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Cervical mucus not only provides energy for sperm but also forms a barrier to block sperm. This paper aims to study the microstructure of cervical mucus in dairy cows during the proestrus, estrus, and metestrus and its effect on sperm permeability. The experiment collected cervical mucus from 60 Holstein cows during these phases, then observed the different shapes of the mucus after crystallization, classified the mucus, and analyzed its proportions. Scanning electron microscopy was used to observe the ultrastructure of the cervical mucus and measure the micro-pore sizes, followed by sperm permeability tests using mucus from different estrous stages and counting the number of permeated sperm. The results indicate that cervical mucus from cows in different estrous phases includes four types (L, S, P, G), with each type constituting a different proportion. During the proestrus, the L type was significantly more prevalent than the other types (p < 0.05); during estrus, the S type was significantly more prevalent than the other types (p < 0.05); and during the metestrus, the p type was significantly more prevalent than the other types (p < 0.05). The micro-pore sizes of the same type of cervical mucus did not show significant differences across different estrous phases (p > 0.05). However, within the same estrous phase, there were significant differences in the micro-pore sizes among the four types (p < 0.05). The number of sperm that permeated the cervical mucus during estrus and metestrus was significantly higher than during the proestrus (p < 0.05). This study provides data support for the research on cervical mucus in dairy cows.

Abstract Anthracnose caused by Colletotrichum species, is one of the most destructive diseases affecting strawberry production worldwide. Fungi of the genus Colletotrichum, being ancient and co-evolved with flowering plants, rank eighth among the top ten economically important fungal pathogens and have diversified to adapt to various hosts. Since it was first reported in United States in 1931, strawberry anthracnose has spread globally, resulting in devastating economic, social, and ecological consequences. In a sense, to produce organic strawberry is impracticable largely because of this disease. Colletotrichum species pathogenic to strawberry have been widely characterized. The review focused on providing an accurate inventory of Colletotrichum species pathogenic to strawberry via revisiting the discovery history scrupulously over 90 years. A total of 23 Colletotrichum species clustered into five species complexes and two singleton taxa were accepted as strawberry pathogens with global occurrence illustrated. The relative prevalence of distinct Colletotrichum species associated with strawberry was systematically analyzed in China. Finally, the host range of the prevailing C. fructicola and C. siamense, also the top two Colletotrichum species causing serious damages on the earth was summarized, with a goal of diagnosing accurately and improving anthracnose management in agricultural ecosystem.

Based on the recently published whole-genome sequence of cultivated strawberry ‘Camarosa’, in this study, 222 FaWRKY genes were identified in the ‘Camarosa’ genome. Phylogenetic analysis showed that the 222 FaWRKY candidate genes were classified into three groups, of which 41 were in group I, 142 were in group II, and 39 were in group III. The 222 FaWRKY genes were evenly distributed among the seven chromosomes. The exon–intron structures and motifs of the WRKY genes had evolutionary diversity in different cultivated strawberry genomes. Regarding differential expression, the expression of FaWRKY133 was relatively high in leaves, while FaWRKY63 was specifically expressed in roots. FaWRKY207, 59, 46, 182, 156, 58, 39, 62 and 115 were up-regulated during achene development from the green to red fruit transition. FaWRK181, 166 and 211 were highly expressed in receptacles at the ripe fruit stage. One interesting finding was that FaWRKY179 and 205 were significantly repressed after Colletotrichum fructicola inoculation in both ‘Benihoppe’ and ‘Sweet Charlie’ compared with Mock. The data reported here provide a foundation for further comparative genomics and analyses of the distinct expression patterns of FaWRKY genes in various tissues and in response to C. fructicola inoculation.

Objective The present study evaluated the preservation of ram semen at 0°C using soybean lecithin with a Tris-fructose extender. Methods Semen was collected by artificial vagina ejaculation from six rams with proven fertility. High quality ejaculates were diluted by soybean lecithin (0.25%, 0.5%, 0.75%, 1.0%, 1.25%) using Tris-fructose extender and control (Tris-fructose egg yolk extender), respectively. The ejaculates were diluted to a concentration of 5×108 sperm/mL, followed by cooling to 0°C in 90 min and maintaining the temperature for 12 days. The diluted semen samples were examined and recorded for sperm progressive motility, acrosome integrity at 0, 24, 72, 144, 216, 288 h, respectively. Two hundred and twenty-three ewes were inseminated for 216 h with optimal soybean lecithin concentrated semen or control via trans-cervical insemination. Results The results showed that there were no differences in sperm progressive motility at 0, 24, 72, and 144 h (p>0.05). After 216 h, the sperm progressive motility in the control group and 0.5% concentration groups was significantly higher when compared to 0.25% concentration (p0.05). At 216 h, the sperm progressive motility of all groups was still above 50%. The acrosome integrity of all groups was decreased with prolongation of storage time, but there was no difference at each time point (p>0.05). There was no significant difference in the lambing rate and pregnancy rate between the 0.5% concentration group and the control group (p>0.05). Conclusion These results suggest that ram sperm is capable of fertilization after preservation at 0°C with 0.5% of soybean lecithin in Tris-based extender substituted for egg yolk and produce normal offspring after insemination.

Colletotrichum fructicola causes strawberry anthracnose, which results in considerable economic losses in quality and production in Asia. However, the molecular mechanism of strawberry infection with C. fructicola is still poorly understood. Here, we employed RNA-Seq technology to mine the strawberry transcriptome for EST-SSRs (Expressed Sequence Tag–Simple Sequence Repeats) following C. fructicola infection. Fortunately, we detected 3019 potential EST-SSRs from the transcriptomic data of cv. Jiuxiang. The seven commonly occurring repeat motifs were A/T (278), GAA/TTC (263), GAG/CTC (215), AAG/CTT(188), GGA/TCC (163), AGA/TCT (153) and AGG/CCT (153). We identified 31, 84 and 129 differentially expressed EST-SSR-containing sequences corresponding to 24, 72 and 96 Hpi, while 13 DE-EST-SSRs -containing sequences were differentially expressed and common for all the three time points. Furthermore, we identified allele polymorphisms for some DE-EST-SSRs among several resistant and susceptible strawberry varieties and wild species. The EST-SSRs in our research were related to the reported QTL resistance loci in strawberry which are differentially expressed after C. fructicola infection and have allele polymorphisms in different strawberry varieties and wild species. These provide a clue for utilisation of the EST-SSRs in marker-assisted breeding to improve anthracnose-resistant breeding in future.

Intragenesis is an all-native engineering technology for crop improvement. Using an intragenic strategy to bring genes from wild species to cultivated strawberry could expand the genetic variability. A robust regeneration protocol was developed for the strawberry cv. ‘Shanghai Angel’ by optimizing the dose of Thidiazuron and identifying the most suitable explants. The expression cassette was assembled with all DNA fragments from F. vesca, harboring a sugar transporter gene FvSTP8 driven by a fruit-specific FvKnox promoter. Transformed strawberry was developed through an Agrobacterium-mediated strategy without any selectable markers. Other than PCR selection, probe-based duplex droplet digital PCR (ddPCR) was performed to determine the T-DNA insert. Four independent transformed shoots were obtained with a maximum of 5.3% efficiency. Two lines were confirmed to be chimeras, while the other two were complete transformants with six and 11 copies of the intragene, respectively. The presence of a vector backbone beyond the T-DNA in these transformants indicated that intragenic strawberries were not obtained. The current work optimized the procedures for producing transformed strawberry without antibiotic selection, and accurately determined the insertion copies by ddPCR in the strawberry genome for the first time. These strategies might be promising for the engineering of ‘Shanghai Angel’ and other cultivars to improve agronomic traits.

Leeches are frequently used in traditional Chinese medicine. However, they are potentially hazardous to human and animal health by transmitting several pathogens. Studies of diseases transmitted by leeches are scarce. The purpose of this study was to analyze the pathogens carried in pond-farmed medicinal leech in China. Leeches were collected from 6 farms in Hubei Province in central China. DNA was extracted from the internal organ of leeches to analyze the origin of blood meal. Leech genera were confirmed through amplification of 18S rRNA and mitochondrial gene cytochrome oxidase I (COI) gene by PCR and host animal species were identified through amplification of mitochondrial cytochrome b gene. Species of Ehrlichia in the leech specimens were screened with PCR using specific primers. PCR amplification and DNA sequencing showed that 620 leeches were Hirudinaria sp. Ehrlichia DNA was detected in 39 specimens from 2 farms. We obtained a total of 65 sequences of the cytB gene from 620 leech internal organ samples including sequences of human (n = 5), rat (n = 1), domestic pig (n = 10), duck (n = 23), goose (n = 12) and buffalo (n = 14). Phylogenetic analysis of the rrs and groEL gene sequences showed that Ehrlichia detected in the study were closely related to Ehrlichia sp. in ticks from Korea and Japan. To the best of our knowledge, this is the first report on Ehrlichia DNA being detected from leeches. Our findings provided new data on Ehrlichia spp. and farmed leech species in China.

Culture of hippocampal neurons in low-Mg2+ medium (low-Mg2+ neurons) results in induction of continuous seizure activity. However, the underlying mechanism of the contribution of low Mg2+ to hyperexcitability of neurons has not been clarified. Our data, obtained using the patch-clamp technique, show that voltage-gated Na+ channel (VGSC) activity, which is associated with a persistent, noninactivating Na+ current (INa,P), was modulated by calmodulin (CaM) in a concentration-dependent manner in normal and low-Mg2+ neurons, but the channel activity was more sensitive to Ca2+/CaM regulation in low-Mg2+ than normal neurons. The increased sensitivity of VGSCs in low-Mg2+ neurons was partially retained when CaM12 and CaM34, CaM mutants with disabled binding sites in the N or C lobe, were used but was diminished when CaM1234, a CaM mutant in which all four Ca2+ sites are disabled, was used, indicating that functional Ca2+- binding sites from either lobe of CaM are required for modulation of VGSCs in low-Mg2+ neurons. Furthermore, the number of neurons exhibiting colocalization of CaM with the VGSC subtypes Nav1.1, Nav1.2, and Nav1.3 was significantly higher in low-Mg2+ than normal neurons, as shown by immunofluorescence. Our main finding is that low-Mg2+ treatment increases sensitivity of VGSCs to Ca2+/CaM-mediated regulation. Our data reveal that CaM, as a core regulating factor, connects the functional roles of the three main intracellular ions, Na+, Ca2+, and Mg2+, by modulating VGSCs and provides a possible explanation for the seizure discharge observed in low-Mg2+ neurons. [ABSTRACT FROM AUTHOR]

The article presents information on a study related to the expression of regulator gene proto-oncogene c-myc in the Tarim deer antler in different growth stages. The method includes examination of gene expression location by immunohitochemistry and quantitative analysis of c-myc expression by fluorescent quantitative polymerase chain reaction. The results show positive expression of c-myc in inner root sheath of hair follicle and hair matrix of dermal layer.

Abstract  The maize genome remains abundant in molecular diversity, and the rich genetic diversity of maize starch-synthesis genes is crucial for controlling various grain traits. To explore the unique mechanism controlling the advantageous waxy trait and characterize the molecular feature of genes relevant to starch composition in two elite waxy inbreds, expression profiling combined with gene organization analysis was performed in them as compared to one normal inbred. Genotype-specific expression patterns were observed for most genes studied. The waxy inbreds were shown to contain mutations in multiple starch-synthesis genes, namely gbssI (wx), gbssIIb and isa2 (potentially isa3 too).The mis-splicing events directly accounted for wx loss of function. Contrarily, disruption of 5′ and 3′ transcript sequence may contribute to the absence of GbssIIb and Isa2 transcripts in waxy inbreds, respectively. Besides, the splicing of Sugary1 transcript was developmentally regulated in the normal inbred, and DNA polymorphisms were detected within SSIIIb-1 gene in waxy inbreds. [ABSTRACT FROM AUTHOR]

We perform an experimental study of the multi-orbital effect on the high-order harmonic generation (HHG) from aligned N2 molecules in both linearly and elliptically polarized intense laser fields. Measured by a home-built extreme ultraviolet (XUV) flat grating spectrometer with the pump–probe method, the angular distributions of different orders of HHG are obtained, which show distinctive behaviors for harmonics in the plateau and the cut-off regions. The ellipticity dependence of HHG is investigated by aligning the molecular axis parallel or perpendicular to the laser polarization. Our results indicate that both the highest occupied molecular orbital (HOMO) as well as the lower one (HOMO-1) contribute to the HHG of N2 molecules, in either linearly or elliptically polarized intense laser field. The study paves the way for understanding the ultrafast electron dynamics of molecules exposed to an intense laser field. [ABSTRACT FROM AUTHOR]

We investigate experimentally multi-orbital effects in high-order harmonic generation (HHG) from aligned CO2 and N2O molecules by intense femtosecond laser fields with linear and elliptical polarizations. For either of the aligned molecules, a minimum in the harmonic spectrum is observed, the position of which shifts to lower-order harmonics when decreasing the intensity or increasing the ellipticity of the driving laser. This indicates that the minimum originates from the dynamic interference of different channels, of which the tunneling ionization and recombination are contributed via different molecular orbitals. The results show that both the highest occupied molecular orbital (HOMO) and low-lying HOMO-2 in CO2 (or HOMO-1 in N2O) contribute to the molecular HHG in both linearly and elliptically polarized strong laser fields. Our study would pave a way for understanding multi-electron dynamics from polyatomic molecules irradiated by strong laser fields. [ABSTRACT FROM AUTHOR]

The article focuses on Shenmei #3, a strawberry cultivar hybridized by the Forestry & Fruit Research Institute (FFRI) of the Shanghai Academy of Agricultural Sciences (SAAS) in China. The plants and fruits of this cultivar are powdery mildew-resistant. The appearance of leaves and fruit of Shenmei #3 are described. The findings on the quality and ascorbic acid content of the fruit are detailed.

Superovulation is a key technique in embryo transfer. This study aimed to investigate the effects of two different sources of FSH (Ningbo Second Hormone Factory – NSHFTM, China and Vetoquinol, Australia), FSH injection methods (decreasing dose and equal dose), embryo collection methods (injection-type flushing tube and three-channel type flushing tube) and seasons (winter and spring) on superovulation response in Belgian Blue cattle. For this purpose, 80 Belgian Blue cattle (aged 2-3 years, parity=1) were randomly divided into four groups, with each group was further divided into two sub-groups. Superovulation experiments were carried out through Cue-Mate+FSH+PGF2α+GnRH, followed by artificial insemination. The effects of the above four factors on superovulation response were determined by comparing the mean numbers of corpora lutea, total embryos and viable embryos. The results showed non-significant difference in the effects of FSH produced by NSHFTM, China and Vetoquinol, Australia on the superovulation response, though the cost of NSHFTM was lower. Moreover, the superovulation response to decreasing-dose injection of FSH was significantly better than that of equal dose injection (P<0.05). Regarding embryo collection methods, the results of injection-type flushing tube were significantly better than those of three-channel type flushing tube (P<0.05). Similarly, superovulation results during spring season were significantly better than those during winter (P<0.05). In summary, the best method of superovulation in Belgian Blue cattle is to inject FSH with reduced dose/s and collect embryos through injection-type flushing tube method during spring season. [ABSTRACT FROM AUTHOR]