Heat shock-induced enhanced susceptibility of barley to Bipolaris sorokiniana is associated with elevated ROS production and plant defence-related gene expression.

Heat stress alters plant defence responses to pathogens. Short-term heat shock promotes infections by biotrophic pathogens. However, little is known about how heat shock affects infection by hemibiotrophic pathogens like Bipolaris sorokiniana (teleomorph: Cochliobolus sativus). We assessed the effect of heat shock in B. sorokiniana-susceptible barley (Hordeum vulgare cv. Ingrid) by monitoring leaf spot symptoms, B. sorokiniana biomass, ROS and plant defence-related gene expression following pre-exposure to heat shock. For heat shock, barley plants were kept at 49 °C for 20 s. B. sorokiniana biomass was assessed by qPCR, ROS levels determined by histochemical staining, while gene expression was assayed by RT-qPCR. Heat shock suppressed defence responses of barley to B. sorokiniana, resulting in more severe necrotic symptoms and increased fungal biomass, as compared to untreated plants. Heat shock-induced increased susceptibility was accompanied by significant increases in ROS (superoxide, H2 O2 ). Transient expression of plant defence-related antioxidant genes and a barley programmed cell death inhibitor (HvBI-1) were induced in response to heat shock. However, heat shock followed by B. sorokiniana infection caused further transient increases in expression of HvSOD and HvBI-1 correlated with enhanced susceptibility. Expression of the HvPR-1b gene encoding pathogenesis-related protein-1b increased several fold 24 h after B. sorokiniana infection, however, heat shock further increased transcript levels along with enhanced susceptibility. Heat shock induces enhanced susceptibility of barley to B. sorokiniana, associated with elevated ROS levels and expression of plant defence-related genes encoding antioxidants, a cell death inhibitor, and PR-1b. Our results may contribute to elucidating the influence of heat shock on barley defence responses to hemibiotrophic pathogens.

First Report of a Septoria sp. on Common Ragweed (Ambrosia artemisiifolia) in Europe.

During a survey for potential biocontrol agents of common ragweed (Ambrosia artemisiifolia var. elatior (L.) Descourt) in 1997, plants exhibiting irregular, brown leaf spots were collected repeatedly from six roadside locations in Pest County, Hungary. Many pycnidia developed in the necrotic tissues on detached leaves after 2 days in moist chambers. Pycnidia were globose to slightly flattened, brown, thin walled, 58 to 100 µm in diameter, with a definite ostiole. Conidia were hyaline, filiform with 2 to 3 septa, and 22.0 to 38.0 × 0.7 to 1.3 µm in size. The fungus was isolated on potato dextrose agar and identified as a Septoria sp. To confirm pathogenicity, potted ragweed seedlings were sprayed with a suspension of 5 × 106 conidia per ml from pure cultures of the Septoria sp., placed in a dew chamber for 72 h, and then grown in a greenhouse at 16 to 24°C. After 2 weeks, inoculated plants developed small, brown lesions on leaves and leaf petioles. Three weeks after inoculation, necrotic lesions had enlarged to 1 to 3 mm in diameter with irregular, distinct margins and light brown centers. The lesions on the lower leaves were larger and more numerous than on leaves nearer the tops of the plants. Pycnidia developed on the senescent leaves after 1 month. Infected leaves became completely necrotic and occasionally entire plants died. The pathogen was reisolated from all inoculated plants, thus satisfying Koch's postulates. A voucher specimen was deposited at the Department of Botany of the Hungarian Natural History Museum in Buda-pest (No. BP 92081). Septoria ambrosiae Hemmi et Naito was described on ragweed in Japan (1), but our isolate is morphologically distinct from that species. This is the first report of a Septoria sp. on A. artemisiifolia in Europe. Reference: (1) N. Naito. Mem. Coll. Agric. Kyoto 47:41, 1940.

First Report of Colletotrichum gloeosporioides on Russian-thistle.

A pathogen identified as Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. was isolated from foliar and stem lesions on Russian-thistle (Salsola tragus Torner ex L.) collected in Bugac, Hungary, in 1996. Symptoms on leaves and stems began as discrete, sunken, 2- to 10-mm-diameter chlorotic spots, followed by formation of circular buff-colored lesions that eventually coalesced, desiccated, and caused plant tissue death above the lesions. Lesions that occurred near ground level usually killed the plant. Salmon-colored spore masses developed in setose acervuli in the center of the necrotic lesions. Conidia were hyaline, one-celled, falcate to nearly straight, and measured 15 to 25 × 5 to 6 µm. The teleomorph stage of the pathogen (Glomerella cingulata (Stoneman) Spauld. & H. Schrenk) was not observed in the field or on inoculated plants. These morphological characteristics of the isolate were consistent with the description of C. gloeosporioides (1). Pathogenicity was proved by completing Koch's postulates in Hungary and the U.S. Inoculation with conidial suspension (106 conidia per ml) sprayed on S. tragus plants in the greenhouse at the three- to four-leaf stage caused severe necrosis and wilting within 6 days and plant death in 2 weeks. Symptoms did not appear on control plants inoculated with sterile, distilled water. Inoculation test was repeated on 6-week-old plants and at the stage of flowering. All treated plants were killed at both stages within 4 weeks. Because of high virulence and host specificity of this isolate of C. gloeosporioides in preliminary pathogenicity tests it is being evaluated for use as a mycoherbicide for Russian-thistle control in the U.S. This is the first report of C. gloeosporioides causing anthracnose on S. tragus. Reference: (1) B. C. Sutton. Pages 1-27 in: Colletotrichum Biology, Pathology and Control. J. A. Bailei and M. J. Jeger, eds. CAB Int., Wallingford, UK, 1992.

First Report of St. John's-Wort Anthracnose Caused by Colletotrichum gloeosporioides in Hungary.

In 1997, we observed high incidence of severe disease symptoms in a plantation of St. John's-wort (Hypericum perforatum L.) that has been cultivated as a medicinal plant in Hungary. Symptoms were characterized by brown, 2- to 15-mm-diameter, sunken lesions that girdled the stems and, when occurring near the base, killed the plants. Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. was consistently isolated from lesions. Acervuli containing conidia and dark setae emerged from the lesions. Perithecia of the teleomorph Glomerella cingulata (Stoneman) Spauld. & H. Schrenk also appeared on stems. Plants were inoculated with conidial suspension of the fungus (2 × 107/ml), then incubated in a moist chamber for 24 h, and subsequently transferred to the greenhouse. The first symptoms appeared on stems and leaves 7 days after inoculation. Chlorotic spots grew into circular and buff-colored lesions that coalesced and then desiccated. The fungus caused severe defoliation and plant death within 2 weeks. Koch's postulates was completed by consistently reisolating the pathogen from inoculated plants. This disease on St. John's-wort has not previously been reported in Hungary. However, an anthracnose disease caused by Glomerella cingulata (Stoneman) Spauld. & H. Schrenk var. migrans Wollenweber (Gloeosporium orbiculare Berk. & Mont.) was reported in Germany in 1949 (1). Reference: (1) H. W. Wollenweber and H. Hochapfel. Z. Parasitenkd. 14:181, 1949.