scholarly journals First report of Bacterial Leaf Streak disease of rice caused by Xanthomonas oryzae pv. oryzicola in Ivory Coast

Plant Disease ◽  
2021 ◽  
Author(s):  
Amadou Diallo ◽  
Sylvain Zougrana ◽  
Mahamadou Sawadogo ◽  
Daouda KONE ◽  
Drissa Silué ◽  
...  
Keyword(s):  
Burkina Faso ◽  
Multiplex Pcr ◽  
Molecular Characterization ◽  
Ivory Coast ◽  
Reference Strain ◽  
Xanthomonas Oryzae ◽  
Sterile Water ◽  
Bacterial Leaf Streak ◽  
First Report ◽  
Gyrb Sequence

Bacterial Leaf Streak (BLS) of rice caused by Xanthomonas oryzae pv. oryzicola (Xoc) is considered as the third emerging infectious disease of rice in Africa. First reported in Africa in the 1980s, the disease is now present in at least eight African countries including Burundi, Burkina Faso, Kenya, Madagascar, Mali, Nigeria, Senegal and Uganda. Yield loss caused by BLS is estimated at 20 to 30% (Sileshi and Gebeyehu, 2021). To our knowledge BLS has so far not been reported in Ivory Coast. While BLS has not been described in the adjacent rice-growing countries Ghana and Liberia, Xoc strains isolated from samples collected between 2003 and 2011 in Burkina Faso and Mali have been characterized (Wonni et al., 2014). Xoc is transmitted through rice seeds which favors its spread through rice trading (Sileshi and Gebeyehu, 2021). Given the extensive rice trade between Burkina Faso, Mali and Ivory Coast, we hypothesized that BLS might also be present in this country. Field surveys were carried out across Ivory Coast in October 2018. Typical symptoms of the disease, e.g. translucent lesions in the form of yellow-brown to black streaks with sometimes visible droplets of exudates on the leaf surface, were observed in the area of Korhogo. 5cm-long leaf pieces were successively disinfected, rinsed in sterile water, and then ground using the Qiagen Tissue Lyser System (QIAGEN, Courtaboeuf, France). Leaf powder was resuspended in 1.5 ml of sterile water and incubated at room temperature for 30 minutes. Then, 10 μl of the suspension was streaked on semi-selective PSA medium and incubated at 28 ° C for 3 to 7 days. Colonies characteristic of Xoc, i.e. round, convex, mucous and straw yellow in color were purified from 6 individual samples from 2 distinct sites in Korhogo. To confirm their identity, isolated strains underwent a pathogenicity and molecular characterization test. The multiplex PCR developed for the identification of X. oryzae pathovars (Lang et al., 2010) revealed for all the isolates the characteristic PCR profile of Xoc (two amplicons of 324 and 691 base pairs). Strains of Xoc BLS256 and Xoo PXO99 were used as controls. The pathogenicity test was performed on 5 weeks-old plants of O. sativa cv. Azucena leaves by infiltration with a needleless syringe of a bacterial suspension at an optical density of 0.5. After 7 days of greenhouse incubation (27 ± 1°C with a 12-hour photoperiod), all infiltration points (2 infiltrations x 3 plants per isolate) developed water-soaked lesions identical to the one challenged with BLS256 while water-infiltrated leaves remained asymptomatic. These lesions were collected and subjected to the isolation and multiplex PCR processes described above, thus fulfilling Koch's postulate. Finally, three of the isolates were subjected to sequencing of the housekeeping gene gyrB by PCR amplification using the primers XgyrB1F and XgyrB1R (Young et al., 2008). Analysis of 780bp of the gyrB sequence of strains CI_k1-1, CI_k2-2 and CI_k3-2 revealed 100% identity with the gyrB sequence of Xoc reference strain BLS256 (Acc. No. CP003057) and 10 polymorphic nucleotides compared to the Xoo reference strain PXO99A (Acc. No. CP000967). To our knowledge, this is the first report of BLS in Ivory Coast supported by molecular characterization methods. New surveys in Ivory Coast and neighboring countries where the disease has not been reported will allow to implement collections and assess disease incidence as part of future control strategies.

scholarly journals First Report of Bacterial Leaf Streak Caused by Xanthomonas oryzae pv. oryzicola on Rice in Burkina Faso

Plant Disease ◽  
2011 ◽  
Vol 95 (1) ◽  
pp. 72-72 ◽  
Author(s):  
I. Wonni ◽  
L. Ouedraogo ◽  
V. Verdier
Keyword(s):  
Burkina Faso ◽  
Multiplex Pcr ◽  
The Philippines ◽  
Xanthomonas Oryzae ◽  
Distilled Water ◽  
Sterile Water ◽  
Bacterial Leaf Streak ◽  
First Report ◽  
Wild Rice Species ◽  
Strain Collection

Bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola is prevalent in Asia where it can decrease yield by as much as 30%. In Africa, BLS has been reported in Madagascar, Nigeria, Senegal, and recently in Mali (1). The pathogen is seed transmitted and rice seeds can be a source of primary inoculum (3). In October 2009, leaf streak symptoms were observed on 3-month-old field rice grown in three regions of Burkina Faso (Haut-Bassin, Cascades, and East Center). Disease was found on cultivated Oryza sativa (varieties TS2, FKR19, and FKR56N), wild rice species (O. longistaminata and O. barthii), and weeds. Symptoms consisted of water-soaked lesions that developed into translucent, yellow streaks with visible exudates at the leaf surface. Yellow-pigmented Xanthomonas-like colonies were isolated on PSA semiselective medium (peptone 10 g, sucrose 10 g, bacto agar 16 g, distilled water 1,000 ml, actidione 50 mg liter–1, cephalexin 40 mg liter–1, and kasugamycin 20 mg liter–1). A multiplex PCR developed for the identification of Xanthomonas oryzae pathovars (2) was used to check the identity of Xanthomonas-like isolates. X. oryzae pv. oryzicola strains BLS256 from the Philippines and CFBP 7331 from Mali were used as positive controls. Three expected DNA fragments (331, 691, and 945 bp) corresponding to X. oryzae pv. oryzicola were obtained from all isolates using the multiplex PCR. No fragment was observed for negative controls (distilled water as the template). Five X. oryzae pv. oryzicola isolates were further analyzed by sequence analysis using portions of the gyrB housekeeping gene together with reference strains. Two sequence types were identified among Burkinabe isolates differing by only one nucleotide. When compared with the nucleotide database with BLAST, three isolates (BAI6, BAI15, and BAI19) were 100% identical to the type culture strain X. oryzae pv. oryzicola BLS256 (gyrB sequence was obtained from GenBank AAQN01000001.1) while the other two (BAI5 and BAI20) demonstrated 99% sequence similarity. The nucleotide sequence of isolate BAI5 was submitted to GenBank (HQ112342). Pathogenicity tests were performed on greenhouse-grown 3-week-old rice plants cv. Nipponbare. Cultures were grown overnight in PSA medium and adjusted in sterile water to 1 × 108 CFU/ml and inoculated into rice leaves with the blunt end of a 1-ml syringe. Four infiltrations were done per isolate per leaf and two leaves were inoculated per plant. Control plants were inoculated with sterile water. After 15 days of incubation in the greenhouse at 27 ± 1°C with a 12-h photoperiod, inoculated leaves exhibited water-soaked lesions with yellow exudates that were identical to symptoms seen in the field. Control plants remained symptomless. Colonies with morphology typical of Xanthomonas were recovered from the symptomatic leaves and typed using multiplex PCR to fulfill Koch's postulates. Three isolates have been deposited in the Collection Française de Bactéries Phytopathogènes (CFBP) and identified as X. oryzae pv. oryzicola strains CFBP7341–43. To our knowledge, this is the first report of X. oryzae pv. oryzicola in Burkina Faso. Further surveys and strain collection will be necessary to evaluate the geographic distribution and prevalence of BLS in Burkina Faso and neighboring countries. References: (1) C. Gonzalez et al. Mol. Plant-Microbe Interact. 20:534, 2007. (2) J. Lang et al. Plant Dis. 94:311, 2010. (3) G. Xie and T. Mew. Plant Dis. 82:1007, 1998.

scholarly journals First Report of Xanthomonas oryzae pv. oryzicola Causing Bacterial Leaf Streak of Rice in Burundi

Plant Disease ◽  
2014 ◽  
Vol 98 (10) ◽  
pp. 1426-1426 ◽  
Author(s):  
O. Afolabi ◽  
B. Milan ◽  
R. Amoussa ◽  
R. Koebnik ◽  
L. Poulin ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Xanthomonas Oryzae ◽  
Gyrb Gene ◽  
Universal Primers ◽  
Dna Fragments ◽  
Sterile Water ◽  
Bacterial Leaf Streak ◽  
First Report ◽  
Rice Plants ◽  
Multiplex Pcr Assay

On May 9, 2013, symptoms reminiscent of bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola were observed on rice plants at the panicle emergence stage at Musenyi, Gihanga, and Rugombo fields in Burundi. Affected leaves showed water-soaked translucent lesions and yellow-brown to black streaks, sometimes with visible exudates on leaf surfaces. Symptomatic leaves were ground in sterile water and the suspensions obtained were subjected to a multiplex PCR assay diagnostic for X. oryzae pathovars (3). Three DNA fragments (331, 691, and 945 bp) corresponding to X. oryzae pv. oryzicola were observed after agarose gel electrophoresis. Single bacterial colonies were then isolated from surface-sterilized, infected leaves after grinding in sterile water and plating of 10-fold dilutions of the cell suspension on semi-selective PSA medium (4). After incubation at 28°C for 5 days, each of four independent cultures yielded single yellow, mucoid Xanthomonas-like colonies (named Bur_1, Bur_2, Bur_6, and Bur_7) that resembled the positive control strain MAI10 (1). These strains originated from Musenyi (Bur_1), Gihanga (Bur_2), and Rugumbo (Bur_6 and Bur_7). Multiplex PCR assays on the four putative X. oryzae pv. oryzicola strains yielded the three diagnostic DNA fragments mentioned above. All strains were further analyzed by sequence analysis of portions of the gyrB gene using the universal primers gyrB1-F and gyrB1-R for PCR amplification (5). The 762-bp DNA fragment was identical to gyrB sequences from the Asian X. oryzae pv. oryzicola strains BLS256 (Philippines), ICMP 12013 (China), LMG 797 and NCPPB 2921 (both Malaysia), and from the African strain MAI3 (Mali) (2). The partial nucleotide sequence of the gyrB gene of Bur_1 was submitted to GenBank (Accession No. KJ801400). Pathogenicity tests were performed on greenhouse-grown 4-week-old rice plants of the cvs. Nipponbare, Azucena, IRBB 1, IRBB 2, IRBB 3, IRBB 7, FKR 14, PNA64F4-56, TCS 10, Gigante, and Adny 11. Bacterial cultures were grown overnight in PSA medium and re-suspended in sterile water (1 × 108 CFU/ml). Plants were inoculated with bacterial suspensions either by spraying or by leaf infiltration (1). For spray inoculation, four plants per accession and strain were used while three leaves per plant and four plants per accession and strain were inoculated by tissue infiltration. After 15 days of incubation in a BSL-3 containment facility (27 ± 1°C with a 12-h photoperiod), the spray-inoculated plants showed water-soaked lesions with yellow exudates identical to those seen in the field. For syringe-infiltrated leaves, the same symptoms were observed at the infiltrated leaf area. Re-isolation of bacteria from symptomatic leaves yielded colonies with the typical Xanthomonas morphology that were confirmed by multiplex PCR to be X. oryzae pv. oryzicola, thus fulfilling Koch's postulates. Bur_1 has been deposited in the Collection Française de Bactéries Phytopathogènes as strain CFBP 8170 ( http://www.angers-nantes.inra.fr/cfbp/ ). To our knowledge, this is the first report of X. oryzae pv. oryzicola causing bacterial leaf streak on rice in Burundi. Further surveys will help to assess its importance in the country. References: (1) C. Gonzalez et al., Mol. Plant Microbe Interact. 20:534, 2007. (2) A. Hajri et al. Mol. Plant Pathol. 13:288, 2012. (3) J. M. Lang et al. Plant Dis. 94:311, 2010. (4) L. Poulin et al. Plant Dis. 98:1423, 2014. (5) J. M. Young et al. Syst. Appl. Microbiol. 31:366, 2008.

scholarly journals First Report of Xanthomonas oryzae pv. oryzicola Causing Bacterial Leaf Streak of Rice in Uganda

Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1579-1579 ◽  
Author(s):  
O. Afolabi ◽  
B. Milan ◽  
L. Poulin ◽  
J. Ongom ◽  
B. Szurek ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Multiplex Pcr ◽  
Dna Sequence ◽  
Disease Incidence ◽  
Xanthomonas Oryzae ◽  
Gyrb Gene ◽  
Sterile Water ◽  
Bacterial Leaf Streak ◽  
Rice Plants ◽  
Multiplex Pcr Assay

In June 2013, symptoms reminiscent of bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola were observed on rice plants at the booting stage in the Doho rice irrigation scheme, Butaleja district, and at the tillering stage in Nambale, Iganga district and Magada, Namutumba district of Uganda. In areas surveyed, disease incidence was about 80, 40, and 30% in Doho, Nambale, and Magada, respectively. Outside the irrigation schemes, it was lower but widespread. Affected leaves showed typical BLS symptoms, such as water-soaked lesions, translucent stripes, and yellow-brown to black streaks, sometimes with visible exudates at the leaf surfaces. To check for the presence of the bacteria, symptomatic leaves were ground in sterile water and the suspension obtained was subjected to a multiplex PCR assay for X. oryzae pathovars, leading to the three diagnostic DNA fragments for X. oryzae pv. oryzicola (3). In parallel, bacterial strains were isolated from surface-sterilized symptomatic leaves. To this end, rice leaves were ground in sterile distilled water and serial dilutions of the cell suspensions were plated on semi-selective PSA medium (4). Each of the three samples yielded yellow, mucoid Xanthomonas-like colonies that resembled the positive control strain MAI10 (1). These isolates were named Ug_1, Ug_10, and Ug_14, which originated from Doho, Magada, and Nambale, respectively. Multiplex PCR on the pure cultures strongly supported that these isolates corresponded to X. oryzae pv. oryzicola. Two isolates, Ug_1 and Ug_14, were further subjected to partial DNA sequence analysis of the gyrB gene upon PCR amplification using the primers XgyrB1F and XgyrB1R (5). The 467-bp DNA sequence was identical to the gyrB sequences from the X. oryzae pv. oryzicola strains BLS256 (Philippines), ICMP 12013 (China), and MAI3 (Mali) (2). The partial nucleotide sequence of the gyrB gene of strain Ug_1 was submitted to GenBank (KJ921786). Pathogenicity tests were performed on greenhouse-grown 4-week-old rice plants of the cultivars Nipponbare, Azucena, IRBB 1, IRBB 2, IRBB 3, FKR 14, PNA64F4-56, TCS 10, Gigante, and Adny 11. For this purpose, bacterial cultures were grown overnight in PSA medium and re-suspended in sterile water at a concentration of 1 × 108 CFU/ml. Bacterial suspensions were sprayed on leaves of rice seedlings. Four seedlings per accession and isolate were inoculated. Fifteen days after incubation in a BSL-3 containment facility (27 ± 1°C with a 12-h photoperiod), inoculated leaves exhibited typical water-soaked lesions with yellow exudates that were similar to the symptoms seen in the fields. Re-isolation of the bacteria from the diseased leaves yielded colonies with the typical morphology of Xanthomonas. Multiplex PCR and sequence analysis of portions of the gyrB gene confirmed that these isolates are X. oryzae pv. oryzicola, thus fulfilling Koch's postulates. One of the three isolates, Ug_1, has been deposited in the Collection Française de Bactéries Phytopathogènes (CFBP) as strain CFBP 8171 ( http://www.angers-nantes.inra.fr/cfbp/ ). Further surveys and strain collections in East and Central Africa will help assess the geographic distribution and importance of BLS. References: (1) C. Gonzalez et al. Mol. Plant Microbe Interact. 20:534, 2007. (2) A. Hajri et al. Mol. Plant Pathol. 13:288, 2012. (3) J. M. Lang et al. Plant Dis. 94:311, 2010. (4) L. Poulin et al. Plant Dis. 98:1423, 2014. (5) J. M. Young et al. Syst. Appl. Microbiol. 31:366, 2008.

scholarly journals Confirmation of Bacterial Leaf Streak Caused by Xanthomonas oryzae pv. oryzicola on Rice in Madagascar

Plant Disease ◽  
2014 ◽  
Vol 98 (10) ◽  
pp. 1423-1423 ◽  
Author(s):  
L. Poulin ◽  
H. Raveloson ◽  
M. Sester ◽  
L.-M. Raboin ◽  
D. Silué ◽  
...  
Keyword(s):  
West Africa ◽  
Multiplex Pcr ◽  
Pcr Amplification ◽  
Rice Fields ◽  
Xanthomonas Oryzae ◽  
Universal Primers ◽  
Bacterial Leaf Streak ◽  
Gyrb Sequence ◽  
Leaf Tissues ◽  
Symptomatic Leaf

Bacterial leaf streak (BLS) caused by Xanthomonas oryzae pv. oryzicola is an important disease of rice. BLS is prevalent in Asia and West Africa, where it was first reported in Nigeria and Senegal in the early 1980s (4). Recently, molecular analysis of strains from Mali (2) and Burkina Faso (5) further confirmed the presence of BLS in West Africa. In Madagascar, BLS symptoms were first reported in the 1980s by Buddenhagen but the causal agent was not unequivocally determined (1). To confirm Buddenhagen's observations using modern molecular typing tools, we surveyed several rice fields in the Antananarivo and Antsirabe districts in March 2013. BLS symptoms were observed on cultivated Oryza sativa grown under both upland and lowland conditions, with a proportion of diseased individuals varying from 30% up to 80%. Symptomatic leaves presenting water-soaked lesions that developed into translucent, yellow streaks with visible exudates at the surface were sampled. One to four centimeter long pieces of diseased leaves were ground using the Qiagen TissueLyser system at 30 rps for 30 s (Qiagen, Courtaboeuf, France). The ground tissue was then macerated in 1 ml of sterile water for 1 h at 4°C. Non-diluted and 10-fold diluted tissue macerates were plated on semi-selective PSA medium (peptone 10 g/liter, sucrose 10 g/liter, glutamic acid 1 g/liter, bacto agar 16 g/liter, actidione 50 mg/liter, cephalexin 40 mg/liter, and kasugamycin 20 mg/liter) and incubated for 3 to 7 days at 28°C. Single, yellow, Xanthomonas-like colonies were isolated on non-selective PSA medium. Diagnostic multiplex PCR was performed on single colonies for pathovar identification (3). Five strains that produced three diagnostic bands corresponding to the X. oryzae pv. oryzicola pattern were further analyzed for pathogenicity on 3-week-old O. sativa cv. Nipponbare plants. Bacteria grown on PSA plates and adjusted to 1 × 108 CFU/ml were infiltrated into rice leaves with a needleless 1-ml syringe (2 × 3 infiltrations per plant and strain). Seven days after incubation in the greenhouse (27 ± 1°C with a 12-h photoperiod), inoculated leaves showed water-soaked lesions that produced yellow exudates corresponding to those initially observed in rice fields and observed for leaves challenged with the X. oryzae pv. oryzicola reference strain BLS256. Symptomatic leaf tissues were ground and plated on non-selective PSA medium, resulting in colonies with typical Xanthomonas morphology that were confirmed as X. oryzae pv. oryzicola by multiplex PCR typing (3), thus fulfilling Koch's postulates. Finally, the five strains were subjected to gyrB sequencing upon PCR amplification using the universal primers XgyrB1F (5′-ACGAGTACAACCCGGACAA-3′) and XgyrB1R (5′-CCCATCARGGTGCTGAAGAT-3′). The 743-bp partial gyrB sequences were 100% identical to the gyrB sequence of strain BLS256. As expected, the gyrB sequence of strains KACC10331, MAFF311018, and PXO99A of the X. oryzae pv. oryzae pathovar respectively showed nine, 16, and 10 mismatches in comparison to the Malagasy strains, thus further supporting that they belong to the pathovar oryzicola. References: (1) I. W. Buddenhagen. Int. Rice Comm. Newsl. 34:74, 1985. (2) C. Gonzalez et al. Mol. Plant Microbe Interact. 20:534, 2007. (3) J. M. Lang et al. Plant Dis. 94:311, 2010. (4) D. O. Niño-Liu et al. Mol. Plant Pathol. 7:303, 2006. (5) I. Wonni et al. Plant Dis. 95:72, 2011.

scholarly journals First Report of Xanthomonas oryzae pv. oryzicola Causing Bacterial Leaf Streak of Rice in Kenya

Plant Disease ◽  
2018 ◽  
Vol 102 (5) ◽  
pp. 1025-1025
Author(s):  
G. Onaga ◽  
R. Murori ◽  
G. Habarugira ◽  
O. Nyongesa ◽  
J. Bigirimana ◽  
...  
Keyword(s):  
Xanthomonas Oryzae ◽  
Bacterial Leaf Streak ◽  
First Report

scholarly journals First Report of Xanthomonas euvesicatoria Causing Bacterial Leaf Spot of Pepper (Capsicum annuum) in Montenegro

Plant Disease ◽  
2021 ◽  
Author(s):  
Tamara Popović ◽  
Jelena Menković ◽  
Marija Pantelić ◽  
Aleksa Obradoviċ
Keyword(s):  
Leaf Spot ◽  
Reference Strain ◽  
The United States ◽  
Bacterial Suspension ◽  
Molecular Characteristics ◽  
Strictly Aerobic ◽  
First Report ◽  
Gyrb Sequence ◽  
Pepper Leaves ◽  
Xanthomonas Euvesicatoria

Leaf spot of pepper was observed on different pepper cultivars in central Montenegro during summer and early autumn in three consecutive growing seasons (2017 - 2019). Necrotic spots were numerous, varying in size, irregular in shape, brown, and surrounded by a weak halo. The most intensive symptoms were observed on lower leaves. In conditions conducive or the infection, the lesions merged resulting in the leaf drop. Symptoms were not observed on pepper stems and fruits. A total of seventeen bacterial strains were isolated from infected pepper leaves collected in seven different localities in the seasons of 2017-19. They formed yellow, convex, and mucoid colonies on yeast extract–dextrose–CaCO3 (YDC) medium and induced hypersensitive reaction in tobacco leaves. They were Gram negative, strictly aerobic, oxidase negative, catalase-positive, hydrolyzed gelatine and esculin and did not reduce nitrate, nor grew on 0.1% TTC and at 37°C. Out of tested 17 strains, eight hydrolyzed starch and three showed pectolytic activity, thus differing in these biochemical traits from Xanthomonas euvesicatoria (Xe) the reference strain KFB 1 (Obradović et al., 2004) used in all tests as a positive control. PCR analysis, with primer pair XeF/XeR, produced a single characteristic band of 173 bp in all 17 strains (Koenraadt et al. 2009). Additionally, the BOX-PCR profile of all the strains produced with the BOX A1R primer (Schaad et al. 2001) showed 100% homology with KFB 1. Based on the locality and year of isolation, nine strains were selected for amplification and partial sequencing of the gyrB gene using sets of primers described by Parkinson et al. (2007). Obtained partial DNA sequences showed that all nine strains (GenBank nos. MZ569011, MZ574079, MZ574080, MZ574081, MZ574082, MZ574083, MZ574084, MZ574085, and MZ574086) share 99.86 to 100% identity of gyrB sequence with Xe type strain ICPM:109 as well as 98.71 to 100 % of gyrB sequence identity with Xe strain LMG930 isolated from pepper in The United States. Pathogenicity of all strains was confirmed by spraying young pepper plants (cv. Slonovo uvo) using a handheld sprayer with the bacterial suspension (108 CFU/ml of sterile tap water), in three replicates. Sterile distilled water and reference Xe strain (KFB 1) were used as negative and positive controls, respectively. The inoculated plants were incubated under plastic bags in the greenhouse providing high humidity conditions for 48h. Symptoms were monitored for two weeks after inoculation. Lesions surrounded by a halo appeared on leaves of all inoculated plants within 10 to 15 days after inoculation, while plants inoculated with SDW remained symptomless. Koch’s postulates were confirmed by reisolation of the pathogen from necrotic tissue and identity check by PCR using primer set of Koenraadt et al. (2009). The pathogen race was determined according to the reaction of cv. Early Calwonder (ECW) and its isogenic lines (ECW-10R, ECW-20R, ECW-30R) (Stall et al. 2009). Obtained results indicated that all tested strains and reference strain Xe (KFB 1) belong to the pepper race P8. Based on pathogenic, biochemical, and molecular characteristics, the strains isolated from pepper leaves in Montenegro were identified as X. euvesicatoria. Pepper production is particularly significant for small farmers in Montenegro. Favorable climate, use of noncertified seed and lack of crop rotation contributes to the disease occurrence and severity. The disease has probably been around for years but the etiology was not confirmed so far. This is the first report of X. euvesicatoria affecting pepper in this country.

Molecular characterization and first report ofCryptosporidiumgenotypes in human population in the Slovak Republic

2015 ◽  
Vol 36 (23) ◽  
pp. 2925-2930 ◽  
Author(s):  
Antónia Petrincová ◽  
Alexandra Valenčáková ◽  
Lenka Luptáková ◽  
František Ondriska ◽  
Jana Kalinová ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Human Population ◽  
Slovak Republic ◽  
First Report

scholarly journals First Report of Leaf Spot caused by Bipolaris oryzae on Switchgrass in Tennessee

Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1654-1654 ◽  
Author(s):  
A. L. Vu ◽  
M. M. Dee ◽  
J. Zale ◽  
K. D. Gwinn ◽  
B. H. Ownley
Keyword(s):  
Leaf Spot ◽  
Panicum Virgatum ◽  
Morphological Characteristics ◽  
Its Region ◽  
Spore Production ◽  
Post Inoculation ◽  
Sterile Water ◽  
Bipolaris Oryzae ◽  
First Report ◽  
Symptomatic Leaf

Knowledge of pathogens in switchgrass, a potential biofuels crop, is limited. In December 2007, dark brown to black irregularly shaped foliar spots were observed on ‘Alamo’ switchgrass (Panicum virgatum L.) on the campus of the University of Tennessee. Symptomatic leaf samples were surface-sterilized (95% ethanol, 1 min; 20% commercial bleach, 3 min; 95% ethanol, 1 min), rinsed in sterile water, air-dried, and plated on 2% water agar amended with 3.45 mg fenpropathrin/liter (Danitol 2.4 EC, Valent Chemical, Walnut Creek, CA) and 10 mg/liter rifampicin (Sigma-Aldrich, St. Louis, MO). A sparsely sporulating, dematiaceous mitosporic fungus was observed. Fungal plugs were transferred to surface-sterilized detached ‘Alamo’ leaves on sterile filter paper in a moist chamber to increase spore production. Conidia were ovate, oblong, mostly straight to slightly curved, and light to olive-brown with 3 to 10 septa. Conidial dimensions were 12.5 to 17 × 27.5 to 95 (average 14.5 × 72) μm. Conidiophores were light brown, single, multiseptate, and geniculate. Conidial production was polytretic. Morphological characteristics and disease symptoms were similar to those described for Bipolaris oryzae (Breda de Haan) Shoemaker (2). Disease assays were done with 6-week-old ‘Alamo’ switchgrass grown from seed scarified with 60% sulfuric acid and surface-sterilized in 50% bleach. Nine 9 × 9-cm square pots with approximately 20 plants per pot were inoculated with a mycelial slurry (due to low spore production) prepared from cultures grown on potato dextrose agar for 7 days. Cultures were flooded with sterile water and rubbed gently to loosen mycelium. Two additional pots were inoculated with sterile water and subjected to the same conditions to serve as controls. Plants were exposed to high humidity by enclosure in a plastic bag for 72 h. Bags were removed, and plants were incubated at 25/20°C with 50 to 60% relative humidity. During the disease assay, plants were kept in a growth chamber with a 12-h photoperiod of fluorescent and incandescent lighting. Foliar leaf spot symptoms appeared 5 to 14 days post-inoculation for eight of nine replicates. Control plants had no symptoms. Symptomatic leaf tissue was processed and plated as described above. The original fungal isolate and the pathogen recovered in the disease assay were identified using internal transcribed spacer (ITS) region sequences. The ITS region of rDNA was amplified with PCR and primer pairs ITS4 and ITS5 (4). PCR amplicons of 553 bp were sequenced, and sequences from the original isolate and the reisolated pathogen were identical (GenBank Accession No. JQ237248). The sequence had 100% nucleotide identity to B. oryzae from switchgrass in Mississippi (GU222690, GU222691, GU222692, and GU222693) and New York (JF693908). Leaf spot caused by B. oryzae on switchgrass has also been described in North Dakota (1) and was seedborne in Mississippi (3). To our knowledge, this is the first report of B. oryzae from switchgrass in Tennessee. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/, 28 June 2012. (2) J. M. Krupinsky et al. Can. J. Plant Pathol. 26:371, 2004. (3) M. Tomaso-Peterson and C. J. Balbalian. Plant Dis. 94:643, 2010. (4) T. J. White et al. Pages 315-322 in: PCR Protocols: a Guide to Methods and Applications. M. A. Innis et al. (eds), Acad. Press, San Diego, 1990.

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