Pleiotropic drug resistance (PDR) is an important subfamily member of the ABC transporter superfamily, which plays important role in response to stress, plant growth and development, but little research has been done in wheat (Triticum aestivum). In order to investigate the response of wheat PDR gene family to stress, genome-wide identification and bioinformatics analysis of TaPDR gene family were performed in this study, and the responses to abiotic stress were verified by qPCR. The results showed that 76 TaPDR genes were mainly distributed on chromosome 3, 5 and 7. The number of coding amino acids was 1 242~1 517, and the molecular weight was 140.69~170.99 kD, pI was between 6.22 and 8.94. According to phylogenetic analysis, TaPDR genes were divided into 5 groups (Class Ⅰ~Ⅴ), most of TaPDRs had highly conserved protein motifs, and TaPDR genes in the same group had more similar gene structure. 10 pairs of tandem repeat genes and 56 pairs of segmental duplication genes were founded with gene duplication analysis method. The tissue-specific analysis showed that TaPDR genes exhibited different expression levels in different tissues. Most TaPDR genes were highly expressed in roots. qPCR results showed that TaPDR genes could respond to salt, drought, cadmium and abscisic acid treatment in different degree. The results provide a reference basis for the mechanism of TaPDR gene family in abiotic stress response.

Southern leaf blight was an important disease in maize planting region of the world. There were significant differences in resistance of maize (Zea mays) cultivar with different genetic backgrounds to the disease. In order to understand response profiles for gene expression in maize to early infection of Cochliobolus heterostrophus, transcriptional data in the resistant cultivar ('Mo17-R') and susceptible cultivar ('Luo31-S') of maize inbred line at 12 and 24 h after inoculation were analyzed using the transcriptome analyses. The results demonstrated that 11 416 and 11 027 differentially expressed genes (DEGs) were found in 'Mo17-R' and 'Luo31-S' after inoculation by C. heterostrophus, respectively. GO enrichment and KEGG metabolic pathway analysis revealed that the DEGs were most rich in the terms of cell, cell part, organelle, membrane, binding, catalytic activity, metabolic process, cellular process and single-organism process, which were also most rich in the pathways of phenylpropanoid biosynthesis, signal transduction, plant-pathogen interaction, starch and sucrose metabolism. Among the enriched pathways associated with resistance, the pathways of phenylpropanoid biosynthesis, plant hormone signal transduction and plant-pathogen interaction were common in 'Mo17-R' and 'Luo31-S', while the DEGs involved in terpenoid biosynthesis pathway were only enriched in 'Luo31-S'. Furthermore, theses DEGs related to peroxidase, cinnamyl alcohol dehydrogenase, ethylene-responsive transcription factor, cyclic nucleotide-gated ion channel, cytochrome P450 were specifically induced in the resistant inbred line ('Mo17-R'), while these DEGs were down-regulated or less induced in the susceptible inbred line ('Luo31-S'). It was speculated that these genes could play an important role in disease resistance for resistant maize at the early stage of C. heterostrophus infection. These results provides a theoretical basis for further exploitation of molecular mechanisms underlying the interaction between maize and C. heterostrophus, and maize resistance to the pathogen.

WRKY family is widely involved in plant growth and development, regulating substance metabolism, responding to environmental signals and so on. To study the function of grape (Vitis vinifera) VvWRKY26, the VvWRKY26 gene (GenBank No. KX 823961.1) was cloned from 'WuHeZaoHong' grape by homologous sequence method, the bioinformatics, spatio-temporal expression and response salicylic acid (SA) expression were analyzed. The results of bioinformatics analysis showed that the CDS region of VvWRKY26 was 1 434 bp length, encoded 477 amino acids, and its secondary structure was composed of random curl, α-helix and β-fold, which was a non-secreted, non-transmembrane protein distributed in the nucleus. It was highly homologous with Petunia hybrid PhPH3 (GenBank No. AMR43368.1), Brassica napus BnTTG2 (Brassica napus transparent testa glabra2, GenBank No. 106423067) and Arabidopsis thaliana AtTTG2 (Arabidopsis thaliana transparent testa glabra2, GenBank No. 818303), and belongs to the WRKY transcription factor family Ⅰ. Promoter analysis showed that VvWRKY26 promoter had response elements such as light signal, resistance to reactive oxygen stress, SA and gibberellin acid (GA) signals, etc.. qPCR analysis showed that VvWRKY26 was expressed in the flowers, stems, buds, tendrils, young leaves, mature leaves and fruit tissues, and the expression levels were higher during the young fruit stage and color changing stage. It participated in SA signal response and the expression level under the SA treatment at 0.5~8 h was significantly higher than that of the control (P<0.05). This study provides a theoretical basis for the further research on the function of VvWRKY26 in SA signal channel.

Grape (Vitis vinifera) often suffers various environmental stresses. The mechanism of tolerance to stresses has been a hot topic of cultivation techniques in grape. Plant stress resistance known to rely on special proteins. DnaJ protein (HSP40) serves as a co-chaperone of heat shock protein 70 (HSP70), playing crucial roles in various stress responses. This study focused on the VvDnaJ05 (GenBank No. OQ102602) and VvDnaJ26 (GenBank No. OQ108519) cloning from DnaJ gene family. The expression patterns of the VvDnaJ05 and VvDna26 in different tissues and under various hormones and stresses were determined by qPCR. Meanwhile, the overexpression and interaction vectors were constructed to explore the protein function and regulatory mechanism of the VvDnaJ05 and VvDnaJ26. The results indicated that the full-length of VvDnaJ05 was 822 bp, encoding 273 amino acids. The full-length of VvDnaJ26 was 1 254 bp, encoding 417 amino acids. VvDnaJ05 and VvDnaJ26 showed the highest expression levels in skin and seed, respectively. Hormones responsive cis-acting elements were found in the promoter region of VvDnaJ05 and VvDna26, significantly changes were observed in the expression level of both under methyl jasmonate and salicylic acid treatment (P<0.05). VvDnaJ05 and VvDnaJ26 may play an essential role when the grape plants were exposed to salt, shade and hot stresses, especially in heat stress, the expression level of both under heat stress (35 ℃) was significantly increased in 6 h after the treatment (P<0.05). Transient expression results showed that the VvDnaJ05 and VvDnaJ26 were able to improve the heat tolerance ability of the tobacco (Nicotiana tabacum), and the malondialdehyde and H₂O₂ significantly decreased (P<0.05) compared with that of the control. The subcellular localization results showed that the VvDnaJ05 was localized in the cytoplasm and nuclear, and the VvDnaJ26 was localized in the cytoplasm. Additionally, the VvDnaJ05 and VvDnaJ26 did not present interaction relationship. The results above indicated that VvDnaJ05 and VvDnaJ26 respectively exert their biological functions and play a key role in stress resistance. The findings of this study could provide a theoretical foundation for the further investigation on the protein functions of the VvDnaJs and breeding of new grape varieties resisted stresses.

Melatonin is widely involved in the regulation of plant stress response, however, the function and regulatory mechanism of melatonin in the sprout tumble of Pinellia ternata under high-temperature are unclear. In this study, the effect of melatonin treatment on the rate of sprout tumble in P. ternata were compared and the corresponding genomic methylation variation in P. ternata were analyzed based on methylation-sensitive amplification polymorphism (MSAP). The results showed that both 50 and 100 μmol/L melatonin significantly reduced the sprout tumble rate of P. ternata. And the effect of 50 μmol/L melatonin treatment on the methylation variation of the P. ternata genome was analyzed. The total methylation rate of the control group was 56.97% and 45.38% at 0 and 6 h high temperature stress, respectively, and the methylation rate of 50 μmol/L melatonin-treated group was 52.11% at 6 h high temperature stress, indicated that melatonin treatment generally delayed the high temperature stress-induced demethylation of the genome. The analysis of differential methylation bands revealed that melatonin treatment mainly affected the modification of methylation in the mitochondrial and chloroplast genomes of P. ternata. Additionally, P. ternata catalase 2 (PtCAT2) was demethylated under melatonin treatment, and the gene expression was up-regulated, indicated that melatonin could regulate the accumulation of reactive oxygen species through methylation modification, and then participated in the regulation of sprout tumble in P. ternata under high temperature stress. This study explored the function of melatonin on the sprout tumble of P. ternata under high-temperature, and analyzed its possible regulatory pathways from an epigenetic perspective, providing a reference for the study of the regulation mechanism of the sprout tumble of P. ternata.

SPL (SQUAMOSA promoter binding protein (SBP)-like) is a plant specific transcription factor, and plays an important role in plant growth, development and metabolism. In this study, SPL family members were screened from the transcriptome data of Pinus massoniana obtained by the previous research. Through bioinformatics methods, the protein physical and chemical properties, subcellular localization prediction, conserved protein motif analysis, and phylogenetic tree analysis of PmSPL gene family were carried out. The results showed that 11 PmSPL family members were identified. All the family members were predicted to locate in the nucleus. The encoded proteins were composed of 190~1 147 amino acids, and the relative molecular mass were 21.6~127.1 kD. All the sequences contained motif1~3, which were conservative sequences of SBP domain. All the 11 PmSPLs contained 2 C2HC zinc finger structures and 1 nuclear localization signal (NLS). The response of PmSPL gene family to low phosphorus stress was detected by qRT-PCR. The results showed that the expression of PmSPL3 and PmSPL8 did not change significantly in the aboveground and underground parts of P. massoniana seedlings treated with low phosphorus at different times, which indicated that they did not respond to low phosphorus stress; PmSPL4 and PmSPL6 expression were significantly up-regulated in both aboveground and underground parts after 12 d of low phosphorus treatment, which might play a role in response to low phosphorus stress. This study provides a reference for further revealing the biological function of PmSPL gene family in response to low phosphorus stress.

Yak (Bos mutus) is an important genetic resource to the plateau region. To study the structure and function of the platelet-derived growth factor D (PDGFD) gene in yaks, and explore the PDGFD gene expression in different tissues at 2 age stages, this study cloned the complete PDGFD gene CDS sequence by using subcutaneous adipose tissue cDNA of 36-month-old Datong yak as the template. Biological software was used for analyzing the PDGFD gene nucleotide sequence and physical and chemical properties of protein. The relative expression of PDGFD in 7 tissues (heart, liver, spleen, lungs, kidney, longissimus muscle and adipose tissue) of embryonic and 36-months old yak was carried out by qPCR. The results showed that the CDS region of PDGFD gene was 1 095bp and encoded 364 amino acids. The most closely genetic relationship of Datong yak was the B. mutus (99.8%), and the most distant was the Sus scrofa (91.2%). The protein analysis of PDGFD showed that PDGFD contains 1 CUB domain of 116 aa, 1 PDGF domain of 93 aa, and no transmembrane structure. PDGFD was predicted to be a stably hydrophilic secretory protein. The PDGFD protein advanced structure was consisted of random coil (48.21%), α-helix (25.62%), extended chain (21.49%), and β-turn (4.68%). In addition, PDGFD mainly interacted with platelet-derived growth factor protein and vascular endothelial growth factor protein. qPCR results showed that PDGFD mRNA was expressed in each tissue at 2 stages. By comparing the 2 periods, PDGFD gene expression was maintained at a high level in the heart. Meanwhile, the expression in adipose tissue changed the most. The mRNA expression level of PDGFD extremely significantly increased in adipose tissue with the growth and development of yaks (P<0.001). The result showed that PDGFD may be closely related to yak's heart, and affected adipose growth and development. This study offers a theoretical foundation for the regulation mechanism of yak fat deposition and provides references for the genetic breeding of yaks.

Leptin (leptin, LP) and its receptor leptin receptor (LEPR) play a key role in food intake and energy consumption control. The role of LP and LEPR in the growth of skin and hair follicles has been gradually proved. In order to explore the distribution characteristics of LP and LEPR in different parts of skin of young yak (Bos grunniens), and their possible roles in different parts of skin, 10 healthy young yaks were selected to collect back, abdomen, axilla and scrotum skin. The exact distribution and expression of LP and LEPR in the skin of back, abdomen, axilla and scrotum of young yaks were studied by qPCR, Western blot and immunohistochemical methods. The results of immunohistochemistry showed that the expression of LP and LEPR was basically the same, mainly distributed in the epidermis, outer root sheath of hair follicles, sweat glands, sebaceous glands, capillary endothelial cells and dermal fibroblasts. The average optical density results showed that the expression levels of LP and LEPR in abdomen and back were significantly higher than those in axilla and scrotum (P<0.05). The qPCR results showed that LP expression in the back and abdomen was significantly higher than that in the axilla and scrotum (P<0.05). The expression of LEPR in the back was significantly higher than that in the other 3 parts (P<0.05); There was no significant difference in abdominal, axilla and scrotum (P>0.05). Western blot results showed that LP expression in abdomen and back was significantly higher than that in axilla and scrotum, and the differences among different parts were significant (P<0.05). The expression of LEPR in the back and abdomen was significantly higher than that in the scrotum and axilla, and the difference was significant among different parts (P<0.05). In this study, the expression of LP and LEPR was detected in young yak skin. The expression of LP and LEPR was basically the same in different parts of skin, mainly in epidermis, outer root sheath of hair follicles, sebaceous glands, sweat glands, capillary endothelial cells and dermal inner layer fibroblasts, but the expression were significant differences in different parts of skin. The results suggest that LP and LEPR may play an important role in the growth of skin and hair follicles. This study provides some basic data for further exploring the regulation mechanism of yak skin and hair follicle growth in plateau environment.

Yes-associated protein (YAP) and TEA domain transcription factor 1 (TEAD1) are downstream transcriptional co-activators of the Hippo signaling pathway that play important roles in female reproduction. Thus, in order to investigate the relationship between YAP and TEAD1 and corpora lutea (CL) formation, development and degeneration in yaks (Bos grunniens), the female yak corpus luteum (CL) tissues were collected, and divided into 5 groups according to the process of corpus luteum formation (corpus rubrum stage and lacunose luteum stage), development (pregnancy luteal phase), and degeneration (degenerative luteal phase and white corpus stage). The relative expression of YAP and TEAD1 genes and proteins in luteal formation, development and degeneration were examined by qPCR and Western blot, and the localization of YAP and TEAD1 in corpus luteum formation, development and degeneration were analyzed by immunohistochemistry. The results showed that YAP and TEAD1 were abundantly expressed in luteal tissues, and there were significantly difference in different stages of luteal tissues, especially the relative expressions of YAP and TEAD1 were the highest in the lacunose luteum stage and significantly difference from the other stages (P<0.05). However, YAP and TEAD1 expressions were the lowest in the degenerated luteal tissues. Immunohistochemical results showed that YAP and TEAD1 were expressed mainly in yak granulosa luteal cells, vascular endothelial cells and fibroblasts. The results suggested that the Hippo signaling pathway mediated by YAP and TEAD1 was involved in the formation, development and function of luteal tissue in female yaks by stimulating the proliferation of luteal cells, and was associated with angiogenesis during this process. The results of this study provide a reference for further exploration the role played by Hippo pathway in the regulation of ovarian function and maintenance of pregnancy in yaks.

Granulosa cells are the key factor affecting the development of ovarian follicles. miRNA affects the ovarian follicular development by controlling the proliferation and apoptosis of follicular granulosa cells. This study aimed to reveal the role of novel-miR-23900 in sheep (Ovis aries) follicular granulosa cells. First, isolated and cultured sheep follicular granulosa cells were treated with follicle-stimulating hormone (FSH), qPCR results showed that the expression levels of CDK2, CDK4, CCND2 and Bcl2 genes significantly increased (P<0.01), the Bax and Casp3 gene expression and novel-miR-23900 level significantly decreased (P<0.01). Then, novel-miR-23900 mimic and mimic negative control (NC) were further transfected into granulosa cells, CCK-8 was used to detect cell proliferation and mitochondrial membrane potential detection kit (JC-1) was used to detect cell apoptosis, and qPCR was used to detect the expression changes of genes related to proliferation and apoptosis. The results showed that the proliferation ability of granulosa cells significantly decreased at 72 and 96 h with transfection of novel-miR-23900 mimic (P<0.05), and extremely significantly decreased (P<0.01) at 48 h. The expression of proliferation-related genes CDK1, CDK4, CCNB1 and CCNB2 extremely significantly decreased (P<0.01), and the expression of CDK2, CCND1 and CCND2 significantly decreased (P<0.05). Apoptosis of granulosa cells was promoted by 48 h transfection with novel-miR-23900 mimic, the expression of apoptosis-related gene Bcl2 significantly decreased (P<0.01), the Casp3 gene expression significantly increased (P<0.01), and Bax gene expression significantly increased (P<0.05). In conclusion, novel-miR-23900 inhibited proliferation and promoted apoptosis in sheep granulosa cells. This study provides basic data for exploring the effects of novel-miR-23900 on sheep granulosa cells and its role in ovary.

Proteomic changes in the semen of dairy sheep (Ovis aries) during cryopreservation may lead to sperm death or fertility decline. In order to explore the differential proteins between fresh and frozen ram semen, the semen of 18 Donghu F₁ dairy sheep was collected by artificial vagina method, and frozen and preserved with glycerol and egg yolk as the main cryoprotectant. The total protein of fresh and frozen ram semen were extracted, purified, compared and analyzed by tandem mass tag (TMT) technology. The results showed that 936 proteins were screened out before and after freezing. Taking fold change (FC)>1.1 and P<0.05 as screening conditions, there were 22 differential proteins between fresh and frozen thawed semen, of which 17 proteins were up-regulated and 5 proteins were down regulated, and all of them were involved in the regulation of sperm metabolism and biological processes. GO enrichment analysis showed that 8 differential proteins were involved in the biological process and cell components. KEGG was used to analyze the signal pathways in which the differentially expressed proteins were significantly enriched. It was found that the differentially expressed proteins were mainly enriched in metabolism, protein metabolism, apoptosis and other related pathways. Two up-regulated and 2 down-regulated proteins were selected to detect their abundances in fresh and frozen semen by Western blot. The results showed that the relative abundance of the 4 proteins were similar to the results of TMT proteomics analysis, indicating that the proteomics data were reliable. This study provides basic data for screening of protein markers of semen freezing injury in Donghu F₁ dairy sheep.

Sequestosome 1 receptor protein (Sqstm1) is a selective autophagic adaptor protein in Nile tilapia (Oreochromis niloticus), which selectively participates in the removal of intracellular damaged organelles and proteins. To analyse the function of sqstm1 gene, and expression distribution of this gene in healthy Nile tilapia tissues and its expression pattern in the process of resisting bacterium and virus. In this study, the ORF of the gene On-sqstm1 (GenBank No. XP_005463852) was cloned, and the characteristics of bioinformatics and the subcellular localization were analysed, its expression patterns in healthy Nile tilapia tissues and after stimulation with different stimulant were analyzed by qPCR. The effect on nuclear factor kappa-B (NF-κB）signal path of On-sqstm1 was examined by using the dual luciferase reporting system. The results revealed that the ORF of the gene On-sqstm1 was successfully cloned and identified. which was 1 287 bp in length and encoded 428 amino acids. The results of subcellular localization showed that On-Sqstm1 was localized in the cytoplasm, the qPCR results revealed that On-sqstm1 was expressed in all tissues of tilapia and the highest in the liver. After challenged with the inactivated Streptococcus agalctiae and Poly I: C virus analogues, the expressions of brain, head kidney, liver, intestine and spleen significantly increased (P<0.05). The results of dual luciferase reporting system revealed that On-Sqstm1 significantly activated the NF-κB signal pathway activity.The above results indicated that On-Sqstm1 protein may participate in the immune response to pathogen infection by mediating NF-κB signaling pathway activity. This study provides a reference for further research on the mechanism of On-Sqstm1.

Methyl jasmonate (MeJA) can induce the biosynthesis of Ganoderma terpenes, in order to explore the relationship between Ganoderma terpene biosynthesis and the regulation of basic helix-loop-helix (bHLH) genes and jasmonic acid (JA) signal regulation. In this study, the bHLH gene fragment related to the biosynthesis of Ganoderma terpenoids was cloned from G. lingzhi by homologous alignment. The full length of the gene was further cloned and analyzed its bioinformatics, transcriptional activity, prokaryotic expression, subcellular localization, overexpression and silencing expression. The results showed that a bHLH gene fragment with high homology with the positively regulated tanshinone synthesis transcription factor was obtained from G. lingzhi. qRT-PCR analysis showed that the gene was significantly indigenous in response to MeJA induction. The expression levels of the 3 key genes in the triterpenoid (GA) synthesis pathway of G. lingzhi were significantly increased under MeJA induction (P<0.05), and the expression pattern was consistented with bHLH. A 1 311 bp GlbHLH1 gene (GenBank No. MW981280.1) was cloned, encoding 436 amino acids with a molecular weight of 46.33 kD. Promoter analysis showed that G-box binding to bHLH transcription factor existed in the promoter regions of the 3 key genes in Ganoderma triterpene synthesis pathway, suggested that GlbHLH1 might be involved in the regulation of Ganoderma triterpene metabolism by responding to JA signal. Phylogenetic analysis showed that GlbHLH1 had the closest relationship with GsPIL37177.1 of G.sinense; the transcriptional activation activity in Saccharomyces cerevisiae showed that the gene had transcriptional activation activity. The prokaryotic expression vector of GlbHLH1 gene was successfully constructed, and the fusion protein of expected size (about 75 kD) was expressed in Escherichia coli BL21. Subcellular localization showed that GlbHLH1 was mainly located in the nucleus; the transgenic results showed that the expressions of GlbHLH1 in the overexpression and silencing strains were significantly up- and down-regulated from that in the wild type (P<0.05), the triterpenoid content in the overexpression strains OE-GlbHLH1-1, OE-GlbHLH1-2 and OE-GlbHLH1-3 increased by 25%, 22% and 38%, respectively, and that in the silencing strains Si-GlbHLH1-1 and Si-GlbHLH1-2 was decreased by about 15% and 30%, respectively. In summary, GlbHLH1 is an important transcription factor that controls the synthesis of triterpenoids in G. lingzhi. The above results provide a theoretical basis for further study on the molecular regulation mechanism of bHLH in G. lingzhi.

Regulator of glycerol channel 2 (RGC2) is a key factor which is down stream in high osmolarity glycerol-mitogen-activated protein kinase (HOG-MAPK) cascade pathway involved in regulation of hyperosmotic stress reaction. In Saccharomyces cerevisiae, ScRGC2 was found to be involved in regulating the concentration of glycerol in the cells and appropriate glycerol content can help cells protect from outside damage. To identify structure characteristic and expression pattern of StRGC2 during its key growth and development periods and under hypertonic stress in Setosphaeria turcica, StRGC2 (GenBank No. XP_008024338) was cloned. The structural analysis based on the gene and its coding protein showed that the gene contained 4 exons and 3 introns, the full length of DNA was 1 857 bp and the full length of cDNA was 1 389 bp. The C-terminal of its coded protein contained a PH conserved domain and was located in the nucleus. The expression patterns of StRGC2 gene at different developmental stages (hypha, conidia, germ tube, invasive nail, appressorium formation) were analyzed in virtue of RNA-seq data obtained in this previous research and the highest expression level appeared in appressorium development stage. Prediction of the promoter element showed that there were various abiotic stress-induced element. Among them, stress-induced element related to hypertonic stress were found. Further, the expression pattern of StRGC2 gene under hypertonic stress showed that the expression level of StRGC2 gene increased sharply under 0.4 mol/L NaCl for 3 h compared to the control, meaning StRGC2 involved in the regulation of hypertonic stress response in S. turcica. This study not only clarified the structural characteristics and expression pattern of StRGC2 gene, but also laid foundation for the function research of StRGC2 gene and its molecular mechanism in the pathogenic process of S. turcica.

Northern corn leaf blight (NCLB) is a crucial foliar fungal disease that seriously affects corn (Zea mays) yield and quality. To determine the genetic diversity and genetic structure of Setosphaeria turcica isolates with different sensitivities to pyraclostrobin in Fujian Province, the sensitivity of S. turcica isolates to pyraclostrobin in Fujian Province was evaluated using the method of measuring the colony growth inhibition on the fungicide-amended plate, and genetic diversity and genetic structure of the S. turcica isolates with different sensitivities to pyraclostrobin was analyzed using inter-simple sequence repeats (ISSR) markers in this study. The results showed that the range of effective concentration for 50% colonial inhibition (EC₅₀) values of 62 S. turcica isolates to pyraclostrobin in Fujian Province was 0.002 1~1.288 4 μg/mL, with the mean value of (0.127 4±0.254 5) μg/mL. The frequency distribution curve of pyraclostrobin was continuous and unimodal, and disobeyed the normal distribution (W=0.742 2, P=0.001 0<0.05). The ranges of EC₅₀ value for S. turcica isolates with pyraclostrobin-sensitivity, -medium and -insensitivity were 0.002 1~0.011 2, 0.020 6~0.100 0 and 0.101 9~1.288 4 μg/mL, respectively. The resistant ratios of pyraclostrobin-insensitive isolates ranged from 3.01 to 38.01. Analysis of variance revealed that the mean EC₅₀ value of pyraclostrobin-insensitive isolates was significant difference compared with those of isolates with pyraclostrobin-medium and -sensitivity, respectively (P<0.05). In conclusion, a subpopulation of S. turcica with decreased sensitivity to pyraclostrobin has been developed in Fujian Province. Genetic diversity analysis indicated that a total of 66 loci were amplified using 11 ISSR primers, and the percentage of polymorphic loci (P_L) reached as high as 98.48%. The lowest DNA polymorphism was detected from pyraclostrobin-sensitive population (P_L=60.61%), whereas the most abundant polymorphism was detected from those in pyraclostrobin-insensitive population (P_L=68.18%). The index values of genetic diversity for the insensitive population were higher than those of the sensitive population, suggesting that genetic diversity in the insensitive population of S. turcica was more diverse than those of sensitive and mediate populations. The results of genetic differentiation (Φ_PT) and gene flow (Nm) analysis indicated that low genetic differentiation (Φ_PT<0.054, P>0.37) with frequent gene exchange (Nm>8.60) were detected among pyraclostrobin-sensitive, -mediate and -insensitive populations. Clustering analysis also indicated that S. turcica isolates with different sensitivities to pyraclostrobin were randomly clustered in the same branch, suggesting no obvious correlation between genetic differentiation and the level of fungicide resistance. Analysis of molecular variance (AMOVA) revealed that the all source of genetic variation in Fujian S. turcica populations with different sensitivities to pyraclostrobin was derived from within populations. Principal coordinates (PCoA) and population genetic structure analysis indicated that the 3 S. turcica populations with different sensitivities to pyraclostrobin could be divided into 2 genetic groups, with a high similar genetic structure being observed among the 3 different populations. The results from this study provide a reference for monitoring of fungicide resistance of S. turcica in the field and for ecological regulation of northern corn leaf blight by using resistant cultivars.

Plectropomus leopardus is a kind of newly farmed fish with high economic value. In recent years, skin ulcer disease occurs in both juvenile and adult fish, resulting in a large number of deaths and great economic losses. It is important to identify the main pathogenic bacteria for disease-resistant breeding. In this study, the diseased fish and normal fish were collected from Hainan, and the skin microbial sequencing and analysis were carried out on 3 generation high-throughput sequencing method. The results showed that there were 403 unique operational taxonomic units (OTUs) in the normal group and 72 unique OTUs in the ulcerated group. The number of OTUs shared by the two groups was 74. In the normal group, there were 16 phyla, 30 classes, 63 orders, 96 families, 145 genera and 168 species were identified on average, while 9 phyla, 12 classes, 27 orders, 38 families, 55 genera and 67 species were identified on average in the diseased fish skin. The difference between two groups indicated that pathogenic bacteria inhibited the growth of normal bacteria, resulting in a significant decrease in the diversity of bacteria structure. There were great differences in dominant bacterium composition between normal group and diseased group. Proteobacteria and Firmicutes were dominant phyla in normal group, while Bacteroidetes and Epsilonbacteraeota were dominant phyla in the diseased fish. ANOVA analysis of variance showed that the content of Vibrio in diseased fish skin was significantly increased, and Vibrio harveyi, V. tubiashii and a specie of uncultured Vibrio were annotated in KRONA analysis. Based on above research, a strain of V. Harveyi was isolated and identified from the ulcer skin of P. leopardus, named as V. harveyi stain Dx21. After the artificial infection with the intraperitoneal injection, the result showed that the V. harveyi stain Dx21 could cause surface ulceration and death in juvenile starfish, which was similar to natural infection. The half-lethal dose (LD₅₀) was 1.0×10⁵ cfu/mL for P. leopardus fries weighing 100 g. This study revealed the flora characteristics of the skin of diseased P. leopardus, suggesting that V. harveyi is one of the main pathogens. This study has certain guiding significance for the skin ulcer disease prevention and control of P. leopardus and the development of vaccine.

Volatile compounds are major impact on total fruit flavor quality, and are critical for promoting consumer consumption and market competitiveness. Fruit aroma is a complex mixture of a large number of volatile compounds whose composition is specific to species, and is determined not only by genetics but also by environmental factors, cultural practices and postharvest handling. Thus, control the synthesis of fruit volatiles from the molecular level is of great significance. Based on their biosynthetic origin, fruit aroma volatiles can be divided into several classes, including terpenoids, fatty acid derivatives and amino acid derivatives, and serve as cues that facilitate a variety of biological functions. Recently, depend on the development of molecular biology technology, the biosynthetic pathways, biosynthesis-related enzymes, transcription factors, genes and important regulatory mechanism were revealed, which enriched the content of breeding aspect. In this paper, the composition, important biological functions and main biosynthesis pathways of fruit volatiles were introduced comprehensively, and the role of some related enzymes, genes, treatments in fruit aroma synthesis were summarized. This review provides scientific theoretical support for fruit aroma quality research, and scientific basis for improving fruit flavor quality based on molecular breeding technology.

The guanine nucleotide-binding protein q polypeptide (GNAQ) is a member of the α subunit encoding of the q-class G protein and is vital in cell signaling. Studies have shown that GNAQ plays an important role in multiple reproductive-related tissues involved in mammalian reproductive regulation. The present review is concerned with the discovery and mechanism of action of GNAQ, the research progress of GNAQ in mammalian hypothalamic hormone secretion, uterine function, ovarian physiology, male testicular development, and spermatogenesis. This review provides a reference for further exploring the mechanism of GNAQ in mammalian reproduction.

Chicken (Gallus gallus) primordial germ cells (PGCs) have a wide application prospect in the fields of transgenic animal preparation and germplasm resources protection. In order to analyze the molecular mechanism of chicken PGCs formation, a key long noncoding RNA-bone morphogenetic protein 4 (LncBMP4) that regulates chicken PGCs formation was identified earlier, and it can encode a small peptide, named EPC5. In this study, the chemical structure and subcellular localization of EPC5 (expression in primordial germ cells chromosome 5) peptides were analyzed by online prediction software. The prokaryotic fusion expression vector of PET-EPC5-His-B2M was constructed by chemical synthesis and transformed into the BL2 competent state of Escherichia coli. After induction by isopropyl-beta-D-thiogalactopyranoside (IPTG), EPC5 antigen was expressed and purified. The purified antigen was used for animal immunization and antiserum (G1480, G1481) was collected. The titer of antiserum was detected by ELISA. The antibody was purified and identified by Western blot. The results indicated that EPC5 mainly localized in cells and had RNA polymerase subunits. The PET-EPC5-His-B2M vector was successfully constructed, and the size of the recombinant protein was about 25 kD after induction. ELISA showed that the titer of G1480 antiserum was 1:5.12×10⁵ (OD_antiserum/OD_{pre-immune blood}≥2.1), and that of G1481 antiserum was 1:1.024×10⁶ (OD_antiserum/OD_{pre-immune blood}≥2.1). After purification of G1480 antiserum, the concentration of EPC5 polyclonal antibody was 10 mg/mL, and the titer was 9.8 ng/mL (OD value>0.2) by ELISA. The results of Western blot showed that the prepared polyclonal antibody could specifically bind to EPC5 protein expressed in vitro and in vivo. In conclusion, the polyclonal antibody against EPC5 was successfully prepared in this study, which laid a foundation for further study on the function and mechanism of EPC5 in the formation of PGCs.

Shallot latent virus (SLV) is one of the major viruses that damage shallot (Allium cepa var. aggregatum) and other Allium crops and prevails in many Allium-growing regions. To develop a SYBR Green Ⅰ real-time fluorescent RT-PCR assay for the sensitive detection of SLV, a pair of specific primers were designed and synthesized based on the conserved region of SLV coat protein (CP) gene registered in GenBank and a series of optimization including primers concentration and annealing temperature were performed. Ct values were linear with the logarithm of the template concentration from standard curve of cDNA. The amplification efficiency was 94.529% and the correlation coefficient was 0.999 6. There was no crossing reaction with Shallot yellow stripe virus (SYSV), Onion yellow dwarf virus (OYDV) and Shallot virus X (SVX). The lowest detection limit was 10^-7 dilution fold, which was 1 000 times higher than that of routine RT-PCR. The coefficients of variation of intra-assay and inter-assay were 0.02%~0.63% and 0.03%~1.24%, respectively, indicating the excellent stability of the method. The SYBR Green Ⅰ real-time fluorescent RT-PCR was used to determine the content of SLV in shallot leaves and bulbs and to detect 50 samples from suspected infectious shallot. The results showed that the content of SLV in leaves was significantly higher than that in bulbs, and 46 out of 50 samples were detected to be positive by this assay, and the detection rate was 6% higher than that of conventional RT-PCR. Therefore, the SYBR Green Ⅰ real-time fluorescent RT-PCR method for detection of SLV provides technical support for the rapid detection of SLV on shallot.