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Glomerella leaf spot (GLS) caused by Glomerella cingulata is a newly emerging disease that results in severe defoliation and fruit spots in apples. In China, the compound of pyraclostrobin and tebuconazole was registered to control GLS in 2018 and has achieved excellent control efficiency. In this study, we showed that the high-level resistant isolates of G. cingulata to pyraclostrobin, caused by the point mutation at codon 143 (GGT→GCT, G143A) in the cytochrome b gene, has appeared in apple orchards in Shandong Province in 2020, and the resistance frequency was 4.8%. Based on the genotype of the resistant isolates, we developed a loop-mediated isothermal amplification (LAMP) assay for detection of the pyraclostrobin resistance. The LAMP assay was demonstrated to have good specificity, sensitivity, and repeatability, and it exhibited high accuracy in detecting pyraclostrobin resistance in the field. This study reported the resistance status of GLS to pyraclostrobin in Shandong Province and developed a molecular tool for the detection of pyraclostrobin resistance, which is of practical significance for the scientific control of GLS.

Nutrient and stress factor 1 (Nsf1), a transcription factor containing the classical Cys2-His2 (C2H2) zinc finger motif, is expressed under non-fermentable carbon conditions and in response to salt stress in Saccharomyces cerevisiae. However, the role of Nsf1 in filamentous fungi is not well understood. In this study, the orthologue of Nsf1 was investigated in Fusarium graminearum (named FgNsf1), a causal agent of Fusarium head blight (FHB). The functions of FgNsf1 were evaluated by constructing a FgNSF1 deletion mutant, designated as ΔFgNsf1, and its functional complementation mutant ΔFgNsf1-C. Gene deletion experiments showed that the mycelial growth rate, asexual and sexual reproduction of ΔFgNsf1 were significantly reduced, but the pigment production of ΔFgNsf1 was remarkably increased compared with the PH-1 and ΔFgNsf1-C. In addition, the tolerance of ΔFgNsf1 to osmotic pressures, cell wall-damaging agents and oxidative stress increased significantly. Sensitivity tests to different fungicides revealed that ΔFgNsf1 exhibited increased sensitivity to carbendazim (MBC) and tebuconazole, and enhanced tolerance to fludioxonil and iprodione than PH-1 and ΔFgNsf1-C. The virulence of ΔFgNsf1 to wheat coleoptiles and flowering wheat heads were dramatically decreased, which was consistent with the decrease in the yield of deoxynivalenol (DON). All of these defects were restored by target gene complementation. These results indicated that FgNsf1 plays a crucial role in vegetative growth, asexual and sexual reproduction, stress responses, fungicide sensitivity, and full virulence in F. graminearum.

Astragali Radix, one of the most widely used traditional Chinese medicines, has been extracted a variety of efficiently ingredients. However, the formation mechanism of the secondary metabolites is still unclear and the genome of Astragali Radix has not been sequenced yet. This study aimed at predicting gene model according to transcriptome data, which can be helpful to the use of Astragalus membranceus in the medicine field.Based on the transcriptome data of Astragalus membranceus and the existing whole genome sequence of its closely related species, the unigene of transcriptome of Astragalus membranceus was predicted and analyzed.Astragalus membranceus had close genetic relationship with Glycine max,Medicago truncatula and Lotus japonicus. Astragalus membranceus shared 2 039 sequences with Medicago truncatula, and their base similarity was 90. 14%. There were 1 948 sequences matched up with Glycine max, and their base similarity was 90. 40%. In the aligement of Astragalus membranceus and Lotus corniculatus, the matched sequences dropped to 1 003,and the base similarity dropped to 89. 77%.The matched gene with three species discributed in chromosome genomes and extrachromosomal genome.

High-resolution-melting analysis (HRM) is a new technology derived from q PCR and is widely used in the study of polymorphism, genotyping, and single nucleotide mutation. Advantages of HRM include cost-effectiveness and time-efficiency over PCR-based genotyping. However, the application of HRM in the authentication of herbal products is still limited with few studies on the classification and identification of herbal products. In this study, Cimicifugae Rhizoma was used as an example to verify the stability and accuracy of HRM technique in identification of Chinese materia medica. HRM assay was established for identification based on ITS2 region of Cimicifugae Rhizomas and its adulterants(including 41 samples). Our findings showed that HRM allows not only the identification of adulteration but also the quantification of the most common admixture. This study is significant for better quality in the verification of the authenticity of herbal medicine. The method is promising for future identification of traditional Chinese medicinal materials.

In this study, Illumina sequencing platform was applied in sequencing rat pancreas, counting expression of target points, analyzing expression differences among blank group, model group and Huangqi Liuyi decoction group and exploring the therapeutic effect and mechanism of Huangqi Liuyi decoction on type 2 diabetes mellitus. According to the result, 24.25% of these genes belonged to the unknown functional class, which was the largest classification unit according to the classification analysis of genes by eggNOG. The rest were classified as energy conversion, amino acid transport and metabolism, nucleotide transport and metabolism, carbohydrate transport and metabolism, coenzyme transport and metabolism, and lipid transport and metabolism, etc.Huangqi Liuyi decoction may play a therapeutic role in the treatment of type 2 diabetes mellitus through four metabolic pathways, namely environmental information processing, cellular process, organismal system and human diseases according to KEGG enrichment analysis. This study shows that, Huangqi Liuyi decoction can significantly improve the fasting blood glucose and glycosylated hemoglobin in type 2 diabetic rats.

In this study, 454/Roche GS FLX sequencing technology was used to obtain the data of the Astragalus membranaceus. Four hundred and fifty-four Sequencing System Software was applied to carry out the transcription of the group from scratch. Using MISA tools, 9 893 unigenes were selected for the sequence of the genome of A. membranaceus, and the information of SSR locus was analyzed. According to the result, the average length of reads was 413 bp, about 86% of the reads was involved in the splicing, the length of the N50 was 1 205 bp, the number of unigenes was measured by the whole transcript. 1 729 SSR loci in the A. membranaceus transcriptome were searched, the occurrence frequency of SSR was 9.24%, the frequency of SSR in the whole transcriptome was 13.42%, the average length of SSR was 7.97 kb. One hundred and twenty-seven kinds of core repeat sequences were found, the dominant type was TG/AC type of dinucleotide, it appeared to account for 4.25% of the total SSR locus. The results of the sequence of the transcription of the A. membranaceus transcriptome revealed the overall expression, and a large number of unigenessequence was obtained, and the SSR locus in the genome of the A. membranaceus is high, and the type is diverse, and the polymorphism of the gene is high.

DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.