Mouse embryo co‐culture with autologous cumulus cells and fetal development post‐embryo transfer

This study was conducted to examine the potential for implantation and sustainable fetal development of mouse embryos cultured from the pronuclear to blastocyst stage. Pronuclear embryos from ICR mice (Harlan Sprague‐Dawley) were cultured in Sydney IVF sequential media (Cook) to the blastocyst stage in medium only or co‐cultured with autologous cumulus cells. We also experimented with co‐culture in 100 µL drops. Drop co‐culture produced blastocyst formation rates with a mean of 47.0%, which was significantly higher (P < 0.05) compared to embryos cultured in identical culture conditions except without cumulus cells at 27.3%. Blastocysts obtained in vitro in Cook medium only and co‐cultured in Cook medium with cumulus cells were transferred to pseudopregnant females of ICR strain. The day of blastocyst transfer into surrogate females was designated as post‐transfer of blastocyst day 1 (PT 1). The implantation and fetal development was compared to embryo transfer of in vivo derived blastocysts, which served as controls. There were no statistical differences for implantation and fetal development rates for blastocysts cultured in vitro in either Cook medium only or co‐culture in Cook medium with cumulus cells compared to in vivo‐derived blastocysts. The advantage of the co‐culture system is in generating more blastocysts available for transfer.     

FTIR-ATR-based prediction and modelling of lignin and energy contents reveals independent intra-specific variation of these traits in bioenergy poplars

Background  

There is an increasing demand for renewable resources to replace fossil fuels. However, different applications such as the production of secondary biofuels or combustion for energy production require different wood properties. Therefore, high-throughput methods are needed for rapid screening of wood in large scale samples, e.g., to evaluate the outcome of tree breeding or genetic engineering. In this study, we investigated the intra-specific variability of lignin and energy contents in extractive-free wood of hybrid poplar progenies (Populus trichocarpa × deltoides) and tested if the range was sufficient for the development of quantitative prediction models based on Fourier transform infrared spectroscopy (FTIR). Since lignin is a major energy-bearing compound, we expected that the energy content of wood would be positively correlated with the lignin content.     

A simple and efficient method for isolating small RNAs from different plant species

Background  

Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species.     

Diofenolan: a novel insect growth regulator in common citrus butterfly, <Emphasis Type="Italic">Papilio demoleus</Emphasis>

Penultimate and last instar larvae of the common citrus swallowtail butterfly, Papilio demoleus L. (Papilionidae: Lepidoptera), were treated with different doses of a novel juvenoid, diofenolan. Several deformities were observed as a result of topical administration of diofenolan which include delay in larval–larval and larval–pupal ecdysis, ecdysial failure, mortality, severe reduction in pupation, deformed pupae and complete inhibition of adult emergence. The juvenoid diofenolan severely hampers the normal growth, development and metamorphosis of P. demoleus and can be incorporated into integrated pest management programs for successful control of this important pest of citrus all over the world.     

Note on <Emphasis Type="Italic">Wahlgreniella nervata</Emphasis> (Gillette, 1908) (Hemiptera: Aphididae): a new pest of Damask rose in Turkey

Alate viviparous females of Wahlgreniella nervata (Gillette) were found on a Damask rose (Rosa damascena Mill.) plantation in Isparta Province, Turkey. The aphid is recorded for the first time from Turkey. Besides, Damask rose is a new host for this species. Brief information about this aphid including a key to the alate females of aphid species known to infest Damask rose in Turkey is provided.     

Resistance of <Emphasis Type="BoldItalic">Spodoptera exigua</Emphasis> to ten insecticides in Shandong,China

Spodoptera exigua (Hübner) collected from three regions (two scallion-producing regions and one ginger-producing region) in Shandong, China and a laboratory colony of Wuhan (WHLC) were evaluated for their susceptibilities to ten insecticides (emamectin benzoate, chlorfenapyr, indoxacarb, spinosyn, tebufenozide, methoxyfenozide, chlorfluazuron, beta-cypermethrin, chlorpyrifos and methomyl) in 2008, 2009 and 2010 using a leaf-dip bioassay method. The results indicate that the resistance ratios of S. exigua to newer insecticides such as emamectin benzoate, chlorfenapyr and indoxacarb were all below 20-fold, with no obvious change in all 3 years. S. exigua exhibits moderate resistance to spinosyn, and its resistance ratios increased from 1.98–5.31-fold in 2008 to 14.31–64.20-fold in 2010 from three regions as compared with WHLC. S. exigua showed moderate to high resistance to insect growth regulators such as tebufenozide, methoxyfenozide, chlorfluazuron in 2008, 2009 and 2010, in which resistance to chlorfluazuron was rapidly increased from 31.49–88.19-fold in 2008 to 1184.39–2789.67-fold in 2010. Resistance of S. exigua to beta-cypermethrin and chlorpyrifos varied greatly among the three regions, ranging from 95.31–437.97-fold and 25.05–40.64-fold in 2008, 951.81–1304.40-fold and 44.91–186.33-fold in 2009, to 27.27–1095.31-fold and 19.12–267.98-fold in 2010. In contrast, S. exigua showed low resistance to methomyl, and the resistance ratio was below 5-fold in 3 years. There are several reasons accounting for varying degrees of resistance, including selection pressure, cropping structure and migration, in which the migration of S. exigua may play an important role.     

A molecular mechanism of resistance to streptomycin in <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzicola</Emphasis>

Xanthomonas oryzae pv. oryzicola, the causal agent of rice leaf streak disease, was found to be sensitive to streptomycin (an aminocyclitol glycoside antibiotic), by inhibition of protein synthesis resulting from interference with translational proofreading. This study aimed to determine the molecular resistance mechanism of X. oryzae pv. oryzicola to streptomycin. Seven streptomycin-resistant mutants were obtained by UV induction or streptomycin selection. These mutants can grow at 100 μg ml⁻¹ of streptomycin while the wild-type strain (RS105) cannot grow at 5 μg ml⁻¹. Sequencing indicated that the rpsL gene encoding ribosomal protein S12 has 375 bp encoding 125 amino acid residues. In all resistant strains, a mutation in which AAG was substituted for AGG (Lys→Arg) occurred either at codon 43 or 88. Two plasmids, pUFRRS and pUFRRX, were constructed by ligating the rpsL gene into the cosmid pUFR034. The plasmids pUFRRS and pUFRRX containing the Lys→Arg mutation of the rpsL gene conferred streptomycin resistance to the sensitive wild-type strain by electroporation. Both transformants, RS1 and RS2, could grow in the medium containing 50 μg ml⁻¹ of streptomycin. A mutation at codon 43 or 88 in rpsL can result in resistance of Xanthomonas oryzae pv. oryzicola to streptomycin.     

Pathogens of bark beetles (Coleoptera: Curculionidae) in Bulgarian forests

The occurrence and prevalence of bark beetle pathogens in forest stands in Bulgaria were investigated in 944 specimens belonging to 21 bark beetle species. Protozoa, microsporidia, fungi and nematodes occurred in 19 of all investigated species. The infections were found in the gut (nematodes, gregarines, microsporidia), gonads (microsporidia) and hemolymph (nematodes) of the infected insects. Protozoan species (Gregarina typographi, Gregarina spp.) were detected in eight bark beetle species. Morphometric data about G. typographi and Gregarina spp. are presented. The prevalence of the gregarines varied between 1.4% and 64.2%. Microsporidia of the genera Nosema and Chytridiopsis were revealed in three bark beetle species. The prevalence of microsporidia ranged between 1.5% and 11.8%. This is the first report of a microsporidium in Taphrorychus villifrons and of gregarines in T. villifrons, Pityogenes bistridentatus, P. conjunctus, and Orthotomicus erosus. The fungus Beauveria bassiana was found in 3.4% of Hylurgops palliatus specimens. Nematodes (in gut and haemolymph) were revealed in 19 bark beetle species and their prevalence varied between 10% and 98.5%.