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High and Low Throughput Screens with Root-knot Nematodes Meloidogyne spp.
Published on: March 12, 2012
First Report of Root-Knot Nematode Meloidogyne arenaria Infesting Roots of Anubias barteri in Guangdong, China
1Laboratory of Plant Nematology, South China Agricultural University, Guangzhou 510642, China.
Abstract:
Anubias spp. are very popular aquatic plants that are native to Africa. Anubias barteri is a broadleaf Anubias and is a popular choice for aquariums because of its robust size. In China, broadleaf Anubias is usually planted in sand, with sponges around the rhizome, in small plastic baskets in aquatic plant nurseries. In December 2010, a survey of phytoparasitic nematodes was conducted in the nurseries in Guangzhou, Guangdong Province. Many second-stage juveniles of a Meloidogyne sp. were detected from the roots of A. barteri, but no root galls were found. To identify these juveniles, molecular identification was performed with PCR. The DNA sequence between CO II and l-rRNA of the mitochondrial gene of single juveniles was amplified with universal primers of Meloidogyne, #C2F3 (5'-GGTCAATGTTCAGAAATTTGTGG-3') and #1108 (5'-TACCTTTGACCAATCACGCT-3') (3). The amplified fragments were approximately 1.1 kb long and could not be digested with restrictive enzyme HinfI. The specific fragments were then sequenced. The blast search result revealed that the DNA sequence (GenBank Accession No. JQ446377) had 99 to 100% identity with submitted sequences of Meloidogyne arenaria (GenBank Accession Nos. EU364879, GQ266686, and AY635610). The other extracted juveniles were inoculated into sterile, potted, water spinach (Ipomoea aquatica) in the greenhouse to obtain more nematodes. After 40 days, root galls and female egg masses were clearly observed, and biochemical, molecular, and morphological identifications were conducted. Isoenzyme phenotype (esterase and malate dehydrogenase (MDH) patterns) and the perineal pattern of several gravid females were the same as M. arenaria (1,2), and PCR amplification of single juveniles produced identical fragments as previously found. Single egg masses were collected and juveniles were hatched out and inoculated onto 10 nematode-free plants of A. barteri in a greenhouse. After 40 days, roots of A. barteri exhibited inconspicuous small galls, and the same identification procedures were conducted as mentioned previously. Isoenzyme phenotypes, perineal patterns of adult females, and amplified fragments of single juveniles were identical to those of M. arenaria. M. arenaria is one of the most important root-knot nematodes and causes great losses in many crops around the world (2). To our knowledge, this is the first record of M. arenaria parasitizing aquatic plants of broadleaf Anubias in China and elsewhere, and A. barteri is a new host of M. arenaria. The economic importance of this nematode to A. barteri production is currently unknown. However, because A. barteri is a commercial aquatic plant, more attention should be given by producers to prevent this nematode from becoming an important pathogen. In addition, this finding is very helpful for relevant plant nematode quarantine work. References: (1) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 17:1, 1985. (2) R. N. Perry et al. Root-Knot Nematodes. CABI. Wallingford, UK, 2009. (3) T. O. Powers and T. S. Harris. J. Nematol. 25:1, 1993.
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