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Petunia hybrida is commonly cultivated for ornamental use in urban parks greening and street embellishment in China. In March 2022, 60% of P. hybrida plants cv. Wave Purple (n≈1800) from an ornamental plant nursery under natural conditions in Tianhe district (N 113°21'21", E 23°9'3.5"), Guangzhou, Guangdong Province, China, were affected with soft rot disease. The distribution of the disease was generally uniform. Infected plants initially exhibit small water-soaked lesions at the base of the stem, which then extended to the leaves. Eventually the diseased plant collapsed and died. Nine diseased plants were collected, and affected tissues cut into small fragments (5 × 5 mm), which were disinfested in 75% ethanol (30 s) and 2% sodium hypochlorite (60 s), followed by three rinses with sterile distilled water. The sterilized sections were macerated in 200 μl sterile water, and streaked on Luria-Bertani (LB) agar medium and incubated at 28°C for 48 h. Single colonies were restreaked three times to obtain purified isolation. Sixteen bacterial strains with similar morphology were isolated, and their colonies were yellowish white, round, and convex with smooth surfaces on LB agar plate. The representative strain BDQ1 was selected for further analyses and the 16S rDNA gene (GenBank Accession ON982467) were amplified using primer pair 27F/1492R, revealed above 99% sequence identity with some Pectobacterium brasiliense isolates (GenBank Accession Nos. CP046380(1421/1422), MN393966(1419/1422), and CP020350(1419/1422)) using BLASTn. A multilocus phylogenetic analysis by neighbor-joining method (1,000 bootstrap values) based on six housekeeping gene sequences of gyrA (GenBank Accession No. ON995454), icdA (ON995455), mdh (ON995456), mtlD (ON995457), proA (ON995458), and rpoS genes (ON995459) (Ma et al. 2007; Waleron et al., 2008). The results of phylogenetic analysis showed BDQ1 strain belong to the P. brasiliense clade. Pathogenicity tests were performed on ten healthy P. hybrida cv. Wave Purple plants by injecting 10 µl of bacterial suspensions of BDQ1 (108 CFU/ml) into the stems; another 10 healthy control plants were injected with 10 µl of sterile water. All plants were grown at 25-30°C and 60% humidity in natural light/dark cycle. After 3 d, all inoculated plants showed soft rot symptoms resembling to those observed in the nursery, while control plants remained healthy. Bacteria were successfully reisolated from the symptomatic tissues and identified to be P. brasiliense by PCR mentioned above. P. brasiliense is considered a very aggressive pathogen, which has been reported in Eurasia and Africa (Oulghazi et al. 2021). To our knowledge, this is the first report of P. brasiliense causing bacterial soft rot on P. hybrida in China. This pathogen may pose threat to P. hybrida production in area with warmand humid climate in China. The current study expands the known host range of P. brasiliense and helped raise attention on controlling pathogen spread.

Background:Testicular aging is known to cause male age-related fertility decline and hypogonadism, but the underlying molecular mechanisms remain unclear.Methods:We survey the single-cell transcriptomic landscape of testes from young and old men and examine age-related changes in germline and somatic niche cells.Results:In-depth evaluation of the gene expression dynamics of germline cells reveals that disturbance of base-excision repair pathway is a major feature of aging spermatogonial stem cells (SSCs), suggesting that defective DNA repair of SSCs may serve as a potential driver for increased de novo germline mutations with age. Further analysis of aging-associated transcriptional changes shows that stress-related changes and apoptotic signaling pathway accumulate in aged somatic cells. We identify age-related impairment of redox homeostasis in aged Leydig cells and find that pharmacological treatment with antioxidants alleviate this cellular dysfunction of Leydig cells and promote testosterone production. Lastly, our results reveal that decreased pleiotrophin (PTN) signaling is a contributing factor for testicular aging.Conclusions:These findings provide a comprehensive understanding of the cell-type-specific mechanisms underlying human testicular aging at a single-cell resolution, and suggest potential therapeutic targets that may be leveraged to address age-related male fertility decline and hypogonadism.Funding:This work was supported by the National Key Research and Development Program of China (2018YFA0107200, 2018YFA0801404), the National Natural Science Foundation of China (32130046, 82171564, 82101669, 81871110, 81971759), the Key Research and Development Program of Guangdong Province (2019B020234001), the Natural Science Foundation of Guangdong Province, China (2022A1515010371), the Major Project of Medical Science and Technology Development Research Center of National Health Planning Commission, China (HDSL202001000), the Open Project of NHC Key Laboratory of Male Reproduction and Genetics (Family Planning Research Institute of Guangdong Province) (KF202001), the Guangdong Province Regional Joint Fund-Youth Fund Project (2021A1515110921), the China Postdoctoral Science Foundation (2021M703736).

Using high throughput method of 3D printing, 20 Inconel 718 alloy samples with different Nb contents (0∼10 wt.%) are conveniently prepared in a single printing process, and then analyzed the microscopic morphology and hardness of these samples. The results show that with the increase of Nb content in the alloy, the content of Laves phase in the alloy gradually increases, and the Laves phase particles also become larger, resulting in a gradual increase in hardness. In conclusion, the new technology breaks disadvantages of traditional new material development such as high cost, long time consuming, single trial and error, and provides the possibility for rapid and accurate development of new materials.

Bacterial soft rot of banana, caused by

The elderly males undergo degenerative fertility and testicular endocrine function that jeopardize the reproductive health and well-being. However, the mechanisms underlying reproductive aging are unclear. Here, we tried to address this by investigating the phenotypes and transcriptomes of seven regions of the male mouse reproductive tract: the testis, efferent ductules, initial segment, caput, corpus and cauda epididymidis, and vas deferens, in adult (3 months) and aged (21 months) mice. Quantitative PCR, immunohistochemistry, immunofluorescent staining, and enzyme-linked immunosorbent assay were performed for the analysis of gene expression in mice, human tissues, and semen samples. Aged male mice showed both systematic and reproductive changes, and remarkable histological changes were detected in the testis and proximal epididymis. Transcriptomes of the male reproductive tract were mapped, and a series of region-specific genes were identified and validated in mouse and/or human tissues, including Protamine 1 (Prm2), ADAM metallopeptidase domain 28 (Adam28), Ribonuclease A family member 13 (Rnase13), WAP four-disulfide core domain 13 (Wfdc13), and Wfdc9. Meanwhile, age-related transcriptome changes of different regions of the male reproductive tract were characterized. Notably, increased immune response was functionally related to the male reproductive aging, especially the T cell activation. An immune response-associated factor, phospholipase A2 group IID (Pla2g2d), was identified as a potential biomarker for reproductive aging in mice. And the PLA2G2D level in human seminal plasma surged at approximately 35 years of age. Furthermore, we highlighted Protein tyrosine phosphatase receptor type C (Ptprc), Lymphocyte protein tyrosine kinase (Lck), Microtubule associated protein tau (Mapt), and Interferon induced protein with tetratricopeptide repeats 3 (Ifit3) as critical molecules in the aging of initial segment, caput, caput, and cauda epididymidis, respectively. This study provides an RNA-seq resource for the male reproductive system during aging in mice, and is expected to improve our understanding of male reproductive aging and infertility.

Gladiolus (

We aimed to compare intracavernosal injection (ICI), tail vein injection (IV), and periprostatic injection (PPI) of adipose-derived stem cells (ADSCs) for their ability to improve erectile function in cavernous nerve injury-induced erectile dysfunction (CNIED) rats and to explore the possible mechanism. Eighty-four male SD rats were divided into the sham group (n = 6), BCNI group (bilateral CN crush injury, n = 6), PBS-ICI group (n = 6), PBS-IV group (n = 6), PBS-PPI group (n = 6), ADSC-ICI group (n = 18), ADSC-IV group (n = 18) and ADSC-PPI group (n = 18). ADSCs were labelled with 5-ethynyl-2'-deoxyuridine (EdU), and six rats each in the ADSC-ICI group, ADSC-IV group, and ADSC-PPI group were sacrificed 2, 7, and 28 days after injection. EdU-labelled ADSCs were tracked by immunofluorescence staining. The intracavernosal pressure (ICP)/mean arterial pressure (MAP) ratio, neuronal nitric oxide synthase (nNOS)-positive nerve fibres in the dorsal penile nerve and the smooth muscle/collagen ratio in the cavernosum between groups were also evaluated. ADSCs can significantly improve erectile function through ICI or IV. The two are similar in efficacy and superior to PPI. The mechanism may be that after CN injury, ADSCs are recruited to around the MPG and secrete a variety of neurotrophic factors that promote the repair of the CN, thereby improving erectile function.

In view of engineering design of printability instrument based on its characteristic, a new method based on CDIO is proposed in this paper. Firstly, the functions of the printability instrument are discussed, and the overall layout of the instrument is designed according to the function to be achieved, and then the parts and assembly are designed to complete the above functions. Finally, the functions of the designed equipment are tested. Using the new approach, the designer could complete a design of a compounded product consisted of different parts easily and logically.

Dickeya zeae is a globally important bacterial pathogen that has been reported to cause severe soft rot diseases in several essential food crops, including bananas, rice, maize, and potatoes. Carvacrol, a hydrophobic terpene component, is found in aromatic plants of the Labiatae family and various essential oils. However, little work has been done on its antimicrobial potential against D. zeae. This study aimed to evaluate the antimicrobial activity and the functional mechanism of carvacrol against D. zeae. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of carvacrol against D. zeae were 0.1 mg/mL and 0.2 mg/mL, respectively. Carvacrol affected the cell membrane of D. zeae, as revealed by decreased intracellular ATP concentration, nucleic acid leakage, and decreased membrane potential. Scanning electron microscopy (SEM) micrographs confirmed that D. zeae cell membranes were damaged by carvacrol. Furthermore, a significant inhibition of D. zeae swimming motility and biofilm formation was observed following treatments with carvacrol at sub-inhibitory concentrations, indicating a significantly negative effect on these virulence factors. Accordingly, the tissue infection test revealed that carvacrol significantly reduced the pathogenicity of D. zeae. In a pot experiment, inoculated banana seedlings displayed remarkably lesser disease symptoms following treatment with carvacrol, and the control efficiency for banana soft rot was 32.0% at 14 days post-inoculation. To summarize, carvacrol exhibits strong antimicrobial activity against D. zeae and great potential applications in the control of D. zeae-associated crop diseases.

We aimed to evaluate the effects of intratunical injection of exosomes derived from human urine-derived stem cells (USC-exo) on plaque formation and erectile function in a transforming growth factor-β1 (TGF-β1) induced Peyronie's disease (PD) rat model. Twenty-four SD rats were randomly assigned equally to three groups: (I) Sham group (50 μl phosphate-buffered saline [PBS] injected into the tunica albuginea [TA]), (II) PD group (0.5 μg TGF-β1 in 50 μl PBS injected into the TA) and (III) USC-exo group (0.5 ug TGF-β1 plus 100 μg USC-exo injected into the TA at the same day). The maximum intracavernous pressure (ICPmax ) and mean arterial pressure (MAP) of each group were evaluated 4 weeks after injection. The plaque formation, fibrosis, matrix metalloproteinases (MMPs) and tissue inhibitor of MMPs (TIMPs) in the TA were evaluated. Four weeks after injection, USC-exo group showed more significantly enhanced ICPmax and ICPmax /MAP than PD group (p < .05). USC-exo could significantly ameliorate the TA fibrosis that could be associated with the inhibition of transdifferentiation of fibroblasts into myofibroblasts, decreased expression of TIMPs (TIMP-1, 2, 3) and increased activity of MMPs (MMP-1, 3, 9) in the TA. According to these findings, USC-exo can be a new candidate for the prevention of PD.

Bacterial soft rot caused by Dickeya zeae MS1 (Erwinia chrysanthemi) is one of the most devastating banana diseases worldwide. However, knowledge of the development and ecological interactions of D. zeae MS1 biofilm is limited. Here, we visualized the development and architecture of D. zeae MS1 biofilm using confocal laser scanning microscopy, and we evaluated the ability of D. zeae MS1 to form biofilms under different environmental conditions (carbon sources, temperatures, pH levels and mineral elements) using a microtiter plate assay. We found that the development of D. zeae MS1 biofilm could be categorized into four phases and that mature biofilm consisted of a highly organized architecture of both bacterial cells and a self-produced matrix of extracellular polysaccharides. Furthermore, sucrose was the most suitable carbon source for supporting the growth of biofilm cells and that 32 °C and pH 7.0 were the most favorable of the temperatures and pH levels examined. Meanwhile, the addition of Ca2+, Fe2+, K+ and Na+ enhanced the formation of biofilm in minimal medium cultures, whereas 2.5 mM Cu2+ and Mn2+ was inhibitory. A better understanding of biofilm formation under different environmental parameters will improve our knowledge of the growth kinetics of D. zeae MS1 biofilm.

MicroRNAs (miRNAs) play an important role in plant growth, development and responses to stresses. Rice blast is one of the most devastating diseases in rice production. However, little is known regarding the effects of miRNAs response to rice blast. Herein, by deep sequencing small RNA from the susceptible line ZhongerRuanzhan and its space-induced blast resistant mutant line H4 under normal conditions and upon Magnaporthe oryzae (M. orzyzae) infection, several known miRNAs were detected and their expression profiles were found to be negatively correlated with their targets. And, a total of 50 novel miRNAs induced by M. oryzae infection were also identified from four small RNA libraries. Moreover, 176 putative targets of 23 novel miRNAs, which are involved in the various functions, were validated by two degradome analysis. Six novel miRNAs were selected for further validation with qRT-PCR analysis and the results showed that their expression levels were associated with blast response. The knowledge obtained in this study will help us understanding the functions of miRNAs and their targets in regulating blast resistance.

Gladiolus (Gladiolus gandavensis Van Houtte) is a perennial plant in the family Iridaceae, which shows sword-shaped leaves and spikes of brilliantly colored irregular flowers arising from corms. It is one of the most important fresh cut flowers and is widely cultivated worldwide, including in China. In September 2020, white pinpoints were first observed on gladiolus leaves in Jiangmen City, Guangdong Province, China. The white spots eventually turned brown. The lesions then developed into oval to circular spots, which were surrounded with an obvious yellow halo. The spots expanded and coalesced, causing leaf blight. These symptoms were observed on approximately 10% of gladiolus plants in fields measuring ca. 70 ha. Symptomatic leaves were sampled from fields, surface sterilized in 75% ethanol for 30 s, submerged in a 2% NaOCl solution for 10 min, and rinsed three times with sterile water. The samples were then cut into pieces (5 × 5 mm) and incubated for 4 d on potato dextrose agar (PDA) at 25°C. A representative fungal colony was subcultured onto new PDA and grown for another 7 d, and its mycelium appeared to be grayish-black and villiform. This strain was named as Cg_TS. Its conidiophores were simple, septate, cylindrical in shape, and moderate brown in color. They occurred singly or in groups. They were straight or slightly flexuous and ranged in size from 57.0 to 80.0 μm × 4.0 to 8.0 μm. Conidia were 3-distoseptate and curved at the third cell from the base. The third cell was swollen to one side and larger than other cells. These conidia ranged in size from 23.5 to 32.0 μm × 11.5 to 16.0 μm. These morphological characteristics were consistent with the description of Curvularia gladioli Boerema & Hamers (Boerema and Hamers 1989). Using primer pair ITS1 and ITS4, PCR was applied to amplify the internal transcribed spacer (ITS) region of rDNA. This sequence (GenBank accession No. MW426196.1) was subjected to BLAST in GenBank, suggesting that it was most similar to C. gladioli sequences, LT631345.1 and HG778987.1, with both of 99.49% of similarity. To fulfill Koch's postulates, healthy two-month-old gladiolus plants were used for pathogenicity testing, and the leaves were wounded by pressing slightly with a pipette tip. Mycelium disks (3 mm diameter) were applied onto wounded leaves of 10 plants. Another 10 healthy plants were inoculated with PDA disks which served as control. Inoculated samples were placed in a greenhouse at 25°C and 90% relative humidity. After 3 d, brown leaf spots appeared on all of pathogen-inoculated leaves. The symptoms were consistent with those initially observed and C. gladioli was re-isolated from the symptomatic tissue. Identification was confirmed by morphological observation and ITS sequencing. Control leaves remained symptomless. The curvularia fungus was firstly reported on gladiolus in Florida in 1947 and spread globally via infected corms (Torre et al. 2015), it was also reported to cause leaf spots on gladiolus in Brazil in 2013 (Torres et al. 2013). Although C. gladioli had been recorded as a Curvularia species occurring in China (Zhang et al. 2006), it was not reported to cause leaf spots on gladiolus in Guangdong Province and elsewhere in China. To our knowledge, this is the first report of Curvularia gladioli causing leaf spots on gladiolus in China. Identification of this pathogen will help develop diagnostic methods for corms and seedlings, and may lead to the development of appropriate chemical management strategies.

Blast resistance and grain quality are major problems in hybrid rice production in China. In this study, two resistance (R) genes, Pi46 and Pita, along with the gene Wxb, which mainly affects rice endosperm amylose content (AC), were introgressed into an elite indica restoring line, R8166, which has little blast resistance and poor grain quality through marker-assisted selection (MAS). Eight improved lines were found to have recurrent genome recovery ratios ranging from 88.68 to 96.23%. Two improved lines, R163 and R167, were selected for subsequent studies. R167, which has the highest recovery ratio (96.23%), showed no significant differences in multiple agronomic traits. In contrast, R163 with the lowest recovery ratio (88.68%) exhibited significant differences in heading date and yield per plant compared with the recurrent parent. At two developmental stages, R163 and R167 had greatly enhanced resistance to blast over the recurrent parent. Similar trends were also observed for agronomic traits and blast resistance in R163- and R167-derived hybrids when compared with the counterparts from R8166. In addition, R163, R167, and their derived hybrids significantly improved the grain quality traits, including amylose content (AC), gel consistency (GC), chalky grain rate (CGR), and degree of endosperm chalkiness (DEC). It confirmed the success of efficiently developing elite restoring lines using MAS in this study.

Plumeria spp. are ornamental trees commonly planted in parks and gardens, and Plumeria rubra cultivars (Frangipani) is most common in Guangdong Province, China. A rust disease of P. rubra was observed on leaves of susceptible plants from August to December 2013. Ten nurseries were surveyed in September 2013, and 91% (220 of 240) of the plumeria plants were infected with rust. Symptoms first appeared as chlorotic spots (about 1 mm in diameter) appearing on adaxial leaf surfaces and then spread to whole leaf, and infection further resulted in leaf necrosis and abscission. Therefore, the ornamental value of diseased trees was greatly diminished. Bright yellow or yellow-orange uredinia were hypophyllous and produced under the epidermis. Urediniospores were catenulate, globose, ovoid or ellipsoid, and sometimes angular in appearance, ranging from 20.0 to 42.0 μm in length by 14.1 to 25.6 μm in width. Their walls were verrucose and 1.3 to 3.2 μm thick. No teliospores were observed. The rust was identified as Coleosporium plumeriae Pat. based on urediniospore morphology (2). Species identity was confirmed with a 1,551-bp sequence (GenBank Accession No. KF879087) of ITS rDNA amplified with rust-specific primers Rust2inv and LR6 (1). The amplicon had a 100% similarity to C. plumeriae (GU145555). Pathogenicity was confirmed by spraying a urediniospores suspension (15,000 spores ml−1) on five plants of P. rubra cultivar. Five leaves of each plant were inoculated and sealed in plastic bags, while five control plants were applied with sterile water. Plants were held at 28°C for 36 h in a dew chamber. All inoculated leaves developed typical rust symptoms with the uredinia appearing after 9 days, no symptoms developed on any control plants. Urediniospores were produced on infected leaves and pathogen identity was confirmed by morphology and re-sequencing of the ITS rDNA. Plumeria rust was first found in Hong Kong (4) and then in Hainan and Yunnan Provinces, China (3). However, this is the first report of plumeria rust in Guangdong Province, China. Frangipani has large, colorful flowers in the summer, and this rapidly spreading disease causes severe damage and affects their aesthetic value in the second half of the year. References: (1) M. C. Aime. Mycoscience 47:112, 2006. (2) N. Patouillard. Bull. Soc. Mycol. Fr. 18:171, 1902. (3) Q. Wang et al. New Dis. Rep. 23, doi:10.5197/j.2044-0588.2011.023.010, 2011. (4) J. Yan et al. Mycosystema 25:327, 2006 (in Chinese).

The aim of this study was to investigate whether intracavernous injection of urine-derived stem cells (USCs) or USCs genetically modified with pigment epithelium-derived factor (PEDF) could protect the erectile function and cavernous structure in a bilateral cavernous nerve injury-induced erectile dysfunction (CNIED) rat model. USCs were cultured from the urine of six healthy male donors. Seventy-five rats were randomly divided into five groups ( n = 15 per group): sham, bilateral cavernous nerve (CN) crush injury (BCNI), USC, USCGFP+, and USCGFP/PEDF+ groups. The sham group received only laparotomy without CN crush injury and intracavernous injection with phosphate-buffered saline (PBS). All of the other groups were subjected to BCNI and intracavernous injection with PBS, USCs, USCsGFP+, or USCsGFP/PEDF+, respectively. The total intracavernous pressure (ICP) and the ratio of ICP to mean arterial pressure (ICP/MAP) were recorded. The penile dorsal nerves, the endothelium, and the smooth muscle were assessed within the penile tissue. The USC and USCGFP/PEDF+ groups displayed more significantly enhanced ICP and ICP/MAP ratio ( p < 0.05) 28 days after cell transplantation. Immunohistochemistry (IHC) and Western blot analysis demonstrated that the protection of erectile function and the cavernous structure by USCsGFP/PEDF+ was associated with an increased number of nNOS-positive fibers within the penile dorsal nerves, improved expression of endothelial markers (CD31 and eNOS) and a smooth muscle marker (smoothelin), an enhanced smooth muscle to collagen ratio, decreased expression of transforming growth factor-β1 (TGF-β1), and decreased cell apoptosis in the cavernous tissue. The paracrine effect of USCs and USCsGFP/PEDF+ prevented the destruction of erectile function and the cavernous structure in the CNIED rat model by nerve protection, thereby improving endothelial cell function, increasing the smooth muscle content, and decreasing fibrosis and cell apoptosis in the cavernous tissue.

Study has been conducted on how, where and why does excessive noise generate on the gear-box of the snowplow, and, measures for improvement have been presented. As well, analysis to the generation mechanism of vibration noise of gearbox and gears has been made, to find out the source of the noise. Next, a precise model of the gearbox, built by using 3D modeling technology, was translated into finite element model to make the modal analysis. Through analyzing characteristics of each vibration mode, positions of the gearbox case on which a vibration is most likely to be generated have been determined. To check the accuracy of theoretical calculation, vibration and noise tests were made to the gearbox. Specific measures for improvement were then put forward by combining results from both finite element analysis and experimental analysis. The comparison results showed that: the improved gearbox case had a significant effect on noise reduction, and the goal of reducing vibration and noise was achieved.

Aims. Cavernosal endothelial dysfunction is one of the factors in developing diabetic erectile dysfunction (DED), but the mechanism of cavernosal endothelial dysfunction is unclear. The present study is aimed at determining the contribution of autophagy in cavernosal endothelial dysfunction of DED rats and explaining the therapeutic effect of urine-derived stem cells (USCs). Methods. After rat corpus cavernosal vascular endothelial cells (CCECs) were isolated and cultured in vitro, CCECs were treated with advanced glycation end products (AGEs) to mimic the diabetic situation. Autophagy flux, proliferation, and apoptosis of CCECs were determined by mRFP-GFP-LC3 adenovirus infection combined with fluorescence observation and western blot analysis. USCs were isolated from the urine of six healthy male donors, and coculture systems of USCs and CCECs were developed to assess the protective effect of USCs for CCECs in vitro. The contribution of autophagy to the cellular damage in CCECs was evaluated by the autophagic inhibitor, 3-methyladenine (3-MA). Then, DED rats were induced by streptozotocin (50 mg/kg) and screened by apomorphine test (100 μg/kg). In DED rats, USCs or PBS as vehicle was administrated by intracavernous injection (n=15 per group), and another 15 normal rats served as normal controls. Four weeks after injection, erectile function was evaluated by measuring the intracavernosal pressure (ICP) and mean arterial pressure (MAP). Cavernosal endothelial function and autophagic activity were examined by western blot, immunofluorescence, and transmission electron microscopy. Results. In vitro, AGE-treated CCECs displayed fewer LC3 puncta formation and expressed less LC3-II, Beclin1, and PCNA but expressed more p62 and cleaved-caspase3 than controls (p<0.05). Coculture of USCs with CCECs demonstrated that USCs were able to protect CCECs from AGE-induced autophagic dysfunction and cellular damage, which could be abolished by 3-MA (p<0.05). DED rats showed lower ratio of ICP/MAP, reduced expression of endothelial markers, and fewer autophagic vacuoles in the cavernosal endothelium when compared with normal rats (p<0.05). Intracavernous injection of USCs improved erectile function and cavernosal endothelial function of DED rats (p<0.05). Most importantly, our data showed that the repaired erectile function and cavernosal endothelial function were the result of restored autophagic activity of the cavernosal endothelium in DED rats (p<0.05). Conclusions. Impaired autophagy is involved in the cavernosal endothelial dysfunction and erectile dysfunction of DED rats. Intracavernous injection of USCs upregulates autophagic activity in the cavernosal endothelium, contributing to ameliorating cavernosal endothelial dysfunction and finally improving the erectile dysfunction induced by diabetes.

Additional file 1: Table S1. The 110 polymorphic markers between the RP and the DPs. Table S2. The information of the remaining 14 markers showing the DPsâ genotype at homozygous or heterozygous state. Table S3. The percentage of parental genome recovery of the 28 BC3F4 plants. Table S4. Performances of main agronomic traits of the tested lines during the early crop season (March to July) of 2016. Figure S1. Genetic background analysis of the seven improved lines by the remaining 14 SSR markers which detected polymorphic between the RP and the BC3F1 sample.

Studies have explored the influence of DNA damage in assisted reproductive technology (ART), but the outcome remains controversial. To determine whether sperm DNA fragmentation index (DFI) has any effect on ART outcomes, we collected detailed data regarding 1,333 IVF cycles performed at our centre, and the data of our retrospective cohort study were extracted for this meta-analysis. We searched PubMed, Web of Science, EMBASE and Google Scholar and performed a systemic review and meta-analysis. Primary meta-analysis of 10 studies comprising 1,785 couples showed that live birth rate was no significantly different between low-DFI group and high-DFI group (p > 0.05). Secondary meta-analysis of 25 studies comprising 3,992 couples showed a higher miscarriage rate in high-DFI group than in low-DFI group (RR=1.57 [1.18, 2.09], p < 0.01). Meta-analysis of eight studies comprising 17,879 embryos revealed a lower good-quality embryo rate (RR=0.65 [0.62, 0.68], p < 0.01). Meta-analysis of 23 studies comprising 6,771 cycles showed that the high-DFI group had a lower clinical pregnancy rate than low-DFI group (RR=0.85 [0.75, 0.96], p < 0.01). Heterogeneity of included studies weakened our conclusions. Our study showed that DFI has adverse effects on ART outcome. More well-designed studies exploring the association between DFI and ART outcome are desired.

Phylogenetic analysis of D. fangzhongdai, D. dadantii and D. solani strains based on the protein sequences of RhsA, RhsB and RhsC in T6SS. (PDF 156 kb)

Marker-assisted selection (MAS) for qualitative traits such as grain quality and resistance to certain diseases has proven to be highly effective. Multiple genes responsible for various quality components and disease resistances can be simultaneously stacked to boost the performance and to lengthen the commercial lifespan of high-yield varieties. Grain quality genes (fgr and Wx) and three disease resistance genes (Pita, Pik and Xa23) have been well characterized and used in MAS breeding. However, stacking all of them together into a single variety has not been reported. We reported here the stacking of the five genes into elite lines in rice. We achieved this through the development of functional markers from causal mutations at fgr, Wx and Pita, a gene-targeted marker at Pik and the use of a linked marker for Xa23. We employed and optimized the high-resolution melting (HRM) analysis method for use as the genotyping platform of fgr, Wx, Pik and Pita. By combining high-throughput DNA isolation, multiplex and nested-PCR methods, we showed that HRM could serve as cost-effective, highly automated, moderate-throughput and reliable non-gel genotyping platform for a small-scale MAS program.

China is one of the world’s largest potato (Solanum tuberosum L.) producing countries, with a production of 5.68 million ha. Among them, winter-grown potato has been developed rapidly in Guangdong Province in recent years, with an area of ​​nearly 66,000 ha and an output value of about 4 billion yuan. In February 2019, soft rot symptoms were observed occurring on potato (cv. Xisen) in a commercial field (15 ha) in Kaipin, Guangdong, China, with a disease incidence of about 10%. Early symptoms included darkened and water-soaked necrotic lesions at the exterior of the infected tubers. The lesions spread quickly, and the inside of the tubers was slimy and emitted a foul smell. Bacterial exudate from water-soaked areas was also observed. Twenty diseased samples were collected, and the margins between the diseased and healthy areas were cut into pieces (5 × 5 mm), which were surface sterilized using 75% ethanol for 30 s and 2% NaOCl for 1 min, followed by three rinses with sterile water. The sterilized sections were macerated in drops of sterile water, and the extract was streaked onto nutrient agar medium and incubated for 24 h at 30°C. Single colonies were chosen after three times of subculturing (Zhang et al. 2014). Fifteen bacterial isolates were obtained, and a representative isolate KP1 was chosen for further analyses. Its 16S rDNA gene sequence (GenBank accession MK674243) amplified by primer pair 27F/1492R showed 99% identity with Pectobacterium carotovorum subsp. brasiliense (Pcb) strains (nos. CP024780 and CP020350) using BLASTn. Phylogenetic analysis (neighbor-joining method and 1,000 bootstrap values) with strains from different Pectobacterium species showed KP1 was grouped into a clade with Pcb strains, based on the multilocus sequences of gapA (GenBank accession no. MK674244), icdA (MK674245), mdh (MK674246), mtlD (MK674247), proA (MK674248), and pel genes (MK674249) that were amplified using the primer pairs provided in previous studies (Darrasse et al. 1994; Ma et al. 2007). Pathogenicity tests were conducted by injecting 100 µl of bacterial suspensions (10⁸ CFU/ml) into the potato tubers (cv. Xisen). For noninoculated controls, another 10 healthy potato tubers were injected with 100 µl of sterile water. Tubers were placed in an incubator at 30°C in darkness at 95% humidity. After 3 days, inoculated tubers showed diseased symptoms similar to those observed in the field. No symptom was observed on the controls. We reisolated the pathogen from the symptomatic tissues and confirmed it to be Pcb using PCR with 16S rDNA primers. This pathogen was previously reported to infect potato in Inner Mongolia Autonomous Region, Northern China (Zhao et al. 2018). To our knowledge, this is the first report of Pcb causing bacterial soft rot on potato in Guangdong Province, Southern China. This study expanded the geographic location of Pcb as the causal agent on potato in China and helped raise attention on controlling the spread of this pathogen to minimize the losses.

The aim of this study was to investigate the prevalence of erectile dysfunction (ED) in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and explore the influence of UPOINT domains, National Institutes of Health-CP symptom index (NIH-CPSI) and other factors on ED prevalence. This was a prospective study of consecutive patients with CP/CPPS seen at 11 tertiary hospitals during January-July 2014. ED was diagnosed as a score of