scholarly journals First Report of Botryosphaeria dothidea Causing Stem Canker on Soybean in China

Plant Disease ◽  
2020 ◽  
Author(s):  
Tangmin Chen ◽  
Xinchi Shi ◽  
Suyan Wang ◽  
Pedro Laborda
Keyword(s):  
Phylogenetic Tree ◽  
Elongation Factor ◽  
Stem Canker ◽  
Phytophthora Sojae ◽  
Likelihood Method ◽  
First Report ◽  
Molecular Phylogenetic Tree ◽  
Molecular Phylogenetic ◽  
Symptomatic Tissue ◽  
Soybean Plants

In September 2020, widespread stem canker on soybean (Glycine max) was detected in southeastern Jiangsu (Nantong municipality; 120.76° E, 32.23° N) in China. Mature plants, 14 weeks of cultivation, exhibited brown necrotic lesions and dried-up stem. The symptoms were observed in eleven soybean fields, 1.6 ha in total, and approximately 80% of the plants were symptomatic. The symptoms were consistent with those previously reported for stem canker on soybean caused by Diaporthe aspalathi, D. caulivora and D. sojae (Ghimire et al. 2019; Mena et al. 2020). Small pieces, approximately 0.4 cm2 in size, of symptomatic tissue were surface sterilized in 1.5% NaOCl for 1 min, and washed twice with sterile ddH2O. The pathogen was isolated and cultured on potato dextrose agar (PDA), containing chloramphenicol (50 µg/mL), under darkness at 28 ºC for 7 days. Amplification of internal transcribed spacer (ITS), elongation factor 1-α (EF1-α) and β-tubulin (TUB2) genes was performed using ITS1/ITS4, EF1-728F/EF1-986R and Bt2a/Bt2b primers, respectively (Jia et al. 2019). Sequences were submitted to GenBank under accession numbers MW130133 (ITS), MW147481 (EF1-α) and MW147482 (TUB2). Blast search revealed that the amplified sequences had 99.65% (ITS; B. dothidea JZB310202, MN945381), 100% (EF1-α; B. dothidea ZB-77, MH726166) and 99.75% (TUB2; B. dothidea ZB-1, MN642587) matches to multiple B. dothidea strains, whereas all reported Diaporthe strains showed no nucleotide identity to the amplified sequences. Molecular phylogenetic tree was constructed using MEGA7 to confirm the identity of the pathogen. ITS, EF1-α and TUB2 sequences were blasted separately in Muscle (https://www.ebi.ac.uk/Tools/msa/muscle/) and then combined together to make the phylogenetic tree. The evolutionary history was inferred by using the Maximum Likelihood method based on the Tamura 3-parameter model, and the tree with the highest likelihood (-4291.3981) is shown in Figure 1. Diaporthe strains found causing stem canker on soybean, some Phytophthora sojae strains (which also cause dried-up stem on soybean) (Yang et al. 2019), and B. dothidea strains found in China in other hosts were included in the phylogenetic tree. To confirm pathogenicity, a sterilized spatula was used to make wounds (3 mm diameter, 1 mm depth) on the stem of 2-week old soybean plants. Mycelial plugs from a 7 day-old culture on PDA were placed on the wounds and covered with Parafilm. Sterilized PDA plugs were used as control. Inoculated plants were maintained in a growth chamber at 28 °C and 60% relative humidity. Typical stem canker symptoms were observed 5 days after inoculation (Figure 2). Microscope observations showed the presence of septate mycelium, fusiform conidia and round conidiomata, and agreed with those previously reported for the morphology of B. dothidea strains (Phillips et al. 2013). During recent months, B. dothidea was reported to cause stem canker and leave wilt on a number of plant species in China (Huang et al. 2020; Ju et al. 2020; Wang et al. 2020a, 2020b, 2020c), confirming the expansion and host promiscuity of this pathogen. Stem canker on soybean has been thoroughly associated to Diaporthe strains; however, this is the first report of B. dothidea causing this disease. We believe that our results will help to better understand the pathogens affecting soybean production in China.

scholarly journals First Report of Colletotrichum brevisporum Causing Soybean Anthracnose in China

Plant Disease ◽  
2020 ◽  
Author(s):  
Xinchi Shi ◽  
Suyan Wang ◽  
Xuchu Duan ◽  
Xing Gao ◽  
Xinyu Zhu ◽  
...  
Keyword(s):  
Phylogenetic Tree ◽  
Likelihood Method ◽  
Soybean Field ◽  
Disease Symptoms ◽  
Molecular Phylogenetic Tree ◽  
Molecular Phylogenetic ◽  
Symptomatic Tissue ◽  
Soybean Plants ◽  
Soybean Anthracnose ◽  
First Time

In March 2020, widespread anthracnose was observed on soybean (Glycine max) in southeastern Jiangsu (Nantong municipality; 120.53° E, 31.58° N) in China. Plants exhibited irregular brown necrotic lesions in stem and leaves, and pronounced wilting. The symptoms were detected in one soybean field, 0.42 ha, surrounded by healthy wheat fields. Approximately 65% of the soybean plants showed the disease symptoms, and crop yield was reduced by 28-35% with respect the yield achieved in previous years, when no symptoms were observed. The symptoms were consistent with those previously reported for anthracnose on soybean caused by Colletotrichum chlorophyti, C. cliviae and C. gloeosporioides (Barbieri et al. 2017; Mahmodi et al. 2013; Yang et al. 2012). Diseased, 3-week old plants were collected. Small pieces, approximately 1 cm2 in size, of symptomatic tissue were surface sterilized in 1.5% NaOCl for 1 min, and washed twice with sterile ddH2O. The pathogen was isolated and cultured on potato dextrose agar (Song et al. 2020), containing chloramphenicol (50 µg/mL), under darkness at 28 °C for 3 days. Sequence of internal transcribed spacer (ITS), actin (ACT), β-tubulin (TUB2) and glyceraldehyde 3-phosphate dehydrogenase (GAP/span>DH) genes was performed as reported by Yang et al. (2015). Sequences were submitted to GenBank under accession numbers MT361074 (ITS) and MT415548-MT415550 (ACT, TUB2 and GAPDH). Blast search revealed that the amplified sequences had 100% (ITS; C. brevisporum TCHD, MH883805), 97.66% (ACT; C. brevisporum S38, KY986905), 99.06% (TUB2; C. brevisporum PF-2, KY705061) and 100% (GAPDH; C. brevisporum LJTJ27, KP823797) matches to multiple C. brevisporum strains, whereas all reported C. chlorophyti, C. cliviae and C. gloeosporioides strains showed no similarity to at least 2 of the studied genes. Molecular phylogenetic tree constructed using MEGA7 confirmed the identity of the pathogen. ACT and ITS sequences were blasted separately in Muscle (https://www.ebi.ac.uk/Tools/msa/muscle/) and then combined together to make the phylogenetic tree. The evolutionary history was inferred by using the Maximum Likelihood method based on the Tamura 3-parameter model, and the tree with the highest log likelihood (-1749.2186) is shown in Figure 1. The Colletotrichum strains previously found causing anthracnoseon soybean, and other relevant strains used in taxonomic analyses were included in the phylogenetic tree. Microscope observations showed the presence of 15-µm-long cylindrical conidia and septate mycelium, and agree with those reported for the morphology of C. brevisporum by Damm et al. (2019). To confirm pathogenicity, the mycelia from a 2 day-old culture on PDA was collected and suspended in sterile ddH2O (≈ 106 cells/mL) to prepare the inoculum. The pathogen was sprayed-inoculated on stem and leaves of healthy soybean plants. In control plants, sterile ddH2O was used. Inoculated plants were maintained in growth chamber at 28 °C and 50% relative humidity. Typical anthracnose symptoms were obsered 20 days after inoculation (Figure 2). C. brevisporum was reported to produce anthracnose on pumpkin, papaya, mulberry, coffee, passion fruit and pepper in China (Liu et al. 2017; Liu et al. 2019; Xue et al. 2019). Here, we report for the first time C. brevisporum causing anthracnose on soybean, an economically-relevant crop in China.

scholarly journals First report of Diaporthe ueckerae causing stem canker on soybean (Glycine max L.) in Colombia

Plant Disease ◽  
2021 ◽  
Author(s):  
Nathali López-Cardona ◽  
YUDY ALEJANDRA GUEVARA ◽  
Lederson Gañán-Betancur ◽  
Carol Viviana Amaya Gomez
Keyword(s):  
Management Strategies ◽  
Elongation Factor ◽  
Morphological Characteristics ◽  
Aerial Mycelium ◽  
Stem Canker ◽  
Growth Stages ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Glycine Max L ◽  
Soybean Plants

In October 2018, soybean plants displaying elongated black to reddish-brown lesions on stems were observed in a field planted to the cv. BRS Serena in the locality of Puerto López (Meta, Colombia), with 20% incidence of diseased plants. Symptomatic stems were collected from five plants, and small pieces (∼5 mm2) were surface sterilized, plated on potato dextrose agar (PDA) and incubated for 2 weeks at 25°C in darkness. Three fungal isolates with similar morphology were obtained, i.e., by subculturing single hyphal tips, and their colonies on PDA were grayish-white, fluffy, with aerial mycelium, dark colored substrate mycelium, and produced circular black stroma. Pycnidia were globose, black, occurred as clusters, embedded in tissue, erumpent at maturity, with an elongated neck, and often had yellowish conidial cirrus extruding from the ostiole. Alpha conidia were observed for all isolates after 30 days growth on sterile soybean stem pieces (5 cm) on water agar, under 25ºC and 12 h light/12h darkness photoperiod. Alpha conidia (n = 50) measured 6.0 – 7.0 µm (6.4 ± 0.4 µm) × 2.0 – 3.0 µm (2.5± 0.4 µm), were aseptate, hyaline, smooth, ellipsoidal, often biguttulate, with subtruncate base. Beta conidia were not observed. Observed morphological characteristics of these isolates were similar to those reported in Diaporthe spp. by Udayanga et al. (2015). DNA from each fungal isolate was used to sequence the internal transcribed spacer region (ITS), and the translation elongation factor 1-α (TEF1) gene, using the primer pairs ITS5/ITS4 (White et al. 1990) and EF1-728F/EF1- 986R (Carbone & Kohn, 1999), respectively. Results from an NCBI-BLASTn, revealed that the ITS sequences of the three isolates (GenBank accessions MW566593 to MW566595) had 98% (581/584 bp) identity with D. miriciae strain BRIP 54736j (NR_147535.1), whereas the TEF1 sequences (GenBank accessions MW597410 to MW597412) had 97 to 100% (330-339/339 bp) identity with D. ueckerae strain FAU656 (KJ590747). The species Diaporthe miriciae R.G. Shivas, S.M. Thomps. & Y.P. Tan, and Diaporthe ueckerae Udayanga & Castl. are synonymous, with the latter taking the nomenclature priority (Gao et al. 2016). According to a multilocus phylogenetic analysis, by maximum likelihood, the three isolates clustered together in a clade with reference type strains of D. ueckerae (Udayanga et al. 2015). Soybean plants cv. BRS Serena (growth stages V3 to V4) were used to verify the pathogenicity of each isolate using a toothpick inoculation method (Mena et al. 2020). A single toothpick colonized by D. ueckerae was inserted directly into the stem of each plant (10 plants per isolate) approximately 1 cm below the first trifoliate node. Noncolonized sterile toothpicks, inserted in 10 soybean plants served as the non-inoculated control. Plants were arbitrarily distributed inside a glasshouse, and incubated at high relative humidity (>90% HR). After 15 days, inoculated plants showed elongated reddish-brown necrosis at the inoculated sites, that were similar to symptoms observed in the field. Non-inoculated control plants were asymptomatic. Fungal cultures recovered from symptomatic stems were morphologically identical to the original isolates. This is the first report of soybean stem canker caused by D. ueckerae in Colombia. Due to the economic importance of this disease elsewhere (Backman et al. 1985; Mena et al. 2020), further research on disease management strategies to mitigate potential crop losses is warranted.

Molecular data allow to elucidate the taxonomic placement of some Umbelliferae from Middle Asia and Afghanistan (Pinacantha, Ladyginia, Peucedanum mogoltavicum)

Phytotaxa ◽  
2018 ◽  
Vol 350 (1) ◽  
pp. 42 ◽  
Author(s):  
GALINA V. DEGTJAREVA ◽  
MICHAEL G. PIMENOV ◽  
TAHIR H. SAMIGULLIN
Keyword(s):  
Phylogenetic Tree ◽  
Dna Sequence ◽  
Sequence Data ◽  
Molecular Data ◽  
Systematic Position ◽  
New Combination ◽  
Dna Sequence Data ◽  
Middle Asia ◽  
Molecular Phylogenetic Tree ◽  
Molecular Phylogenetic

The systematic position of three Apiaceae-Apioideae taxa, Pinacantha porandica, Ladyginia bucharica and Peucedanum mogoltavicum, from Middle Asia and Afghanistan, is clarified based on nrITS DNA sequence data. In the molecular phylogenetic tree, the monotypic Pinacantha is placed in unresolved position within the Ferulinae. Although there is no morphological information on essential characters, we propose a new position of Pinacantha porandica within the genus Ferula. As a result a new combination Ferula porandica is proposed, with a new section Pinacantha to accommodate it. The attribution of Peucedanum mogoltavicum to Ferula has been confirmed, its correct name being Ferula lithophila. The genus Ladyginia should not be included in Ferula, its closest relatives being Mozaffariania and Glaucosciadium from the Glaucosciadium Clade.

scholarly journals DNA Barcoding and Molecular Phylogeny of Drosophila lini and Its Sibling Species

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Yi-Feng Li ◽  
Shuo-Yang Wen ◽  
Kuniko Kawai ◽  
Jian-Jun Gao ◽  
Yao-Guang Hu ◽  
...  
Keyword(s):  
Molecular Phylogeny ◽  
Phylogenetic Tree ◽  
Dna Barcoding ◽  
Sibling Species ◽  
Phylogenetic Trees ◽  
The Other ◽  
Species Delineation ◽  
Molecular Phylogenetic Tree ◽  
Limited Applicability ◽  
Molecular Phylogenetic

Drosophila lini and its two sibling species, D. ohnishii and D. ogumai, are hardly distinguishable from one another in morphology. These species are more or less reproductively isolated. The mitochondrial ND2 and COI-COII and the nuclear ITS1-ITS2 regions were sequenced to seek for the possibility of DNA barcoding and to reconstruct the phylogeny of them. The character-based approach for DNA barcoding detected some diagnostic nucleotides only for monophyletic D. ogumai, but no informative sites for the other two very closely species, D. lini and D. ohnishii, of which strains intermingled in the molecular phylogenetic trees. Thus, this study provides another case of limited applicability of DNA barcoding in species delineation, as in other cases of related Drosophila species. The molecular phylogenetic tree inferred from the concatenated sequences strongly supported the monophyly of the cluster of the three species, that is, the lini clade. We propose some hypotheses of evolutionary events in this clade.

scholarly journals First Report of Root Rot of Soybeans Caused by Corynespora cassiicola in Wisconsin

Plant Disease ◽  
1999 ◽  
Vol 83 (7) ◽  
pp. 696-696 ◽  
Author(s):  
S. J. Raffel ◽  
E. R. Kazmar ◽  
R. Winberg ◽  
E. S. Oplinger ◽  
J. Handelsman ◽  
...  
Keyword(s):  
Root Rot ◽  
Growth Stage ◽  
Lateral Roots ◽  
Seedling Emergence ◽  
Fluorescent Light ◽  
Corynespora Cassiicola ◽  
First Report ◽  
Seedling Roots ◽  
Symptomatic Tissue ◽  
Soybean Plants

Corynespora cassiicola (Berk. & M. A. Curtis) C. T. Wei was isolated from diseased soybean plants (Glycine max) collected in two fields near Racine and Arlington, WI. Plants sampled at seedling emergence (VC), late vegetative (V5), and mid-reproductive (R5) stages exhibited reddish to dark brown longitudinal lesions on the exterior of the tap root extending vertically on the hypocotyl to the soil line, and extensive necrosis of lateral roots. Sample size at each growth stage was 144 plants per site. Roots were surface sterilized in 0.5% sodium hypochlorite for 2 min and sections of symptomatic tissue placed on water agar (12 g/liter) containing 100 μg of streptomycin per ml. Sporulation occurred on lesions and on mycelium that had grown out from the plant tissue onto the water agar following a 2-week incubation at 24°C under fluorescent light (280 μmol s-1 m-2). Incidence of isolation of C. cassiicola at both sites was 40% of plants sampled at growth stage VC, 67% at V5, and 78% at R5. Conidia characteristic of C. cassiicola were particularly abundant on the surface of necrotic lateral root tissue. Elongated conidia produced on water agar were 151 ± 5 μm × 15 ± 0.5 μm with an average of 13 ± 0.4 cells separated by hyaline pseudosepta (1). To confirm pathogenicity, a 1-cm lateral slice into each of four 5-day-old soybean seedling roots was made and a plug of agar taken from the margin of a colony of C. cassiicola grown on potato dextrose agar was placed in each wound and incubated for 14 days at 24°C in a growth chamber. Symptoms similar to those of diseased field plants were observed and C. cassiicola was reisolated from all plants inoculated with C. cassiicola; all controls treated with agar alone had no symptoms and C. cassiicola was recovered from none of the noninoculated controls. This is the first report of root rot caused by C. cassiicola on soybean in Wisconsin. Reference: (1) W. L. Seaman and R. A. Shoemaker. Can. J. Bot. 43:1461, 1965.

scholarly journals First report of Nigrospora sphaerica causing fruit dried-shrink disease in Akebia trifoliata from China

Plant Disease ◽  
2021 ◽  
Author(s):  
Xiujing Hong ◽  
Shijia Chen ◽  
linchao Wang ◽  
Bo Liu ◽  
Yuruo Yang ◽  
...  
Keyword(s):  
Elongation Factor ◽  
Morphological Characteristics ◽  
Its Region ◽  
Average Diameter ◽  
Ethanol Solution ◽  
Likelihood Method ◽  
Conidial Suspension ◽  
Multiple Sequence ◽  
First Report ◽  
Pure Cultures

Akebia trifoliata, a recently domesticated horticultural crop, produces delicious fruits containing multiple nutritional metabolites and has been widely used as medicinal herb in China. In June 2020, symptoms of dried-shrink disease were first observed on fruits of A. trifoliata grown in Zhangjiajie, China (110.2°E, 29.4°N) with an incidence about 10%. The infected fruits were shrunken, colored in dark brown, and withered to death (Figure S1A, B). The symptomatic fruits tissues (6 × 6 mm) were excised from three individual plants, surface-disinfested in 1% NaOCl for 30s and 70% ethanol solution for 45s, washed, dried, and plated on potato dextrose agar (PDA) containing 50 mg/L streptomycin sulfate in the dark, and incubated at 25℃ for 3 days. Subsequently, hyphal tips were transferred to PDA to obtain pure cultures. After 7 days, five pure cultures were obtained, including two identical to previously reported Colletotrichum gloeosporioides causing leaf anthracnose in A. trifoliata (Pan et al. 2020) and three unknown isolates (ZJJ-C1-1, ZJJ-C1-2, and ZJJ-C1-3). The mycelia of ZJJ-C1-1, ZJJ-C1-2 and ZJJ-C1-3 were white, and formed colonies of approximate 70 mm (diameter) in size at 25℃ after 7 days on potato sucrose agar (PSA) plates (Figure S1C). After 25 days, conidia were formed, solitary, globose, black, shiny, smooth, and 16-21 μm in size (average diameter = 18.22 ± 1.00 μm, n = 20) (Figure S1D). These morphological characteristics were similar to those of N. sphaerica previously reported (Li et al. 2018). To identify species of ZJJ-C1-1, ZJJ-C1-2 and ZJJ-C1-3, the internal transcribed spacer (ITS) region, β-tubulin (TUB2), and the translation elongation factor 1-alpha (TEF1-α) were amplified using primer pairs including ITS1/ITS4 (Vilgalys and Hester 1990), Bt-2a/Bt-2b (Glass and Donaldson 1995), and EF1-728F/EF-2 (Zhou et al. 2015), respectively. Multiple sequence analyses showed no nucleotide difference was detected among genes tested except ITS that placed three isolates into two groups (Figure S2). BLAST analyses determined that ZJJ-C1-1, ZJJ-C1-2 and ZJJ-C1-3 had 99.73% to N. sphaerica strains LC2705 (KY019479), 100% to LC7294 (KY019397), and 99.79-100% to LC7294 (KX985932) or LC7294 (KX985932) based on sequences of TUB2 (MW252168, MW269660, MW269661), TEF-1α (MW252169, MW269662, MW269663), and ITS (MW250235, MW250236, MW192897), respectively. These indicated three isolates belong to the same species of N. sphaerica. Based on a combined dataset of ITS, TUB2 and TEF-1α sequences, a phylogenetic tree was constructed using Maximum likelihood method through IQ-TREE (Minh et al. 2020) and confirmed that three isolates were N. sphaerica (Figure S2). Further, pathogenicity tests were performed. Briefly, healthy unwounded fruits were surface-disinfected in 0.1% NaOCl for 30s, washed, dried and needling-wounded. Then, three fruits were inoculated with 10 μl of conidial suspension (1 × 106 conidia/ml) derived from three individual isolates, with another three fruits sprayed with 10 μl sterilized water as control. The treated fruits were incubated at 25℃ in 90% humidity. After 15 days, all the three fruits inoculated with conidia displayed typical dried-shrink symptoms as those observed in the farm field (Figure S1E). The decayed tissues with mycelium and spores could be observed on the skin or vertical split of the infected fruits after 15 days’ inoculation (Figure S1F-H). Comparably, in the three control fruits, there were no dried-shrink-related symptoms displayed. The experiment was repeated twice. The re-isolated pathogens were identical to N. sphaerica determined by sequencing the ITS, TUB2 and TEF-1α. Previous reports showed N. sphaerica could cause postharvest rot disease in kiwifruits (Li et al. 2018). To our knowledge, this is the first report of N. sphaerica causing fruits dried-shrink disease in A. trifoliata in China.

Diversification of Stem Morphological and Anatomical Characteristics in Eleocharis and Related Genera (Cyperaceae, Monocotyledoneae)

2021 ◽  
Vol 46 (4) ◽  
pp. 935-950
Author(s):  
Katsuhiro Yashiro ◽  
Yasuhiko Endo
Keyword(s):  
Phylogenetic Tree ◽  
Evolutionary Process ◽  
Fiber Bundles ◽  
Running Waters ◽  
Anatomical Characteristics ◽  
Molecular Phylogenetic Tree ◽  
Molecular Phylogenetic ◽  
Anatomical Characters

Abstract— The genus Eleocharis (Cyperaceae, monocotyledons) is characterized by bladeless leaves, which are leaves having only leaf-sheaths. To study the evolutionary process through which Eleocharis species lost their leaf blades, we analyzed the outer morphological and anatomical characters of the representative Eleocharis species, plus species from nine phylogenetically related genera. From the analysis, we recognized eight characters and we optimized their character states on a recent molecular phylogenetic tree. As a result, we recognized five characteristics shared by Eleocharis species as follows: (1) the most apical internode is more than seven times longer than the next apical internode; (2) bladeless leaves having only leaf sheaths; (3) transversely septate aerenchyma in culms; (4) densely and peripherally located, slender, and square timber-shaped fiber bundles in culms; and (5) palisade chlorenchyma in culms. In these characteristics, (2) and (4) are synapomorphies of Eleocharis. These two apomorphic characteristics seemed to be adaptations for inhabiting running waters.

scholarly journals 2M1700 Constructing a molecular phylogenetic tree of chemical evolution in hydrothermal environments.

2002 ◽  
Vol 42 (supplement2) ◽  
pp. S140
Author(s):  
S. Yokoyama ◽  
N. Kikuchi ◽  
A. Koyama ◽  
E. Imai ◽  
H. Honda ◽  
...  
Keyword(s):  
Phylogenetic Tree ◽  
Chemical Evolution ◽  
Molecular Phylogenetic Tree ◽  
Molecular Phylogenetic ◽  
Hydrothermal Environments

Species richness in the genus Microplana (Platyhelminthes, Tricladida, Microplaninae) in Europe: as yet no asymptote in sight

2017 ◽  
Vol 31 (3) ◽  
pp. 269 ◽  
Author(s):  
Eduardo Mateos ◽  
Ronald Sluys ◽  
Marta Riutort ◽  
Marta Álvarez-Presas
Keyword(s):  
New Records ◽  
Elongation Factor ◽  
Sequence Information ◽  
Major Step ◽  
Molecular Sequence ◽  
Accumulation Curve ◽  
Molecular Phylogenetic Tree ◽  
Molecular Phylogenetic ◽  
Morphological Species ◽  
Elongation Factor 1

This study forms a major step towards a comprehensive morphological and molecular analysis of the species diversity of European microplanid land planarians. It presents a molecular phylogenetic tree on the basis of information from the genes Cox1, 18S, 28S and elongation factor 1-α, and applies molecular and morphological species delimitation methodologies for Microplana specimens sampled over a wide geographic range within Europe. The study suggests that as yet there is no plateau or asymptote in the accumulation curve for European Microplana species, as our results facilitated an integrative delimitation of seven new species as well as the diagnosis of new populations of three already known species and one doubtful or problematic species. In some cases, the new records considerably enlarged the known range of a species. An integrative account is provided of each of these species. Molecular sequence information on newly collected land flatworms may quickly point the planarian systematist to taxa that need to be examined morphologically and thus may considerably reduce laborious and time-consuming histological analyses.

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