Cryphodera sinensis n. sp. (Nematoda: Heteroderidae), a non-cyst-forming parasitic nematode from the root of ramie Boehmeria nivea in China.

Cryphodera sinensis n. sp. is described from ramie (Boehmeria nivea) based on the morphology and molecular analyses of rRNA small subunit (SSU), D2D3 expansion domains of large subunit (LSU D2D3) and internal transcribed spacer (ITS). This new species is characterized by oval females with a distinct subcrystalline layer and pronounced and protruding vulval lip, distinctly concave vulva-anus profile and a vulva-anus distance of 29.5-35.8 µm. Males possess two annuli in the lip region, a stylet 27-32.5 µm in length with round knobs sloping slightly posteriorly, lateral fields with three lines, spicules 20-28 µm long and the presence of a short cloacal tube. Second-stage juveniles possess three lip annuli, a stylet 28-31 µm in length with well-developed knobs projected anteriorly and three lines along the lateral field. The pointed tail, 52-65 µm long, possesses a mucro-like tip and a hyaline region, 24.5-35 µm long. Large phasmids with a lens-like structure are located 2-6 annuli posterior to the anus. Phylogenetic analysis shows that the species has unique SSU, LSU D2D3 and ITS rRNA sequences. Phylogenetic relationships of the three rDNA sequences of C. sinensis n. sp. and other cystoid/cyst nematodes are analysed together with a comparison of other species within the genus Cryphodera.

First Report of Meloidogyne enterolobii on Cotton and Soybean in North Carolina, United States.

Stunted cotton plants (Gossypium hirsutum L. cvs. PHY 375 WR and PHY 565 WR) from two separate fields near Goldsboro in Wayne County, North Carolina were collected by the NCDA&CS Agronomic Division nematode lab for nematode assay and identification in December 2011. The galls on cotton plants were very large in comparison with those commonly associated with Meloidogyne incognita Kofoid and White (Chitwood) infected cotton. In August 2012, the lab also received heavily galled roots of soybean (Glycine max (L.) Merr. cv. 7732) from Wayne and Johnston counties. Population densities of the 2nd-stage juveniles ranged from 150 to 3,800 per 500 cc soil. Female perineal patterns were similar to M. incognita, but PCR and DNA sequencing matched that of M. enterolobii Yang and Eisenback (4). DNA sequences of ribosomal DNA small subunit, internal transcribed spacer, large subunit domain 2 and 3, intergeneric spacer, RNA polymerase II large subunit, and histone gene H3, were found to be 100% homologous when comparing populations of M. enterolobii from North Carolina and China. Species identification was also confirmed using PCR by a species-specific SCAR primer set MK7-F/MK7-R (2). M. enterolobii Yang & Eisenback was described in 1983 from a population causing severe damage to pacara earpod tree (Enterolobium contortisiliquum (Vell.) Morong) in China (4). In 2004, M. mayaguensis Rammah & Hirschmann, a species described from Puerto Rico, was synonymized with M. enterolobii based on esterase phenotype and mitochondrial DNA sequence (3). M. enterolobii is considered to be a highly pathogenic species and has been reported from vegetables, ornamental plants, guava, and weeds in China, Africa, Central and South America, the Caribbean, and Florida in the United States (1,3,4). Of particular concern is its ability to develop on crop genotypes carrying root-knot-nematode resistance genes (Mi-1, Mh, Mir1, N, Tabasco, and Rk) in tobacco, tomato, soybean, potato, cowpea, sweet potato, and cotton. Consequently, this species was added to the European and Mediterranean Plant Protection Organization A2 Alert list in 2010. Two populations of M. enterolobii one from soybean and one from cotton were reared on tomato (Solanum lycopersicum L. var. lycopersicum) in a greenhouse setting. Eggs were extracted using NaOCl and inoculated, at a rate of 7,000 per 15-cm-diameter clay pot, into a sandy soil mixture (1:1 washed river sand and loamy sand). Tomato, peanut (Arachis hypogaea L.), cotton, watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai), pepper (Capsicum annuum L.), and root-knot-susceptible and -resistant tobacco (Nicotiana tabacum L. cvs. K326 and NC 70, respectively) were transplanted immediately into the infested soil with four replications. Root galls on the host differentials were evaluated after 90 days. Reproduction occurred on all hosts except for peanut, which is consistent with reports for M. enterolobii and M. incognita race 4 (4). Adult females from pepper plants used in the host differential test were sequenced on partial 18S and ITS1 region and confirmed to be M. enterlobii. To our knowledge, this is the first report of a natural infection of North Carolina field crops with M. enterolobii. References: (1) J. Brito et al. J. Nematol. 36:324, 2004. (2) M. S. Tigano et al. Plant Pathol. 59:1054, 2010. (3) J. Xu et al. Eur. J. Plant Pathol. 110:309, 2004. (4) B. Yang and J. D. Eisenback. J. Nematol. 15:381, 1983.

Multiplex PCR for the simultaneous identification and detection of Meloidogyne incognita, M. enterolobii, and M. javanica using DNA extracted directly from individual galls.

Meloidogyne incognita, M. enterolobii, and M. javanica are the most widespread species of root-knot nematodes in South China, affecting many economically important crops, ornamental plants, and fruit trees. In this study, one pair of Meloidogyne universal primers was designed and three pairs of species-specific primers were employed successfully to rapidly detect and identify M. incognita, M. enterolobii, and M. javanica by multiplex polymerase chain reaction (PCR) using DNA extracted from individual galls. Multiplex PCR from all M. incognita, M. enterolobii, and M. javanica isolates generated two fragments of ≈500 and 1,000, 500 and 200, and 500 and 700 bp, respectively. The 500-bp fragment is the internal positive control fragment of rDNA 28S D2/D3 resulting from the use of the universal primers. Other Meloidogyne spp. included in this study generated only one fragment of ≈500 bp in size. Using this approach, M. incognita, M. enterolobii, and M. javanica were identified and detected using DNA extracted directly from individual galls containing the Meloidogyne spp. at various stages of their life cycle. Moreover, the percentage of positive PCR amplification increased with nematode development and detection was usually easy after the late stage of the second-stage juvenile. The protocol was applied to galls from naturally infested roots and the results were found to be fast, sensitive, robust, and accurate. This present study is the first to provide a definitive diagnostic tool for M. incognita, M. enterolobii, and M. javanica using DNA extracted directly from individual galls using a one-step multiplex PCR technique.

First Report of Meloidogyne enterolobii on Arrowroot in China.

The rhizome of arrowroot (Maranta arundinacea L.) is used as a source of edible starch in many tropical/subtropical areas. In July 2009, cultivated arrowroot plants in a field in Haikou, Hainan Province, China were observed to be exhibiting symptoms of decline, including stunting and yellowing. Roots of affected plants were found to have severe root galling similar to that caused by root-knot nematodes. Meloidogyne sp. females were dissected from the galls. Morphological characteristics of the females fit the description of M. enterolobii Yang & Eisenback (4). The perineal patterns were variable, with moderately high to high dorsal arch and mostly lacking lateral lines, but when present, the lines were not conspicuous, similar to those in the original description.(4). For further confirmation of species identity, isoenzyme patterns of malate dehydrogenase (Mdh) and esterase (Est) phenotypes were analyzed and partial sequences of mtDNA were obtained. Enzyme electrophoresis showed that the phenotypes of Mdh and Est were N1a and VS1-S1 respectively, which were consistent with those of M. enterolobii (1). Amplification and sequencing of the mtDNA between COII and the lRNA gene was accomplished with primers C2F3 (5'-GGTCAATGTTCAGAAATTTGTGG-3') and 1108 (5'-TACCTTTGACCAATCACGCT-3') (2). A DNA fragment of 705 bp was obtained and the sequence (GenBank Accession No. GQ870255) was compared with those in GenBank. A BLAST search indicated the sequence was 100% identical to the sequences of M. mayaguensis (GenBank Accession Nos. AY446969 and AY446970), a synonym of M. enterolobii (3). On the basis of these results, the root-knot nematodes isolated from arrowroot in Hainan were confirmed as M. enterolobii. This species has a high reproduction rate and a wide host range; moreover, it can induce severe galling and is able to overcome the resistance gene Mi-1 in tomato and pepper (4). M. enterolobii has become increasingly important because it has been found in many countries on diverse hosts. In China in recent years, the nematode has been reported on approximately 20 plant species belonging to five families, including Fabaceae, Cucurbitaceae, Solanaceae, Myrtaceae, and Annonaceae. To our knowledge, this is the first record of M. enterolobii parasitizing a plant (i.e., arrowroot) in the family Marantaceae in China. References: (1) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 17:6, 1985. (2) T. O. Powers and T. S. Harris. J. Nematol. 25:1, 1993. (3) J. Xu et al. Eur. J. Plant Pathol. 110:309, 2004. (4) B. Yang and J. D. Eisenback. J. Nematol. 15:381, 1983.

Partial molar volumes of some alpha-amino acids in aqueous sodium acetate solutions at 308.15 K.

The apparent molar volumes V(2,phi) have been determined for glycine, DL-alpha-alanine, DL-alpha-amino-n-butyric acid, DL-valine and DL-leucine in aqueous solutions of 0.5, 1.0, 1.5 and 2.0 mol kg(-1) sodium acetate by density measurements at 308.15 K. These data have been used to derive the infinite dilution apparent molar volumes V(0)(2,phi) for the amino acids in aqueous sodium acetate solutions and the standard volumes of transfer, Delta(t)V(0), of the amino acids from water to aqueous sodium acetate solutions. It has been observed that both V(0)(2,phi) and Delta(t)V(0) vary linearly with increasing number of carbon atoms in the alkyl chain of the amino acids. These linear correlations have been utilized to estimate the contributions of the charged end groups (NH(3)(+), COO(-)), CH(2) group and other alkyl chains of the amino acids to V(0)(2,phi) and Delta(t)V(0). The results show that V(0)(2,phi) values for (NH(3)(+), COO(-)) groups increase with sodium acetate concentration, and those for CH(2) are almost constant over the studied sodium acetate concentration range. The transfer volume increases and the hydration number of the amino acids decreases with increasing electrolyte concentrations. These facts indicate that strong interactions occur between the ions of sodium acetate and the charged centers of the amino acids. The volumetric interaction parameters of the amino acids with sodium acetate were calculated in water. The pair interaction parameters are found to be positive and decreased with increasing alkyl chain length of the amino acids, suggesting that sodium acetate has a stronger dehydration effect on amino acids which have longer hydrophobic alkyl chains. These phenomena are discussed by means of the co-sphere overlap model.

Activity coefficients for NaCl-monosaccharide (D-glucose, D-galactose, D-xylose, D-arabinose)-water systems at 298.15 K.

Electrochemical cells with a sodium ion selectivity electrode (Na-ISE) versus a chloride ion selectivity electrode (Cl-ISE) as a reference electrode were used to determine the activity coefficients for NaCl-monosaccharide (D-glucose, D-galactose, D-xylose, and D-arabinose) systems in water at 298.15 K. A comparison of the results thus obtained was made with those determined by another electromotive force (emf) method. It is shown that agreement is excellent. The Gibbs free energy parameters of the interactions between these sugars and NaCl in water were evaluated together with the parameter C1(CHOH, exo), indicating the interaction of the exocyclic CHOH group of saccharide molecules and NaCl. The results suggested that the interactions of these monosaccharides with NaCl are controlled mostly by the dominant conformer of their molecules in water.

Volumetric properties for the monosaccharide (D-xylose, D-arabinose, D-glucose, D-galactose)-NaCl-water systems at 298.15 K.

Densities have been measured for monosaccharide (D-xylose, D-arabinose, D-glucose and D-galactose)-NaCl-water solutions at 298.15 K. These data have been used to determine the apparent molar volumes of these saccharides and NaCl in the studied solutions. Infinite-dilution apparent molar volumes for the saccharides (V0(phi,S)) in aqueous NaCl and those for NaCl (V0(phi,E)) in aqueous saccharide solutions have been evaluated, together with the standard transfer volumes of the saccharides (delta(t) V0S) from water to aqueous NaCl and of NaCl (delta(t) V0E) from water to aqueous saccharide solutions. It is shown that the delta(t) V0S and delta (t) V0E values are positive and increase with increasing co-solute molalities. Volumetric parameters indicating the interactions of NaCl with saccharides in water have been obtained, respectively, by using transfer volumes of the saccharides and NaCl, and the resulting values are in good agreement with each other within experimental error. The interactions between saccharides and NaCl are discussed in terms of the structural interaction model and the stereochemistry of the saccharide molecules in water.

[An IR study of ion solvation and association of lithium perchlorate in some organic solvents].

The infrared (IR) spectra of propylene carbonate (PC), gamma-butyrolactone (BL) and diethyl carbonate (DEC) in the presence of LiClO4 have been investigated. It is shown that the interactions between Li+ and these solvents occur on the oxygen atoms of the carbonyl of solvent molecules. On the other hand, spectral curve fitting for band shape of perchlorate anion shows the presence of ion association in these solutions.