Drought and diseases represent major challenges to achieve high yield in crops, underscoring the urgent need to explore drought- and disease-resistant genetic resources and breed crop varieties with multi-stress resistance. Here, using a genome-wide association study combined with function analyses, PibH8, a homologous of the blast resistance gene Pib encoding a nucleotide-binding leucine-rich repeat receptor (NLR) is identified, which plays a crucial role in rice drought resistance. PibH8 interacts with phenylalanine ammonia-lyase OsPAL1, a rate-limiting enzyme in phenylpropanoids biosynthesis pathway in rice. It protects OsPAL1 from degradation by competitively binding to E3 ubiquitin ligase OsFBK16, which facilitates OsPAL1 degradation. This protective mechanism enhances PAL activity, leading to increased lignin and flavonoids content and improved drought resistance. Genetic evidence indicates that PibH8 acts upstream of OsPAL1 in conferring drought resistance. Furthermore, a causal variation in the PibH8 promoter that is associated with drought resistance is identified. Introgression of a superior haplotype, which exhibits high PibH8 expression, into the elite rice variety Kongyu131 significantly improved drought and blast resistance. This research not only elucidates a regulatory mechanism of NLR protein in drought resistance, but also highlights a promising breeding value of PibH8 for simultaneously improving drought and blast resistance.

Background/Aim With the pharmaceutical innovation and clinical knowledge updating, the continuing education and on-the-job training are extremely important for improving community pharmacists’ professional competence. Previous training often adopted traditional lecture-based teaching, and the efficacy was limited. The aim of this study is to develop a new strategy for community pharmacist training. Methods Based on the BOPPPS (Bridge-in, Objective, Pre-assessment, Participatory Learning, Post-assessment and Summary) teaching model and workshop method, a continuing on-the-job training program was constructed. Participates were randomly and evenly divided into two groups by random number table method. Twenty-four community pharmacists in total completed all training contents and evaluation components in this study. Twelve pharmacists in experimental group were trained via this new BOPPPS-based workshop, while others still adopted traditional didactic lecture-based approaches. Results After training, quantitative examination combined with clinical pharmacy practice tests were carried out to evaluate the effectiveness and outcomes of two training groups. For written exam, the total scores from the BOPPPS-based workshop group (82.67 ± 4.70) was higher than that of traditional lectured-base group (73.75 ± 6.15) ( P < 0.001). Encouragingly, compared with the results of practical ability assessment from traditional training group (71.75 ± 4.75), the pharmacists receiving BOPPPS-based workshop training presented more excellent performance (78.25 ± 5.03), which displayed statistically significant differences ( P < 0.01). In addition, an anonymous questionnaire was used to survey trainees’ feelings after completing this continuing education program. The results revealed that the BOPPPS-based workshop can bring a better learning experience than traditional lecture-based training, and the percentages of positive response to each item were more than 91.7%. Conclusions Through multi-dimensional evaluation, it was suggested that our BOPPPS-based workshop achieved desired training effects. Moreover, our research also demonstrated that this strategy had advantages of stimulating inspiration, autonomous learning, team-work spirit and pharmacy practice improvement. It may provide a reference of innovative training method for community pharmacists.

【Objective】The use of artificial inoculation to ferment cigar tobacco leaves is an inevitable trend in the industrial production of cigar tobacco leaves, and the neutral aroma components are the important quality components of cigar tobacco leaves. The study aims to investigate the effect of Bacillus safensis GL2 on the neutral aroma components of cigar tobacco leaves.【Method】GL2 was screened from Cuban cigar Cohiba Robert and used to ferment cigar leaves. The neutral aroma components in tobacco leaves were determined by distillation extraction-gas chromatography-mass spectrometry, and multivariate statistical methods were used for analysis and statistics.【Result】The total aroma and the contents of most neutral aroma components in cigar tobacco leaves were significantly increased after GL2 fermentation (P < 0.05), and the increased aroma substances mainly included olefins, ketones and alcohols. A total of 13 differentially neutral aroma components were selected by VIP > 1 and P < 0.05, and 9 of them were significantly up-regulated and positively correlated with tobacco leaf aroma. There were 7 species with significant positive correlation (P < 0.05), which were the key differential aroma components of 2, 6, 6-trimethyl-2-cyclohexene-1, 4-dione, 2, 6, 6-trimethyl-1, 4-cyclohexanedione, geraniyl acetone, acanthyl acetone, neophydiene, solanone and phytoalcohol. In addition, 4 different aroma components were significantly decreased and negatively correlated with tobacco leaf aroma, among which 2, 4-di-tert-butyl-6-nitrophenol was significantly negatively correlated with cigar tobacco aroma (P < 0.05). 2, 6, 6-trimethyl-2-cyclohexene-1, 4-dione, geranyl acetone, solanone and phytol contributed significantly to the aroma of fermented cigar tobacco leaves, resulting in a stronger overall dry and floral fragrances. As a result, they enriched the aroma and improved the sensory quality of cigar tobacco leaves.【Conclusion】Inoculation of B. safensis during cigar tobacco fermentation is beneficial for improving the quality of cigar tobacco leaves, which provide theoretical support for improving the fermentation quality of cigar tobacco leaves with biotechnology.

[目的]探明烟草源微生物发酵对雪茄烟叶品质和挥发性致香成分的影响,为人工接菌发酵雪茄烟叶工业化生产提供菌种资源和理论依据.[方法]利用从进口雪茄烟中筛选的1株地衣芽孢杆菌(Bacil-lus licheniformis)FZ1发酵雪茄烟叶,对其进行感官评价,通过GC-MS检测烟叶中性致香成分,利用SIM-CA进行主成分分析(PCA)和正交偏最小二乘分析(OPLS-DA),使用变量投影重要度(Variable Importance in Projection,VIP)＞1和独立t检验中P＜0.05筛选差异致香成分,并结合感官评分进行相关性分析与ROAV值变化,明确地衣芽孢杆菌(B.licheniformis)FZ1提升雪茄烟叶感官品质的关键致香成分及其对香气的贡献.[结果]雪茄烟叶经FZ1发酵后香气明显提升、刺激性减少;香气总量提高18.84％,以烯烃类、酮类和醇类增加为主;共筛选出15种差异中性致香成分,其中,11种差异致香成分显著上调,新植二烯、金合欢基丙酮、茄酮、香叶基丙酮、2,4-二叔丁基苯酚和植物醇6种与雪茄烟叶香气呈显著正相关关系,是FZ1发酵雪茄烟叶的关键差异致香成分;另有正十五烷、邻苯二甲酸二丁酯、苯乙醛和2,4-二叔丁基-6-硝基苯酚共4种差异致香成分显著下调,其中,2,4-二叔丁基-6-硝基苯酚和苯乙醛与雪茄烟叶的刺激性呈显著正相关关系;茄酮、香叶基丙酮、巨豆三烯酮A、巨豆三烯酮B和植物醇对香气的贡献增大.[结论]FZ1发酵雪茄烟叶可明显改善香气品质,增加关键差异致香成分.

Abstract Background Tuberculosis (TB) is a zoonotic disease that affects humans and domesticated and wild animals. Animals in zoos are potentially an important source of TB for humans; however they are often neglected in routine disease surveillance programs. This investigation reports an outbreak of TB in milu deer and northern pig‐tailed macaques in a zoo in Wuhan, China, which highlighted the need for improved prevention and control of TB in China. Methods Between 24 November and 9 December 2020 two milu deer and a northern pig‐tailed macaque that were displaying signs of wasting died. Post‐mortem, histopathological diagnosis and acid fast staining were used for the dead animals. Multiple PCR for Mycobacterium tuberculosis complex (MTBC) was performed to identify the bacterial in both milu deer and northern pig‐tailed macaque. The serum antibody iELISA for MTBC was then performed for all the surviving milu deer and northern pig‐tailed macaques. Six seropositive milu deer and a seropositive northern pig‐tailed macaque were subsequently euthanised and, along with two other dead milu deer, necropsied. DNA from these tissue samples was extracted and detected MTBC using PCR and Real‐time PCR. Subsequently bacterial isolation was used to confirm the infection. Results The lungs of the dead animals displayed gross and histological TB‐like lesions and changes, and red staining bacilli were detected in smears of the lesions by microscopy after acid fast staining. Mycobacterium bovis (M. bovis) was detected in the two milu deer and Mycobacterium tuberculosis (M. tb) in the northern pig‐tailed macaque using multiple PCR for MTBC. 35.3% surviving milu deer and 50% surviving northern pig‐tailed macaques MTBC serologically positive. Six of the euthanised milu deer were also positive on a DNA test for M. bovis and the euthanised northern pig‐tailed macaque was positive to M. tb. Conclusions This is the first report of tuberculosis in the endangered species, milu deer and northern pig‐tailed macaques, in China, and warrants urgent attention by researchers and conservation authorities. These cases highlight the need for expanding surveillance for MTBC to zoos in China.

Rice ratooning, the fast outgrowth of dormant buds on stubble, is an important cropping practice in rice production. However, the low ratooning ability (RA) of most rice varieties restricts the application of this cost-efficient system, and the genetic basis of RA remains unknown. In this study, we dissected the genetic architecture of RA by a genome-wide association study in a natural rice population. Rice ratooning ability 3 (RRA3), encoding a hitherto not characterized nucleoredoxin involved in reduction of disulfide bonds, was identified as the causal gene of a major locus controlling RA. Overexpression of RRA3 in rice significantly accelerated leaf senescence and reduced RA, whereas knockout of RRA3 significantly delayed leaf senescence and increased RA and ratoon yield. We demonstrated that RRA3 interacts with Oryza sativa histidine kinase 4 (OHK4), a cytokinin receptor, and inhibits the dimerization of OHK4 through disulfide bond reduction. This inhibition ultimately led to decreased cytokinin signaling and reduced RA. In addition, variations in the RRA3 promoter were identified to be associated with RA. Introgression of a superior haplotype with weak expression of RRA3 into the elite rice variety Guichao 2 significantly increased RA and ratoon yield by 23.8%. Collectively, this study not only uncovers an undocumented regulatory mechanism of cytokinin signaling through de-dimerization of a histidine kinase receptor-but also provides an eximious gene with promising value for ratoon rice breeding.

Brucellosis has re-emerged in China in recent years, resulting in an increasing health burden and economic losses for humans and the livestock industries. This study integrated data from human and livestock brucellosis surveillance systems to explore the changing epidemiology of brucellosis from 2004 to 2018 in China. A total of 524,980 human cases of brucellosis were reported, with the average annual incidence in humans being significantly higher for the period 2012-2018 than for 2004-2011 (3.3 vs. 1.9 per 100,000 residents). An autoregressive integrated moving average (ARIMA) model predicted an upward trend in the monthly incidence of brucellosis in humans in 2019 and 2020. Characteristics including being male, aged 45-54 years, working in the livestock industries, and residing in the northern provinces of China increased the risk of people contracting brucellosis. The percentage of provinces with infected people increased from 67.7% (21/31) in 2004 to all provinces in 2018. A total of 29,115 outbreaks were reported in livestock from 2004 to 2018, with 443,883 seropositive animals although only 381,224 (85.9%) of these were culled. The monthly incidence of brucellosis in humans was strongly positively correlated (r = .539, p < .001) with the number of outbreaks of brucellosis in livestock reported 3 months prior to the human cases. At the provincial level, the annual incidence of brucellosis in humans was significantly positively correlated with the sheep population (r = .786, p < .01). In conclusion, brucellosis in humans and livestock has been spreading in mainland China in the past decade. A more active surveillance of brucellosis in both livestock and humans in China should be coordinated and adjusted by adopting an evidence-based 'One Health' approach, particularly in high-risk regions and livestock industries.

Tuberculosis (TB), a contagious disease mainly caused by Mycobacterium tuberculosis ( M . tb ), Mycobacterium bovis ( M . bovis ), and Mycobacterium caprae ( M . caprae ), poses a major global threat to the health of humans and many species of animals. Developing an ante-mortem detection technique for different species would be of significance in improving the surveillance employing a One Health strategy. To achieve this goal, a universal indirect ELISA was established for serologically detecting Mycobacterium tuberculosis complex infection for multiple live hosts by using a fusion protein of MPB70, MPB83, ESAT6, and CFP10 common in M . tb , M . bovis , and M . caprae as the coating antigen (MMEC) and HRP-labeled fusion protein A and G as a secondary antibody. After testing the known positive and negative sera, the receiver operating characteristic curves were constructed to decide the cut-off values. Then, the diagnostic sensitivity and specificity of MMEC/AG-iELISA were determined as 100.00% (95% CI: 96.90%, 100.00%) and 100.00% (95% CI: 98.44%, 100.00%) for M . bovis infection of cattle, 100.00% (95% CI: 95.00%, 100.00%) and 100.0% (95% CI: 96.80%, 100.00%) for M . bovis infection of sheep, 90.74% (95% CI: 80.09%, 95.98%) and 98.63% (95% CI: 95.14%, 99.76%) for M . bovis infection of cervids, 100.00% (95% CI: 15.81%, 100.00%) and 98.81% (95% CI: 93.54%, 99.97%) for M . bovis infection of monkeys, 100.00% (95% CI: 86.82%, 100.00%) and 94.85% (95% CI: 91.22%, 97.03%) for M . tb infection of humans. Furthermore, this MMEC/AG-iELISA likely detects M . caprae infection in roe deer. Thus this method has a promising application in serological TB surveillance for multiple animal species thereby providing evidence for taking further action in TB control.

Clarification of the genome composition of the potato + eggplant somatic hybrids cooperated with transcriptome analysis efficiently identified the eggplant gene SmPGH1 that contributes to bacterial wilt resistance. The cultivated potato is susceptible and lacks resistance to bacterial wilt (BW), a soil-borne disease caused by Ralstonia solanacearum. It also has interspecies incompatibility within Solanaceae plants. Previously, we have successfully conducted the protoplast fusion of potato and eggplant and regenerated somatic hybrids that showing resistance to eggplant BW. For efficient use of these novel germplasm and improve BW resistance of cultivated potato, it is essential to dissect the genetic basis of the resistance to BW obtained from eggplant. The strategy of combining genome composition and transcriptome analysis was established to explore the gene that confers BW resistance to the hybrids. Genome composition of the 90 somatic hybrids was studied using genomic in situ hybridization coupled with 44 selected eggplant-specific SSRs (smSSRs). The analysis revealed a diverse set of genome combinations among the hybrids and showed a possibility of integration of alien genes along with the detection of 7 smSSRs linked to BW resistance (BW-linked SSRs) in the hybrids. Transcriptome comparison between the resistant and susceptible gene pools identified a BW resistance associated gene, smPGH1, which was significantly induced by R. solanacearum in the resistant pool. Remarkably, smPGH1 was co-localized with the BW-linked SSR emh01E15 on eggplant chromosome 9, which was further confirmed that smPGH1 was activated by R. solanacearum only in the resistant hybrids. Taken together, the identified gene smPGH1 and BW-linked SSRs have provided novel genetic resources that will aid in potato breeding for BW resistance.

Bacterial wilt (BW) caused by Ralstonia solanacearum is an important disease of many plant species especially Solanaceae. To compensate for lack of BW resistance in cultivated potato, we fused UV-treated protoplasts of a resistant eggplant variety with protoplasts of a susceptible potato clone to obtain 32 somatic hybrids. Although asymmetric protoplast fusion has the potential to transfer traits from distant species, introgression frequency and preference of alien fragments remain obscure, as well as the genetic basis for control of a trait. In the present research, the genome components of 32 somatic hybrids were determined by parent-specific SSRs. Each hybrid had integrated from one to eight alien chromosome fragments, providing a foundation for selection of BW resistance transmitted from eggplant. When the selected eggplant sequences were aligned with potato genome sequence it showed a similarity of 46.7 %, suggesting a large genetic distance between these two species. The results also revealed that introgression of eggplant fragments is non-selective, which may allow any part of alien chromosomes to be integrated. Distribution of eggplant loci in individual hybrids suggested a possible relationship between markers emk03O04, emi04P17 and emd13E02a and BW resistance, which are potential loci that control target traits and therefore deserve further investigation. With genome-wide selection of parent-specific molecular markers and sequence alignment, the present research substantiated interspecific introgression of a trait lacking in potato. Moreover, an efficient strategy was established to estimate genomic components of the somatic hybrids, and to explore the candidate loci that associate with target traits.

KEY MESSAGE:Bacterial wilt resistant somatic hybrids were obtained via protoplast fusion between potato and eggplant and three types of nuclear genomes were identified in the hybrids through GISH and SSR analysis.ABSTRACT:Cultivated potato (Solanum tuberosum L.) lacks resistance to bacterial wilt caused by Ralstonia solanacearum. Interspecific symmetric protoplast fusion was conducted to transfer bacterial wilt resistance from eggplant (S. melongena, 2n = 2x = 24) into dihaploid potato (2n = 2x = 24). In total, 34 somatic hybrids were obtained, and of these, 11 rooted and were tested for genome components and resistance to race 1 of R. solanacearum. The hybrids exhibited multiple ploidy levels and contained the dominant nuclear genome from the potato parent. Three types of nuclear genomes were identified in the hybrids through genomic in situ hybridization (GISH) and simple sequence repeat (SSR) analysis, including (1) the potato type of the tetraploids in which eggplant chromosomes could not be detected by GISH but their nuclear DNA was confirmed by SSR, (2) the biased type of the hexaploids in which the chromosome dosage was 2 potato:1 eggplant, and (3) the chromosome translocation type of the mixoploids and aneuploids that was characterized by various rates of translocations of nonhomologous chromosomes. Cytoplasmic genome analysis revealed that mitochondrial DNA of both parents coexisted and/or recombined in most of the hybrids. However, only potato chloroplast DNA was retained in the hybrids speculating a compatibility between cpDNA and nuclear genome of the cell. The pathogen inoculation assay suggested a successful transfer of bacterial wilt resistance from eggplant to the hybrids that provides potential resistance for potato breeding against bacterial wilt. The genome components characterized in present research may explain partially the inheritance behavior of the hybrids which is informative for potato improvement.