Fungus Colletotrichum Lindemuthianum Research Articles

Chitin Deacetylase as a Biocatalyst for the Selective N-Acylation of Chitosan Oligo- and Polymers

Chitosans, derived from the abundant natural resource chitin, are among the most versatile and promising functional biopolymers due to their unique physicochemical properties and biological activities. These can be further improved or modified by various functionalizations, targeting both the hydroxy and amino groups. However, the chemical routes used for functionalization typically use harsh chemicals, increasing the ecological footprint of the products, tend to yield low degrees of substitution, and often lack chemoselectivity. We here report on an alternative, biocatalytic route of chitosan N-acylation using a recombinant chitin deacetylase (CDA). These enzymes are known for their ability to chemo- and regioselectively N-acetylate glucosamine oligomers, and they were recently shown to also exhibit this reverse activity toward polyglucosamine, yielding partially acetylated chitosan polymers with a nonrandom pattern of acetylation. As chitin deacetylases can possess a certain cosubstrate promiscuity, we explored the ability of a CDA from the fungus Colletotrichum lindemuthianum (ClCDA) to N-acylate glucosamine tetramers and polyglucosamines using a range of small carboxylic acids as cosubstrates. The resulting tetramers were analyzed using ultra-high-performance hydrophilic liquid chromatography-tandem mass spectrometry (UHPLC-HILIC-MS), and the kinetic parameters of the acylation reactions thus determined gave deeper insight into the limitation of the cosubstrate scope of ClCDA. Using polyglucosamines as substrates, we obtained N-propiolated chitosan polymers with high fractions of substitution of 0.7. Copper-catalyzed azide–alkyne cycloaddition (CuAAC) then yielded a fluorescence-labeled polymer, providing proof-of-principle for click functionalization of chitosans using this chemoenzymatic approach. Given the known regioselectivity of chitin deacetylases, which is retained during reverse N-acetylation, this process might give access to a broad variety of functionalized chitosans with nonrandom substitution patterns.

Submerged culture fermentation of Colletotrichum lindemuthianum DSM 12250 as biotechnological strategy for fungal chitin biotransformation

Chitin and its derived products have great potential in various industries, such as biomedical, pharmaceutical, food, among others, due to its biodegradability, biocompatibility, null toxicity, and versatility. At present, these compounds are obtained by extraction with physical, chemical, or enzymatic processes. The fungus Colletotrichum lindemuthianum DSM 12250 which produces the enzyme chitin deacetylase (ClCDA), is proposed as an alternative for the biotransformation of fungal chitin. Chitin with a % N-Acetylation of 63.6% was obtained from the dry mycelium of Aspergillus niger that, based on analysis of Fourier transform infrared spectroscopy (FT-IR) and Differential Scanning Calorimetry (DSC), is structurally similar to commercial chitin. During fermentation in submerged culture, chitin presence was not found to generate inhibition in microbial growth or the activity of the ClCDA. The maximum enzymatic activity was 0.910 ± 0.002 U/mL and deacetylation of 20.9% were achieved. This was possible due to the enzymatic machinery of the fungus that includes CDA, endo-chitinases, and N-acetyl hexosaminidases. These compounds contribute to decreasing the substrate complexity, obtaining chitosan with a DDA of 57.3%. Submerged culture fermentation of Colletotrichum lindemuthianum DSM 12250 with fungal chitin as substrate represents a potential alternative for integrating different steps in the extraction of chitin-derived products while contributing to a cleaner and more efficient production.

Evaluation of plant extracts for the management of Cowpea anthracnose disease caused by Colletotrichum lindemuthianum

The fungus Colletotrichum lindemuthianum is both seed and soil borne which is the causative of cowpea anthracnose. This causes a significant loss of the crop and becoming a major hindrance in its bumper productivity in all countries where such crop is grown. In order to find out an environment friendly, sustainable control protocol, experiments were conducted by taking eight different plant extracts against the pathogen. The extracts were applied to the plant samples after the 25th day of germination in the field. Application of neem followed by ginger, onion, garlic and tulsi extracts effectively reduce the percent disease incidence by 90%, 85%, 82%, 79% and 76%, respectively. Similarly, the percent green pod yield increased by 134,127, 114, 93 and 84, respectively over control as well as the cost-benefit ratio which can be recommended to the farming community.

Climbing bean breeding for disease resistance and grain quality traits

AbstractCommon bean (Phaseolus vulgaris) is grown in two growth types, bush and climbing beans. The latter are preferred in several regions in East and Southern African as well as in Latin America (dominant in Rwanda and Colombia), due to higher yields and resilience compared with bush type. Common bean productivity is reduced by several pests and diseases between them. Bean common mosaic virus (BCMV), which is the most common and destructive poty‐virus affecting bean production worldwide, and anthracnose, caused by the fungus Colletotrichum lindemuthianum, can cause yield losses as high as 95% in susceptible cultivars. Further important traits in common bean are high micro mineral contents to alleviate malnutrition and grain quality traits such as the canning quality of bean varieties, which is important for farmers to access the processing market. In this study, new climbing bean populations were generated (coded ENF/CGA) to combine high seed iron (SdFe) and multiple diseases resistance. Double and triple crosses between parents with virus and anthracnose resistance, high SdFe, and good agronomic traits were employed. In trials in Darien and Popayan, lines were identified that combine BCMNV/BCMV and anthracnose resistance with seed yields above 4000 kg/ha. Phenotypic evaluations validated the usefulness of SNP markers tagging the genes bc‐3 and I for BCMN and Co‐3 for anthracnose as a selection tool for field resistance. These results show the genetic potential of the lines that are now being tested in target regions to be delivered to smallholder farmers.

Inhibition of the fungus Colletotrichum lindemuthianum in the presence of different potassium sorbate concentrations

One of the major problems of in vitro plant cultivation is caused by contaminations such asfungal, which represent great losses for plant cultivation laboratories. In view of the problem about contamination and gaps in efficient disinfection protocols, the present study aimed to evaluate the inhibition of the fungus Colletotrichum. lindemuthianum in the presence of the potassium sorbate food preservative at different concentrations (0 g L-1; 0.076 g. L-1; 0.15 g. L-1; 0.30 g. L-1; 0.45 g. L-1; 0.60 g. L-1 and 0.75 g. L-1). The analyses of the Percentage of Micelal Growth Inhibition (ICP) and the growth at 24 and 72 hours at different concentrations were quantified. It was observed that Potassium Sorbate was effective in the analysis of ICP at different concentrations in relation to control, with greater effectiveness for T4 to T7. The highest percentages of inhibition (T5 to T7) presenting an inhibition rate of mean mycelial growth of 17.5 % in relation to the control for the 72h period, indicating, therefore, that with further studies this preservative can be used to combat fungal microorganisms in vitro culture.

Genome-wide association mapping reveals race-specific SNP markers associated with anthracnose resistance in carioca common beans.

Brazil is the largest consumer of dry edible beans (Phaseolus vulgaris L.) in the world, 70% of consumption is of the carioca variety. Although the variety has high yield, it is susceptible to several diseases, among them, anthracnose (ANT) can lead to losses of up to 100% of production. The most effective strategy to overcome ANT, a disease caused by the fungus Colletotrichum lindemuthianum, is the development of resistant cultivars. For that reason, the selection of carioca genotypes resistant to multiple ANT races and the identification of loci/markers associated with genetic resistance are extremely important for the genetic breeding process. Using a carioca diversity panel (CDP) with 125 genotypes and genotyped by BeadChip BARCBean6K_3 and a carioca segregating population AM (AND-277 × IAC-Milênio) genotyped by sequencing (GBS). Multiple interval mapping (MIM) and genome-wide association studies (GWAS) were used as mapping tools for the resistance genes to the major ANT physiological races present in the country. In general, 14 single nucleotide polymorphisms (SNPs) showed high significance for resistance by GWAS, and loci associated with multiple races were also identified, as the Co-3 locus. The SNPs ss715642306 and ss715649427 in linkage disequilibrium (LD) at the beginning of chromosome Pv04 were associated with all the races used, and 16 genes known to be related to plant immunity were identified in this region. Using the resistant cultivars and the markers associated with significant quantitative resistance loci (QRL), discriminant analysis of principal components (DAPC) was performed considering the allelic contribution to resistance. Through the DAPC clustering, cultivar sources with high potential for durable anthracnose resistance were recommended. The MIM confirmed the presence of the Co-14 locus in the AND-277 cultivar which revealed that it was the only one associated with resistance to ANT race 81. Three other loci were associated with race 81 on chromosomes Pv03, Pv10, and Pv11. This is the first study to identify new resistance loci in the AND-277 cultivar. Finally, the same Co-14 locus was also significant for the CDP at the end of Pv01. The new SNPs identified, especially those associated with more than one race, present great potential for use in marker-assisted and early selection of inbred lines.

A common bean truncated CRINKLY4 kinase controls gene-for-gene resistance to the fungus Colletotrichum lindemuthianum.

Identifying the molecular basis of resistance to pathogens is critical to promote a chemical-free cropping system. In plants, nucleotide-binding leucine-rich repeat constitute the largest family of disease resistance (R) genes, but this resistance can be rapidly overcome by the pathogen, prompting research into alternative sources of resistance. Anthracnose, caused by the fungus Colletotrichum lindemuthianum, is one of the most important diseases of common bean. This study aimed to identify the molecular basis of Co-x, an anthracnose R gene conferring total resistance to the extremely virulent C. lindemuthianum strain 100. To that end, we sequenced the Co-x 58 kb target region in the resistant JaloEEP558 (Co-x) common bean and identified KTR2/3, an additional gene encoding a truncated and chimeric CRINKLY4 kinase, located within a CRINKLY4 kinase cluster. The presence of KTR2/3 is strictly correlated with resistance to strain 100 in a diversity panel of common beans. Furthermore, KTR2/3 expression is up-regulated 24 hours post-inoculation and its transient expression in a susceptible genotype increases resistance to strain 100. Our results provide evidence that Co-x encodes a truncated and chimeric CRINKLY4 kinase probably resulting from an unequal recombination event that occurred recently in the Andean domesticated gene pool. This atypical R gene may act as a decoy involved in indirect recognition of a fungal effector.

POTASSIUM FERTILIZATION MANAGEMENT IN COMMON BEAN PROTECTION AGAINST COLLETOTRICHUM LINDEMUTHIANUM

Anthracnose of common bean caused by the fungus Colletotrichum lindemuthianum is considered one of the most important fungal diseases that attacks the aerial part of the common bean for causing significant losses in final productivity. Potassium is one of the elements that is involved in the defense of the plant. Thus, the objective was to evaluate the effect of potassium doses in the control of anthracnose in three common bean cultivars. The design used was completely randomized with five treatments in three repetitions. The severity of the disease was obtained at 15 days after inoculation using a diagrammatic scale of notes and by counting the number of lesions in the central and lateral leaflets and the length of the lesions in the midrib of the central leaflet.

Genetic resistance of common bean cultivar Beija Flor to Colletotrichum lindemuthianum

Anthracnose, which is caused by the fungus Colletotrichum lindemuthianum, is one of the most widespread and important diseases of the common bean (Phaseolus vulgaris L.) in the world. The objective of the present study was to characterize the genetic resistance of the Beija Flor cultivar by inheritance and to conduct allelism tests. The inheritance test was conducted in the F2 population derived from the Beija Flor (resistant) x TU (susceptible) cross inoculated with race 2047 of C. lindemuthianum. Furthermore, allelism tests exhibited a fitted segregation ratio of 15R:1S, thereby indicating the independence of the Beija Flor gene from the following previously characterized genes: Co-1, Co-2, Co-4, Co-42, Co-6, Co-12, Co-14, Co-15, and Co-Pe. Based on the aforementioned results, we are proposing the symbol Co-Bf to designate the new anthracnose resistance gene in the Brazilian Andean common bean cultivar Beija Flor. This cultivar is an important source of resistance to C. lindemuthianum that should provide a valuable contribution to the common bean breeding program for anthracnose resistance.

Identification of anthracnose races in Manitoba and Ontario from 2005 to 2015 and their reactions on Ontario dry bean cultivars

Anthracnose, caused by the fungus Colletotrichum lindemuthianum (Sacc. & Magnus) Briosi & Cavara, is one of the most destructive diseases of dry bean (Phaseolus vulgaris L.) in the world. Between 2005 and 2015, commercial fields of dry beans in Manitoba and Ontario were surveyed to determine the frequency of occurrence of races of the anthracnose fungus. Throughout the study, race 73 was most prevalent in Manitoba and Ontario. However, three anthracnose races not previously reported in Canada also were identified. These three new races and four previously identified anthracnose races were used to screen 52 dry bean cultivars, as well as a mung bean and azuki bean cultivar from Ontario, for their seedling reactions to determine their patterns of race resistance. The dry bean cultivars were classified into a total of 19 resistance spectra based on the pattern of seedling reactions to the seven anthracnose races. The most common resistance spectrum was susceptible to the majority of the anthracnose races and no cultivar was resistant to all of the races. Many bean cultivars produced intermediate anthracnose ratings to races 31 and 105 and tests of 16 dry bean cultivars against those races indicated that all cultivars with intermediate ratings to a specific race were segregating in their seedling reactions and none of the cultivars produced plants with only intermediate anthracnose severity ratings. This study provides new information on the anthracnose reactions of common bean cultivars in Canada, which should be useful for the development of new bean cultivars with durable resistance.

Evaluation of Common Bean (Phaseolous vulgaris L.) Varieties for the Reaction of Common Bean Anthracnose (Colletotrichum lindemuthianum) at Sirinka, Eastern Amhara, Ethiopia

Common bean (Phaseolous vulgaris L.) is the most important food legume consumed as source of protein and cash crop in Ethiopia. The production of the crops is constrained by biotic and abiotic factors. Common bean anthracnose caused by the fungus Colletotrichum lindemuthianum is a major production constraint in common bean growing regions of Ethiopia. Field experiment was conducted at Sirinka Agricultural research center during 2017/18 main cropping season to evaluate reactions of common bean varieties to the disease. The experiment consisted of twenty two common bean varieties evaluated for the reaction to anthracnose under natural infestation conditions. The highest disease severity (58%) was recorded from Awash-1 variety while the lowest disease severity (45%), was recorded from Awash Melka variety at final assessment day. The highest (3.03 t ha-1) yield was recorded from Awash Melka variety while the lowest (0.97 t ha-1) yield was recorded from KAT-B1 variety. From the present study, it is possible to conclude that, the advantage of screening resistant verities increases the opportunity to select for a broad range of anthracnose resistance and help to know the variability of the common bean anthracnose disease.

Antifungal Activity of Arginine-Based Surfactants

Background:Amino acid based surfactants constitute an important class of surface active biomolecules showing remarkable biocompatible properties. Antimicrobial activity is one of the most remarkable biological properties of this kind of surfactants, which have been widely studied against a broad spectrum of microorganisms. However, the antifungal activity of this kind of compound has been less well investigated. The aim of this work is the study of the antifungal activity of two novel argininebased surfactants (Nα-benzoyl-arginine decylamide, Bz-Arg-NHC10 and Nα-benzoyl-arginine dodecylamide, Bz-Arg-NHC12), obtained by an enzymatic strategy, against phytopathogenic filamentous fungi and dermatophyte strains.Methods:Four phytopathogenic fungi (Fusarium oxysporum, Fusarium solani, Colletotrichum gloeosporioides and Colletotrichum lindemuthianum) and two human pathogenic fungi (dermatophytes Trichophyton rubrum and Trichophyton mentagrophytes) were tested. Inhibition of vegetative growth and conidia germination was investigated for the phytopathogenic fungi. In order to elucidate the possible mechanism of biocide action, membrane integrity, as well as the production of reactive oxygen species (ROS) were evaluated. Additionally, the inhibition of germination of dermatophyte microconidia due to both arginine-based surfactants was studied. Minimum inhibitory concentration, as well as the concentration that inhibits 50% of germination were determined for both compounds and both fungal strains.Results:For the vegetative growth of phytopathogenic fungi, the most potent arginine-based compound was Bz-Arg-NHC10. All the tested compounds interfered with the conidia development of the studied species. Investigation of the possible mechanism of toxicity towards phytopathogenic fungi indicated direct damage of the plasma membrane and production of ROS. For the two strains of dermatophyte fungi tested, all the proved compounds showed similar fungistatic efficacy.Conclusion:: Bz-Arg-NHC10 and Bz-Arg-NHC12 were demonstrated to have broad biocidal ability against the proliferative vegetative form and the asexual reproductive conidia. Results suggest that both membrane permeabilization and induction of oxidative stress are part of the antifungal mechanisms involved in the interruption of normal conidia development by Bz-Arg-NHCn, leading to cell death.

Survival of the bean anthracnose fungus (Colletotrichum lindemuthianum) on crop debris in Canada

Anthracnose caused by the fungus Colletotrichum lindemuthianum (Sacc. & Magnus) Lams.-Scrib. has been a major constraint to dry bean (Phaseolus vulgaris L.) production in Manitoba and Ontario, as well as elsewhere in the world. There is a lack of consensus on the length of time the anthracnose fungus can survive on infected crop debris as well as the effectiveness of crop rotation to control the disease under temperate climatic conditions. A multi-year study was established at field sites near Morden, MB and Exeter, ON to examine the survival of C. lindemuthianum on seed, pod and stem tissue samples of infected beans on the soil surface and buried in the soil over different 2-year periods. The survival of the anthracnose fungus was influenced by location, type of infected tissue and burial in the soil. On average, C. lindemuthianum could persist for 18 months at Morden and about 9 months at Exeter, which was long enough to serve as a source of primary infection to subsequent crops of dry beans. At both field sites, the anthracnose fungus overwintered longer on infected stems and pods than it did on infected bean seed. The burial of infected bean tissue reduced the viability of C. lindemuthianum in all tissue types compared with samples that remained on the soil surface. A 3-year crop rotation study at Morden in which either a wheat, fallow or bean treatment were grown in the second year, confirmed that the anthracnose fungus could survive long enough under zero-tillage conditions to infect a bean crop in the third year of the study and cause considerable downgrading of the seed. This study demonstrated the importance of tillage and crop rotation in the integrated control of bean anthracnose.

The repertoire of effector candidates in Colletotrichum lindemuthianum reveals important information about Colletotrichum genus lifestyle.

The fungus Colletotrichum lindemuthianum is the causal agent of anthracnose in the common bean (Phaseolus vulgaris), and anthracnose is one of the most devastating diseases of this plant species. However, little is known about the proteins that are essential for the fungus-plant interactions. Knowledge of the fungus' arsenal of effector proteins is of great importance for understanding this pathosystem. In this work, we analyzed for the first time the arsenal of Colletotrichum lindemuthianum effector candidates (ClECs) and compared them with effector proteins from other species of the genus Colletotrichum, providing a valuable resource for studying the infection mechanisms of these pathogens in their hosts. Isolates of two physiological races (83.501 and 89 A2 2-3) of C. lindemuthianum were used to predict 353 and 349 ClECs, respectively. Of these ClECs, 63% were found to be rich in cysteine, have repetitive sequences of amino acids, and/or possess nuclear localization sequences. Several conserved domains were found between the ClECs. We also applied the effector prediction to nine species in the genus Colletotrichum, and the results ranged from 247 predicted effectors in Colletotrichum graminicola to 446 in Colletotrichum orbiculare. Twelve conserved domains were predicted in the effector candidates of all analyzed species of Colletotrichum. An expression analysis of the eight genes encoding the effector candidates in C. lindemuthianum revealed their induction during the biotrophic phase of the fungus on the bean.

Gene/QTL discovery for Anthracnose in common bean (Phaseolus vulgaris L.) from North-western Himalayas.

Common bean (Phaseolus vulgaris L.) is one of the most important grain legume crops in the world. The beans grown in north-western Himalayas possess huge diversity for seed color, shape and size but are mostly susceptible to Anthracnose disease caused by seed born fungus Colletotrichum lindemuthianum. Dozens of QTLs/genes have been already identified for this disease in common bean world-wide. However, this is the first report of gene/QTL discovery for Anthracnose using bean germplasm from north-western Himalayas of state Jammu & Kashmir, India. A core set of 96 bean lines comprising 54 indigenous local landraces from 11 hot-spots and 42 exotic lines from 10 different countries were phenotyped at two locations (SKUAST-Jammu and Bhaderwah, Jammu) for Anthracnose resistance. The core set was also genotyped with genome-wide (91) random and trait linked SSR markers. The study of marker-trait associations (MTAs) led to the identification of 10 QTLs/genes for Anthracnose resistance. Among the 10 QTLs/genes identified, two MTAs are stable (BM45 & BM211), two MTAs (PVctt1 & BM211) are major explaining more than 20% phenotypic variation for Anthracnose and one MTA (BM211) is both stable and major. Six (06) genomic regions are reported for the first time, while as four (04) genomic regions validated the already known QTL/gene regions/clusters for Anthracnose. The major, stable and validated markers reported during the present study associated with Anthracnose resistance will prove useful in common bean molecular breeding programs aimed at enhancing Anthracnose resistance of local bean landraces grown in north-western Himalayas of state Jammu and Kashmir.

Identifying the markers and tagging a leucine-rich repeat receptor-like kinase gene for resistance to anthracnose disease in vegetable cowpea [Vigna unguiculata (L.) Walp.

ABSTRACTAnthracnose caused by the fungus Colletotrichum lindemuthianum is the most destructive disease of cowpea. Field-type cowpeas show various levels of resistance, whereas pole-type vegetable cowpeas are highly susceptible. Transfer of resistance available in field types to vegetable types is a major breeding objective in cowpea. This paper details the development of an F2 mapping population by crossing field-type cowpea variety Kanakamony (Vigna unguiculata ssp. cylindrica) with pole-type vegetable cowpea variety Sharika (Vigna unguiculata ssp. sesquipedalis), screening this population with artificial inoculation and Bulked Segregant Analysis (BSA) with random marker systems Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeats (ISSR); the objective is to identify the markers linked with major resistance-contributing genes. RAPD primer OPA02 has yielded marker at 850 bp in susceptible genome, whereas ISSR primers UBC810 and UBC811 have yielded markers at 1.4 kb and 1.5 kb respectively in resistant genomes. These markers were reproducible and their linkage with resistance and suitability in marker assisted selection (MAS) were confirmed through co-segregation analysis in F3 population. UBC811 marker was eluted, cloned on pGEM-T, and sequenced. The sequence had shown that this marker is anchored on LRR receptor-like serine/t\\hreonine protein kinase gene which could be involved in the resistance mechanism.

Methyl and p-Bromobenzyl Esters of Hydrogenated Kaurenoic Acid for Controlling Anthracnose in Common Bean Plants.

Kaurenoic acid derivatives were prepared and submitted to in vitro assays with the fungus Colletotrichum lindemuthianum, which causes anthracnose disease in the common bean. The most active substances were found to be methyl and p-bromobenzylesters, 7 and 9, respectively, of the hydrogenated kaurenoic acid, which presented a minimum inhibitory concentration (MIC) of 0.097 and 0.131 mM, respectively, while the commercial fungicide methyl thiophanate (MT) presented a MIC of 0.143 mM. Substances 7 (1.401 mM) and 9 (1.886 mM) reduced the severity of anthracnose in common bean to values statistically comparable to MT (2.044 mM). According to an in silico study, both compounds 7 and 9 are inhibitors of the ketosteroid isomerase (KSI) enzyme produced by other organisms, the amino acid sequence of which could be detected in fungal genomes. These substances appeared to act against C. lindemuthianum by inhibiting its KSI. Therefore, substances 7 and 9 are promising for the development of new fungicides.

Differential expression of genes during the interaction between Colletotrichum lindemuthianum and Phaseolus vulgaris

The fungus Colletotrichum lindemuthianum is the causal agent of anthracnose, one of the most severe diseases of the common bean (Phaseolus vulgaris). The infection process begins with the adhesion of conidia to the plant’s surface. Appressoria are then formed, allowing penetration of the fungus. Next, the biotrophic phase begins, followed by the necrotrophic phase. Due to the peculiar nutrition mode of the fungus, including both of the previously mentioned stages, it is of great interest to determine which genes are involved in the transition between the two phases during the infection process. To determine this, suppression subtractive hybridization (SSH) was used in association with qRT-PCR in the present study. These methods allowed for the identification of genes that were differentially expressed at each developmental stage of the fungus in the plant. This is the first report on the use of the cited techniques to evaluate the infectious cycle of the fungus. A total of 175 sequences exhibited significant identity (e ≤ 10−5) with sequences present in the sequenced genomes of P. vulgaris and C. lindemuthianum; approximately 41 % of those were determined to belong to the fungus, and 59 % were determined to belong to the plant. Of the predicted sequences, 68 % were of unknown function or were not found in the databases. Among the analyzed expressed sequence tags (ESTs), sequences were found that encode proteins related to: primary and secondary metabolism; the transport of different compounds; the degradation/modification of proteins; cell regulation and signaling; cellular stress response; and the degradation of exogenous compounds. The obtained results allowed for the identification of sequences encoding proteins that are essential for the progression of anthracnose. Furthermore, it was possible to identify new genes, the functions of which have not yet been described, and even to identify unique genes of C. lindemuthianum that are involved in the pathogenicity and virulence of this fungus.

Genetic resistance to Colletotrichum lindemuthianum in the Andean cultivar Jalo Pintado 2 of common bean

The anthracnose caused by fungus Colletotrichum lindemuthianum (Sacc. & Magnus) Briosi & Cavara is the most widespread disease and economically important fungal disease of common bean (Phaseolus vulgaris L.). The use of resistant cultivars is considered as one of the most effective methods in controlling this disease. The present study had as aim to characterize the genetic resistance of the Andean common bean cultivar Jalo Pintado 2 to the C. lindemuthianum through inheritance and allelism tests. The experiment was conducted under greenhouse conditions at Laboratório de Melhoramento do Feijão Comum e de Biologia Molecular do Núcleo de Pesquisa Aplicada a Agricultura (Nupagri) at Universidade Estadual de Maringá, Paraná, Brazil. The results of the F2 population from the crossing 'Jalo Pintado 2' (R) × Cornell 49-242 (S), inoculated with race 73 of C. lindemuthianum, adjusted to the ratio of 3R: 1S, demonstrating the action of a dominant gene in the cultivar Jalo Pintado 2. The allelism tests evidenced that the gene in the 'Jalo Pintado 2' is independent from those previously characterized: Co-1, Co-2,Co-3, Co-34, Co-4, Co-42, Co-43, Co-5, Co-6, Co-11, Co-12, Co-13, Co-14, Co-15 and Co-16. This gene is also independent from those genes not yet named present in Paloma, Perla and Amendoim Cavalo cultivars. The authors propose the Co-

Genome-Wide Association Study of Anthracnose Resistance in Andean Beans (Phaseolus vulgaris).

Anthracnose is a seed-borne disease of common bean (Phaseolus vulgaris L.) caused by the fungus Colletotrichum lindemuthianum, and the pathogen is cosmopolitan in distribution. The objectives of this study were to identify new sources of anthracnose resistance in a diverse panel of 230 Andean beans comprised of multiple seed types and market classes from the Americas, Africa, and Europe, and explore the genetic basis of this resistance using genome-wide association mapping analysis (GWAS). Twenty-eight of the 230 lines tested were resistant to six out of the eight races screened, but only one cultivar Uyole98 was resistant to all eight races (7, 39, 55, 65, 73, 109, 2047, and 3481) included in the study. Outputs from the GWAS indicated major quantitative trait loci (QTL) for resistance on chromosomes, Pv01, Pv02, and Pv04 and two minor QTL on Pv10 and Pv11. Candidate genes associated with the significant SNPs were detected on all five chromosomes. An independent QTL study was conducted to confirm the physical location of the Co-1 locus identified on Pv01 in an F4:6 recombinant inbred line (RIL) population. Resistance was determined to be conditioned by the single dominant gene Co-1 that mapped between 50.16 and 50.30 Mb on Pv01, and an InDel marker (NDSU_IND_1_50.2219) tightly linked to the gene was developed. The information reported will provide breeders with new and diverse sources of resistance and genomic regions to target in the development of anthracnose resistance in Andean beans.