First Record of Nematode Longidorus attenuatus on Soybean in Poland.
Instytut Ochrony Roślin - Państwowy Instytut Badawczy
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Soybean (Glycine max) is one of the most important agricultural crops worldwide. Due to the economic value of this crop, soybean is being introduced to areas where it has not been traditionally cultivated. An example is Poland, where the total area of cultivated soybean has increased more than 10 times between 2008 and 2012. The plant-parasitic nematode Longidorus attenuatus Hooper, 1961 is known to parasitize many plant species and it is associated with docking disorder of sugarbeet (Whitehead and Hooper 1970). Additionally, it is a vector of Tomato black ring virus (TBRV) (Harrison 1964). In September 2014, 10 soil samples were collected to a depth of 10 to 45 cm from the rhizosphere of the soybean variety Lissabon at a field in Winna Góra Village, Wielkopolska region, Poland. Nematodes were extracted from 100 cm3 of soil using the decant and sieve method (Brown and Boag 1988), with final separation on a 100-µm sieve. Longidorus attenuatus was present in all samples, with an average density of 11 specimens/100 cm3 of soil. Measurements of females (n = 7) include: a mean body length (L) = 6869 ± 468.8 (6257 to 7341) µm; width at lips 13.8 ± 0.5 (13 to 14) µm; width at vulva 44.5 ± 1.91 (42 to 46) µm; odontostylet length 83 ± 2.2 (80 to 85) µm; tail 55 ± 7.8 (48 to 64) µm; L/maximum body width= 154 ± 12.1 (142 to 169); anterior end to vulva/L (in %) = 47.7 ± 0.7 (46.9 to 48.6); tail length/width at beginning of tail = 1.68 ± 1.19 (1.5 to 1.88). Identification was confirmed after DNA was extracted from 6 specimens using QIAamp DNA Micro Kit (Qiagen, Hilden, Germany) and 28S large ribosomal D2-D3 expansion segment was amplified and sequenced as described by Kornobis et al. (2015). The sequenced product was subjected to a database search using BLAST. The sequence from 28S region was identical (100% BLAST similarity) to a sequence from Belgium (GenBank Accession No. AY601572). The sequence was deposited under EMBL/GenBank Accession No. KR911851. To demonstrate that soybean is a host for L. attenuatus, seven replicate plastic boxes were filled with 800 cm3 of soil. The soil used in the experiment was taken from a place with no L. attenuatus and additionally sterilized (121°C, 205 kPa, 30 min) using a Systec DX-90 2D autoclave. In each box, two soybean Lissabon seeds were planted. When seedlings were about 5 cm high, 20 L. attenuatus females were added to five boxes (two remaining being a control with no nematodes) and subsequently kept in a greenhouse (22°C, 15 h of daylight per day). After twelve weeks, nematodes were extracted using the decant and sieve method from the whole volume of soil in each box. A mean of 184 nematodes/box (23 nematodes/100cm3 soil) were recovered. No nematodes were recovered from controls. Although no obvious symptoms were observed on the plants, at the end of the experiment those with nematodes had visibly more yellowing leaves compared with plants from the control. To our knowledge, this is the first record of L. attenuatus from soybean. Additionally, due to the fact that this species occurs in many European countries it should be considered as a potentially important pathogen of soybean.
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