Due to its accuracy, sensitivity and high throughput, real time quantitative PCR (RT-qPCR) has been widely used in analysing gene expression. The quality of data from such analyses is affected by the quality of reference genes used. Expression stabilities for nine candidate reference genes widely used in soybean were evaluated under different stresses in this study. Our results showed that EF1A and ACT11 were the best under salinity stress, TUB4, TUA5 and EF1A were the best under drought stress, ACT11 and UKN2 were the best under dark treatment, and EF1B and UKN2 were the best under virus infection. EF1B and UKN2 were the top two genes which can be reliably used in all of the stress conditions assessed.

Callose is a β-l,3-glucan with diverse roles in the viral pathogenesis of plants. It is widely believed that the deposition of callose and hypersensitive reaction (HR) are critical defence responses of host plants against viral infection. However, the sequence of these two events and their resistance mechanisms are unclear. By exploiting a point inoculation approach combined with aniline blue staining, immuno-electron microscopy and external sphincters staining with tannic acid, we systematically investigated the possible roles of callose deposition during viral infection in soybean. In the incompatible combination, callose deposition at the plasmodesmata (PD) was clearly visible at the sites of inoculation but viral RNA of coat protein (CP-RNA) was not detected by RT-PCR in the leaf above the inoculated one (the upper leaf). In the compatible combination, however, callose deposition at PD was not detected at the site of infection but the viral CP-RNA was detected by RT-PCR in the upper leaf. We also found that in the incompatible combination the fluorescence due to callose formation at the inoculation point disappeared following the injection of 2-deoxy-D-glucose (DDG, an inhibitor of callose synthesis). At same time, in the incompatible combination, necrosis was observed and the viral CP-RNA was detected by RT-PCR in the upper leaf and HR characteristics were evident at the inoculation sites. These results show that, during the defensive response of soybean to viral infection, callose deposition at PD is mainly responsible for restricting the movement of the virus between cells and it occurs prior to the HR response.

The Sox (Sry-type HMG box) genes encode a group of proteins characterized by the existence of an SRY (sex-determining region on Y chromosome) box, a 79 amino acid motif that encodes an HMG (high mobility group) domain which can bind and bend DNA, which is the only part in SRY that is conserved between species. The Sox gene family functions in many aspects in embryogenesis, including testis development, CNS neurogenesis, oligodendrocyte development, chondrogenesis, neural crest cell development and other respects. The Sox gene family was originally identified through homology with Sry. The Sry gene is the mammalian testis-determining gene. It functions to open the testis determination pathway directly and close the ovary pathway indirectly. Sry and Sox9 are the most important two genes expressed during testis determination. Besides, researchers have found that Sox8 and Sox9 have functions in the male fertility maintenance after birth. In this review, information was evaluated from mouse or from human if not mentioned otherwise.

Along with the increasing application of nanoparticles (NPs) in many walks of life, environmental exposure to NPs has raised considerable health concerns. When NPs enter a pregnant woman’s body through inhalation, venous injection, ingestion or skin permeation, maternal toxic stress reactions such as reactive oxygen species (ROS), inflammation, apoptosis and endocrine dyscrasia are induced in different organs, particularly in the reproductive organs. Recent studies have shown that NPs disturb the developing oocyte by invading the protective barrier of theca cells, granulosa cell layers and zona pellucida. NPs disrupt sex hormone levels through the hypothalamic–pituitary-gonadal axis or by direct stimulation of secretory cells, such as granule cells, follicle cells, thecal cells and the corpus luteum. Some NPs can cross the placenta into the fetus by passive diffusion or endocytosis, which can trigger fetal inflammation, apoptosis, genotoxicity, cytotoxicity, low weight, reproductive deficiency, nervous damage, and immunodeficiency, among others. The toxicity of these NPs depend on their size, dosage, shape, charge, material and surface-coating. We summarize new findings on the toxic effect of various NPs on the ovary and on oogenesis and embryonic development. Meanwhile, we highlight the problems that need to be studied in the future. This manuscript will also provide valuable guidelines for protecting the female reproductive system from the toxicity of NPs and provide a certain reference value for NP application in the area of ovarian diseases.

In this study, melatonin (MEL)‐mediated plant resistance to tobacco mosaic virus (TMV) was examined to study local infection in Nicotiana glutinosa and systemic infection in Solanum lycopersicum. Exogenous application of 100 µm MEL increased anti‐virus infection activity to 37.4% in virus‐infected N. glutinosa plants. The same treatment significantly reduced relative levels of virus RNA analysed by qRT‐PCR and virus titres measured by dot‐ELISA, and increased the relative expression levels of the PR1 and PR5 genes analysed by qRT‐PCR, in virus‐infected S. lycopersicum. MEL treatment induced considerable accumulations of salicylic acid (SA) and nitric oxide (NO) but did not significantly affect production of hydrogen peroxide (H2O2) in the virus‐infected S. lycopersicum plants. Transgenic nahG N. tabacum was used to determine whether MEL‐induced TMV resistance was dependent on the SA pathway. The results showed that the relative RNA level of the TMV analysed by qRT‐PCR and virus titres analysed by dot‐ELISA were not reduced by the MEL treatment in the nahG transgenic N. tabacum seedlings treated twice with 100 µm MEL. The increased relative expression levels of PR1 and PR5 were greatly reduced when cPTIO, an NO scavenger, was included in the MEL treatment. A working model of MEL‐mediated plant resistance to TMV is proposed. MEL‐mediated plant resistance to viruses provides a new avenue to control plant viral diseases.