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【Objective】The present paper aimed to investigate the important role of NAC gene in the growth and development of Cinnamomum camphora and its role in salt stress, and to clarify the response mechanism of C. camphora under salt stress. 【Method】The genome-wide NAC genes of C. camphora were identified and analyzed by bioinformatics to investigate the mode of response to abiotic stress in C. camphora. By identifying the members of NAC family of C. camphora,the paper aonducteda systematic analysis in terms of physicochemical properties, phylogenetic analysis, conserved motifs and gene structures, chromosomal localization and inter-species covariance, cis-acting element prediction and expression patterns.【Result】A total of 103 CcNAC genes were identified, the amino acid counts ranged from 137 to 1136, the isoelectric points from 4.54 to 10.00, the relative molecular masses from 15 644.40 to 129 859.29 u, and the mean hydrophobicity of all proteins was negative, indicating hydrophilicity. Subcellular localization was predicted to be in the nucleus and cytoplasm. A phylogenetic evolutionary tree was constructed with Arabidopsis thaliana NAC gene sequences, which classified CcNAC genes into 16 subfamilies. The NAM subfamily had the most members, with 17 CcNAC genes, while 25 CcNAC genes were not included. Conservative motif and gene structure analysis revealed that CcNAC genes were found to contain 2-8 introns and 10 motifs. Among these, motif3 appeared to be particularly stable in the genetic evolution of C. camphora genes, while motif9 and motif10 were distributed across the NAM subfamily. Chromosome localization and replication event analysis revealed that 103 genes were distributed across 12 chromosomes, with the highest number of genes（13） found in CcNAC genes distributed across chromosomes Chr02, with the lowest number of genes found on Chr10 and Chr11, which contained two genes each; Fragment duplication was the main mode of amplification for the CcNAC family. Multi-species co-linkage analysis revealed that CcNAC genes formed 162 pairs of co-linkage genes with CcNAC genes, indicating a closer relationship than with other species. The CcNAC genes were more closely related to the CcNAC genes and to those of other species than to any other. The CcNAC gene promoters contained several abiotic stress, growth and development, and hormone response elements. The results of the qRT-PCR showed that in leaves, the expression levels of the CcNAC058 and CcNAC092 genes were maximised under low salt stress. In roots, the expression of CcNAC007, CcNAC033, CcNAC058 and CcNAC092 genes increased with increasing salt concentration.【Conclusion】The NAC gene family may be involved in the process of salt stress response during the growth and development of C. camphora. The CcNAC092 and CcNAC058 genes exhibite significant responses in both leaves and roots under salt stress. It is hypothesised that CcNAC058 and CcNAC092 are the key genes contributing to the ability of the CcNAC to produce self-defensive behaviours under salt stress. [ABSTRACT FROM AUTHOR]

(Objective] The present paper aimed to explore the relationship between fatty acid desaturase gene ADS2 (19 fatty acid desaturase) and cold resistance in Brassica rapa L. (Method) Based on the previous transcriptome results, the BrADS2 gene was obtained by onestep directional cloning. GFP fusion expression vector was constructed and transiently transformed into tobacco lower epidermal cells by injection method. At the same time, Zhongshuang 11' was infected by Agrobacterium infection method. The expression pattern of ADS2 gene in the leaves of winter rapeseed varieties with different cold resistance was detected by qRT-PCR, and the morphological changes, antioxidant enzyme activity and conductivity content of winter rapeseed under low temperature were analyzed. (Result] The BrADS2 gene could encode 307 amino acids, and the protein encoded by this gene had hydrophilic properties. The results of qRT-PCR analysis showed that the expression of BrADS2 gene increased first and then decreased, and the expression level reached the peak at 4 C. The results of subcellular localization showed that BrADS2 protein was expressed in cytoplasm and cell membrane. Under low temperature treatment, the relative conductivity and antioxidant enzyme activity of the leaves of the three varieties increased significantly. The antioxidant enzyme activity of the strong cold-resistant varieties was higher than that of the weak cold-resistant varieties, but the conductivity was the opposite. The hypocotyls of Zhongshuang 11' were infected by Agrobacterium tumefaciens, and 15 transgenie positive seedlings were obtained. The expression of BrADS2 gene in the leaves of transgenic rape positive plants was significantly higher than that of wild type. [Conclusion】 It is speculated that BrADS2 gene plays an important role in the regulation of membrane lipid unsaturation in winter rapeseed, revealing the relationship between ADS2 gene and crop resistance to low temperature stress [ABSTRACT FROM AUTHOR]

【Objective】The paper aimed to screen the suitable reference genes and to analyze and verify the expression of photosynthesis-related genes in Chenopodium album responding to Aureobasidium pullulans PA-2.【Method】The leaves of C. album that were treated for 0, 1, 3, 5 and 7 days under PA-2 infection and the root, stem and leave of normal growth C. album were used as materials, and qRT-RCR technology and Ct value were used to assess the expression stability of three candidate reference genes (GAPDH, β-ACTIN and β-TUBULIN) . The expression levels of photosynthesis-related genes (LFRN, MDH, PetC, psaA, CAB40, psaN, BHY and MO2) were quantitatively analyzed by the optimal selected reference genes.【Result】The results indicated that GAPDH was the optimal reference gene of C. album responding to A. pullulans PA-2, and can be used for analyzing the related genes expression level to photosynthesis pathway. Using GAPDH as internal reference, the expression levels of 8 photosynthesis-related genes showed varying degrees of down-regulation, except for MDH, which reached the lowest level at 5 days. Among them, PetC, psaN, CAB40, BHY and MO2 was down-regulated C. album infected 0-1 day, reaching a high level at 3 days, and increased in the later stage. The expression levels of pasA was firstly up-regulated and then down-regulated during the infection. The expression level of LFRN was down-regulated continuously after PA-2 infection. The expression level of MDH was significantly down-regulated at first, reached the lowest level at 3 days, and increased in the later stage. In different tissues infected with PA-2, 8 photosynthesis-related genes were highly expressed in leaves, among which MDH gene was highly expressed in stems, while PctC and CAB40 were the lowest expression levels in roots. Except for MDH gene, the expression levels of 7 photosynthesis-related genes in different tissues were ranked as: leaves＞stems＞roots.【Conclusion】The study provides foundation for the analysis of C. album genes by using qRT-RCR and the infection mechanism of A. pullulans PA-2. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to reveal the structural characteristics of UGT family members in Aquilegia oxysepala and A. oxysepala f. pallidiflora, and screen the key UGT genes for anthocyanin biosynthesis.【Method】Based on previous floral color transcriptome of A. oxysepala and A. oxysepala f. pallidiflora, members of UGT family were selected and identified. Their physicochemical properties, conserved motifs, phylogenetic evolution and cis-acting elements were analyzed. Combining with expression patterns of UGT genes in the sepals of two A. oxysepala species at different periods, candidate genes for anthocyanin synthesis were screened out.【Result】A total of 154 AoUGT gene members were identified. Their encoded proteins contained 52-1293 amino acids, relative molecular weight was 5.897-144.174 kDa, theoretical isoelectric points were 3.21-7.14, and most of proteins were acidic and hydrophilic.The secondary structures were dominated by α-helix and random crimp, both of which had UGT domains. The UGT gene family members of A. oxysepala were divided into 14 subfamilies. The number of UGT gene members in group A was the largest, and the number of UGT gene members in group G and group J was the least. MEME conserved motif analysis showed that the UGT gene family contained two highly conserved motifs Motif 1 and Motif 4, which together formed the PSPG conserved domain of the UGT gene family. The analysis of cis-acting elements of promoter showed that the promoter regions of AoUGT family members involved in light responsive, hormone responsive, stress responsive and flavonoid MYB binding sites. According to expression patterns and cognate relationship, AoUGT147/127/45/143 significantly increased during pre-flowering period in A. oxysepala, however, these gene expressions were continuously sustained at low levels in A. oxysepala f. pallidiflora. Therefore, these members played important roles during floral color variation, which were determined to be candidate genes.【Conclusion】AoUGT147/127/45/143 are key genes for flower color variation in A. oxysepala and play an important role in the formation of flower color. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to analyze the salt tolerance mechanism and molecular mechanism of giant rice by analyzing the expression specificity of differential genes in response to salt stress at seedling stage, and to provide a theoretical basis for genetic improvement of salt tolerance of giant rice. [Method] Xiangju 1 was used as the material, and the control and one salt concentration level (0.5% NaCl) were set up and treated for 72 hours. The dynamic changes and main enrichment pathways of response genes under salt stress were analyzed by transcriptome sequencing. [Result] After salt stress, seedling height and fresh weight of Xiangju 1 were damaged by less than 30% stress. The total clean reads per each sample were 58.93 G Clean bases, the average GC content per each sample was 53.51%, the average Q20 and Q30 per each sample were 97.84% and 93.89% respectively. Gene differential expression analysis identified 2327 differentially expressed genes, including 1363 up-regulated genes and 964 down-regulated genes. The differentially expressed genes were analyzed by functional annotation, GO enrichment analysis showed that they were significantly enriched in 166 GO terms, among which the most significantly enriched terms were biological process of oxidation reduction process, nuclear metabolism process, transcriptional regulation; The cellular component of the cell component; The molecular function of the oxidoreductase activity, monooxygenase activity, transcription factor activity, etc. KEGG analysis showed that differential genes were significantly enriched in secondary metabolite synthesis, plant hormone signal transduction, phenylpropanoid biosynthesis, and glutathione metabolism. [Conclusion] Xiangju 1 has strong salt tolerance, GO analysis shows that Xiangju 1 responds to salt stress by regulating antioxidant enzyme activity and transcription factor expression. KEGG analysis shows that plant hormone signal transduction, phenylpropanoid biosynthesis, glutathione metabolism and other pathways play a leading role in response to salt stress. In the study, RNA-Seq sequencing technology is used to screen candidate genes of rice in response to salt stress, which provides a basis for further study of salt tolerance regulation mechanism of giant rice, and also provides an important theoretical basis for the development and cultivation of new varieties. [ABSTRACT FROM AUTHOR]

[Objective] The paper aimed to isolate and identify the Exosomes (Exos) of macrophages infected with Listeria monocytogenes (LM) in monocyte proliferation mice, and analyze their miRNA expression profile, laying a solid research foundation for revealing the regulatory mechanism of macrophage Exos miRNA in LM infection. [Method] Macrophages infected with LM were used as the experimental group, while macrophages not infected with LM were used as the control group. Exos in the supernatant of macrophages were isolated and extracted using ultracentrifugation. The morphology, size and surface marker characteristics of Exos were identified by transmission electron microscopy, nanoparticle tracking technology and Western blot. Small RNA-seq sequencing of miRNA in macrophage Exos from the two groups was performed using the Illumina SE50 sequencing platform to screen for significantly differentially expressed miRNA. Targetscan, miRDB and miRWalk databases were used to predict the target genes of differentially expressed miRNA. GO and KEGG-Pathway functional enrichment analysis were conducted on the different miRNA target genes. The miRNA-mRNA regulatory network diagram of the top 20 Hub genes was drawn using Cytoscape software. [Result] The diameter of Exos ranged from 30 to 150 nm, with a typical double-layer membrane structure, and the surface marker molecules CD9, CD63 and TSG101 were positively expressed. High-throughput sequencing results showed that compared to the control group, 9 significantly differentially expressed miRNA were screened in the experimental group Exos, including 8 significantly down-regulated genes (mmu-miR-7a-5p, mmu-miR-365-2-5p, г u-miR-365-2-5p, mmu-miR-122-5p, mmu-miR-122b-3p, mmu-let-7i-5p, mmu-miR151-3p, mmu-miR-182-5p and mmu-miR-1198-5p) and 1 significantly up-regulated gene (mmu-miR-192-5p), with a total of 1064 predicted miRNA-regulated target genes. GO enrichment analysis results showed that the different miRNA target genes were mainly enriched in processes such as axon formation, cell junction assembly, actin binding and metal ion transmembrane transport; KEGG analysis showed that the target genes were significantly enriched in the cGMP-PKG signaling pathway and the FeyR-mediated phagocytosis pathway. [Conclusion] The miRNA expression profile of macrophage Exos in mice infected with LM underwent significant changes, and the potential target genes regulated by them were mainly involved in signaling pathways related to infection and immune response. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to explore the role of AcinJHBP gene in the growth, development and diapause of Apocheima cinerarius. [Method] Based on the transcriptome data (non-diapause pupa/diapause pupa and diapause pupa/diapause depupa), JHBP gene was cloned and named as AcinJHBP (GenBank entry number: OR860357). The sequence and physicochemical properties of AcinJHBP gene were analyzed by bioinformatics software. The temporal and spatial expression of Acin JHBP (different developmental stages and tissues) was detected by qPCR. [Result] The complete CDS sequence of Acin JHBP gene of A. cinerarius was 726 bp, encoding 241 amino acids, the predicted molecular weight of protein was 26.97 kDa, and the predicted isoelectric point was 5. 13. The predicted molecular formula of Acin JHBP gene was C121 H1915 N313 0364 S9, the total charge of negative charge residues (Asp + Glu) was 34, the total charge of positive charge residues (Arg + Lys) was 26, the half-life and aliphatic index were 30 hours and 97.51, respectively. The average hydrophilicity of the protein was 0.098, and the predicted instability coefficient was 27. 13. Therefore, the protein was a hydrophilic stable protein, and the N-terminal amino acid was methionine (Met, M). AcinJHBP contained a signal peptide of 17 amino acid residues at the N-terminus, but had no transmembrane region. Phosphoric acid sites were found in serine, threonine and tyrosine phosphate sites were 9,6 and 3, respectively, and no glycosylation sites were found. Subcellular localization results showed that it was mainly located outside the cell. Phylogenetic analysis showed that AcinJHBP had the closest relationship with the hemolymph juvenile hormone binding protein JHBP of Operophtera brumata. The results of gene expression showed that the expression of Acin JHBP was lower at different developmental stages (except the 3rd instar larvae), and there was no significant difference between the egg and the 5th instar larvae (P>0.05), for all that the expression of Acin JHBP showed a decreasing trend in non-diapause pupa-adult stage. In different tissues, the expression of AcinJHBP gene was the highest in the male head of A. cinerarius. [Conclusion] Acin JHBP gene is closely related to the growth, development and diapause of A. cinerarius. The results lay a theoretical foundation for further revealing the function of the gene in the summer and winter diapause of the pupae in A. cinerarius. [ABSTRACT FROM AUTHOR]

[Objective) The present paper aimed to obtain the complete sequence of SsREMO-la gene, conduct tissue-specific expression, qPCR expression analysis, sub-cellular localization study, and the phenotypic changes of Arabidopsis thaliana with SsREMO-la gene transfer and the expression pattern of CAT-1 gene. (Method) The complete sequence of SsREMO-la gene was obtained by PCR, purification, cloning and sequencing from Hainan sugarcane wild species (Sp-24). Drought stress was applied to the normal growth of the seedlings. Young leaves, roots and stems at 0, 1, 2, 3, 4 and 5 days after the stress treatment were collected for tissue-specific expression analysis and qPCR analysis. The tissue location and function of SsREMO-1a were analyzed by tobacco sub-cellular mapping and gene transformation techniques. PB1221-SSREMO-1a-GFP fusion expression vector was constructed and transformed into tobacco for cell localization analysis. The over-ex- pression vector pCAMBIA 1304-SsREMO-la was constructed, and SsREMO-1a was introduced into A. thaliana by agrobacterium transformation. The phenotypes of plants at 0, 1, 2, 3, 4 and 5 days after drought stress and the expression of CAT-1 gene were observed. [Result] The full length of SsREMO-1a gene was 738 bp, and the open code frame (ORF) was 564 bp, encoding 187 amino acids. Tissue-specific expression showed that SsREMO-1a was expressed in roots, stems and leaves. qPCR results showed that the gene was induced by drought stress and showed significantly up-regulated expression. The results of sub-cellular localization showed that SsREMO-1a protein was localized on the cell membrane. A total of 11 transgenic A. thaliana lines were obtained, and the drought resistance identification showed that the transgenic plants could improve the drought resistance of the plants. The expression of CAT-1 gene in transgenic plants was significantly in- creased. [Conclusion] SsREMO-1a can improve the drought resistance of plants and regulate CAT-1 gene to some extent. [ABSTRACT FROM AUTHOR]

[Objective] The paper aimed to explore the function of the phosphoproteinase 1 (EbPP1) gene related to the number of leaves in Erigeron breviscapus, which lays the foundation for revealing the molecular mechanisms of E. breviscapus leaf number and flowering time. [Method) The EbPP1 gene from the leaves of E. breviscapus was cloned and constructed into the plant overexpression vector RP101. The dip- ping flower method was used to infect newly bolted Arabidopsis thaliana plants for heterologous expression, several T1, T2 and T, generations of A. thaliana positive plants were obtained through screening. Finally, the number of leaves and flowering time of T3 generation positive plants were counted. The expression level of EbPPI was detected by RT-qPCR, and the endogenous hormone content of A. thaliana plants was measured. [Result] The full-length EbPP1 gene was successfully cloned and overexpression vector RP101-GFP EbPP1 was constructed for this gene, cloned stable genetic transformation had been achieved in A. thaliana, and 24 positive EbPPI transgenic plants were obtained in the T3 generation. Real time fluorescence quantitative analysis was performed on the T3 generation positive plants, and RT-qPCR results showed that the EbPP1 gene was overexpressed in the transgenic A. thaliana, with its expression level increasing from 1.034 in the wild type to 136.33; Transgenic A. thaliana exhibited phenotypic characteristics of increased leaf number and delayed flowering time; Quantitative determination of endogenous hormone content in plants by LC-MS showed that the content of various hormones such as indole-3-carboxylic acid (ICA) and gibberellin-24 (GA24) in T3 transgenic A. thaliana were significantly higher than that in the wild type, with the precursor substance of growth hormone L-tryptophan (TRP) being particularly significant, reaching three times that of the wild type. [Conclusion] Suc- cessfully, the heterologously EbPPI gene is expressed in A. thaliana plants. Compared with wild A. thaliana plants, transgenic A. thaliana plants have an increased number of leaves, increase its expression of EbPPI gene, and increase endogenous hormone content. Based on the above results, it is suggested that EbPP1 may affect the number of leaves and flowering time of E. breviscapus by regulating plant hormone levels. The study lays the foundation for further analyzing the mechanisms related to the development of leaf number and flowering molecular mechanisms of E. breviscapus, and also provides a certain genetic basis for the breeding of high-quality E. breviscapus varieties. [ABSTRACT FROM AUTHOR]

[Objective] The paper aimed to comprehend the expression characteristics of cinnamyl alcohol dehydrogenase from Pinus yunnanensis and its involvement in lignin biosynthesis. (Method) The relative concentrations of 18 specific small molecules within the lignin pathway were assessed in the stems, buds, and needles of one-year-old P. yunnanensis. Additionally, the CAD genes of P. yunnanensis were identified, cloned and bioinformatics analyzed by comprehensive analysis of transcriptome data. RT-qPCR was employed to examine their expression patterns across various tissues. (Result] The findings revealed that among the 18 lignin pathway molecules, the relative con- tent of 5-0-caffeoylshikimic acid, 5-0-p-coumaroyl shikimic acid, and caffeic acid were higher in the buds, while the relative content of 3- O-p-coumaroyl quinic acid, p-coumaric acid, ferulic acid, coniferylaldehyde, L-phenylalanine, coniferyl alcohol, p-coumaryl alcohol, L- (-)-tyrosine, sinapinaldehyde were higher in the stems. In addition, the relative content of coniferin, p-coumaraldehyde, caffeic aldehyde, and syringin were higher in the needles. Four members of the CAD family were screened from the transcriptome of P. yunnanensis, and phylogenetic analysis showed that PyCAD-3 of P. yunnanensis was homologous to AtCADI, and PyCAD-1, PyCAD-2 and PyCAD-4 were closely related to CAD of Pinus, AtCAD4 and ALCAD5. The results of correlation analysis showed that the PyCAD-1 and PyCAD-2 were closely related to small molecules in the lignin pathway upstream and downstream. The open reading frame of PyCAD-1 and PyCAD-2 was 570 and 1083 bp, respectively, and the molecular weights of PyCAD-1 and PyCAD-2 were 20.38 and 39.30 kDa, respectively. The results of RT-qPCR showed that the relative expression of PyCAD-1 was the highest in bud and tender stem, and PyCAD-2 was the highest in tender stem. [Conclusion] It is speculated that PyCAD-2 may be directly involved in lignin biosynthesis, while PyCAD-1 may have some functional redundancy. The study provides a theoretical basis for studying CAD genes family and genes related to the lignin metabolism pathway of P. yunnanensis. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to study VdActin7 gene expression pattern in different tissues of Vaccinium dunalianum Wight.【Method】The cDNA and DNA of VdActin7 gene were successfully cloned by PCR technique based on the transcriptome database of 3 generations of V. dunalianum. The expression of VdActin7 gene in young leaves, mature leaves, flower buds, flowers, green fruits, red fruits, green fruit stems and red fruit stems of V. dunalianum was further detected and analyzed using qRT-PCR technique. 【Result】The full length of VdActin7 gene and cDNA were 1657 and 1387 bp, respectively, which contained 3 introns, namely, intron1 of 93 bp, intron2 of 89 bp and intron3 of 88 bp. It had an open reading frame length of 1134 bp, encoding 377 amino acids. The protein VdActin7 with a relative molecular weight of 41.70 KD and a theoretical isoelectric point of 5.31, was a stable hydrophilic protein, without signal peptide and transmembrane domain. Phylogenetic analysis revealed that the VdActin7 was closest to V. ashei Actin7, which also belonged to the Vaccinium family. The codon usage bias analysis of the VdActin7 gene showed that GC contents of codon(GC) and 3rd nucleotide in codon GC3s were 48.2% and 44.1%, respectively, the effective number of codon(ENC) was 49.28%, a total of high-frequency codons was 31. The codon bias was similar to that of Ziziphus, both with the strongest bias for CUU, followed by GUU, and the bias for CUU was significantly stronger in V. dunalianum(3.43) than that in Ziziphus(2.57). qRT-PCR analysis showed that the VdActin7 gene expression showed obvious tissue specificity, with the highest in young leaves while lower expression in mature leaves and flowers, meanwhile no detectable expression or no expression in fruits, suggesting that the VdActin7 gene might play an important role in the development of young leaves. 【Conclusion】In the paper, we reported the Actin7 gene sequence and its characteristics in V. dunalianum for the first time, and found that the VdActin7 gene had tissue expression specificity. This laid the foundation for further study of VdActin7 gene function, expression regulation and mechanism of action. [ABSTRACT FROM AUTHOR]

【Objective】 The present paper aimed to overexpress and purify rice cold stress response protein OsCML16 in Escherichia coli, and prepare OsCML16 protein antibody to provide antigen, and provide reference for further study of biological function of rice OsCML16 protein by using the antibody. 【Method】 At first, the transmembrane region of the target protein OsCML16 was analyzed online using TMHMM 2.0 software, and then the fragment of target gene was amplified by high fidelity PCR, the prokaryotic expression vector pET30a-OsCML16 was constructed by gene recombination method, and the recombinant plasmid was transformed into expression E.coli line BL21 (DE3). OsCML16 protein expression was induced by IPTG, and purified by affinity chromatography, then the authenticity of the protein sequence was verified by mass spectrometry. 【Result】 By analyzing the transmembrane region of OsCML16 protein, it was found that the protein did not contain transmembrane region, and it was suitable for heterologous expression of the full-length protein. The prokaryotic expression vector PET30A-OSCML16 was successfully constructed by linking OsCML16 gene to pET30a vector. The result showed that the target gene had no mutation site and could be used for expression of the target protein. Exogenous expression OsCML16 protein was analyzed by SDS-PAGE electrophoresis, and results showed that OsCML16 protein could be induced expression at 28 ℃.The fusion protein His6-OsCML16-His6 could be purified by affinity chromatography, and the 30 kD fusion protein was obtained successfully. The purified protein was validated by mass spectrometry, and a total of three peptides were identified as OsCML16 protein, indicating that the exogenously expressed protein was the OsCML16 protein. 【Conclusion】 The rice cold stress response gene OsCML16 was expressed in E.coli by prokaryotic expression method. The fusion expression protein His6-OsCML16-His6 was purified by affinity chromatography according to the histide-carrying label of the fusion protein, and the sequence of the protein was verified by protein spectroscopy. Rice OsCML16 protein can be used for antibody preparation and protein function study in the future. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to identify CBF family members using whole genome data of Prunus sibirica, and to analyze the characteristics and functions of the conserved domains and their expression patterns after low temperature stress of CBF transcription fac- tors. [Method] By the P. sibirica whole genome database, the CBF genes of P. sibirica were searched using BLASTP and HMM, and their conserved structural domains were verified by CDD and MEME. Bioinformatics tools such as Prot-Param, WOLF PSORT, MAGA X, Phyre2 and PlantCARE were used to analyze protein physicochemical properties, subcellular localization, phylogeny, tertiary structure, chromosom- al localization and cis-elements, respectively. Quantitative real-time PCR (RT-qPCR) was used to analyze the expression pattern of CBF gene after low temperature stress in P. sibirica. [Result] In the study, four PSCBF's were identified, all of which possessed the conserved structural domains PKKPAGR and DSAWR sequences of the AP2 gene family. The PSCBFs contained 206-246 amino acids, with a mean molecular weight of 25. 80 KDa and a mean isoelectric point of 5.92. PSCBF3 and PSCBF4 were similar in gene structure and had closer ge- netic relationship. The subcellular localization of 4 PSCBFs was located in the nucleus. Promoter cis-acting element analysis showed that CBF family members could participate in plant hormone and abiotic stress responses. Four CBF members were all acidic and hydrophilic pro- teins. The expression of PSCBF's was higher mainly in the early stage (15 min) after low temperature stress. [Conclusion] The study shows more complete information on the CBF gene family of P. sibirica, which offers new insights into the complex mechanisms and pathways of PSCBF in response to low-temperature stress at different stages. It lays an important theoretical foundation for further investigations into the biological functions of CBF family members. [ABSTRACT FROM AUTHOR]

[Objective] The paper aims to explore whether calmodulin-like CMLs participate in the sexual differentiation process in Cucumis sativus L., so as to provide reference for improving the regulatory network of sexual differentiation furtherly. [Method] In the study, CMLS, ACSIG, ACS2 and ACS11 genes of cucumber were cloned, qPCR was used to detect the expressions of CMLS gene in different types of the flowers, and the interactions between CMLS and proteins encoded by major genes of cucumber sex differentiation were detected by the method of yeast two-hybrid. [Result】 CML8 was cloned from cucumber which contained a complete ORF of 441 bp, encoding 146 aa without signal peptide. It was an extracellular protein without transmembrane domain and introns, and contained 4 EF-hand domains with Ca2+ binding ca- pacity, and subcellular was located in cell membrane and cytoplasm. ACSIG, ACS2 and ACSII, three key genes of cucumber sexual differ- entiation were cloned, and sequence alignment showed that the nucleotide sequences and encoded protein sequences of the three genes were consistent with those in the cucumber database. There were significant differences in the expression levels of three genes in the female flow- ers, male flowers and bisexual flowers, and extremely significant differences in male flowers. The yeast two-hybrid experiment showed that CMLS could interact with ACSIG and ACS2, but not with ACS11. [Conclusion] Calmodulin-like 8 (CMLS) was cloned, and CMLS could interact with ACSIG and ACS2, which might participate in the sexual differentiation of cucumber, and the study was the first time to explore the involvement of calmodulin in the process of plant sexual differentiation, and expect to provide reference for calcium signaling system to participate in the process of plant sexual differentiation. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed tl Exolease I(EX01) is a ultifunctionnal3’→5’exolease that was mainly used to digest single-stranded segments in double-stated DNA or RNA samples.Most of existing conimercialized EXO1 are induced from Escherichia coli.But low expression level and complex purification steps limit its application. The present paper aimed to synthesize sbcB gene, highly express in E.coli and purify the expressed product. [Method] The sbcB gene was synthesized by overlapping PCR and overexpressed by pET-30 a system in E.coli BL21(DE3), which was induced by IPTG, and the expression conditions were optimized to obtain the high expression of EXO1. The recombinant protein was purified by affinity chromatography and the enzyme activity was studide. [Result] The product EXO1 was consistent with the predicted 54.5 KD. Further functional verification also showed that EXO1 can digest single chains but can not digest double chains. The enzyme activity of purified interest protein was well. [Conclusion] EXO1 prepared by this method showed high yield, high purity, and was suitable for pre-treatment of PCR products in laboratory-sequencing. [ABSTRACT FROM AUTHOR]

【Objective】The paper aimed to effectively solve the problem of pollen pollution and cultivate male sterile Lilium using molecular biology technology. The role of ATPase3 gene, a member of the ATPase family, in the male sterility Lilium was studied, which would provide a theoretical basis for the application of this gene in the pollen-free molecular breeding of Lilium.【Method】Anthers of fertile lines and sterile lines of Liliums were used as experimental materials, and a significantly up-regulated ATPase3 gene was screened from the results of transcriptome sequencing of fertile lines. Relative expression of ATPase3 gene in different parts of Lilium was detected by fluorescence quantitative qRT-PCR during the period of microsporocyte. Full-length cDNA sequence of anther ATPase3 gene of Lilium was amplified by qRT-PCR and Race technology, and the structure and physicochemical properties of Lilium ATPase3 were analyzed by various online bioinformatics software（Prot Param, Prot Scale, NCBI Conserved Domain, SWISS-MODEL）. 【Result】The expression of ATPase3 gene in Lilium was the highest in flower organs, and the expression in anthers of fertile lines was 7 times higher than that of sterile lines. Full-length ORF of Lilium ATPase3 gene was 327 bp, encoding 109 amino acids and containing a typical conserved ATPase3 domain; Relative molecular weight of the protein was 12.14 KD and theoretical isoelectric point was 7.71. It was a hydrophilic protein with transmembrane structure, and the secondary and tertiary structures of the protein were predicted. Bioinformatics analysis showed that the predicted protein had typical characteristics of ATP synthase. Homologous sequence analysis showed that the protein had the closest homology with Arabidopsis thaliana plasma membrane ATPase3 and H[+]ATPase1, H[+]ATPase2, H[+]ATPase3, H[+]ATPase8. This protein was a plasma membrane ion transporter that provided energy in plants. It was speculated that the expression level of this protein was down-regulated in the anthers of Lilium sterile lines, which led to the male sterility of Lilium.【Conclusion】In the study, ATPase3 gene was verified to be related to male sterility in Lilium, and ATPase3 gene was successfully cloned and analyzed by bioinformatics. It provided a theoretical basis for the occurrence of male sterility in Lilium caused by the loss of plasma membrane ion transporter function which was encoded by ATPase3 gene. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to explore the effects of different proportions of red and blue light on flower bud differentiation of ‘Red Globe’ grape in facilities, and to solve the problem of poor flower bud differentiation of ‘Red Globe’ grape caused by low light environment in facilities.【Method】Taking ‘Red Globe’ grape as experimental material, without light supplement as control（CK）, the effects of red and blue light on flower bud differentiation were explored by LED light treatment of red（R）, blue（B）, red and blue 2∶1（R2B1）, red and blue 4∶1（R4B1） and red and blue 6∶1（R6B1）, together with the study of histology, physiological indexes and floral gene expression.【Result】Different proportions of red and blue significantly affected flower bud differentiation and flower related gene expression of facility ‘Red Globe’ grape.R4B1 and R6B1 treatments significantly increased the flowering rate of ‘Red Globe’ grape in greenhouse.Among them, under R4B1 treatment, the flower bud differentiation rate was fast, and the flowering rate was the highest, reaching 66% in the fifth section. The contents of carbohydrate, soluble protein and flavonoid in leaves increased, and the flowering genes VvFUL, VvFT, VvAG, VvAP1, VvAP2, VvSOC1 and VvSPL10 were up-regulated, while VvLFY and VvFLC were down-regulated.The correlation analysis between flowering genes and flowering rate showed that the flowering rate under R4B1 treatment was positively correlated with the expression levels of VvFUL, VvFT, VvAG and VvSPL10, and VvAP1 was positively correlated with VvFT, and negatively correlated with VvAP2 and VvAP3.VvFLC was negatively correlated with VvAG, and VvFT was positively correlated with VvAP2, VvSOC1 and VvSPL10.Network interaction analysis showed that the presentation and total correlation of VvAP1, VAP2 and VvSOC1 were high, and VvAP1, VvAP2, VvSOC1 and VvLFY were the critical nodes of flowering gene interaction.【Conclusion】R4B1 treatment significantly increased leaf nutrient accumulation and floral gene expression in the ‘Red Globe’ grape and promoted flower bud differentiation. These results can provide a reference for further understanding the physiological and molecular mechanisms of red and blue light regulating flower bud differentiation of ‘Red Globe’ grape, and provide a theoretical basis for improving flower bud differentiation of ‘Red Globe’ grape. [ABSTRACT FROM AUTHOR]

【Objective】 The present paper aimed to study the saline-alkaline stress on the content of lignin, chlorophyll, as well as the genes expression levels in lignin and chlorophyll biosynthetic pathways, to provide a theoretical reference for revealing the molecular response mechanism of maize in response to saline-alkaline stress and cultivating new saline-alkaline-resistant maize varieties.【Method】Corn seedlings were treated with saline-alkali stress, and high-throughput transcriptome sequencing was conducted.The chlorophyll and lignin content were detected.【Result】In total, 39 722 genes were identified, including 5659 differentially expressed genes.The differentially expressed genes were mainly annotated in carbon metabolism, phenylpropanoid biosynthesis, starch and sucrose metabolism pathways.Under saline-alkaline stress, the chlorophyll content and lignin content were reduced by 30.45% and 31.26%, both H-type and S-type monolignol reduced, while G-type monolignol increased.The analysis of the relationship between gene expression level and component content showed that, the down-regulation of chlorophyll/bacteriochlorophyll synthase, and up-regulation of chlorophyllase and chlorophyll（ide） b reductase led to the reduction in chlorophyll content.The 161 DEGs in ‘phenylpropanoid biosynthesis’ pathway may cause the variation in lignin and monolignol content.【Conclusion】Saline-alkalines reduce the total lignin content mainly by inhibiting the biosynthesis of H-type and S-type monolignol.Saline-alkaline reduces the chlorophyll content probably by down-regulated chlorophyll/bacteriochlorophyll synthase genes, and up-regulated chlorophyllase and chlorophyll（ide） b reductase genes.Overall, the study provides a theoretical basis for improving saline-alkaline resistance and controlling lignin content of maize by molecular breeding. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to enrich the salt tolerance gene resources of Arundo donax and provide help for molecular breeding.【Method】The ‘LZ＿ No.1’ A. donax with consistent growth were treated with 0, 50, 100, 150 and 200 mmol/L NaCl salt solutions for 36 hours, and each treatment consisted of 3 replicates. Stems from the same site were collected for transcriptome sequencing and bioinformatics analysis. 5 differentially expressed genes were randomly selected for qRT-PCR to verify the expression pattern.【Result】De novo assembly was performed on the sequencing data after quality control, and a total of 971 309 transcripts were generated, with a GC content of 47.14%, and the N50 of Unigene contigs reached 1126 bp, indicating a good assembly effect. The 7305 differentially expressed genes identified were subjected to GO and KEGG enrichment analysis, and the results showed that stems of ‘LZ＿ No.1’ responded to salt stress mainly through nitrogen metabolism, ion transport, terpenoid synthesis, zeatin synthesis, linoleic acid metabolism, salicylic acid biosynthesis and osmotic regulator substances. The differentially expressed genes were screened, and a total of 104 core differentially expressed genes were obtained, of which 88315＿c0＿g1＿i1, 1200＿c2＿g1＿i1, 13330＿c0＿g1＿i8, 46571＿c0＿g1＿i7, 3434＿c0＿g1＿i11 and 713＿c0＿g1＿i32 genes with large differential multiples may play an important role in the response to the salt. The identification of transcription factors indicated that AP2/ERF-ERF family, MYB-related family and WRKY family were the most important gene families in salt response, and most of these three transcription factor family members were up-regulated in response to salt stress, indicating that they mainly worked by increasing the expression of genes. The detection results of the five differentially expressed genes by qRT-PCR were basically consistent with the RNA-seq results, which also demonstrated the reliability of the transcriptome data.【Conclusion】The study identifies important genes and transcription factors of A.donax in response to salt stress, which can lay a solid foundation for its future genetic improvement work. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to clone the transcription factor gene BoWRKY10 of kale (Brassica oleracea var. acephala DC) and study the drought tolerance regulation mechanism of the gene.【Method】The gene BoWRKY10 was amplified from the cDNA of kale leaves by RT-PCR and transferred into tobacco. The expression pattern of the gene under drought stress was detected by real-time fluorescence quantitative PCR.【Result】The survival rate of transgenic lines were significantly higher than that of non-GMO wild types (WT) after 10 days of drought. There was no obvious difference between transgenic lines (OE3, OE4 and OE6) and WT in terms of germination rate in 1/2 MS culture medium, however, in medium containing 15% PEG4000, germination rate of OE3, OE4 and OE6 were 65.9%, 75.4% and 84.3%, respectively, while the one of WT was 22.5%. In medium containing 15% PEG4000, the root length of transgenic lines was significantly higher than that of WT after 14 days. Meanwhile, ABA content, proline content, soluble sugar content, POD, CAT and SOD activities were higher, while MDA and H2O2 content were significantly lower than those of WT. The expression level of drought responsive gene in OE3, like NtSOD, NtAPX, NtCAT, NtGST, NtSPSA and NtNCED1, was significantly higher than that of WT.【Conclusion】Expression of BoWRKY10 from kale remarkably improved the tolerance of tobacco to drought stresses in which the pleiotropy could be beneficial gene resource of gene engineering about resistance to stress in plant. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to analyze the expression differences of ZmGly1 gene in different tissues in responsive to drought and high temperature stress, so as to find out its possible role in heat and drought tolerance, and finally to provide useful genetic resources for the upcoming breeding of maize varieties with higher stress tolerance.【Method】The structure, physicochemical properties and genetic relationship of ZmGly1 protein were explored using bioinformatics software such as DNAMAN 8.0, ProtParam tool, MEGA 7.0. The transcript levels of ZmGly1 was performed using qRT-PCR method.【Result】ZmGly1 was composed of 222 amino acids with a theoretical molecular weight of 24.98 kD and an isoelectric point of 6.6. The total number of amino acid residues with negative charges（Asp + Glu） and residues with positive charges（Arg + Lys） was 30 and 29, respectively. The hydrophobic coefficient was 0.448, suggesting that it was a hydrophilic protein. ZmGly1 was not a transmembrane protein and contained a Zn2+ metal binding site on the A chain and B chain, located at Gln68,Glu134 and His162,Glu208, respectively. The phylogenetic tree analysis demonstrated that ZmGly1 and SbGly1 had the highest sequence identity（92.51%）, indicating that they were closely related to each other. The qRT-PCR analysis exhibited that the transcript levels of ZmGly1 was the highest in leaves, but relatively low in roots and stems. Compared with the control, the expression pattern of ZmGly1 in the leaves under drought treatment showed a tendency of first increasing and then decreasing, while under high temperature treatment a tendency of first decreasing, then increasing and finally decreasing with the extension of time. 【Conclusion】ZmGly1 is a Zn2+ enzyme, which is closely related to SbGly1. ZmGly1 participates in the stress response of plant to drought and high temperature. The study lays the foundation for the in-depth analysis of its biological functions in the near future. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to explore the molecular mechanism of Apocheima cinerarius response to stress and explore the main associated genes about low temperature and starvation. 【Method】This study cloned and obtained the chitinase gene of A. cinerarius, named AcinCht10（GenBank accession No. : OK504624）, the molecular characteristics and basic physicochemical properties of amino acids of this gene were analyzed, sequence alignment was carried out and phylogenetic tree was constructed. And the expression profile of this gene under stress（low temperature and starvation） was analyzed. 【Result】The predicted molecular formula of AcinCht10 gene was C5017H7689N1349O1490S39, the length of coding protein region was 2967 bp, encoding a total of 988 amino acids, the predicted molecular weight of AcinCht10 was 111. 99 kDa, and the predicted isoelectric point was 6. 25. The hydrophilic coefficient of the protein was-0. 547, and the predicted instability coefficient was 37. 19, indicating that it was stable hydrophilic protein. One signal peptide was found, but no transmembrane structure was found. Two catalytic regions and one chitin binding domain were found. There were 48 serine, 43 threonine and 14 tyrosine phosphoric acid sites, no glycosylation sites were detected. Four phosphoric acid sites were detected. Sequence alignment results showed that the consistency of chitinase gene between AcinCht10 and other Lepidopteran insects was higher than 91%. Phylogenetic results showed that the chitinase genes were grouped into 8 groups（type I-VIII）. AcinCht10 was grouped with Manduca sexta Cht10, Trichoplusia ni Cht10, and Ostrinia furnacalis Cht10, which were also lepidoptera. The results of gene expression analysis showed that AcinCht10 gene expression decreased with the increase of starvation treatment time, and there was no significant difference between adjacent treatments. The expression of AcinCht10 gene was the highest under 25 ℃, while the expression of AcinCht10 gene was at a low level under other temperature stresses with no significant difference. 【Conclusion】AcinCht10 gene was closely related to the response to low temperature and starvation stress. The results are helpful to reveal the molecular function of chitinase gene of A. cinerarius under stress, and lay a foundation for further revealing the molecular mechanism of A. cinerarius response to stress. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to identify and analyze the KUP/HAK/KT gene family in sweet potato and provided a theoretical basis for further study on the function of IbHAK gene. 【Method】Based on bioinformatics, the KUP/HAK/KT gene family of sweet potato was identified at the whole genome level by the genome data of Taizhong No.6, and the basic characteristics, phylogenetic evolution, gene replication and expression patterns of family members were further analyzed. 【Result】A total of 22 IbHAK genes were identified in sweet potato. The length of IbHAK protein ranged from 227 aa to 1252 aa, the molecular weight ranged from 18 297.95 to 140 640.46 u, and the isoelectric point ranged from 5.54 to 9.31, all IbHAK genes were located on the plasma membrane, and the complete secondary structure of IbHAK protein had 4 to 13 transmembrane regions. According to the phylogenetic relationship, it could be divided into four evolutionary clusters: Cluster Ⅰ, Cluster Ⅱ, Cluster Ⅲ and Cluster Ⅳ. Chromosome mapping showed that 22 genes were distributed on 11 chromosomes, among which Chr4 chromosome was the most distributed, with 4 IbHAK genes, and 8 replication genes were produced by 22 IbHAK genes. All of the 22 IbHAK genes contained TATA and CAAT elements related to plant growth and development as well as light-responsive elements. All of the IbHAK genes contained at least one stress-responsive element and one hormone-responsive element. IbHAK genes showed tissue expression specificity, among which IbHAK6, IbHAK8, IbHAK9 and IbHAK22 had the highest expression levels in roots, and the expression levels of many IbHAK genes changed under stress. 【Conclusion】Twenty-two KUP/HAK/KT gene family members were identified from the whole genome of sweet potato. IbHAK6, IbHAK8, IbHAK9 and IbHAK22 gene family members were conserved in evolution. Most of IbHAK genes were able to respond to stress. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to provide theoretical basis for studying the molecular mechanism of betacyanin accumulation and improving the nutritional value of Hylocereus polyrhizus, the betacyanin content in fruits, cloned HpCYP76 AD1 gene are detected, and the relationship between gene expression and betacyanin content are analyzed.[Method] The betacyanin content during fruit maturation was measured using the colorimetric method. HpCYP76 AD1 gene was obtained based on fruit transcriptome sequencing data and RT-PCR clone. The physicochemical properties and phylogenetic relationship of protein encoding by HpCYP76 AD1 were analyzed and the gene expression pattern during fruit maturation was detected by qRT-PCR.[Result] The betacyanin accumulated rapidly during fruit ripening, whose content in mature fruit（30 days after pollination） was about 150 times higher than the one 20 days after pollination. The open reading frame of HpCYP76 AD1 gene was 1521 bp, which encoded a protein sequence of 506 amino acids. The relative molecular weight of HpCYP76 AD1 protein was 57.5 kDa and the predicted isoelectric point was 8.86. The HpCYP76 AD1 amino acid sequence had a highest similarity with pear cactus OfCYP76 AD8, which up to 89.96%. HpCYP76 AD1 and Beta vulgaris BvCYP76 AD1, Mirabilis jalapa MjCYP76 AD3 and Amaranthus cruentus belonged to the α class of CYP76 AD1 family, which suggested that HpCYP76 AD1 had a catalytic activity in the two hydroxylation reactions of betacyanin accumulation in fruits. The expression of HpCYP76 AD1 gene was significantly up-regulated during fruit ripening, which was positively correlated with the betacyanin accumulation pattern in fruits.[Conclusion] HpCYP76 AD1 gene from cytochrome P450 family member of H. polyrhizus belonged to the α class, which indicated that it may have a catalytic activity in the two-step hydroxylation reaction of betacyanin accumulation. The expression of HpCYP76 AD1 gene was greatly up-regulated during fruit ripening, which was involved in betacyanin accumulation. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to have a broader and deeper understanding of the regulatory factors of capsaicinnoid biosynthesis, and provide molecular basis for biotechnology breeding of pepper. [Method] The transcriptome of fruits in rapid growth stage and green mature stage from two pepper materials with different spicy degrees, '754-3-1-1-1' and 'Yujiao 5', were sequenced. The gene differential expression was analyzed by using the software cufflinks, and was verified by RT-PCR. The difference expression genes were obtained by comparing the transcriptome of two materials during two development stages, and GO enrichment analysis of the differential expression genes were carried out. [Result] A total of 139, 1903, 5550 and 3220 genes were differentially expressed in the pairwise comparisons of L754LS_vs_L754KS, YJ5LS_vs_YJ5KS, L754KS_vs_YJ5KS and L754 LS_vs_YJ5LS. There were 2667 (48%) unique differentially expressed genes in rapid growth stage and 871 (27%) in green maturity stage. In addition, a total of 18 CapCyc model genes were identified from four pairwise comparisons. Among them, PAL, CCoAOMT, CCR and other structural genes in capsaicin synthesis pathway were expressed more than 3 times differently in different samples. Moreover, 3 6 MYB and 4 ERF family transcription factors that may be involved in capsaicinnoid synthesis were identified. [Conclusion] Capsaicinnoid content was more affected by the expression level of genes in the early stage of fruit development. PAL and other structural genes may have the role of regulating the biosynthesis of capsaicinnoid. The study provides more candidate genes for future studies on capsaicinnoid biosynthesis regulatory network. [ABSTRACT FROM AUTHOR]

[ Objective] The present paper aimed to analyze whether the accumulation of glycoside alkaloids in different potato tissues has influence on human use and food processing, the accumulation levels of glycoside alkaloids in different potato tissues and organs under different storage conditions were summarized. [ Method ] In the present study, the contents of glucoside alkaloids in tuber and bud tissues of potato resources 1-76 and 6-36 (purple bark), 5-36 and 6-1 ( white bark) were determined by HIGH performance liquid chromatography-tandem mass spectrometry after 60 days of storage in light and cellars. Real-time fluorescence RCR was used to analyze the gene expression of glycoside alkaloid biosynthesis. [ Result ] The detection of glycoalkaloids showed that the content of glycoalkaloids in four kinds of potato sprouts was significantly higher than that in tubers after storage under different conditions, the content of glycoalkaloids in tubers exceeded the safe edible standard ( 0. 2 mg/ g, fresh quality), but whether it was safe to eat and food processing needs more experimental verification ; The detection in the expression of glycoalkaloid biosynthesis-related genes showed that the expression levels of some genes ( StCAS, StSSR2, StGAME4, StSGT I, StSG12) in the glycoalkaloid biosynthesis pathway of different potato resources were closely related to the content of glycoalkaloids, and the expression of other genes in potato had certain tissue specificity. [ Conclusion] By simply and quickly detecting the content of glycoalkaloids in potato tubers and sprouts, this test found that the content of glycoalkaloids in tubers of potato resources ( 1-76, 6-1 ) after storage did not exceed the safety standard, but it was not recommended. The results of this study provided a theoretical reference value for human safe consumption of potatoes. [ABSTRACT FROM AUTHOR]

[ Objective]This present paper aimed to clarify the effect of kisspeptin on reproductive endocrinology in pre-pubertal female mice and its regulatory mechanism. [ Method] Tested mouses were seperately injected with different concentrations of kisspeptin, subsequently, FSH and LH concentrations of peripheral blood, ovarian weight, ovarian diameter, vaginal opening rate and pituitary gene expression were respectively detected. [ Result]The 3 µ,g kisspeptin not only significantly increased mouse ovarian weight, diameter and vaginal opening rate ( P < 0. 05), but also significantly increased FSH and LH concentrations of peripheral blood ( P <0. 05); Meanwhile, 3 µ,g kisspeptin treat ment could significantly up regulate GnRH-R, FSH/3 and LH/3 gene expression in pituitary gland ( P < 0. 05) . [Conclusion] Kisspeptin treat ment could advance the start of puberty in mice; Kisspeptin might increase the concentration by up-regulating the expression of FSH/3 and LH/3 mRNA or by indirectly up-regulating the expression of GnRH-R mRNA in the adenohypophysis of pre-pubertal mice. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to study the function of PPO gene related to heat browning chestnut kernels metabolic mechanism.【Method】A PPO unigenes was obtained from transcriptome data based on transcriptome sequencing and bioinformatics analysis. [Result] The full length of open reading frame was 1794 bp，belonged to the PPO gene family，named CmPPO, encoding 597 amino acids with a molecular weight of 66 335. 38, containing the (common central region and the (conserved intermediate domain of the PPO, which had a theoretical pl of 6.45, including common central domain of tyrosinase and PPO middle binding domain. The secondary structure of the CmPPO protein contained 21.22% alpha-helix, 59.13% random coils, 15.91% extended strands and 3.85% beta-turns. The three-dimensional structure of the CmPPO protein was similar to apple complexase (MdPPO1), sequence identity was 72.75%, sequence similarity was 53%, and amino acid sequence identity was 68.43%, suggesting that they had similar functions. Based on RT-PCR, the expression of CmPPO gene in difference cultivated varieties and tissues of chestnut kernels was different from the degree of fruit browning, but the highest expression was found in weak browning varieties. [Conclusion] This indicated that the browning was not controlled by single gene, but by multiple genes and metabolic pathways. [ABSTRACT FROM AUTHOR]

【Objective】This paper aimed to isolate and conduct bioinformatics analysis of the sucrose: sucrose-1-fructosyltransferase(1-SST) gene from Jerusalem artichoke. 【Method】 The Jerusalem artichoke 1-SST by Tail-PCR was isolated. A sequence of 1816 bp upstream of the gene start codon(SSTP). Bioinformatics analysis of SSTP sequences was performed by PLANTCARE. 【Result】 A total of 140 cis-acting elements were detected in the SSTP sequence. In addition to the 44 TATA-box and 31 CAAT-box elements of the promoter, there were 65 cis-independent expressions of 1-SST gene expression. Acting element of the 65 cis-acting elements, 23 cis-acting elements were capable of predicting their function. There were 13 light-regulating, two components of abscisic acid, anaerobic process and low temperature stress, and one component of salicylic acid, stress resistance, meristem expression and endosperm expression. The identification of these cis-acting elements provided a theoretical basis for 1-FFT participation in the corresponding biological processes. The CaMV 35 S promoter of pCAMBIA1301 vector was replaced with the sequence of SSTP, 1/2 SSTP and 1/4 SSTP, respectively, and transiently expressed in tobacco leaves. After GUS staining, SSTP, 1/2 SSTP and 1/4 SSTP were all driven to drive GUS. Promoter activity of gene expression, and activity is comparable to CaMV35 S. 【Conclusion】 It is preliminarily estimated that the promoter core region of 1-SST gene is within 400 bp upstream of the start codon. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to obtain the transcriptome dataset of roots of Angelica dahurica and understand the differentially expressed gene(DEG) of roots of Angelica dahurica in different areas. 【Method】The root transcriptome dataset of Angelica dahurica was obtained using the high-throughput sequencing platform Illumina HiSeq X Ten, followed by differentially expressed genes analyses.【Result】The number of differentially expressed gene between cbz, bbz and cbz vs bbz were 2686, 1704, and 11 549, respectively, and annotated by GO with 1713,1112, and 4418. And the functions of differentially expressed genes were related to cell composition, cells, organelles, binding, catalytic activity, cell processes, and metabolic processes. The KEGG pathway classification showed that the differentially expressed gene was divided into 4 categories and 24 subcategories, and the pathway was mainly related to the metabolic process.The expression of genes encoding 4-coumarate CoA ligase, Phenylalanine ammonia-lyase 3 and 3-hydroxy-3-methylglutaryl-coenzyme A reductase increased in cbz, while, the expression of genes encoding dehydrodolichyl diphosphate synthase and probable 1-deoxy-D-xylulose-5-phosphate synthase 2 decreased. 【Conclusion】In this study, the differentially expressed gene of Angelica dahurica root was analyzed by transcriptional group sequencing, which provided theoretical basis for the formation of quality of Angelica dahurica in the future. [ABSTRACT FROM AUTHOR]

[Objective] In order to study salt tolerant pepper varieties and improve the salt tolerance and drought resistance of dried pepper, the CaNAC61 gene was cloned, bioinformatics and expression level were analyzed. [Method]The CaNAC61 gene was cloned by RT-PCR, and the physicochemical properties, secondary and tertiary structure of protein and phylogenetic tree were analyzed. Quantitative real-time PGR ( qRT-PCR) was used to analyze the expression of the gene under stress. [Result] NAG transcription factor gene CaNAC61 (Genebank: XM_016719693. 1) was obtained from dried Pepper, the open reading frame length of the gene was 885 bp, encoding 294 amino acids, and the relative molecular weight of protein was 33.67 kD , the theoretical isoelectric point was 6. 26. CaNAC61 was aligned with the homology of NAG family members in different plants and the phylogenetic tree analysis showed that, CaNAC61 belongs to the ATAF subfamily. Quantitative RT-PGR analysis showed that CaNAC61 responded under 6 abiotic stress treatments. The expression was up-regulated more significantly under NaCI treatment than other stress treatment, followed by drought treatment. [Conclusion]In this paper, CaNAC61 gene was cloned successfully, which provided a theoretical basis lor studying the function of NAG transcription factors in dried pepper. [ABSTRACT FROM AUTHOR]

[Objective] Phospholipase D(PLD) is an important protein family in rice, which works essentially for growth, development and response for abiotic stresses. The present paper aimed to acquire the basic biological information of rice PLD. [Method]The bioinformatics studies of 16 PLD-family members were performed on gene structure, protein motif, spatio-temporal expression patterns and hormone-induced expression patterns. [Result]The rice PLD genes varied in the length from 2982 to 11 305 bp, and they contained different exon numbers from 3 to 21 while it remained almost the same numbers for those genes belonging to a subtype. Moreover, this family presented a high similarity on protein sequence, and exhibited same motifs except for OsPLDφ and OsPLDκ. According to the database of RiceXPron, PLD-family genes were found expressed higher in roots, stems, leaves, anthers and ovary, and lower in germ, endosperm and other parts. In addition, they behaved different response to honnones, many of which were up-regulated by jasmonic acid(JA) and abscisie acid(ABA), repressed by brassinolide(BR) and cytokinin(CTK), while insensitive to gibberellin(GA) and auxin(IAA). [Conclusion] Members contain the same type of motifs, indicating that they are functionally conserved and consistent;each member exhibits a different response pattern after different hormone treatments, suggesting that rice PLD members perform differenl functions in stress response. These findings provided a basis for further studies on biological functions of rice PLD family members. [ABSTRACT FROM AUTHOR]

[Objective] The present paper aimed to study the expression difference of the six melanin-related genes of goat ASIP,EDN3,PAX3,TYR,TYRP1andDCT in the skin tissues of adult goats with different skin color and the expression pattern in B16-F10 cells.[Method] The skin tissue of Youzhou dark-skin goats and Banjiao goats and the B16-F10 cells of proliferation phase and differentiated phase were used as experimental materials, then the expression levels of six melanin-related genes were detected by qRT-PCR and the correlation of six genes was also analyzed.[Result] The results of skin tissue test showed that compared to Banjiao goats, the expression of ASIP in Youzhou dark-skin goats was significantly down-regulated(P<0.01),but PAX3, TYR, TYRP1 and DCT were significantly up-regulated(P<0.05 or P <0.01).The results of vitro experiments showed that in proliferation phase of melanocytes, expression patterns of ASIP, EDN3, TYRP1 and DCT were similar, while PAX3 and TYR were similar, indicating that they played different regulatoiy roles in pigment cell proliferation phase. In differentiation phase, ASIP had similar expression patterns with TYR, TYRP1 and DCT, and the expression levels were significantly correlated, suggesting that they were involved in the differentiation of melanocytes and the regulation of melanin synthesis. [Conclusion] The down-regulation of ASIP, the up-regulation of PAX3, TYR, TYRP1 and DCT were closely related to the abnormal depositional phenotype of skin melanin of Youzhou dark-skin goaLs. [ABSTRACT FROM AUTHOR]

Objective] This paper aimed to investigate the effects of different nitrogen levels on the expression of 0sAAP6 gene in rice and further explore the effects of OsAAP6 gene on the contents of protein (including glutenin, prolamin, globulin and albumin) and amino acids in rice grains under different nitrogen fertilizer conditions so as to utilize OsAAP6 gene for improving the nutritional quality of rice in the future. [Method] Five nitrogen concentration gradients were set up to detect the expression level of OsAAP6 gene from the genetic materials in rice, and the protein contents and amino acid contents in the mature seeds by near infrared grain analyzer and Coomassie Brilliant Blue G-250 method were analyzed. [Result]With the increase of nitrogen fertilizer concentration in the field, the expression level of OsAAP6 gene increased significantly, which could promote the accumulation of glutenin in rice grain and then increased the total content of grain protein in rice. Simultaneously, with the increase of nitrogenous fertilizer application, OsAAP6 gene could significantly increase the total content of amino acids in rice grain. [Conclusion] OsAAP6 gene expression level increased with the increase of nitrogen fertilizer concentration in the field, and could promote the nutritional quality in rice, such as the protein content and amino acids content in the grain, which provided an important reference for the future utilize of OsAAP6 gene to improve the nutritional quality of rice grain. [ABSTRACT FROM AUTHOR]

[Objective] Steroidal saponins is one of the main active compounds of Liriope spicala var. prolifera Y. T. Ma (LSVP). For improving its content in tuber by gene expression regulation, it was very necessary to screen the proteins or enzymes involved in steroidal saponins synthesis. [Method] In the present paper, the suppression subtractive hybridization (SSII) libraries were constructed using the cDNA of LSVP tuber from different growth period; the beginning growth stage tuber (SJQ) and the harvest stage tuber (CSQ). [Result]308 effective EST sequences, which 586. 99 bp average length and mainly distributed in 250 - 1000 bp, were obtained from SJQ and CSQ SSI1 cDNA library. In addition, the sequences that could match the genes in the database of NR, SWISS PROT, TREMRL, CDD, PFAM were 65. 26 % , 34. 42 % , 63. 96 % , 65. 26 % and 14. 94 % ,respectively. And the 23 and 103 Unigenes were found that could be classified into 12 COG classifications and 19 KOG classifications respectively as well. The KEGG analysis showed that there were 81 unigenes which could be noted into 48 enzymes and mapped into 92 KEGG pathways. Gene Ontology analysis was also conducted and the unigene ’ s distribution in different functional category was found that in biological process, mainly focused on cellular process and metabiolie process, and in cellular component, mainly related to cell, cell part and organelle, and in molecular function, were binding and catalytic activity. [Gonelusion] In this experiment, the suppression subtractive hybridization (SSII) libraries were successfully constructed based on steroidal saponins, and several differentially expressed genes were screened. [ABSTRACT FROM AUTHOR]