in arid sandy grasslands with strong drought resistance to water stress (Valença et al. 2020). In August 2021, brown or dark tan spots on the leaves of S. viridis were observed on natural grassland in Qingyang city, Gansu Province, China (36.55°N, 107.32°E). Leaf disease incidence was around 13%, and the infected area of each leaf was about 20%. For isolation, 30 tissue pieces (5×5 mm) from 10 symptomatic leaf samples were surface-sterilized with 70% ethanol for 30 seconds and rinsed three times with steriled distilled water. Then these tissues were placed on the potato dextrose agar (PDA) at 25 °C, and incubated in dark from 2 to 5 days. Sixteen single-spored cultures with consistent colony characteristics were obtained by single spore isolation as described in Manamgoda et al. (2012). Colonies on PDA were irregular shapes with black color and white edge; conidia were dark brown with septate, cylindrical, straight or flexuous geniculate at the upper end, 22.7 to 69.38×9.2 to 17.6 µm (n = 50; average size 40.8×13.2 µm) under microscope (40X). The internal transcribed spacer (ITS) and glyceraldehyde-3-phosphate dehydrogenase (GPD) regions of the representative isolate 83HXGWC, 84HXGWC and 85HXGWC were amplified using the primers of ITS1/ITS4 and GPD1/GPD2 (Manamgoda et al. 2012). All the amplified gene sequences were deposited into GenBank with accession numbers ITS (OP703331, OQ978850, and OQ978851) and GPD (OQ559683, OQ992505, and OQ992506). BLAST analysis of both the two segments showed 100% identity with those reported sequences of ex-type isolate BRIP 11512 (KJ415538 and KJ415408) of B. zeae Sivan (Tan et al. 2014; Bhunjun et al. 2020), respectively. In the maximum-likelihood phylogenetic tree, 83-HXGWC, 84HXGWC and 85HXGWC strains were grouped with the reference sequences of B. zeae with a high bootstrap supporting values of 100%. Five healthy green bristlegrass (6 weeks) plants in pots were sprayed with conidial suspensions (1×106 conidia ml-1). Another five pots were sprayed with sterilized distilled water (controls). Then all pots were individually covered with transparent polyethylene bags for 5 days to maintain high relative humidity and placed in a greenhouse maintained at 18 to 25 °C. After incubation for 14 days, the typical symptoms of leaf spots developed from brown to dark brown on the plants inoculated with conidial suspension, whereas no symptoms were observed on the control plants. The measurement of pathogenicity was carried out three times. The same pathogens were consistently reisolated from inoculated leaves, and was confirmed as B. zeae based on morphological and molecular analyses, fulfilling Koch's postulates. This study provides a new record of B. zeae on green bristlegrass on natural grassland in China, which can potentially impose disease challenges to other crops. Since, B. oryzae has been known to cause disease in cereal crops (Manamgoda et al. 2014), this can be a potential alternate source of survival of this pathogen.

First Report of Pseudopithomyces palmicola Causing Leaf Spot on Italian Ryegrass in China.

Italian ryegrass (Lolium multiflorum) is widely cultivated for the production of forage, hay and silage worldwide, due to its high nutritional value and palatability (Feng et al. 2021). It has been infected by a number of foliar fungal diseases caused by different fungal pathogens (Xue et al. 2017, 2020; Victoria Arellano et al. 2021; Liu et al. 2023). In August 2021, three Pseudopithomyces isolates with similar colony characteristics were isolated from fresh samples of leaf spots on Italian ryegrass that was collected from the Forage Germplasm Nursery, Maming town of Qujing city, Yunnan province, China (25.53833°N, 103.60278°E). For specific isolation, tissue pieces (about 0.5 cm × 1 cm) from symptomatic leaves were surface-disinfested in 75% ethanol solution for 40s, rinsed three times in sterilized distilled water and air dried, plated on potato dextrose agar (PDA), and incubated at 25°C for 3 to 7 days in the dark. Following initial isolation, a representative isolate, KM42, was selected for further study. Colonies growing on PDA were cottony, white to grey, 53.8 to 56.9 mm diameter after 6 days in the dark at 25°C, with white and regular edge; in reverse, colonies were brown to dark brown in the center, and pale yellow toward the periphery with concentric rings. Under near-UV light at room temperature (20 ± 3°C), colonies were incubated on PDA for ten days to produce conidia. Conidia were globose or ellipsoid to amygdaloid, 1- to 3-transversely septate, 0- to 2-vertical septate, light brown to brown, measuring 11.6 to 24.4 × 7.7 to 16.8 µm (avg. 17.3 × 10.9 µm). The internal transcribed spacer regions 1 and 2 and 5.8S nuclear ribosomal RNA(ITS), large subunit nrRNA (LSU), and partial DNA-directed RNA polymerase II second largest subunit (RPB2) genes were amplified with primers described by Chen et al. (2017). Sequences were deposited in GenBank (OQ875842 for ITS, OQ875844 for LSU, and OQ883943 for RPB2). BLAST analysis of all these three segments showed 100% (ITS: MF804527), 100% (LSU: KU554630), and 99.4% (RPB2: MH249030) identity with sequences of reported isolate CBS 143931 (= UC22) of Pseudopithomyces palmicola (Lorenzini et al. 2016; Liu et al. 2018). To fulfill Koch's postulates, four 12-week-old healthy Italian ryegrass plants were spray-inoculated separately with a mycelial suspension of about 5.4 × 102 CFU/ml of an isolate of P. palmicola. In addition, four control plants were sprayed with sterilized distilled water. All plants were individually covered with transparent polyethylene bags for 5 days to maintain high relative humidity and placed in a greenhouse at 18 to 22°C. Small brown to dark brown spots developed on leaves 10 days after inoculation; symptoms were not observed on control plants. Pathogenicity tests were conducted three times by the same method. The same fungus was re-isolated from the lesions, and confirmed by morphological and molecular methods as described above. To the best of our knowledge, this is the first report of P. palmicola causing leaf spot on Italian ryegrass in China or worldwide. This information will be useful to forage grass managers and plant pathologists in recognizing the disease and developing effective control measures.

Differences in the Characteristics and Pathogenicity of Pyrenophora Species Associated with Seeds of Italian Ryegrass.

Pyrenophora is a genus of pathogens that cause leaf damage and a common seedborne fungus of Italian ryegrass (Lolium multiflorum Lam.). To elucidate the main seedborne Pyrenophora species, 36 seed samples of Italian ryegrass were collected; in total, 113 strains, representing 12.3% of all isolated strains (921), were identified as Pyrenophora species using the identity of ITS sequences in NCBI and the similarity of morphological characteristics. P. dictyoides (97) was the most frequent species. By pure culture technique, 24 representative pure isolates were obtained for further study. Based on DNA analysis of multiple loci (ITS, LSU, GPDH, CHS-1, and RPB1) and morphological characters, eight Pyrenophora species were identified, P. avenicola, P. chaetomioides, P. dictyoides, P. lolii, P. nobleae, P. teres, P. triseptata, and P. tritici-repentis; among them, P. avenicola, P. tritici-repentis, and P. triseptata were newly reported on Italian ryegrass worldwide. Seed inoculation showed that P. dictyoides, P. lolii, and P. teres remarkably decreased the final germination percentages and germination indexes compared with control treatments (P ≤ 0.05); and plant inoculation showed that P. dictyoides, P. lolii, and P. nobleae could cause typical brown spot in vivo with a higher infection rate (P ≤ 0.05). In conclusion, pathogenicity tests showed that all Pyrenophora species could both inhibit seed germination and infect Italian ryegrass to different degrees; among them, P. dictyoides was the most important seedborne pathogen based on the combination of its isolation and infection rate, followed by P. lolii and P. nobleae. The data generated in this study are helpful for the accurate identification of Pyrenophora species and the development of seedborne disease management strategies.

First Report of Didymella sinensis Causing Leaf Blight on Italian Ryegrass in China.

Italian ryegrass (Lolium multiflorum Lam.) is a high-yield, high-quality forage grass and is cultivated widely in southern China. In April 2021, small black spots were observed on leaves of Italian ryegrass in the field of about 300 ha located in DuShan county, Guizhou province, China (25.62056°N, 107.53139°E). Approximately 1 to 3% of plants were affected. For isolation, eleven tissue pieces (about 0.5 × 1 cm) from four symptomatic leaves were surface-disinfested in 75% ethanol solution for 40s, rinsed thrice in sterilized distilled water and air dried; then these tissues were plated on potato dextrose agar (PDA), and incubated at 25°C for 4 days in the dark. Nine fungal isolates with similar colony characteristics were obtained, and three representative isolates (LMDS1, LMDS2 and LMDS3) were selected for further study. Colonies on PDA were 47 to 57 mm diam after 5 days, margin regular, dark gray in the center surrounded by white to gray, with floccose aerial mycelia on the upper side, and dark brown to black on the reverse side. There was no fungal sporulation when these isolates were incubated under continuous ultraviolet light on PDA, oatmeal agar (OA), malt extract agar (MEA) and potato carrot agar (PCA). ITS-rDNA, LSU-rDNA, and two other protein-coding genes (RPB2 and TUB2) were amplified with primers described by Chen et al. (2017). Sequences were deposited in GenBank (ON692740 to ON692742 for ITS, ON692775 to ON692777 for LSU, ON704660 to ON704662 for RPB2, and ON704657 to ON704659 for TUB2). BLAST analysis of all these four segments showed >99.7% identity with those sequences of ex-type isolate CGMCC 3.18348 of D. sinensis (Chen et al. 2017; Hou et al. 2020). Maximum likelihood (RAxML) phylogenetic tree based on the combined ITS, LSU, RPB2 and TUB2 alignments also showed these three isolates and the other two reported D. sinensis isolates formed a subclade with 100% bootstrap support. Referring to our previous method (Xue et al. 2020), five 8-week-old healthy plants of Italian ryegrass were spray-inoculated separately with a mycelial suspension of about 1.5 × 104 CFU/ml. In addition, five plants considered as non-inoculated controls were sprayed with sterilized distilled water. All plants were individually covered with transparent polyethylene bags for 5 days to maintain high relative humidity and placed in a greenhouse at 23 to 26°C. The small black spots similar to those observed on infected plants in the field developed on leaves fifteen days after inoculation. The symptoms consisted of brown to dark brown spots when leaves were severely infected; however, symptoms were not observed on non-inoculated plants (controls). Pathogenicity tests were carried out three times. The same fungus was re-isolated from the lesions, and confirmed by morphological characterization and molecular technique as described above, thus fulfilling Koch's postulates. To the best of our knowledge, this is the first report of D. sinensis causing leaf blight on Italian ryegrass in China. The accurate identification of this pathogen would be useful for the prevention and control of leaf spot on Italian ryegrass in the future.

The First Report of Leaf Spot Caused by Alternaria alternata on Italian Ryegrass (Lolium multiflorum) in China.

Italian ryegrass (Lolium multiflorum Lam.) is one of the most important forage crops in southwestern China. In 2018, a leaf spot was observed in a field of Italian ryegrass in Mengyang, Sichuan province, China (30.96925°N, 104.10223°E). From January to early March, this leaf spot developed sporadically and appeared as brown to dark brown lesions. In late May, this disease reached a peak with incidence up to 80% and appeared as reddish-brown necrotic spots with a grayish white to brown center. To isolate the pathogen, sections (0.5 × 1 cm2) of 30 diseased leaves collected from 10 plants were surface-disinfested in 70% ethanol solution for 30 s, 5% NaOCl solution for 5 min, rinsed thrice in sterilized distilled water, air dried, plated on potato dextrose agar (PDA), and incubated at 25°C in the dark for 4 days. To obtain pure isolates, the single-spore isolation technique (Cai et al. 2009) was used. The conidial suspensions were diluted to a reasonable concentration, spread onto PDA, and incubated at 25°C in dark for 24 to 48 h, and then single germinated conidia were transferred onto new PDA plates (Cai et al. 2009). Nine pure isolates showing similar morphology were obtained for further study. Colonies on PDA were dark gray in the center surrounded by white to gray, with gossypine mycelia on the upper side, and red to dark red on the reverse side. Conidia were obclavate or pyriform, olivaceous to dark brown, with 0 to 6 transverse septa and 0 to 4 longitudinal septa, 13.2 to 55.0 (27.9) × 6.3 to 12.5 (9.8) µm. Conidiophores were septate, hyaline to olivaceous brown, either branched or unbranched, geniculate at the tip, 2.5 to 5.9 µm wide and up to 70 µm long. These morphological and cultural characteristics were consistent with the descriptions of Alternaria alternata (Fr.) Keissl. isolated from Apple (Elfar et al. 2018). To confirm the pathogenicity on Italian ryegrass, healthy plants (8-week-old) of cultivar Splendor grown in five pots filled with potting soil were spray-inoculated with conidial suspension (1 × 106 conidia/ml). Plants in another, five pots were sprayed with sterilized distilled water as controls. All pots were individually covered with transparent polyethylene bags for 5 days to maintain high relative humidity and placed in a greenhouse at 18 to 25°C. At 14 days post inoculation, symptoms typical of brown to dark brown leaf spots developed on the plants inoculated with conidial suspension, whereas no symptoms on the control plants. The pathogenicity tests were carried out three times. The same pathogen was consistently re-isolated from inoculated leaves and confirmed by morphological characterization as described above. To further identify this pathogen, isolate HMCH-9 (=CGMCC 3.19924) was selected as a representative for molecular characterization. Following Woudenberg et al. (2015), the internal transcribed spacer regions 1 and 2 and intervening 5.8S rDNA (ITS), glyceraldehyde-3-phosphate dehydrogenase (GPD), translation elongation factor 1-alpha (TEF), RNA polymerase second largest subunit (RPB2), and Alternaria major allergen (Alt) genes were partially amplified and sequenced. Sequences were deposited in GenBank (accession nos. MH567106 for ITS, MH567107 for GPD, MH567109 for TEF, MH567110 for RPB2, and MH567108 for Alt). BLAST analysis of all these five segments showed >99.8% identity with those sequences of ex-type isolate CBS 916.96 of A. alternata (Woudenberg et al. 2015). To our knowledge, this is the first report of A. alternata causing leaf spot on Italian ryegrass in China. The accurate identification of this pathogen would be useful for the prevention and control of leaf spot on Italian ryegrass in the future.

First Report of Seedling Blight of Oat (Avena sativa) Caused by Microdochium nivale in China.

Oat (Avena sativa) is extensively planted as a fodder crop on the vast ranges of northern and northwestern China, and it has become an important supplementary feed for grazing livestock (Yang et al. 2010). Microdochium nivale has been reported associated with seedling blight in many temperate regions (Imathiu et al. 2010) and the damage can result in serious loss of oat production. In August 2018, a serious seedling blight of oat (cv. Baiyan 7; about 30-day-old) was observed in the field in Shandan County, Zhangye City, Gansu Province (38.22° N, 101.22° E). More than 20% of oat plants were severely affected. Symptoms included leaf chlorosis and wilt. The root systems of infected plants were black and severely rotted, often with only a small amount of fine root remaining after removal from the soil. Twenty isolations were made from blackened roots on potato dextrose agar (PDA) and five isolations (TM-1, TM-2, TM-3, TM-4 and TM-5) were further purified by a single-spore method (Choi et al. 1999). Each isolate was identical based on preliminary molecular analyses of their DNA sequences of ITS by blast in the NCBI GenBank. The representative isolate TM-2 was selected for sequencing of the RNA polymerase II subunit (RPB2) gene. The isolated colonies were grown on PDA and formed colonies of approximately 62 mm (diameter) in 5 days at 25 ± 1 °C. Colonies exhibited entire margins, the color varied from white to pale yellow, and the sparse aerial mycelium were villous-floccose and cottony. The conidia were falcate, straight to curved, apex pointed or obtuse to subacute, lacking basal differentiation, 0-3-septate, most one-septate, 2.2 to 3.1 × 12.3 to 22.6µm (av.= 2.8 ×17.6; n=50). These morphological characteristics were consistent with previous descriptions of Microdochium (Zhang et al. 2010). Molecular identity was confirmed by sequencing partial sequences of ITS gene (ITS1 and ITS4 primers) (White et al. 1990) and RPB2 regions (RPB2-5f2 and RPB2-7cr) (O'Donnell et al. 2010). Sequences were deposited in GenBank under accessions MN428647 (RPB2) and MN428646 (ITS). Blast search revealed that both of the ITS and RPB2 sequences to be 99% similar to the corresponding sequences of M. nivale(CBS 116205) accession numbers KP859008.1 and KP859117.1. For pathogenicity tests, millet seed-based inoculum of M. nivale was prepared using a modified procedure of Fang et al. (2011). Three-week-old healthy oat seedlings of cv. Baiyan 7 were transplanted into potting mix containing millet seed-based inoculum of M. nivale at a rate of 3%. Control seedlings for comparison were transplanted into pots containing uninoculated potting mix. After 10 days, all the inoculated plants had developed seedling blight symptoms and that were similar to those observed in the field; while control plants remained healthy. The pathogen was reisolated from inoculated plants and identified as M. nivale based on morphological characteristics and the molecular methods described above. To our knowledge, this is the first report of seedling blight of oat caused by M. nivale in China.

Characterization and Pathogenicity of Colletotrichum Species on Philodendron tatei cv. Congo in Gansu Province, China.

In recent years in China, leaf spot caused by Colletotrichum species has been an emerging disease of Philodendron tatei cv. Congo. From 2016 to 2019, typical symptoms, appearing as circular or ovoid, sunken, and brown lesions with a yellow halo, were commonly observed on P. tatei cv. Congo in and around Lanzhou, Gansu Province, China. Conidiomata were often visible on infected leaf surfaces. Leaf disease incidence was approximately 5 to 20%. A total of 126 single-spored Colletotrichum isolates were obtained from leaf lesions. Multilocus phylogenetic relationships were analyzed based on seven genomic loci (ITS, ACT, GAPDH, HIS3, CAL, CHS-1, and TUB2) and the morphological characters of the isolates determined. These isolates were identified as three Colletotrichum species in this study. A further 93 isolates, accounting for 74% of all Colletotrichum isolates, were described as new species and named as Colletotrichum philodendricola sp. nov. after the host plant genus name, Philodendron; another two isolates were named as C. pseudoboninense sp. nov. based on phylogenetic and morphological relativeness to C. boninense; the other 31 isolates, belonging to the C. orchidearum species complex, were identified as a known species-C. orchidearum. Both novel species C. philodendricola and C. pseudoboninense belong to the C. boninense species complex. Pathogenicity tests by both spray and point inoculations confirmed that all three species could infect leaves of P. tatei cv. Congo. For spray inoculation, the mean infection rate of leaves on the three species was only 4.7% (0 to 12%), and the size on lesions was mostly 1 to 2 mm in length. For point inoculation, 30 days after nonwounding inoculation, the infection rate on leaves was 0 to 35%; in wounding inoculation, the infection rate of leaves was 35 to 65%; wounding in healthy leaves greatly enhanced the pathogenicity of these three species to P. tatei cv. Congo; however, the sizes of lesions among the three species were not significantly different. To our knowledge, this is the first report of Colletotrichum species associated with anthracnose diseases on P. tatei cv. Congo. Results obtained in this study will assist the disease prevention and appropriate management strategies.

Characterization of Pyrenophora Species Causing Brown Leaf Spot on Italian Ryegrass (Lolium multiflorum) in Southwestern China.

Drechslera leaf spot (DLS) caused by Pyrenophora (Drechslera) species is one of the most serious diseases affecting Italian ryegrass (Lolium multiflorum) in China. Between 2015 and 2018, this disease was observed in three Italian ryegrass fields in the province of Sichuan, China. Average leaf disease incidence was approximately 1 to 12% but could range up to 100%. Symptoms appeared as brown or tan spots surrounded by a yellow halo, or brown to dark brown net blotch; subsequently, spots increased in number and size, and they later covered a large area of leaf, eventually causing leaf death. In this study, 86 strains of Pyrenophora fungi were isolated from leaf lesions of Italian ryegrass. Coupled with phylogenetic analysis of the internal transcribed spacer region, partial 28S ribosomal RNA gene, and glyceraldehyde-3-phosphate dehydrogenase gene, morphological characteristics showed that Pyrenophora dictyoides and P. nobleae are associated with Italian ryegrass in southwest China. Pathogenicity tests confirmed that both species can infect Italian ryegrass, causing leaf spot, whereas the virulence of the two species differed; P. nobleae showed lower pathogenicity to Italian ryegrass. This is the first time that these two Pyrenophora species were formally reported on Italian ryegrass based on both morphological and molecular characters. Overall, this study improves knowledge of the Pyrenophora species associated with Italian ryegrass and provides a foundation for control of this disease in the future.

Characterization, Phylogenetic Analyses, and Pathogenicity of Colletotrichum Species on Morus alba in Sichuan Province, China.

Brown spot disease caused by Colletotrichum species was found on leaves of mulberry (Morus alba L.) in Dujiangyan, Sichuan Province, China. Fungal isolates from leaf lesions were identified as six Colletotrichum species based on morphological characteristics and DNA analysis of the combined sequences ITS, GAPDH, ACT, CHS-1, TUB2, and GS. These included Colletotrichum fioriniae, C. fructicola, C. cliviae, C. karstii, C. kahawae subsp. ciggaro, and C. brevisporum. Results showed that the most important causal agent of mulberry anthracnose was C. fioriniae, causing typical brown necrotic spots or streaks, followed by C. brevisporum, C. karstii, and C. kahawae subsp. ciggaro, whereas the two other species (C. fructicola and C. cliviae) showed no pathogenicity to mulberry. This study is the first report of these species associated with mulberry in China.

Physiological and biochemical responses of Festuca sinensis seedlings to temperature and soil moisture stress.

The interaction effects between temperature and soil moisture on Festuca sinensis Keng ex E.B.Alexeev were analysed to determine how F. sinensis responds to these environmental conditions. A pot experiment was conducted in a greenhouse under simulated growth conditions with four soil moisture contents (80, 65, 50 and 35% relative saturation moisture content) and three temperature conditions (15, 20 and 25°C). Physiological (relative water content and root activity) and biochemical parameters (chlorophyll, peroxidase (POD), malondialdehyde (MDA), soluble protein, soluble sugar and free proline) were evaluated at the seedling stage. Results showed that with a decrease in soil water content, the POD activities, MDA content, soluble protein content, soluble sugar content and free proline content of plants under the 15°C and 20°C treatments initially decreased and then increased, whereas they increased with a decrease of soil water content at 25°C. The relative water contents of plants under the three temperature treatments decreased with a decreasing soil moisture content, but then increased temperature significantly reduced the relative water content of the seedlings under low soil water content. The chlorophyll contents of plants under the 25°C treatment decreased with a decrease of soil moisture content, but those of plants under the 15°C and 20°C treatments initially increased and then decreased. The root activities of plants under the 15°C and 20°C treatments increased with a decreasing soil moisture content; however, those of plants under the 25°C treatment initially increased and then decreased. Thus, results indicated that changes of temperature and soil moisture content had significant and complicated effects on the physiological-biochemical characteristics of F. sinensis; the conditions of 20°C and 65% RSMC had positive effects on F. sinensis seedling growth and the appropriate drought stress could promote the growth of seedling roots under the three different temperature conditions. In conclusion, F. sinensis seedlings could adapt to certain changes in the ecological environment by regulating their physiological and biochemical reactions.