First Report of Leaf Brown Spot Caused by Diaporthe phoenicicola on Pachira glabra in China.

Pachira glabra is an increasingly important ornamental landscape tree in southern China. In August 2022, brown spots were observed on P. glabra leaves in Xiangtan City, Hunan Province, China (27.932°N, 113.020°E), affecting up to 40% of the 792 trees surveyed. On each diseased tree, nearly 30% leaves had symptoms, with an average severity of 21.2 ± 5.8% (n=100). The disease initially started as small yellow lesions along leaf margins, which later progressed to pale brown to brown with dark brown borders, eventually coalescing into large necrotic areas. Thirty symptomatic leaf samples (2 × 2 mm) were surfaced-sterilized in 75% ethanol for 10 s, 2% NaOCl for 30 s, rinsed in sterile water three times, placed on potato dextrose agar (PDA), and incubated at 25°C for 5 to 7 days in dark. Eight morphologically similar isolates were obtained from diseased leaf samples through single-spore isolation. On PDA, colonies initially appeared white, turning gray, while the reverse developed a pale yellowish hue. Aerial mycelia were white, cottony, and developed visible black pycnidia with oil droplets at maturity. The α-conidia were unicellular, hyaline, aseptate, oval or fusiform, usually with 1 or 2 guttule(s) and rounded at each end. These conidia were 5.3-8.6 × 1.7-2.5 µm (avg. 6.7 × 2.2 µm, n = 100) and present more frequently than ß-conidia.The ß-conidia were unicellular, hyaline, aseptate, filiform, straight or hamate, eguttulate, 14.6-23.3 × 0.4-1.3 µm (avg. 18.4 × 0.9 µm, n = 30). Morphologically, the fungi were identified as Diaporthe sp. (Udayanga et al. 2014). For molecular identification, the internal transcribed spacer region (ITS), translation elongation factor 1α (EF1-α), calmodulin (CAL), tubulin 2 (TUB2), and histone H3 (HIS3) sequences of all isolates were amplified from genomic DNA, using primers ITS4/ITS5 (White et al. 1990), TEF-2/728F and CALD-38F/CALD-752R (Carbone and Kohn 1999), Bt2a/Bt2b and H3-1a/H3-1b (Glass and Donaldson 1995; Crous et al. 2004), respectively. The GenBank accession numbers for a representative isolate gpg2023-1 were OR533573 (ITS), OR570887 (EF1-α), OR570888 (TUB2), OR570890 (CAL), and OR570889 (HIS3). BLAST results showed that the ITS, EF1-α, TUB2, HIS, and CAL sequences were 99%, 99%, 99%, 99%, and 98% identity, respectively, with those of Diaporthe phoenicicola (GenBank: KC343032.1, KC343758.1, KC344000.1, KC343516.1, and KC343274.1). To confirm the pathogen's identity, phylogenetic analysis using MEGA7.0 based on Maximum Likelihood was constructed. Isolate gpg2023-1 clustered with D. phoenicicola. Based on morphological and molecular data, the fungus was identified as D. phoenicicola. Next, pathogenicity tests were performed three times on one-year-old potted P. glabra plants. For each isolate, twelve healthy leaves on each of three plants were either wounded by a sterile needle or left unwounded, and then sprayed with a conidial suspension (1×106 conidia/ml) for each isolate. Control plants received with sterile water only. Plants were kept in a greenhouse at 25°C, 80% relative humidity, with a 12-h photoperiod. All wounded, inoculated leaves developed brown spot symptoms similar to those observed in the field with six days, while unwounded leaves and control plants remained symptom-free. The fungus was reisolated from all diseased leaves, fulfilling Koch's postulates and proving D. phoenicicola as the causative agent of this brown spot disease on P. glabra. While D. pachirae has been reported to cause leaf spot on P. glabra in Brazil (Milagres et al. 2018), this study marks the first report of D. phoenicicola causing leaf brown spot on P. glabra in China. This finding can help develop control strategies for this disease.

First Report of White Leaf Spot Caused by Hypomontagnella monticulosa on Pachira glabra in China.

Pachira glabra Pasq. is an ornamental tree widely distributed in tropical and subtropical regions of China. In August 2021, an unknown leaf spot was observed on P. glabra in Xiangtan County, Hunan, China (27.976°N, 113.041°E). Over 1,200 plants were evaluated, and up to 20% of the plants were diseased. In moderately diseased plants, approximately one third of leaves had symptoms with the disease severity estimated to be 31.6 ± 9.4% (n=100). The symptoms first appeared as pale yellow to yellow small dots often confined between leaf veins. These dots gradually enlarged, and coalesced into large pale or white spots with brown borders and yellow halo. In severe infections, early leaf death and defoliation occurred. Thirty lesions (2 × 2 mm) collected from ten trees were sterilized in 75% ethanol for 15 s, 5% sodium hypochlorite for 15 s, rinsed in sterile water three times, placed on oatmeal agar medium (OA) plate with lactic acid (3 ml/liter), and incubated at 28°C for 15 days. After incubation, five isolates with a similar morphology were obtained by single-spore culture. Colonies on OA were white and then turned pale grey. Pigments on the reverse side were pale brown. Conidiophores were hyaline, smooth to finely roughened, usually with virgariella-like branching patterns. Conidiogenouscells were hyaline, smooth, and measured 13.9 to 53.8 long and 1.5 to 2.3 µm wide (average 30.8 × 2.0, n=50). Conidia were single-celled, transparent, smooth, ellipsoid to obovoid, 2.3 to 4.6 × 1.7 to 3.1 µm (average 3.1 × 2.3, n=100) in measurement. For further molecular identification, internal transcribed spacer (ITS), ß-tubulin (TUB2), RNA polymerase II (RPB2), and large ribosomal subunit (LSU) genes of a representative isolate TT422 were amplified from genomic DNA, using primers ITS1/ITS4 (Mills et al. 1992), T1/T22 (O'Donnell et al. 1997), RPB2-5F/7cR (Liu et al. 2000), and LROR/LR7 (Rehner et al. 1994), respectively. Sequences of ITS (accession no. OM070368), TUB2 (OM201746), LSU (OM070369), and RPB2 (OM141478) from the isolate TT422 showed >98% identity where sequences overlapped to the reference strain of Hypomontagnella monticulosa MUCL 54604 (KY610404, KX271273, KY624305, and KY610487). Concatenated sequences were used for a phylogenetic analysis based on Maximum Likelihood using MEGA7. Based on morphological and molecular data, the isolate TT422 was identified as H. monticulosa (Ju & Rogers 1995; Lambert et al. 2019). Pathogenicity tests were performed three times on healthy leaves using the isolate TT422. Three leaves on one-year-old plants were slightly wounded by a sterile needle, and sprayed with conidial suspension (1×106 conidia/ml, containing 0.05% Tween 20) . Control leaves were sprayed with sterile water containing 0.05% Tween 20. All plants were kept in a greenhouse for 24 h at 28°C and 80% relative humidity, with a 16-h photoperiod and then transferred to natural conditions. All inoculated leaves developed white leaf spot symptoms after 7 days similar to those observed in the field, whereas no visible symptoms appeared on the control leaves. H. monticulosa strains were reisolated from all symptomatic leaves, fulfilling Koch's postulates. H. monticulosa isolated from marine or endophytic origin has been reported to produce bioactive metabolites with anticancer and microbial activities (Leman-Loubière et al. 2017; Lutfia et al. 2021; Zhang et al. 2021), but not as a phytopathogen. To our knowledge, this is the first report of H. monticulosa causing white leaf spot on P. glabra in China and worldwide.

First Report of Colletotrichum siamense Causing Leaf Anthracnose on Pachira glabra in China.

Pachira glabra Pasq.is an important landscape tree in southern China due to its ornamental value. Between March and April - 2021, anthracnose-like symptoms on P. glabra leaves were found in the botanical garden (27.904°N, 112.918°E) of Hunan University of Science and Technology located in Xiangtan of Hunan Province. Over 700 plants were evaluated, and up to 30% of the plants were symptomatic. On each plant, approximately 22% leaves had symptoms. Disease severity was estimated to be 15.6 ± 6.1% (n=100) in moderately diseased plants. Initially, subcircular or irregular shaped, water-soaked spots with pale green to yellow centers appeared mostly along leaf margins. Later, theses spots turned light brown to dark brown with black borders, gradually enlarged, and often coalesced into large sunken, necrotic areas, leading to early leaf death and abscission. Thirty lesions (2 × 2 mm) collected from ten trees were sterilized in 75% ethanol for 10 s, 2% sodium hypochlorite for 30 s, rinsed in sterile water three times, placed on potato dextrose agar (PDA) with lactic acid (3 ml/liter), and incubated at 28°C for 5 days. After incubation, six isolates with a similar morphology were obtained by single-sporing. Colonies on PDA were white and with age produced a light brown pigmentation on the underside of the colony. Acervuli present in aged cultures, brown to black, circular to subcircular and measured 31.9 to 108.7 µm (71.4 ± 6.2 µm, n=30). Conidia were single-celled, transparent, smooth, fusiform to cylindrical with obtuse to slightly ronded ends, and measured 7.8 to 11.1 µm long and 2.5 to 3.1 µm wide (9.3 ± 1.0 × 2.9 ± 0.7, n=100). For further molecular identification, Internal transcribed spacer (ITS), actin (ACT), glyceraldehyde-3-phosphate (GAPDH), calmodulin (CAL), and beta-tubulin (TUB2) genes of the isolates were amplified from genomic DNA, using primers ITS1/ITS4 (Mills et al. 1992), GDF/GDR (Cannon et al. 2012), ACT-512F/ACT-783R, CL1CF/CL2CR (Weir et al. 2012), and T1F/T22R (O'Donnell et al. 1997), respectively. Sequences of ITS (accession no. OM074029), ACT (OM190777), GAPDH (OM190778), CAL (ON210110), and TUB2 (ON210109) from CS-1 showed >98% identity where sequences overlapped to the reference strain of Colletotrichum siamense CBS 130420 (JX010259.1, JX009549.1, JX009974.1, JX009713.1 and JX010415.1). Concatenated sequences were used for a phylogenetic analysis based on Maximum Likelihood using MEGA-X. Based on morphological and molecular data, isolate CS-1 was identified as C. siamense (Cannon et al. 2012). . Pathogenicity tests were performed three times on healthy leaves using isolate CS1. Ten leaves on one-year-old plants were either slightly wounded by a sterile needle or unwounded, and inoculated with 10 µl of conidial suspension (1×106 conidia/ml, containing 0.05% Tween 20) per wound. The control plants were treated with sterile water. All plants were kept in a greenhouse for 24 h at 28°C and 80% relative humidity, with a 12-h photoperiod and then transferred to natural conditions. All wounded, inoculated leaves developed leaf spot symptoms after 14 days similar to those observed in the field, whereas no visible symptoms appeared on the intact and noninoculated leaves. C. siamense strains were reisolated from all symptomatic leaves, fulfilling Koch's postulates. C. siamense has been reported as a causal agent of anthracnose associated with diverse species (Udayanga et al. 2013), but not including P. glabra. To our knowledge, this is the first report of C. siamense causing anthracnose on P. glabra.

First Report of Alternaria alternata Causing Leaf Spot of Tartary Buckwheat (Fagopyrum tataricum) in China.

Buckwheat (Fagopyrum tataricum) is recognized as a healthy food with abundant nutrients and high levels of rutin. In April and May of 2020, an unknown tartary buckwheat leaf spot distinct from Nigrospora leaf spot (Shen et al. 2020) was observed in Xiangxiang, Hunan, China (27°49'54″N, 112°span style="font-family:'Times New Roman'; color:#0000ff">18'48″E.). Disease incidence was 60-70% within three fields (totally 7, 000 m2). The disease occurred after plants emerged. Initial symptoms began as circular, or ellipsoid, chlorotic, water-soaked spots, mostly on leaf apexes or leaf margins. The small spots gradually enlarged and often coalesced to form large circular or irregular, pale to light brown lesions, and the infected leaves eventually withered and fell off. Thirty 2 × 2 mm infected tissue pieces collected from five locations were sterilized in 70% ethanol for 10 S, in 2% NaClO for 30 S, rinsed in sterile water for three times, dried, and placed on PDA with lactic acid (3 ml/L). After 3-5 days at 28°C in the dark, 17 fungal isolates were purified using single-spore isolation method. Almost all fungal isolates had similar morphology. Colonies were initially olive green with white margin and later turned dark olive or black with profuse sporulation. Conidia were borne in long chains, tawny to brownish green, with 1-3 longitudinal and 1-7 transverse septa, pyriform, and measured 9.5-39.6 µm long, and 5.1-12.6 µm wide (n=50). Based on morphological characteristics, the fungus was identified as Alternaria alternata (Simmons 2007). Partial internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), translation elongation factor 1-α(TEF) and Alternaria major allergen (Alt a1) genes of isolate BLS-1 were amplified using ITS1/ITS4 (Mills et al. 1992), EF1-728F/EF1-986R (Carbone and Kohn 1999), Gpd1/Gpd2 and Alt-4for/Alt-4rev (Lawrence et al. 2013), respectively. Sequences were deposited into GenBank with acc. nos MW453091 (ITS), MW480219 (GAPDH), MW480218 (TEF), and MW480220 (Alt a1). BLASTn analysis showed 99.8% (ITS, MH854758.1), 100% (GAPDH, KP124155.1), 99.8% (TEF, KP125073.1) and 100% (Alt a1, KP123847.1) identity with reference strain CBS 106.24 of A. alternata, confirming isolate BSL-1 to be A. alternata. A neighbor-joining phylogenetic tree constructed by MEGA7.0 based on concatenated sequences of the four genes indicated that BSL-1 formed a distinct clade with A. alternata CBS 106.24 with 100% bootstrap values. Pathogenicity test was triplicately performed on healthy leaves. Twenty leaves of five 20-day-old plants (cv. Pinku1) were sprayed with conidial suspension (1×106 conidia/ml) collected from PDA cultures with 0.05% Tween 20. An equal number of control leaves were sprayed with sterile water to serve as the controls. Treated plants were kept in a greenhouse at 28±3 °C with relative humidity of 80±5% for 24 h and transferred to natural conditions (22-30°C, RH 50-60%). After 4 to 6 days, all inoculated leaves developed symptoms similar to those observed in the fields, while the control leaves remained healthy. A. alternata was re-isolated from all infected leaves. Occasionally-isolated Diaporthe isolates were not pathogenic. A. alternata causes leaf spot of oat (Zhao et al. 2020) and leaf blight of F. esculentum (Lu et al. 2019). To our knowledge, this is the first report of A. alternata causing leaf spot on F. tataricum in China and the world. Effective strategies should be developed to manage the disease.

First Report of Nigrospora osmanthi Causing Leaf Spot on Tartary Buckwheat in China.

Buckwheat (Fagopyrum tataricum) is a traditional short-season pseudocereal crop originating in southwest China and is cultivated around the world. Antioxidative substances in buckwheat have been shown to provide many potential cardiovascular health benefits. Between August and November in 2019, a leaf spot was found in several Tartary buckwheat cv. Pinku1 fields in Xiangxiang County, Hunan Province, China. The disease occurred throughout the growth cycle of buckwheat after leaves emerged, and disease incidence was approximately 50 to 60%. Initially infected leaves developed a few round lesions, light yellow to light brown spots. Several days later, lesions began to enlarge with reddish brown borders, and eventually withered and fell off. Thirty lesions (2×2 mm) collected from three locations with ten leaves in each location were sterilized in 70% ethanol for 10 sec, in 2% sodium hypochlorite for 30 sec, rinsed in sterile water for three times, dried on sterilized filter paper, and placed on a potato dextrose PDA with lactic acid (3 ml/L), and incubated at 28°C in the dark for 3 to 5 days. Fungal colonies were initially white and later turned black with the onset ofsporulation. Conidia were single-celled, black, smooth, spherical to subspherical, and measured 9.2 to 15.6 µm long, and 7.1 to 11.6 µm wide (n=30). Each conidium was terminal and borne on a hyaline vesicle at the tip of conidiophores. Morphologically, the fungus was identified as Nigrospora osmanthi (Wang et al. 2017). Identification was confirmed by amplifying and sequencing the ITS region, and translation elongation factor 1-alpha (TEF1-α) and partial beta-tublin (TUB2) genes using primers ITS1/ITS4 (Mills et al. 1992), EF1-728F/EF-2 (Carbone and Kohn 1999; O'Donnell et al. 1998) and Bt-2a/Bt-2b (Glass et al. 1995), respectively. BLAST searches in GenBank indicated the ITS (MT860338), TUB2 (MT882054) and TEF1-α (MT882055) sequences had 99.80%, 99% and 100% similarity to sequences KX986010.1, KY019461.1 and KY019421.1 of Nigrospora osmanthi ex-type strain CGMCC 3.18126, respectively. A neighbor-joining phylogenetic tree constructed using MEGA7.0 with 1,000 bootstraps based on the concatenated nucleotide sequences of the three genes indicated that our isolate was closely related to N. osmanthi. Pathogenicity test was performed using leaves of healthy F. tataricum plants. The conidial suspension (1 × 106 conidia/ml) collected from PDA cultures with 0.05% Tween 20 buffer was used for inoculation by spraying leaves of potted 20-day-old Tartary buckwheat cv. Pinku1. Five leaves of each plant were inoculated with spore suspensions (1 ml per leaf). An equal number of control leaves were sprayed with sterile water to serve as a control. The treated plants were kept in a greenhouse at 28°C and 80% relative humidity for 24 h, and then transferred to natural conditions with temperature ranging from 22 to 30°C and relative humidity ranging from 50 to 60%. Five days later, all N. osmanthi-inoculated leaves developed leaf spot symptoms similar to those observed in the field, whereas control leaves remained healthy. N. osmanthi was re-isolated from twelve infected leaves with frequency of 100%, fulfilling Koch's postulates. The genus Nigrospora has been regarded by many scholars as plant pathogens (Fukushima et al. 1998) and N. osmanthi is a known leaf blight pathogen for Stenotaphrum secundatum (Mei et al. 2019) and Ficus pandurata (Liu et al. 2019) but has not been reported on F. tataricum. Nigrospora sphaerica was also detected in vegetative buds of healthy Fagopyrum esculentum Moench (Jain et al. 2012). To our knowledge, this is the first report of N. osmanthi causing leaf spot on F. tataricum in China and worldwide. Appropriate strategies should be developed to manage this disease.

Rice WRKY4 acts as a transcriptional activator mediating defense responses toward Rhizoctonia solani, the causing agent of rice sheath blight.

WRKY transcription factors have been implicated in the regulation of transcriptional reprogramming associated with various plant processes but most notably with plant defense responses to pathogens. Here we demonstrate that expression of rice WRKY4 gene (OsWRKY4) was rapidly and strongly induced upon infection of Rhizoctonia solani, the causing agent of rice sheath blight, and exogenous jasmonic acid (JA) and ethylene (ET). OsWRKY4 is localized to the nucleus of plant cells and possesses transcriptional activation ability. Modulation of OsWRKY4 transcript levels by constitutive overexpression increases resistance to the necrotrophic sheath blight fungus, concomitant with elevated expression of JA- and ET-responsive pathogenesis-related (PR) genes such as PR1a, PR1b, PR5 and PR10/PBZ1. Suppression by RNA interference (RNAi), on the other hand, compromises resistance to the fungal pathogen. Yeast one-hybrid assay and transient expression in tobacco cells reveal that OsWRKY4 specifically binds to the promoter regions of PR1b and PR5 which contain W-box (TTGAC[C/T]), or W-box like (TGAC[C/T]) cis-elements. In conclusion, we propose that OsWRKY4 functions as an important positive regulator that is implicated in the defense responses to rice sheath blight via JA/ET-dependent signal pathway.

Development of a TaqMan Real-Time PCR for Early and Accurate Detection of Anthracnose Pathogen Colletotrichum siamense in Pachira glabra.

Anthracnose, caused by Colletotrichum siamense, is a destructive disease of Pachira glabra in southern China. Early and proper monitoring and quantification of C. siamense is of importance for disease control. A calmodulin (CAL) gene-based TaqMan real-time PCR assay was developed for efficient detection and quantification of C. siamense, which reliably detected as low as 5 pg of genomic DNA and 12.8 fg (5800 copies) of target DNA. This method could specifically recognize all tested C. siamense isolates, while no amplification was observed in other closely related Colletotrichum species. The assay could still detect C. siamense in plant mixes, of which only 0.01% of the tissue was infected. A dynamic change in the amount of C. siamense population was observed during infection, suggesting that this real-time PCR assay can be used to monitor the fungal growth progression in the whole disease process. Moreover, the method enabled the detection of C. siamense in naturally infected and symptomless leaves of P. glabra trees in fields. Taken together, this specific TaqMan real-time PCR provides a rapid and accurate method for detection and quantification of C. siamense colonization in P. glabra, and will be useful for prediction of the disease to reduce the epidemic risk.

Constitutive expression of rice WRKY30 gene increases the endogenous jasmonic acid accumulation, PR gene expression and resistance to fungal pathogens in rice.

WRKY transcription factors are crucial regulatory components of plant responses to pathogen infection. In the present study, we report isolation and functional characterization of the pathogen-responsive rice WRKY30 gene, whose transcripts accumulate rapidly in response to salicylic acid (SA) and jasmonic acid (JA) treatment. Overexpression of WRKY30 in rice enhanced resistance to rice sheath blight fungus Rhizoctonia solani and blast fungus Magnaporthe grisea. The enhanced resistance in the transgenic lines overexpressing WRKY30 was associated with activated expression of JA synthesis-related genes LOX, AOS2 and pathogenesis-related (PR)3 and PR10, and increased endogenous JA accumulation under the challenge of fungal pathogens. WRKY30 was nuclear-localized and had transcriptional activation ability in yeast cells, supporting that it functions as a transcription factor. Together, our findings indicate that JA plays a crucial role in the WRKY30-mediated defense responses to fungal pathogens, and that the rice WRKY30 seems promising as an important candidate gene to improve disease resistance in rice.

Up-regulation of chloroplastic antioxidant capacity is involved in alleviation of nickel toxicity of Zea mays L. by exogenous salicylic acid.

A pot experiment was carried out to investigate the effect of exogenous salicylic acid (SA) on the growth, photosynthesis, oxidative stress and responses of chloroplastic antioxidant defense system of maize (Zea mays L.) plants grown in a nickel (Ni)-contaminated soil. The results indicate that exogenous SA significantly decreased the reduction in dry weight, chlorophyll and beta-carotene contents, and net photosynthetic rate of the Ni-stressed maize, demonstrating an alleviating effect of SA on Ni toxicity of plants. Superoxide anion generation rate, H(2)O(2) and malondialdehyde (MDA) contents, and lipoxygenase (LOX, EC 1.13.11.12) activity significantly increased in the chloroplasts of maize exposed to Ni stress, revealing an oxidative damage occurred in maize chloroplasts, whereas, the values of these parameters were markedly lowered in the SA-treated plants under Ni stress. Application of SA significantly enhanced the activities of superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.6.4.2), and the poll of reduced ascorbate and glutathione in chloroplasts of the Ni-stressed maize. Accordingly, the fact that SA up-regulates the capacity of antioxidant defense system in chloroplasts, thus reducing the oxidative damage, is involved in the SA-induced alleviation of Ni toxicity in maize.

OsWRKY80-OsWRKY4 Module as a Positive Regulatory Circuit in Rice Resistance Against Rhizoctonia solani.

BACKGROUND: Plant WRKY transcription factors play pivotal roles in diverse biological processes but most notably in plant defense response to pathogens. Sheath blight represents one of the predominant diseases in rice. However, our knowledge about the functions of WRKY proteins in rice defense against sheath blight is rather limited. RESULTS: Here we demonstrate that the expression of Oryza sativa WRKY80 gene (OsWRKY80) is rapidly and strongly induced upon infection of Rhizoctonia solani, the causal agent of rice sheath blight disease. OsWRKY80 expression is also induced by exogenous jasmonic acid (JA) and ethylene (ET), but not by salicylic acid (SA). OsWRKY80-GFP is localized in the nuclei of onion epidermal cells in a transient expression assay. Consistently, OsWRKY80 exhibits transcriptional activation activity in a GAL4 assay in yeast cells. Overexpression of OsWRKY80 in rice plants significantly enhanced disease resistance to R. solani, concomitant with elevated expression of OsWRKY4, another positive regulator in rice defense against R. solani. Suppression of OsWRKY80 by RNA interference (RNAi), on the other hand, compromised disease resistance to R. solani. Results of yeast one-hybrid assay and transient expression assay in tobacco cells have revealed that OsWRKY80 specifically binds to the promoter regions of OsWRKY4, which contain W-box (TTGAC[C/T]) or W-box like (TGAC[C/T]) cis-elements. CONCLUSIONS: We propose that OsWRKY80 functions upstream of OsWRKY4 as an important positive regulatory circuit that is implicated in rice defense response to sheath blight pathogen R. solani.

[Effects of manganese on antioxidant system of manganese-hyperaccumulator Phytolacca americana L.].

A hydroponic experiment was conducted to study the growth, manganese (Mn) accumulation, lipid peroxidation, H2O2 concentration, and antioxidant system of Phytolacca americana L. exposed to different concentration Mn. With increasing Mn concentration in the medium, the plant Mn content increased significantly, and the Mn accumulation was in the sequence of leaf > stem > root. Comparing with the control, low concentration (5 mmol x L(-1)) Mn promoted the plant growth, decreased the leaf H2O2 concentration, and had less effects on the leaf malondialdehyde (MDA) content, while high concentration (> or = 10 mmol x L(-1)) Mn led to a remarkable increase of leaf H2O2 and MDA contents, indicating an evident oxidative damage occurred in leaves. The activities of ascorbate peroxidase, dehydroascorbate reductase and glutathione reductase and the content of reduced ascorbate increased with increasing Mn concentration, while the SOD activity was inhibited significantly at 5 mmol x L(-1) of Mn but enhanced at > or = 10 mmol x L(-1) of Mn. The activities of catalase and peroxidase and the content of reduced glutathione increased at 5-10 mmol x L(-1) of Mn but dropped markedly at 20 mmol x L(-1) of Mn. All the results suggested that the Mn-induced oxidative damage and Mn accumulation might be responsible for the growth inhibition of P. americana plants at high Mn exposure, and the increase of antioxidative enzyme activities and low molecular antioxidant contents was, at least partly, contributed to the Mn tolerance and hyperaccumulation of P. americana. However, due to their different Mn concentration-dependent change modes, these antioxidants played different roles in the Mn tolerance of P. americana.