Sweet cherry is an important fruit crop with high economic and ornamental value in China. However, cherry fruit anthracnose, caused by Colletotrichum species, greatly impacts cherry yield and quality. Here, we surveyed cherry anthracnose in Guizhou, China from 2019–2020. Necrotic sweet cherry fruits were collected from different areas in Guizhou and examined. A total of 116 Colletotrichum strains were isolated from these symptomatic fruits. Based on the morphological characteristics of the isolates and phylogenetic analyses of concatenate internal transcribed spacer (ITS) region and ACT, CHS-1, GAPDH, TUB2, and HIS3 genes, the pathogen responsible for causing sweet cherry anthracnose was identified as Colletotrichum godetiae. Pathogenicity tests were conducted by inoculating healthy sweet cherry fruits with spore suspensions of the fungal pathogen, and Koch’s postulates were confirmed by pathogen re-isolation and identification. The Q-1 isolate showed different sensitivities to 13 fungicides, exhibiting seven different modes of action, and its EC₅₀ values ranged from 0.04 to 91.26 μg ml⁻¹. According to that, the sensitivity of 20 isolates from different samples to ten fungicides with better performance, were measured. The results showed that 6 of the 10 fungicides (difenoconazole, propiconazole, prochloraz-manganese, pyraclostrobin, trifloxystrobin-tebuconazole, and difenoconazole-azoxystrobin) all showed higher sensitive to the 20\u00B0C. godetiae isolates, and no resistance groups appeared. Its EC50 values ranged from 0.013 to 1.563 μg ml⁻¹. In summary, this is the first report demonstrating that C. godetiae causes sweet cherry anthracnose and the results of this study provide insights into how sweet cherry anthracnose could be effectively controlled in China.

In animals, malectin is well known to play an essential role in endoplasmic reticulum quality control (ERQC) by interacting with ribophorin I, one unit of the oligosaccharyltransferase (OST) complex. However, the functions of malectin in plants remain largely unknown. Here, we demonstrate the rice OsMLD1 is an ER- and Golgi-associated malectin protein and physically interacts with rice homolog of ribophorin I (OsRpn1), and its disruption leads to spontaneous lesion mimic lesions, enhanced disease resistance, and prolonged ER stress. In addition, there are many more N-glycosites and N-glycoproteins identified from the mld1 mutant than wildtype. Furthermore, OsSERK1 and OsSERK2, which have more N-glycosites in mld1, were demonstrated to interact with OsMLD1. OsMLD1 can suppress OsSERK1- or OsSERK2-induced cell death. Thus, OsMLD1 may play a similar role to its mammalian homologs in glycoprotein quality control, thereby regulating cell death and immunity of rice, which uncovers the function of malectin in plants.

Research has demonstrated that strigolactones (SLs) mediate plant disease resistance; however, the basal mechanism is unclear. Here, we provide key genetic evidence supporting how SLs mediate plant disease resistance. Exogenous application of the SL analog, rac-GR24, increased Arabidopsis thaliana resistance to virulent Pseudomonas syringae. SL-biosynthetic mutants and overexpression lines of more axillary growth 1 (MAX1, an SL-biosynthetic gene) enhanced and reduced bacterial susceptibility, respectively. In addition, rac-GR24 promoted bacterial pattern flg22-induced callose deposition and hydrogen peroxide production. SL-biosynthetic mutants displayed reduced callose deposition but not hydrogen peroxide production under flg22 treatment. Moreover, rac-GR24 did not affect avirulent effector-induced cell death between Col-0 and SL-biosynthetic mutants. Furthermore, rac-GR24 increased the free salicylic acid (SA) content and significantly promoted the expression of pathogenesis-related gene 1 related to SA signaling. Importantly, rac-GR24- and MAX1-induced bacterial resistance disappeared completely in Arabidopsis plants lacking both callose synthase and SA. Taken together, our data revealed that callose and SA are two important determinants in SL-mediated plant disease resistance, at least in Arabidopsis.

Magnolia grandiflora is a widely cultivated ornamental tree in China. In June 2020, a leaf blight disease was observed on M. grandiflora in Guizhou University (26° 44' 57'' N, 106° 65' 94'' E) in Guiyang, China. The initial symptoms on leaves were expanding round necrotic lesions with a grey center and dark brown edge, and twigs were withered when the disease was serious. Of the 100 plants surveyed 65% had symptoms. To isolate the potential causal pathogen, diseased leaves were collected from an M. grandiflora tree at Guizhou University. Isolations from made form the junction between healthy and symptomatic tissue and disinfested by immersing in 75% ethanol for 30 seconds, 3% NaOCl for 2 minutes, and then washed 3 times in sterile distilled water. Symptomatic tissue was then plated on potato dextrose agar (PDA) and incubated at 25ºC with 12-hour light for 3-5 days. Three isolates (GUCC 21235.1, GUCC 21235.2 and GUCC 21235.3) were obtained. Colonies on PDA after 7 d were dark brown, pycnidia embedded in the mydelium were dark brown to black, single and separated. Conidiophores were transparent measuring 7-12.5 × 2.5-4.5 µm (mean = 9.5 × 3.6 µm, n = 30) in length. Conidia were transparent becoming brown when mature with a diaphragm, with round ends measuring, 21-27 × 10-15 µm (mean = 23.6 × 12.6 µm, n = 30). To confirm the pathogen by molecular characterization, four genes or DNA fragments, ITS, LSU, tef1 and β-tubulin, were amplified using the following primer pairs: ITS4-F/ ITS5-R (White et al., 1990), LR0R/ LR5 (Rehner & Samuels, 1994), EF1-688F/ EF1-986R (Carbone & Kohn, 1999) and Bt2a/ Bt2b (O'Donnell & Cigelnik, 1997). The sequences of four PCR fragments of GUCC 21235.1 were deposited in GenBank, and the accession numbers were MZ519778 (ITS), MZ520367 (LSU), MZ508428 (tef1) and MZ542354 (β-tubulin). Bayesian inference was performed based on a concatenated dataset of ITS, LSU, tef1 and β-tubulin gene using MrBayes 3.2.10, and the isolates GUCC 21235.1 formed a single clade with the reference isolates of Diplodia mutila (Diplodia mutila strain CBS 112553). BLASTn analysis indicated that the sequences of ITS, LSU, tef1 and β-tubulin revealed 100% (546/546 nucleotides), 99.82% (568/569 nucleotides), 100% (302/302 nucleotides), and 100% (437/437 nucleotides) similarity with that of D. mutila in GenBank (AY259093, AY928049, AY573219 and DQ458850), respectively. For confirmation of the pathogenicity of this fungus, a conidial suspension (1×105 conidia mL-1) was prepared from GUCC 21235.1, and healthy leaves of M. grandiflora trees were surface-disinfested by 75% ethanol, rinsed with sterilized distilled water and dried by absorbent paper. Small pieces of filter paper (5 mm ×5 mm), dipped with 20 µL conidial suspension (1×105 conidia mL-1) or sterilized distilled water (as control), were placed on the bottom-left of the leaves for inoculation. Then the leaves were sprayed with sterile distilled water, wrapped with a plastic film and tin foil successively to maintain high humidity in the dark dark. After 36 h, the plastic film and tin foil on the leaves was removed, and the leaves were sprayed with distilled water three times each day at natural condition (average temperature was about 25 °C, 14 h light/10 h dark). After 10 days of inoculation, the same leaf blight began to appear on the leaves inoculated with conidial suspension. No lesion was appeared on the control leaves. The fungus was re-isolated from the symptomatic tissue. Based on the morphological information and molecular characterization, the isolate GUCC 21235.1 is D. mutila. Previous reports indicated that D. mutila infects a broad host range and gives rise to a canker disease of olive, apple and jujube (Úrbez-Torres et al., 2013; Úrbez-Torres et al., 2016; Feng et al., 2019). This is the first report of leaf blight on M. grandiflora caused by D. mutila in China.

Coix ( Coix lacryma-jobi L.) is an important cereal and medicinal crop threatened by smut disease ( Ustilago coicis ). However, the unavailability of rapid and efficient functional genomics tools has impeded progress in understanding gene function and host-pathogen interactions in Coix. Here, we report the first virus-induced gene silencing (VIGS) system for Coix using a Tobacco Rattle Virus (TRV) vector. Using the phytoene desaturase gene ( ClPDS ) as a visual reporter, we optimized vacuum infiltration parameters and discovered that the immunosuppressant methylglyoxal dramatically enhances silencing efficiency. Supplementation of the infiltration buffer with 200 μM methylglyoxal boosted silencing efficiency from 25% to 57%, whereas tenoxicam showed no effect. To validate the TRV-VIGS system for studying plant-pathogen interactions, we first developed a U. coicis inoculation protocol for Coix seedlings to evaluate smut disease resistance. We then demonstrated the system’s utility by silencing the host defense regulator ClNPR1 , which increased susceptibility to U. coicis , and by silencing the pathogen effector gene UcPep1 via host-induced gene silencing (HIGS), which enhanced resistance. These findings provide an effective tool for functional genomics studies in Coix and have significant implications for screening and validating potential gene functions under various stresses.