Survey of nematodes associated with sugarcane in the state of Paraná, Brazil

Sugarcane-associated nematodes (Saccharum spp.) can reduce productivity up to 50%. Through the survey, it was possible to identify the main nematodes that occur in a certain region as a tool for designing the best management and control strategies. The aim of this study was to characterize the population of nematodes associated with sugarcane in the North Central, North Pioneiro and Northwest mesoregion of the state of Paraná, Brazil, quantify the nematode genera associated with the crop and identify the species of Pratylenchus and Meloidogyne. A total amount of 89 soil and root composite samples were collected in nine municipalities. Nematodes were extracted and counted in a Peters counting chamber under an optical light microscope. Morphological description followed identification keys. Pratylenchus spp. were identified by morphological characteristics; Meloidogyne spp. were identified by morphological characteristics and isoenzyme electrophoresis. Twelve genera of nematodes associated with sugarcane were identified: Pratylenchus, Meloidogyne, Helicotylenchus, Xiphinema, Mesocriconema, Trichodorus, Aphelenchus, Hoplolaimus, Tylenchus, Tylenchorhynchus, Ditylenchus, and Paratrichodorus. The genera Pratylenchus and Meloidogyne were found with the highest frequencies in the roots. Among the species of Pratylenchus, P. zeae and P. brachyurus were found, with P. zeae being the most frequent. Among the Meloidogyne species, only M. javanica was found. These results are essential to aid decision making in the management of phytonematodes, mainly in the development of new control strategies and in directing genetic breeding programs for development of sugarcane cultivars for the Paraná state.

A molecular approach reveals Tranzschelia discolor as the causal agent of rust on plum and peach in Brazil.

Plum and peach are important crops in the southernmost regions of Brazil and in the majority, fresh fruit producers are small producers, which guarantee their family income. Tranzschelia discolor and T. pruni-spinosae are the etiological agents of rust on Prunus domestica (plum) and P. persica (peach) in Brazil (Mendes and Urben, 2020). The molecular characterization of Tranzschelia specimens revealed different clades that are not attributed to known species, showing the need for taxonomic evaluation of Tranzschelia species in the tropics (Scholler et al. 2014; 2019). As Tranzschelia species reported in Brazil were identified only by morphological characteristics, this study aimed to carry out a survey to verify the etiology of rust on plum and peach based on molecular data. In 2018, rust symptoms in peach and plum trees were observed with maximum severity of 30% and 35%, respectively, in three Brazilian states. Symptoms of plum and peach rust are yellowish-green spots visible on the adaxial side of the leaves and uredia/uredinial sori releasing the brown urediniospores on the abaxial side (Supplementary figure 1). Symptomatic leaves of plum and peach were collected at Curitiba in the states of Paraná (lat. 25°25'47" S and long. 49°16'19" W, altitude of 935 meters) in a research station, Videira in Santa Catarina (lat. 27°00'30" S and long. 51°09'06" W, altitude of 750 meters) in a research station and Paranapanema in São Paulo (lat. 23º23'19" S and long. 48º43'22" W, altitude of 610 meters) in a farmer field, and deposited in the herbarium of the Municipal Botanical Museum of Curitiba (MBM 429790 to 429795). Urediniospores collected on plum and peach leaves were all echinulate, obovoid, orange-brown, and measured 18.0 - 33.5 µm × 10.5 - 20.5 µm (n=150) and 22.5 - 40.0 µm × 11.5 - 20.5 µm (n=150), respectively. The genomic DNA of the urediniospores was extracted for amplification and sequencing of the internal transcribed spacer region (ITS) using primers ITS5-u and ITS4-u (Pfunder et al. 2001). The sequences were deposited (Accession Nos. MT786213 to MT786218) and compared to sequences in the GenBank repository using the BLASTn algorithm. The sequences of ITS showed a high percentage of identity (>99%) with sequences from T. discolor (Accession Nos. AB097449, EU014071, KU712078, KY764179, MH599069, MN545867, DQ995341, DQ354542, and KX985768). Additionally, our isolates clustered with others T. discolor in a Bayesian phylogenetic tree based on ITS sequences (study S26663 deposited in TreeBASE) (Supplementary figure 2). A pathogenicity test was carried out on plants by inoculation of a 1.5 × 105 urediniospores mL-1 suspension on the abaxial side of the leaves. Leaves sprayed with sterile water were used as controls. The plants were incubated in a growth chamber (GC) in the dark for 48 h at 23 °C and maintained with 100% RH to establish infections. The inoculated plants were afterwards kept in the GC at a photoperiod of 12 h under same conditions until 14 days when the symptoms and pathogen structures were observed to all six isolates. Control leaves remained symptomless. Tranzschelia discolor infect plants in the genus Prunus, including almond, apricot, nectarine, cherry, peach, and plum (Farr and Rossman 2021). As T. pruni-spinosae was not found, T. discolor is probably the prevalent species in the main regions of Brazil. This information reveals T. discolor as the causal agent of plum and peach rust in Brazil and helps to understand the distribution of this disease in tropics or worldwide.

Occurrence of Plasmopara destructor causing Downy Mildew on Impatiens walleriana in Brazil.

Impatiens walleriana (Balsaminaceae), popularly known as Impatiens, is an African succulent and a popular ornamental plant worldwide (GBIF, 2019). In Brazil it is broadly grown indoors and outdoors, including in public parks of Curitiba, State of Paraná (Viezzer et al. 2018). In September 2018, I. walleriana plants showing typical downy mildew symptoms were observed in wastelands and gardens in Curitiba. The symptoms included adaxial chlorotic leaf spots with abundant white sporulation on abaxial side (Supplementary figure 1). The disease led to severe defoliation of the plants and the incidence of the plant disease varied from 20 to 80% of plants in an area ranging from 400 to 40,000 m2. A representative sample was deposited in herbarium of the Museu Botânico Municipal de Curitiba (MBM 331601). The following morphology was observed: Sporangiophores (n = 30), hyaline, thin walled, emerging through stomata, 407.3 to 551.1 µm long, slightly swollen base, first branch at 165.8 to 324.7 µm from base, end branches 5.1 to 13.1 µm long, sporangia (n = 50) hyaline, thin-walled subglobose to ovoid, from 12.8 to 21.9 µm x 12.5 to 17.9 µm, slightly papillate. Due to morphological and genetic variations within the species Plasmopara obducens, Görg et al. (2017) proposed the new species P. velutina and P. destructor. The morphology of the Curitiba specimen was equivalent to that described for P. destructor (Görg et al. 2017). DNA was extracted from LEMIDPRTf-19-02 isolate and the ITS1 and cox2 regions were PCR amplified as described in Görg et al. (2017). The resulting sequences were deposited in GenBank (ITS1, MT680628; cox2, MT952335). A BLASTn analysis of the sequences revealed 100% homology with ITS (MF372742) and cox2 (MF372728) sequences of type strain of P. destructor (GLM-F107554). A Bayesian phylogenetic analysis was performed to compare the sequences from this study with reference sequences for P. obducens, P. destructor and P. velutina (Görg et al. 2017; Salgado-Salazar et al. 2018). The oomycete from Curitiba grouped in a reliable clade with P. destructor (Supplementary figure 2). Pathogenicity was carried out by ex vivo and in vivo tests. For ex vivo, stems with approximately four healthy leaves of I. walleriana (n = 10) were embedded in aluminum grid inside of gerbox with the stem bases immersed in distilled water. The inoculation of five stems was carried out by spraying a suspension with 6 x 104 sporangia mL-1 on the abaxial side of the leaves. Five stems with leaves inoculated with sterile water were used as controls. They were incubated in a growth chamber in the dark for 48 h at 20 °C and another 12 days in a 12 h light photoperiod. The confirmation of pathogenicity in plants (in vivo) was obtained with the inoculation of I. walleriana seedlings (one-month old) grown in 2 dm3 aluminum pots. The inoculation methodology and number of plants were the same as the stems test. After the inoculation, plants were incubated in a growth chamber for 48 h in the dark at 20 °C with 100% RH with nebulization, and another 10 days at a photoperiod of 12 hours of light. For both tests, abundant sporulation was observedwith morphology equivalent to Plasmopara destructor described by Görg et al. (2017). No disease developed on control plants. To our knowledge, this is the first report of P. destructor on I. walleriana in Brazil (Farr and Rossman 2019, Silva et al. 2019) representing a potential loss to flower production and a reduction in flowering period in public gardens and parks.

Magnetic Field Measurements during Magnetic Pulse Welding Using CMR-B-Scalar Sensors.

The possibility of applying CMR-B-scalar sensors made from thin manganite films exhibiting the colossal magnetoresistance effect as a fast-nondestructive method for the evaluation of the quality of the magnetic pulse welding (MPW) process is investigated in this paper. This method based on magnetic field magnitude measurements in the vicinity of the tools and joining parts was tested during the electromagnetic compression and MPW of an aluminum flyer tube with a steel parent. The testing setup used for the investigation allowed the simultaneous measurement of the flyer displacement, its velocity, and the magnitude of the magnetic field close to the flyer. The experimental results and simulations showed that, during the welding of the aluminum tube with the steel parent, the maximum magnetic field in the gap between the field shaper and the flyer is achieved much earlier than the maximum of the current pulse of the coil and that the first half-wave pulse of the magnetic field has two peaks. It was also found that the time instant of the minimum between these peaks depends on the charging energy of the capacitors and is associated with the collision of the flyer with the parent. Together with the first peak maximum and its time-position, this characteristic could be an indication of the welding quality. These results were confirmed by simultaneous measurements of the flyer displacement and velocity, as well as a numerical simulation of the magnetic field dynamics. The relationship between the peculiarities of the magnetic field pulse and the quality of the welding process is discussed. It was demonstrated that the proposed method of magnetic field measurement during magnetic pulse welding in combination with subsequent peel testing could be used as a nondestructive method for the monitoring of the quality of the welding process.

Novel Approach and Interpretation for the Determination of Electromagnetic Forming Limits.

A new method to determine electromagnetic forming limits curves (EM-FLCs) for sheet metals is proposed. The different strain paths (between uniaxial and biaxial tension) are achieved by specific tool coil and specimen designs. It is ensured that the apex of the specimen deforms on a constant strain path, and excess bending at the apex is avoided. This is done so that the determined EM-FLCs are comparable to their quasi-static counterparts. The method determines the EM-FLCs for the aluminum alloys AA-1050a-H24 and EN AW-5083-H111 and the magnesium alloy Mg AZ31-O. Overall, it is observed that the necking limits in electromagnetic forming (EMF) are higher compared to quasi-static forming. The fracture surfaces of electromagnetically deformed specimens are examined to reveal the existence of out-of-plane shear stresses. A numerical analysis corroborates this observation and their variation with strain rate. The presence of such stresses is proposed as a possible reason for the increased necking limits in EMF. As reasons for higher forming limits, previous research has identified inertial stabilization, strain rate hardening, die impact, and change in deformation mechanism. The current study reaffirms the positive effect of inertial stabilization and makes key observations in the increase of twinning in EMF of Mg AZ31-O.