Identification and pathogenicity of Fusarium species associated with pokkah boeng of sugarcane in Brazil

Brazil is the world’s biggest producer of sugarcane (Saccharum spp. hybrids). Pokkah boeng is an important fungal disease in this crop caused by members of the Fusarium fujikuroi species complex (FFSC) and characterized by deformation of the aerial part of the plant and stem rot. While the occurrence of symptoms has been reported in plantations in Brazil, no official reports of the disease exist. In this study, species of the FFSC were identified that are associated with sugarcane plants with symptoms of pokkah boeng in Brazil. This was achieved using two-loci molecular phylogeny, sexual compatibility and analysis of morphological markers. The ability of strains to cause disease in plants of sugarcane, maize, sorghum and millet was also evaluated. The 39 isolates studied were identified as F. sacchari, F. proliferatum and another, still unknown, phylogenetic lineage that is sister to F. andiyazi. Crossing field isolates of F. sacchari and F. proliferatum with their respective tester strains produced fertile perithecia and viable ascospores. All three species induced symptoms of pokkah boeng on inoculated sugarcane plants and caused stem rot in maize, sorghum and millet. Symptoms on sugarcane are chlorosis and necrosis of leaves, punctured lesions, twisted leaves, reduction of the total leaf area, death of the top of the plant and stalk rot. The findings confirmed the aetiology of the disease in Brazil, generated basic knowledge for the development of strategies for diagnosis and monitoring of the disease and support breeding programmes for selecting resistant germplasm.     

Isolation and pathogenic variability of Colletotrichum falcatum causing red rot in sugarcane

Journal of Plant Diseases and Protection - Red rot of sugarcane caused by the fungus Colletotrichum falcatum Went is the most destructive and severe disease worldwide. The present study identifies...     

安徽省玉米穗腐病主要致病镰孢菌的分离与鉴定

为明确安徽省玉米穗腐病主要致病镰孢菌的种类,采用单孢分离、形态学鉴定和分子生物学鉴定的方法,对2017年采集自安徽省6个玉米主产区——合肥市、蚌埠市、淮北市、阜阳市、亳州市和宿州市的玉米穗腐病病样进行了分离鉴定。在获得的455株镰孢菌中,拟轮枝镰孢菌、禾谷镰孢复合种、层出镰孢菌、尖孢镰孢菌和黄色镰孢菌的分离频率分别为59.13%、21.28%、13.68%、5.12%和0.79%。其中拟轮枝镰孢菌在各地的分布最广,属于安徽省的优势致病镰孢菌。     

Fusarium rot of melon is caused by several Fusarium species

Fusarium rot of melon, caused by species of the genus Fusarium, has become an important postharvest disease for many Brazilian producers. Due to the delayed onset of symptoms, this disease is often only detected when fruits arrive at the importing country, thus generating economic loss for the exportation of the fruit. This study was developed with the aim of investigating which Fusarium species cause fruit rot in melon and to evaluate any differences in aggressiveness and development of symptoms. Species were identified through phylogenetic analysis of two loci and morphological markers. The 28 isolates obtained from diseased melon fruits of different commercial cultivars were identified as Fusarium falciforme (FSSC), F. sulawesiense, F. pernambucanum (FIESC), and F. kalimantanense (FOSC). Three isolates belong to a new phylogenetic lineage within the F. fujikuroi species complex (FFSC). All isolates were tested for pathogenicity, and first symptoms of rot in Canary melon were observed 2 days after inoculation. Isolates of F. falciforme and F. sulawesiense were shown to be more aggressive. Our results extend information on Fusarium species that cause fruit rot in melon and support the development of management strategies, as there is currently no efficient control for this disease. To our knowledge, this is the first report of the occurrence of species of the FSSC, FOSC, and FFSC from muskmelon fruits in Brazil.     

Diversity and pathogenicity of Fusarium graminearum species complex from maize stalk and ear rot strains in northeast China

The Fusarium graminearum species complex (FGSC) is an important group of pathogens distributed in maize‐producing areas worldwide. This study investigated the genetic diversity and pathogenicity of 40 FGSC isolates obtained from stalk rot and ear rot samples collected from 42 locations in northeastern China during 2013 and 2014. A phylogenetic tree of translation elongation factor (EF‐la) sequences designated the 40 isolates as F. graminearum sensu stricto (67.5%) and F. boothii (32.5%). By using inter‐simple sequence repeat analysis (ISSR), it was shown that the isolates were divided into two clades, which corresponded to the species identity of the isolates. However, the isolates from the two different diseases could not be distinguished in pathogenicity. The disease severity index of seedlings inoculated with stalk isolates was slightly higher than that of seedlings inoculated with isolates from infected ears, whereas the pathogenicity of the stalk and ear isolates were identical.     

Fusarium commune associated with wilt and root rot disease in rice

Wilt and root rot disease in plants has been caused mainly by Fusarium species. Previous studies reported that members of the Fusarium oxysporum species complex (FOSC) were usually associated with this disease, but there has been no report of it being caused in rice by specific Fusarium species. However, in this study, Fusarium commune was identified and characterized as a causal agent of wilt and root rot disease of rice. Four Fusarium isolates (BD005R, BD014R, BD019R, and BD020R) were obtained from different parts (root, stem, and seeds) of diseased rice plants. In morphological studies, these isolates produced key characteristics of F. commune, such as long and slender monophialides, polyphialides, and abundant chlamydospores. In molecular studies, the isolates were identified as F. commune based on sequences of the translation elongation factor 1-α (TEF1) gene that had 99.7%–100% sequence identity with the reference strain F. commune NRRL 28058. The phylogenetic tree showed that all four isolates belonged to the F. commune clade. A mating type test determined that three isolates carried MAT1-2. Their teleomorph stage was still unknown. Pathogenicity assays showed that all the isolates produced wilt and root rot symptoms and the isolate BD019R was observed as the most virulent among the isolates. To our knowledge, this is the first report of F. commune causing wilt and root rot disease on rice.     

Molecular identification and pathogenicity of Fusarium and Alternaria species associated with root rot disease of wolfberry in Gansu and Ningxia provinces,China

Wolfberry (Lycium barbarum) ranks in the top 10 best-selling medicinal plants in China and it has been used for centuries as a medicine and a food supplement. It is suggested to have benefits on human health due to the rich content of polysaccharides, carotenoids, flavonoids, and alkaloids contained in its fruits, leaves, and root bark. Recently, severe root rot diseases have been causing plant losses in major growing areas. Here, we report fungi causing root rot disease in Chinese wolfberry plants. The analysis of nucleotide sequences of the internal transcribed spacer (ITS) region revealed a total of 92 isolates isolated from both soil and plant material samples. Fusarium spp. were the most abundant (58%), followed by Penicillium spp. (9%), and Alternaria spp. (5%). Fusarium spp. included F. oxysporum (36%), F. solani (30%), F. chlamydosporum (9%), F. nematophilum (9%), and F. tricinctum (8%). Sequences from the translation elongation factor 1-α gene (TEF-1α) were used to confirm the identity of Fusarium spp. and showed the predominance of F. oxysporum and F. solani. To confirm the pathogenicity of isolates, four isolates belonging to Fusarium spp. and one isolate belonging to Alternaria spp., isolated from wolfberry root tissues with root rot symptoms, were tested in outdoor and laboratory conditions. Results revealed that the five tested isolates were pathogenic with varying degrees of aggressiveness and ability to induce symptoms of root rot in wolfberry seedlings. The five isolates were recovered from inoculated seedlings, completing Koch's postulates. This is the first report on causative agents of root rot in Chinese wolfberry.     

玉米穗腐病和茎基腐病镰孢菌间相关性的RAPD分析

 利用随机扩增多态性DNA (RAPD)技术对玉米穗腐病和茎基腐病镰孢菌之间的相互关系进行了研究,证实了引起2种病害的串珠镰孢菌同源性很高,在遗传上具有较高的相似性,病菌不易受地域或环境和寄生部位选择作用的影响,同一类型的串珠镰孢菌可以是穗腐病和茎腐病的共同病原菌。禾谷镰孢菌间的遗传变异性很强,易受地域、环境和寄生部位选择作用的影响,出现明显的分化现象,穗腐病和茎基腐病可以由同一禾谷镰孢菌分化类型侵染所致,也可由不同分化类型侵染所致。     

中国芒果炭疽病菌复合种对苯醚甲环唑敏感性测定

芒果炭疽病是芒果上最严重的病害之一?本文采用生长速率法测定了来自不同芒果产区的芒果炭疽病菌对苯醚甲环唑的敏感性?结果表明, 苯醚甲环唑对供试的136个菌株的平均EC50为(1.46±0.14)mg/L, 其中, 最小值为0.09 mg/L, 最大值为9.74 mg/L, 其敏感性频率不符合正态分布?苯醚甲环唑对不同采集地区菌株的平均EC50无显著差异, 对贵州的菌株平均EC50最高, 对海南的最低?供试的芒果炭疽病菌包含13个种, 其中, 苯醚甲环唑对Colletotrichum cordylinicola的平均EC50最低, 对C.gloeosporioides 的平均EC50最高?     

Multilocus identification and thiophanate-methyl sensitivity of Colletotrichum gloeosporioides species complex associated with fruit with symptoms and symptomless leaves of mango

Colletotrichum spp. are known causal agents of anthracnose in a broad host range, causing severe losses. Currently, the most effective way to reduce disease is by fungicide application, which could give rise to resistant populations. This study aimed to determine the Colletotrichum species present in conventional and organic mango orchards and to evaluate their pathogenicity and sensitivity to the benzimidazole fungicide thiophanate-methyl. Seventy-one isolates from fruit with symptoms and symptomless leaves were obtained. From these, 20 representative morphotypes were analysed based on glyceraldehyde-3-phosphate dehydrogenase partial gene sequencing. A subset of 10 isolates based on different species, isolation source, and fungicide sensitivity was used for morphological and multilocus phylogenetic analysis. Colletotrichum queenslandicum was only identified in conventional production systems, C. chrysophilum only in organic systems, and C. asianum and C. siamense in both. Pathogenicity tests showed all species were pathogenic, and only C. asianum caused symptoms via both unwounded and wounded inoculation methods. Overall, 25.3% of isolates (n = 18) that belong to C. siamense, isolated from a conventional orchard, grew on thiophanate-methyl amended media at 1,000 µg/ml, suggesting high resistance. Resistance was not correlated with any common point mutations at positions 198 and 200 of the β-tubulin 2 protein, as commonly found in other fungal pathogens resistant to benzimidazole. The 74.7% remaining isolates (n = 53) belonging to the other species were sensitive, reaching 100% inhibition at <10 µg/ml. Even with benzimidazole application, anthracnose symptoms persist due to the emergence of pathogenic Colletotrichum subpopulations that are resistant to thiophanate-methyl.     

A new Fusarium lineage within the Gibberella fujikuroi species complex is the main causal agent of mango malformation disease in Brazil

Mango malformation is a serious disease in tropical and subtropical areas of the world and has been attributed to various Fusarium spp., including F. mangiferae , F. proliferatum , F. sacchari , F. sterilihyphosum and F. subglutinans . Isolates of Fusarium associated with mango malformation from Brazil, Egypt, India, South Africa and the United States were evaluated through amplified fragment length polymorphisms (AFLPs) and partial DNA sequences of the genes encoding β-tubulin ( tub2 ) and translation elongation factor 1-α ( tef1 ). These techniques were used to delimit species and to estimate the genetic and phylogenetic relatedness of the isolates. In the AFLP analysis, most of the Brazilian isolates formed a unique cluster. Additionally, one small cluster was formed by isolates of F. sterilihyphosum from Brazil and South Africa, and another by isolates of F. mangiferae from Egypt, India, South Africa and the United States. In the phylogenetic analysis, most of the Brazilian isolates represented a new phylogenetic lineage in the Gibberella fujikuroi species complex, where they formed a sister clade to F. sterilihyphosum. Representatives of both clades were pathogenic to mango (cv. Tommy Atkins) and Koch's postulates were completed for isolates belonging to the new lineage and to F. sterilihyphosum . Thus, most of the mango malformation disease in Brazil is due to a distinct phylogenetic lineage of Fusarium , and to a lesser extent by F. sterilihyphosum. The new phylogenetic lineage identified in this study, together with F. mangiferae and F. sterilihyphosum , are the only known taxa of Fusarium proven to be capable of causing mango malformation.     

Molecular identification and prevalence of Acidovorax avenae subsp. avenae causing red stripe of sugarcane in China

Red stripe caused by the bacterium Acidovorax avenae subsp. avenae (Aaa) is a disease of sugarcane that is distributed worldwide. In this study, 108 sugarcane leaf samples were collected in 2013–2016 from nine sugarcane‐growing regions in China. Aaa was detected by PCR with specific and novel primers from the 16S–23S rDNA internal transcribed spacer region in 81 of 84 (96%) leaves with red stripe symptoms and in 20 of 24 (83%) leaves without symptoms. Furthermore, Aaa was detected in all nine sampling locations representing six sugarcane‐producing provinces in China. The 101 amplified fragments were cloned and sequenced. The size of the nucleotide sequences varied from 436 to 454 bp and the sequence identity ranged from 89.2% to 100%, suggesting a significant genetic variation among Aaa strains from China. Five major restriction fragment length polymorphism (RFLP) profiles were obtained by in silico and polyacrylamide gel electrophoresis analyses of the PCR products digested with HindIII and EcoRI. The causal agent of sugarcane red stripe was also successfully isolated from a diseased plant and its pathogenicity confirmed by inoculation of healthy sugarcane plantlets and reproduction of disease symptoms. The data showed that Aaa is currently widespread in China, suggesting that control methods should be implemented to limit the impact of red stripe on sugarcane production.     

Fusarium species associated with stalk rot and head blight of grain sorghum in Queensland and New South Wales,Australia

Historical records report Fusarium moniliforme sensu lato as the pathogen responsible for Fusarium diseases of sorghum; however, recent phylogenetic analysis has separated this complex into more than 25 species. During this study, surveys were undertaken in three major sorghum‐producing regions in eastern Australia to assess the diversity and frequency of Fusarium species associated with stalk rot‐ and head blight‐infected plants. A total of 523 isolates were collected from northern New South Wales, southern Queensland and central Queensland. Nine Fusarium species were isolated from diseased plants. Pathogenicity tests confirmed F. andiyazi and F. thapsinum were the dominant stalk rot pathogens, whilst F. thapsinum and species within the F. incarnatum–F. equiseti species complex were most frequently associated with head blight.     

Colletotrichum species causing grape ripe rot disease in Vitis labrusca and V. vinifera varieties in the highlands of southern Brazil

Ripe rot is one of the most important bunch diseases of grapes in the highlands of southern Brazil and a matter of concern for winegrowers. Sixty-one strains isolated from berries of Vitis labrusca and V. vinifera varieties with symptoms were classified by phylogenetic analysis of the sequences of β-tubulin, glyceraldehyde-3-phosphate dehydrogenase, D1/D2 domain of 28S rDNA, and rDNA internal transcribed spacer. They were also characterized by morphology, and their pathogenicity was evaluated. The combined molecular data allowed identification of six Colletotrichum species: C. fructicola, C. kahawae and C. viniferum (gloeosporioides complex), C. limitticola, C. nymphaeae (acutatum complex), and C. karstii (boninense complex). This is the first report of C. karstii and C. limitticola associated with the ripe rot of grapes. Morphological characteristics varied within and among species, confirming their separation at the complex level. Pathogenicity tests on V. vinifera berries showed that the most prevalent species, C. viniferum (37.8%) and C. fructicola (36.1%), were more virulent than the less prevalent species C. limitticola and C. karstii. Our findings indicate that there is a high diversity of Colletotrichum species associated with ripe rot disease of grapes in Brazil. There were no clear differences in the distribution of Colletotrichum species between V. labrusca and V. vinifera varieties. The determination of fungal species responsible for grape ripe rot in Brazilian vineyards may contribute to further epidemiological studies and the development of more efficient prophylactic methods for ripe rot management.     

Lack of host specificity of Colletotrichum spp. isolates associated with anthracnose symptoms on mango in Brazil

The aim of the present study was to analyse the genetic and pathogenic variability of Colletotrichum spp. isolates from various organs and cultivars of mango with anthracnose symptoms, collected from different municipalities of São Paulo State, Brazil. Colletotrichum gloeosporioides isolates from symptomless citrus leaves and C. acutatum isolates from citrus flowers with post‐bloom fruit drop symptoms were included as controls. Sequencing of the ITS region allowed the identification of 183 C. gloeosporioides isolates from mango; only one isolate was identified as C. acutatum. amova analysis of ITS sequences showed larger genetic variability among isolates from the same municipality than among those from different populations. fAFLP markers indicated high levels of genetic variability among the C. gloeosporioides isolates from mango and no correlation between genetic variability and isolate source. Only one C. gloeosporioides mango isolate had the same genotype as the C. gloeosporioides isolates from citrus leaves, as determined by ITS sequencing and fAFLP analysis. Pathogenicity tests revealed that C. gloeosporioides and C. acutatum isolates from either mango or citrus can cause anthracnose symptoms on leaves of mango cvs Palmer and Tommy Atkins and blossom blight symptoms in citrus flowers. These outcomes indicate a lack of host specificity of the Colletotrichum species and suggest the possibility of host migration.     

A novel lineage in the Fusarium incarnatum-equiseti species complex is one of the causal agents of fusarium rot on melon fruits in Northeast Brazil

Fusarium rot on melon fruits is a postharvest disease and its importance in Brazil is increasing since its first report in 1999. Initially this disease was attributed to the fungus Fusarium semitectum based on pathogen morphology. However, there is controversy regarding the aetiology of this disease because, in the current species concept based on phylogenetic analysis, F. semitectum is regarded as a synonym to different Fusarium species. With the objective of elucidating the disease aetiology in Northeast Brazil, Fusarium isolates were obtained from melon fruits showing rot symptoms in the main producing areas of the country. From the phylogenetic analyses of TEF1 and RPB2 gene sequences, the isolates were identified as belonging to two phylogenetic species of the Fusarium incarnatum-equiseti species complex (FIESC), one in the Incarnatum clade (Fusarium sulawense) and the other in the Equiseti clade, which corresponds to a new lineage. The newly identified lineage is close to Fusarium lacertarum. Isolates from the two species showed morphological characteristics typical of the Incarnatum and Equiseti clades, agreeing with the molecular identification, and were pathogenic when inoculated on melon fruits. This is the first report of F. sulawense on melon fruits. The data generated in this study are potentially useful for a better management of the fusarium rot on melon.     

Distinguishing characteristics and vegetative compatibility of Colletotrichum kahawe in comparison with other related species from coffee

On the basis of pathogenicity tests on green berries or hypocotyls of coffee and by morphological and biochemical characteristics in culture, 31 isolates of Colletotrichum were classified into C. kahawe (24 isolates), C. gloeosporioides (six isolates) or C. acutatum (one isolate). Within these groups of isolates, vegetative compatibility groups (VCGs) were determined by complementation tests with mutants in the nitrate assimilation pathway. There were distinct incompatibility barriers between the three species. Among the C. gloeosporioides group, the three isolates tested were self-compatible but incompatible with each other. Within C. kahawe , 18 isolates were self-compatible and only one main VCG was detected. However, partial compatibility in C. kahawe was also indicated by variation in the intensity of heterokaryon formation between different pairs of isolates and between different types of mutant. The existence of only one VCG in C. kahawe is consistent with the low level of variation found in previous work on DNA polymorphism.     

Elimination of Sugarcane yellow leaf virus from infected sugarcane plants by meristem tip culture visualized by tissue blot immunoassay

Sugarcane yellow leaf virus (SCYLV), a member of the Luteoviridae , is implicated in the sugarcane disease known as yellow leaf syndrome (YLS), which is characterized by yellowing of the leaf midrib followed by leaf necrosis and possible growth suppression. YLS is distributed worldwide and susceptible cultivars are commonly infected with SCYLV. However, not all cultivars infected with SCYLV show symptoms of YLS and some cultivars that show symptoms do so sporadically. Since it is difficult to obtain virus-free plants of susceptible cultivars, it has not been possible to study the factors involved in SCYLV infection nor the effects of infection on plant growth and yield. A tissue blot immunoassay was used to visualize in vivo presence of the virus so that virus-infected and virus-free plants could be distinguished. Meristem tip cultures were used to produce virus-free plantings of six SCYLV-susceptible sugarcane cultivars. Nearly all of the regenerated sugarcane lines remained virus-free over a period of up to 4 years, whether grown in isolated fields or in the glasshouse. Experimental re-infection of the virus-free plants by viruliferous aphids demonstrated that meristem tip culture did not affect susceptibility of sugarcane to SCYLV. Improved diagnosis and production of virus-free plants of SCYLV-susceptible cultivars will facilitate research to quantify the effect of the virus on yield and to analyse the processes involved in disease development.     

New Leaf Spot Disease of Cymbidium Species Caused by Fusarium subglutinans and Fusarium proliferatum

Fusarium species were consistently isolated from yellow, swollen spots with reddishbrown centers and small black spots on leaves of Cymbidium plants in the greenhouse. Fusarium subglutinans caused the yellow spots and Fusarium proliferatum caused either the yellow or the black spots. We propose the name “yellow spot” for the new disease. To denote differences in their pathogenicity to orchid plants, we designate the population causing yellow spot as race Y and that causing black spot as race B of F. proliferatum. Received 29 October 1999/ Accepted in revised form 10 March 2000     

Morphology,phylogeny and pathogenicity of Fusarium species from Sansevieria trifasciata in Malaysia

Leaf blight is a common disease affecting Sansevieria trifasciata in many countries, including Malaysia. In the present study, Fusarium isolates were consistently recovered from the diseased leaves collected from various locations throughout the country. Based on morphology and multigene phylogenetic analysis using mitochondrial small subunit (mtSSU), intergenic spacer region (IGS) and translation elongation factor 1-α (TEF1-α) gene sequences, seven Fusarium species were identified, with F. oxysporum being the most prevalent (67.6%) among 34 isolates. Pathogenicity tests resulted in the discovery of pathogenic isolates that belonged to F. oxysporum, F. proliferatum, and F. pseudocircinatum, whereas all isolates of F. brachygibbosum, F. concentricum, F. mangiferae, and F. solani were nonpathogenic. The results suggest that several Fusarium species are accountable for causing disease on S. trifasciata in Malaysia.