OsAFB4 gene encodes a kind of rice auxin receptor protein, and its deletion can make rice resistant to 2, 4-dichlorophenoxyacetic acid (2, 4-D) and picloram. In order to study its function, characteristics and interaction with auxin herbicides, it is necessary to establish a prokaryotic expression system for OsAFB4 protein. In this study, OsAFB4 gene was cloned into prokaryotic expression vectors pET-32a, pGEX-4T, and pMAL-C6T, respectively. Then, pET-32a-OsAFB4 was transformed into Escherichia coli strain competent cells of OverExpress C43 (DE3), Rosseta2 (DE3) and ER2566, pGEX-4T-OsAFB4 was transformed into E. coli strain competent cells of OverExpress C43 (DE3) and Rosseta2 (DE3), and pMAL-C6T-OsAFB4 was transformed into E. coli strain competent cells of OverExpress C43 (DE3), Rosseta2 (DE3) and TB1. The expression of fusion protein was induced by isopropylthio-β-D-galactoside (IPTG), and the induction conditions were optimized using different induction times, temperatures and IPTG concentrations. The protein gel electrophoresis results indicated that the optimal induction condition for prokaryotic expression of OsAFB4 protein was 200 μmol/L IPTG at 23 ℃ for 12 h. In conclusion, the establishment of the OsAFB4 prokaryotic expression system provides a foundation for research on auxin receptor function and the development of auxin herbicide resistance sites.

Wheat is relatively long from germination to seed maturity, and normally only one generation can be produced annually. Shortening the whole growth period, consequently producing seeds of multiple generations per year, is beneficial for improving wheat breeding efficiency. Low temperature (vernalization response) and light length (photoperiod response) are the two key factors affecting the processes of wheat growth and development, or the whole growth period. On the basis of the relevant characteristics, we conducted different photoperiod and low-temperature vernalization treatments on wheat plants with different growth habits. Through observations of the growth stages, we found that super-long lighting (22 h light/2 h darkness) significantly shortened the wheat growth period and had a compensatory vernalization effect. No significant differences in panicle length, number of effective tillers per plant, or thousand grain weight were detected between the super-long lighting treatment and the control (16 h light/8 h darkness). Immature embryo culture in vitro at different time after pollination revealed that the 12-day-old young embryo had a high germination rate. Accordingly, the combination of super-long lighting and immature embryo culture in vitro may obtain 6-7 and 3-4 generations per year for spring- and winter-habit wheats, respectively. In conclusion, the method of speeding breeding developed in this study can be used in wheat breeding programs to improve breeding efficiency.

Ethylene is closely related to the floral organ development and yield of soybeans, and ethylene insensitive 3 gene (EIN3) and its homologous gene EIN3-like 3 (EIL3) may be key node genes involved in soybean yield regulation; however, there is still a lack of sufficient evidence. The aim of this study was to prepare EIL3 antibodies, verify their effectiveness and preliminarily explore the biological function of EIL3 in the development of soybean seedlings. EIL3 was expressed in a prokaryotic strain, followed by the preparation of polyclonal antibodies and immunohistochemical detection of EIL3 in soybean seedlings. The results showed that the EIL3 protein (Glyma.15G031800) coding region (486-590 amino acids) was cloned into Escherichia coli Rosetta strain, and the EIL3 protein was expressed in the form of inclusion bodies. Two Japanese male large-ear white rabbits were immunized with purified EIL3 protein, and two EIL3 polyclonal antisera were obtained. When they were diluted 81 000 times, the absorbance values were much greater than 0.4, indicating that they had high-quality serum titers. The lower detection limit for the antigen was both 1 ng. The expression of the EIL3 protein was strongly induced by 1-aminocyclopropane-1-carboxylic acid (ACC), and eil3 mutant reduced the accumulation and growth inhibition of EIL3, which improved the plant height, hypocotyl elongation, and lateral root development of the seedlings. This study provides an important means for exploring the biological function of soybean EIL3 at the protein level.

To explore the function and potential application of NUDIX (nucleoside diphosphates linked to other moieties X) gene family in Rosa chinensis ‘Old Blush’, 41 NUDIX gene family members of Rosa chinensis ‘Old Blush’ were identified using bioinformatics methods, and their phylogenetic relationships, gene structures, and expression patterns were analyzed as well. Phylogenetic tree of NUDIX was constructed from Rosa chinensis ‘Old Blush’, Rosa multiflora, Rosa rugosa, Rosa wichuraiana ‘Basye’s Thornless’, Fragaria vesca, Malus domestica, Arabidopsis thaliana and Oryza sativa. It was found that the NUDIX can be classified into 13 subfamilies, among which the number of members of NUDIX1, NUDIX2, NUDIX4, and NUDIX11 subfamilies was greater than that of the other subfamilies. Collinearity analysis of genes revealed that the whole genome duplication event occurred in NUDIX gene family of Rosa chinensis ‘Old Blush’, resulting in two pairs of collinear genes. The chromosome localization results indicated that NUDIX genes of Rosa chinensis ‘Old Blush’ underwent multiple tandem duplication event, most of which went through purifying selection. Transcriptomic data analysis revealed that RcNUDIX1a/1b/1d/1e were specifically expressed in petals. RcNUDIX1b/1e/4c/9/11d might be involved in the stress response in leaf, RcIDI might participate in regulating young root and early floral development in plant tissues, and RcNUDIX2b/2d/2h might participate in the regulation of flower organ senescence. In conclusion, the quantity amplification of NUDIX genes in Rosa chinensis ‘Old Blush’ was caused mainly by tandem duplication event. The NUDIX gene family might play an important role in physiological processes such as secondary metabolism, growth and development, and stress response in leaf of Rosa chinensis ‘Old Blush’, which provides a theoretical basis for further research on the function of NUDIX genes in Rosa chinensis ‘Old Blush’.

This study aims to elucidate the lambda-cyhalothrin stress response and biological function of the glutathione-S-transferase (GST) gene AiGSTe3 from the black cutworm Agrotis ipsilon. The cDNA sequence of AiGSTe3 was retrieved from the transcriptome database of A. ipsilon by the homology search method. The sequence characteristics and expression level of AiGSTe3 were analyzed, together with the enzymatic properties and lambda-cyhalothrin detoxification and antioxidant activities of the protein encoded by AiGSTe3. The results showed that AiGSTe3 contained an open reading frame of 654 bp and encoded a protein consisting of 217 amino-acid residues. Phylogenetic analysis revealed that the protein encoded by AiGSTe3 belonged to the epsilon class. The highest expression level of AiGSTe3 was detected in the larval midgut and at the pupal stage. The expression level of AiGSTe3 was significantly upregulated at 12 h after treatment with the median lethal concentration (LC₅₀) of lambda-cyhalothrin. The AiGSTE3 recombinant protein was successfully expressed in Escherichia coli. AiGSTE3 was capable of catalyzing the conjugation of reduced glutathione (GSH) to 1-chloro-2, 4-dinitrobenzene (CDNB) with a maximum velocity (v_max) of 4.33 μmol/(min?mg) and a Michaelis constant (K_m) of 0.33 mmol/L. The results of the high performance liquid chromatography analysis indicated that AiGSTE3 was able to metabolize lambda-cyhalothrin in vitro. In addition, E. coli cells overexpressing AiGSTE3 displayed significant tolerance to oxidative injury. Taken together, AiGSTe3 can respond to the stress induced by lambda-cyhalothrin. AiGSTE3, encoded by AiGSTe3, can not only metabolize lambda-cyhalothrin directly, but also enhance the tolerance of A. ipsilon to lambda-cyhalothrin by alleviating the oxidative damage induced by insecticide exposure.

Citronella essential oil was extracted by steam distillation method, and its components were determined by headspace gas chromatography-mass spectrometry (HS-GC-MS) technology. The effects of different varieties (No. 1, No. 7, and No. 12) of citronella essential oils and different concentrations of No. 12 citronella essential oil on the fresh-keeping quality of Shine Muscat grapes during room temperature storage were studied by fine fruit rate, weight loss rate, firmness, soluble solid content, titratable acid content and vitamin C (Vc) content of fruit as measuring indicators. The results showed that the yield of No. 12 citronella essential oil was the highest, with citronellal as the main volatile component. The main volatile component of No. 1 citronella essential oil was citral, and the main volatile components of No. 7 citronella essential oil were β-myrcene and citral. The sensory quality and fine fruit rate of fruit treated with No. 12 citronella essential oil were significantly better than those of No. 7 and No. 1 citronella essential oils. Compared with those in control group, different concentrations of No. 12 citronella essential oil could obviously reduce fruit decay, significantly inhibit the weight loss of fruit and slow down the decline of fruit firmness, titratable acid content and Vc content. 0.125, 0.250 and 0.500 μL/L essential oil treatments had better effects on inhibiting fruit decay; 0.250 and 2.000 μL/L essential oil treatments had better effects on maintaining the soluble solid content of fruit; 0.250 μL/L essential oil treatment had the best effect on maintaining the sensory quality of fruit, inhibiting the weight loss and slowing down the decline of titratable acid content of fruit; 2.000 μL/L essential oil treatment had the best effect on slowing down the decline in firmness and Vc content of fruit. In summary, 0.250 μL/L No. 12 citronella essential oil treatment is more suitable for the preservation of Shine Muscat grapes. This study provides a theoretical basis for the future application of citronella essential oil in the preservation of fruits and vegetables.

The basic fruit quality indicators and characteristic aroma of the main Chinese bayberry variety ‘Biqi’ (‘BQ’) and a new selection ‘2010-70’ were analyzed. Headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) was used to determine the volatile components of both ‘BQ’ and ‘2010-70’ and the relative contents of their distilled liquors made from primary and secondary distillations. Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to compare and analyze volatile components in different Chinese bayberry distilled liquors. The results showed that the characteristic aroma β-caryophyllene in the volatile compounds of ‘2010-70’ and ‘BQ’ juices had the highest content. The relative contents of terpenoids and esters were the highest in the Chinese bayberry distilled liquor. For terpenoids, α-caryophyllene, β-caryophyllene and (－)-isocaryophyllene in Chinese bayberry juice were retained, and the contents of ethyl decanoate, benzyl benzoate and ethyl laurate were relatively high in esters. The results of PCA and OPLS-DA showed a significant difference in the volatile components of the distilled liquors from the two Chinese bayberry varieties, with 2-hexenal being the most significant component. Combined with the sensory evaluation, for both varieties, the volatile components of the first distillation were generally better than those of the second distillation, and the volatile components of ‘BQ’ primary and secondary distillated liquor were generally better than those of ‘2010-70’. However, the ‘2010-70’ primary distilled liquor had the best sensory evaluation. In conclusion, different Chinese bayberry varieties and fermentation and distillation times strongly influence the volatile component content and flavor of Chinese bayberry distilled liquor. Primary fermentation and distillation can better preserve the characteristic fruity flavor of Chinese bayberry in distilled liquor.

The plant microbiome is one of the key determinants of plant health and productivity, providing host plants with functions such as access to low-abundance nutrients, resistance to pathogen invasion, and adaptation to the environment. In this study, the flavor and physicochemical indexes of white loquat were comprehensively determined, and 16S rRNA amplicon technology was used to analyze the composition and function of microorganisms in the roots, leaves and bulk soil environments of white loquat, in order to reveal the relationships between the microbial community and fruit quality. The results showed that the microbial communities in soil environment covered most of the microorganisms in the root and leaf environments, but there were obvious differences in community structure and microbial function, indicating that there was obvious niche differentiation of endophytic microorganisms in different organs of plants. An analysis of the core microbiome and non-core microbiome revealed that functional redundancy existed in all three environments. On the basis of the correlation analysis between fruit quality and microorganisms, it was found that microorganisms in roots had an obvious regulatory effect on fruit quality but were not found in leaves. This study clarifies the microbiome-host interaction in white loquat and identifies the microorganisms associated with fruit quality, laying an important foundation for the study of precision fertilization and efficiency enhancement in white loquat.

Accurate identification of the species of shark fins has been a significant challenge for customs and other law enforcement agencies to fulfill Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). In this study, we established a DNA barcode identification method for cytochrome c oxidase subunitⅠ (COⅠ) gene of shark species, applied it to 113 shark fin samples intercepted at Zhuhai-Macao Port, and identified a total of 19 species, dominated by Carcharhinidae family species such as Prionace glauca and Carcharhinus falciformis. 56.64% of the samples were listed in the 2019 version of the CITES Appendix Ⅱ, and 90.27% were listed in the 2023 version of the CITES Appendix Ⅱ. The analysis results showed that the established method produced specific amplifications and obtained valid sequences for all samples. The intra-specific genetic distance of the measured species was ≤2.08% (average of 0.63%), and the inter-specific genetic distance ranged from 2.42% to 27.27% (average of 15.37%). The smallest inter-specific genetic distance was greater than the largest intra-specific genetic distance. The average inter-specific genetic distance was 23 times greater than intra-specific genetic distance. All species clustered in the phylogenetic tree based on their degree of kinship. Species from the same family further clustered in sub-branches. There was no cross-distribution of species among sub-branches. In summary, the DNA barcode technique based on theCOⅠ gene could accurately identify the species of shark fins, which is important for the diversity conservation of sharks.

To investigate the regulatory mechanisms of silicon (Si) application on phosphorus (P) activation and uptake in the paddy soil-rice system, pot experiments were conducted using three waterloggogenic paddy soils from Changchun City of Jilin Province, Kunming City of Yunnan Province, and Hangzhou City of Zhejiang Province (designated as paddy soil 1, paddy soil 2, and paddy soil 3, respectively). The effects of different application amounts of Si (0, 30, and 60 mg/kg) on the P content in different parts of rice throughout the growth cycle, as well as yield, root iron plaque, and P availability and phosphatase activity in the rhizosphere soil were studied. The results showed that in paddy soil 1 and paddy soil 3, Si application significantly increased rice yield by 33.1%-115.0%, and enhanced P translocation to aboveground parts and grains. In paddy soil 1 and paddy soil 2, Si application significantly increased rice grain P levels by 11.0%-136.9%. Si application reduced the amount of P bound in root iron plaque by 6.3%-43.0% and decreased iron content in the plaque, with the most pronounced reduction (33.6%-47.6%) observed in the high-iron paddy soil 2. Furthermore, Si application increased alkaline phosphatase activity in paddy soil 3 by 95.9%-122.7%, enhancing the soil’s P mineralization potential. Overall, Si application improves rice growth, alters iron content in root iron plaques, mitigates iron toxicity to rice, particularly to the roots, and enhances soil P mineralization, thereby activating P in the paddy soil-rice system and increasing the efficiency of P uptake by rice plants.

In this study, single nucleotide polymorphism (SNP) scans were performed on the single copy gene regions of Y chromosome genome of 110 individuals from 9 cattle breeds including 25 Qaidam cattle. The paternal genetic diversity, population structure differences, differentiation, and clustering of these cattle breeds were analyzed. The results showed that a total of 14 Y chromosome genome haplotypes were identified in 9 breeds, 4 of which (H7, H12, H13, and H14) were identified in Qaidam cattle. Genomic diversity analysis showed that the paternal genetic diversity of Qaidam cattle was relatively rich (H_d=0.690±0.080), but the level of paternal genetic diversity was lower than that of Mongolian cattle, Yanbian cattle, Tibetan cattle and Kazakh cattle. Phylogenetic analysis showed that the paternal inheritance of Qaidam cattle was composed of Y1, Y2a, and Y2b haplogroups/sub-haplogroups, and the Y2b sub-haplogroup was dominant, indicating that Qaidam cattle was of Bos taurus origin. The genetic differentiation analysis showed that the differentiation degrees between Qaidam cattle and Mongolian cattle or Tibetan cattle were low, and the differentiation degree between Qaidam cattle and Yanbian cattle was moderate, and the differentiation degrees between Qaidam cattle and Kazakh cattle, Simmental cattle or Angus cattle were high. Cluster analysis showed that Qaidam cattle had the closest clustering relationship with Mongolian cattle, which was closer to that of Tibetan cattle and Yanbian cattle but far from that of Kazakh cattle, Simmental cattle and Angus cattle. The results of this study provide a reference for in-depth understanding of the paternal genomic diversity, population structure and genetic relationships between Qaidam cattle and other yellow cattle breeds, enrich the genome database/dataset of Qaidam cattle, and lay a foundation for the subsequent rational protection and molecular breeding practices of Qaidam cattle.

Reproductive traits are one of the important economic traits in the sheep industry. The purpose of this study was to explore the non-genetic factors affecting the reproductive traits of Tianmu multiparous sheep and their genetic parameters. In this study, the reproductive performance records of 5 335 Tianmu multiparous sheep from 2014 to 2022 were collected from Zhejiang Sino Ecological Agriculture Co., Ltd., including litter size at birth (LSB), litter mean weight per lamb born (LMWLB), total litter weight at birth (TLWB) and litter survival lamb numbers (LSLN). SAS 8.1 and DMU softwares were used to analyze the effects of non-genetic factors on reproductive traits, estimate genetic parameters and breeding values, and further analyze genetic trends and phenotypic trends. The results showed that birth parity, year of lambing and type of lambing had extremely significant effects on all traits (P＜0.01). The direct heritability estimates of LSB, LMWLB, TLWB and LSLN were 0.016, 0.063, 0.049 and 0.034, respectively, indicating that these were low heritability traits. The LSB was negatively correlated with LMWLB both genetically and phenotypically. The genetic correlation between LMWLB and LSLN was positive, but their phenotypic correlation was negative. The genetic and phenotypic correlations of the remaining traits were both positive. The average breeding value of LSB first increased and then decreased, and the average phenotypic value showed an overall upward trend. The average breeding values of LMWLB and TLWB first decreased and then increased, and the average phenotypic value first increased and then decreased. The average breeding value and average phenotypic value of LSLN first increased and then decreased. This study has laid a foundation for the genetic improvement of reproductive traits of Tianmu multiparous sheep, and also provided guidance for breeding work.

Glycoside hydrolases such as glucanase and xylanase are important components of enzyme preparations that can hydrolyze and ablate non-starch polysaccharide antinutritional factors in feed. According to the metatranscriptomic data obtained previously from rumen microbiota in Hu sheep, a novel bifunctional glucanase/xylanase gene, IDSGH5-23, was cloned from cDNA of rumen microbiota. The gene was heterologously expressed in Escherichia coli, and the recombinant enzyme was functionally characterized. The results showed that the theoretical molecular weight and isoelectric point of IDSGH5-23 were 45.8 kDa and 4.44, exhibiting 99.51% similarity to amino acid sequence of GH5 protein (GenBank accession number: WP_013499238.1) from Ruminococcus albus. The IDSGH5-23 had the optimum temperature of 30 ℃ and the optimum pH of 6.0, and remained stable ranging of pH 4.0-7.0. Moreover, the IDSGH5-23 was robust in catalyzing barley β-glucan, lichenan, konjac gum and beechwood xylan. The enzyme IDSGH5-23 mainly released cellotriose from lichenan or cellooligosaccharides with a degree of polymerization beyond 3 from barley β-glucan. Besides, the enzyme IDSGH5-23 was capable of generating xylotriose, xylotetraose, xylopentaose and xylohexaose from beechwood xylan. In summary, a bifunctional glucanase/xylanase protein derived from rumen microbes, IDSGH5-23, is obtained and characterized in this study, laying a foundation for feed enzyme development and oligosaccharides preparation.