Alternative methods for angular stain control in mangoes (Mangifera indica L.)

The conventional methods used for angular stain control are generally chemical methods, however the use of these products can cause high environmental impact and damage to consumer health if it is used in large quantities and undiluted and applied correctly. Based on this problem, this work aimed to evaluate in vitro alternative forms of control using Saccharomyces yeast (with probiotic potential), ethanolic extracts of Mauritia flexuosa (Buriti) and Miconia albicans (Cinnamon-old) plants. To evaluate four GRAS substances in angular leaf spot control caused by Xanthomonas campestris pv. Mangifera indica, during the postharvest period in mangoes. In vitro results using antagonist yeasts showed no inhibitory effect against X. campestris. However, the extracts of the plants Miconia albicans and Mauritia flexuosa showed a significant inhibition. Thus, as two GRAS substances, 1%, 1.5% and 3% sodium carbonate and 3% sodium bicarbonate inhibited X. campestris growth 100%. Given the results obtained, the plant extracts and the GRAS substances tested were effective in controlling phytobacteria and proved to be an alternative in controlling angular leaf spot, thus avoiding economic losses during the mango postharvest phase.

Draft Genome Sequences of Two Xanthomonas fragariae Strains

Xanthomonas fragariae is the causal agent of angular leaf spot of strawberry. Short-read sequences were generated for two X. fragariae strains with different virulence phenotypes on the Illumina HiSeq 2000 platform. The genome sequences will contribute to a better understanding of pathogen evolution and the genes contributing to virulence in X. fragariae .

Angular Leaf Spot Resistance Loci Associated With Different Plant Growth Stages in Common Bean

Angular leaf spot (ALS) is a disease that causes major yield losses in the common bean crop. Studies based on different isolates and populations have already been carried out to elucidate the genetic mechanisms of resistance to ALS. However, understanding of the interaction of this resistance with the reproductive stages of common bean is lacking. The aim of the present study was to identify ALS resistance loci at different plant growth stages (PGS) by association and linkage mapping approaches. An BC2F3 inter-gene pool cross population (AND 277 × IAC-Milênio – AM population) profiled with 1,091 SNPs from genotyping by sequencing (GBS) was used for linkage mapping, and a carioca diversity panel (CDP) genotyped by 5,398 SNPs from BeadChip assay technology was used for association mapping. Both populations were evaluated for ALS resistance at the V2 and V3 PGSs (controlled conditions) and R8 PGS (field conditions). Different QTL (quantitative trait loci) were detected for the three PGSs and both populations, showing a different quantitative profile of the disease at different plant growth stages. For the three PGS, multiple interval mapping (MIM) identified seven significant QTL, and the Genome-wide association study (GWAS) identified fourteen associate SNPs. Several loci validated regions of previous studies, and Phg-1, Phg-2, Phg-4, and Phg-5, among the 5 loci of greatest effects reported in the literature, were detected in the CDP. The AND 277 cultivar contained both the Phg-1 and the Phg-5 QTL, which is reported for the first time in the descendant cultivar CAL143 as ALS10.1UC. The novel QTL named ALS11.1AM was located at the beginning of chromosome Pv11. Gene annotation revealed several putative resistance genes involved in the ALS response at the three PGSs, and with the markers and loci identified, new specific molecular markers can be developed, representing a powerful tool for common bean crop improvement and for gain in ALS resistance.

First Report of Xanthomonas fragariae strain YL19 causing crown infection pockets in strawberry in Liaoning Province, China

Strawberry (Fragaria × ananassa Duch.) is an important fruit crop in China. Typical crown infection pockets symptoms were observed on the infected strawberry in Liaoning province, China (121°60′E, 38°90′N) in the autumn of 2017. The disease incidence was estimated to be around 5 to 10 %, but could reach 30 to 40 % in some heavily affected plastic tunnels. The infected plants early displayed water-soaked symptoms on the abaxial leaf surface and subsequently developed reddish-brown shaped stripes and coalesced lesions on the adaxial leaf surface around the main veins (Fig. 1-A, 1-B). Several variable-size (0.3-0.8 mm in diameter) pockets were observed inside the crown tissues after dissection (Fig. 1-C). The diseased plants rarely reached fruiting and were easily broken between the crown tissue and the stem, and would eventually die. To identify the causal agent of this disease, the several surface-disinfested infected main veins and crown tissues were individually ground in sterile water and plated on sucrose peptone agar(SPA) medium (Hayward 1960) with 10-fold serial dilutions and incubated at 25℃. A number of yellow colonies grew on the medium at the 10-4 dilution 7 days after plating (Fig. 1-D) in all specimens. The colonies were aerobic, yellow, viscous, smooth, and gram-negative, which is a typical characteristic of Xanthomonas. To confirm identity of the causal bacteria, 18 colonies selected randomly were subjected to polymerase chain reactions (PCR) for the amplification of the cpn60 (Sahin et al. 2010), gyrB, rpoD, and fyuA (C Manceau et al. 2011), respectively. The results showed that the 18 colonies are identical. The cpn60, gyrB, rpoD, and fyuA sequences of this isolate were deposited in GenBank with accession numbers MT513132.1, MW233896, MW233897, and MW233895, respectively. BLAST searches with sequences of this isolate cpn60, gyrB, rpoD, and fyuA revealed 97.7%, 96.4%, 97.8%, and 97.3% similarity with the corresponding sequences of X. fragariae strain NBC2815 (LT853880.1), respectively. The resulting concatenated data set of cpn60-gyrB-rpoD-fyuA was used to build a Multilocus Sequence Analysis (MLSA) by maximum likelihood criteria (Fig. 2). The cpn60-gyrB-rpoD-fyuA sequences of the isolate from Liaoning clustered in the clade containing the type strain of X. fragariae NBC2815, indicating that it belongs to X. fragariae. Thus, the bacterial strain from Liaoning was designated as X. fragariae strain YL19. To fulfill Koch’s postulates, the base of leaf petioles of disease-free strawberry plants were syringe-infiltrated inoculated with bacterial suspension (2×108 CFU) prepared from colonies of X. fragariae YL19 washed from SPA plates. The inoculated and control (treated with sterile water) were placed in a chamber (25/20℃day/night,≥90% relative humidity(RH), 12/12 h photoperiod) for three months. After one month, water-soaked symptoms were observed in the crown tissues of all X. fragariae YL19-inoculated plants. Two months after inoculation, a significant crown pocket similarly to initial symptoms observed in the field was developed on all inoculated plants. No symptoms were observed in the control plants. The bacteria were re-isolated from the symptomatic leaves, petioles and crowns, and confirmed as X. fragariae YL19 by the above mentioned morphological and molecular analyses. Pathogenicity tests were conducted three times and the same results were obtained. It was reported that X. fragariae usually causes angular leaf spot, a serious bacterial disease in many strawberry production regions worldwide. The typical symptoms of angular leaf spot caused by X. fragariae include reddish-brown, irregular spots on the upper leaf surface, water-soaked lesions developed along leaf veins. Although angular leaf spot caused by X. fragariae has been reported in Tianjin and Taiwan province, China (Wang et al. 2017; Wu et al. 2020), there is no report about the symptoms that infection pockets on crowns caused by X. fragariae strain YL19 as described above. This result indicated that YL19 is different from the other two X. fragariae strains reported in China or the disease caused by YL19 could be a severe case of angular leaf spot and vascular decline or collapse in strawberry (Bradbury, 1977). . To the best of our knowledge, these results showed a previously unreported new strain YL19 of X. fragariae is the causal agent of crown infecton pocket in strawberry in China, it may lead to serious losses to the local strawberry industry. This report will assist in developing management measures for this disease promptly.

The Use of Aerated Steam as a Heat Treatment for Managing Angular Leaf Spot in Strawberry Nursery Production and Its Effect on Plant Yield

Xanthomonas fragariae, the bacterium causing angular leaf spot (ALS) of strawberry, is found routinely on strawberry nursery stock. Although ALS can be a serious disease in production fields, it can be particularly problematic in the sale and trade of commercial nursery stock because of international trade regulations. Heat treatment has been shown to be an effective treatment for managing ALS on nursery stock in small-scale experimental trials. The objective of this research was to design, build, and trial precision thermotherapy units for managing ALS for both research and commercial applications and to test a new thermotherapeutic protocol on strawberry nursery stock that combines a conditioning thermal treatment with an eradicative thermal treatment. Small-plot trials were conducted to evaluate the efficacy of precision thermotherapy on control of ALS using known sources of infected nursery stock. Additionally, trials were conducted in cooperation with commercial nurseries to determine the impact of thermotherapy on plant health and yield and on the natural development of ALS. In the small-plot trials, ALS incidence was significantly lower in plots treated with precision thermotherapy. In the commercial trials, precision thermotherapy had a variable, but negligible, effect on plant growth and yield. ALS, when it occurred, was always lower in thermotherapy-treated plots. Heat is a near-universal biocide. Thus, in addition to managing ALS, the commercial application of precision thermal therapy to strawberry nursery stock may be effective for managing a wide range of pest and disease threats to strawberry while simultaneously reducing pesticide usage. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CCO “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.

First Report of Xanthomonas fragariae Causing Angular Leaf Spot on Strawberry (Fragaria x ananassa) in Taiwan

Angular leaf spot of strawberry, considered an A2 quarantine pest by the European and Mediterranean Plant Protection Organization (EPPO 2019), is an important bacterial disease in many regions. Since 2017, symptoms similar to angular leaf spot were observed in several strawberry cultivars including ‘Taoyuan No. 1’ and ‘Xiang-Shui’. Early symptoms were angular, water-soaked lesions on the abaxial leaf surface, and later, reddish-brown irregular spots and coalesced lesions developed on the adaxial surface. In the humid conditions, sticky bacterial ooze exuding from lesions was observed. To isolate the causal agent, leaves showing water-soaked lesions were surface sterilized, cut into small pieces and soaked in 5 ml sterile water for at least 15 min. The supernatant from the cut-up pieces was serially diluted followed by spreading on sucrose peptone agar (SPA) (Hayward 1960). After incubating at 20°C for 4-5 days, single colonies grown on SPA were transferred to a new SPA plate and cultured at 20°C until colonies appeared. The yellow, glossy and mucoid colonies, which resembled the colony morphology of Xanthomonas fragariae, were selected as candidates for further confirmation. First, bacterial DNA of four candidate isolates, B001, B003 and B005 from Miaoli County and B004 from Taoyuan City, was PCR amplified with X. fragariae-specific primers: XF9/XF12 (Roberts et al. 1996) and 245A/B and 295A/B (Pooler et al. 1996). All four isolates could be detected by XF9/XF12 primer. Furthermore, isolates B003 and B004 could be detected by both 245A/B and 295A/B primers, while B001 and B005 could be detected by 295A/B only. Next, DNA gyrase subunit B (gyrB) was PCR amplified with the primers XgyrB1F/XgyrB1R (Young et al. 2008). The gyrB sequences of these four isolates were deposited in GenBank with accession numbers MT754942 to MT754945. The gyrB phylogenetic tree was constructed based on Bayesian inference analysis and maximum likelihood analysis. The gyrB sequences of the four isolates from Taiwan clustered in the clade containing the type strain of X. fragariae ICMP5715, indicating that they belong to X. fragariae. B001 and B005 formed a sub-group separated from B003 and B004, suggesting genetic differences between these isolates. To fulfill Koch’s postulates, the abaxial surface of strawberry leaves were syringe infiltrated (KJP Silva et al., 2017) or wounded inoculated (Wang et al., 2017) with bacterial suspensions (final OD600 = 1.0-2.0) prepared from colonies of B001 and B003 washed from SPA plates. Inoculated plants were enclosed in a plastic bag (> 90% RH) at 25/20°C (day/night) under a 12-h/12-h photoperiod. After 7-14 days, water-soaked lesions similar to those observed in the field were developed on all inoculated leaves. The bacteria were successfully re-isolated from lesions of inoculated leaves and confirmed by specific primers XF9/XF12, 245A/B and 295A/B. We also found that the disease commonly occurs in the strawberry fields/nurseries with sprinkler irrigation during winter or early spring, and was particularly serious in the windward side or near riverside. To our knowledge, this is the first report of X. fragariae causing angular leaf spot on strawberry in Taiwan. Currently, the disease only occurs severely in certain regions, but establishment of effective management strategies will be needed to prevent spreading of this disease and potential economic loss in the future.

Genetics of angular leaf spot (ALS) resistance in South African market class dry bean (Phaseolus vulgaris) cultivars

Angular leaf spot (ALS) caused by Pseudocercospora griseola (Sacc.) is one of the serious fungal diseases affecting dry bean in Africa, including South Africa. Host plant resistance is the best management strategy, of which its effectiveness requires knowledge of the genetics underlying the resistance in guiding breeding efforts. In this study, the inheritance of ALS resistance was studied through the generation mean analysis biometrical procedure. Six generations consisting of the two parents Ukulinga and Gadra, and its cross derived filial progenies (F1), second-generation (F2), and backcrosses of F1 to Ukulinga (BCP1) and Gadra (BCP2) were planted in a net-house and later inoculated using a mixture of P. griseola isolates. Leaf lesions (% disease severity) were rated using a CIAT 1-9 scale and analysed using SAS macros in Proc GLM of SAS version 9.3. Results of ANOVA for a full model displayed significant additive effects (P<0.05) and highly significant (P<0.001) additive x dominance effects. Segregation analysis indicated 9:7 ratio, implying the involvement of complementary gene effects. The number of genes was estimated to be 1.79; however, due to epistasis two or more genes possibly govern the resistance in this cross. Broad and narrow sense heritabilities were 0.40 and 0.33, respectively. Therefore, the estimated narrow-sense heritability, additive gene effects, and epistatic interaction imply that it is prudent to delay selections until later stages, in which homozygosity could be achieved and additive effects fixed.