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Averrhoa carambola, a key tropical and subtropical economic tree in the Oxalidaceae family, is distinguished by its unique pentagram-shaped fruit. This study investigates the developmental processes shaping the polarity of A. carambola fruit and their underlying hormonal and genetic mechanisms. By analyzing the Y1, Y2, and Y3 developmental stages—defined by the fruit diameters of 3–4 mm, 4–6 mm, and 6–12 mm, respectively—we observed that both cell number and cell size contribute to fruit development. Our findings suggest that the characteristic pentagram shape is established before flowering and is maintained throughout development. A hormonal analysis revealed that indole-3-acetic acid (IAA) and abscisic acid (ABA) show differential distribution between the convex and concave regions of the fruit across the developmental stages, with IAA playing a crucial role in polar auxin transport and shaping fruit morphology. A transcriptomic analysis identified several key genes, including AcaGH3.8, AcaIAA20, AcaYAB2, AcaXTH6, AcaYAB3, and AcaEXP13, which potentially regulate fruit polarity and growth. This study advances our comprehension of the molecular mechanisms governing fruit shape, offering insights for improving fruit quality through targeted breeding strategies. [ABSTRACT FROM AUTHOR]

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Copyright of Acta Prataculturae Sinica is the property of Acta Prataculturae Sinica Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This study proposed a two-step process optimization for enhanced cumulative isoprene production. This involves establishing an isoprene biosynthetic pathway in Escherichia coli BL21 DE-3 via up-regulation of native deoxy xylulose 5-phosphate (DXP) synthase (Ec DXS), isopentenyl pyrophosphate isomerase (Ec IPPI /Ec IDI) and introducing isoprene synthase (Pm IspS) from kudzu (Pueraria montana), followed by the process conditions (incubation temperature, time, and inducer concentration) optimization using the Box-Behnken design (BBD) approach. BBD showed the maximum cumulative isoprene production (160.15 mg L-1), productivity (11.18 mg L-1 h-1), and yield (7.69 mg gdcw-1) at the optimized process conditions (incubation temperature 27.32 ºC, incubation time 14.25 h, and inducer concentration 0.453 mM). The transgenic E. coli isoprene production, -productivity, and -yield were 1.52-, 1.46-, and 1.24-fold higher than the unoptimized condition (incubation temperature 30 ºC, incubation time 16 h, and inducer concentration 0.10 mM). This work demonstrates that fine tuning of MEP pathway in addition with process conditions optimization is an efficient strategy for improving isoprene production from engineered E. coli. [Display omitted] • Isoprene, a high-value natural product, entirely being produced by petroleum route. • Recombinant E. coli has potential for sustainable production of isoprene. • DXS, IPPI, and IspS were overexpressed under IPTG inducible promoter. • RSM based statistical model was used for optimizing culture growth conditions. • Engineered E. coli produced 160.15 mg L-1 of isoprene at optimal values of variables. [ABSTRACT FROM AUTHOR]

Drought is a significant environmental limiting factor that restricts the growth of Quercus wutaishanica Mayr. The MYB transcription factor plays a wide role in controlling the growth of plants. In this study, the QwMYB108 gene was cloned and the bioinformatics was analyzed, and we examined how QwMYB108 responded to various gradient drought stresses. The results demonstrated that QwMYB108 encoded 275 amino acids using an 828 bp open reading frame. Subcellular localization indicated that the gene was located in the nucleus. Phylogenetic analysis showed that QwMYB108 was close to Q. robur, and that the highest level of expression was found in leaves, which was significantly different from other tissues. The expression of QwMYB108 increased as the stress degree rose when drought stress was present, and there was a significant difference between severe drought stress and other gradient stress. In this study, the function of QwMYB108 in drought stress response was investigated, and the drought response function gene of Q. wutaishanica was further explored to provide a theoretical basis. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

【Objective】The purpose of this study was to clone the FLS2 gene from wild eggplant species (Solanum sisymbriifolium Lam.) in Yunnan province, and to analyzed the physicochemical properties, subcellular localization and phylogenetic evolution of its encoded protein, as well as the expression, in order to preliminarily explore the biological functions of the wild eggplant FLS2 gene under the stress of verticillium wilt.【Method】In this study, the FLS2 gene, named SsFLS2, was cloned based on the transcriptome data (S. sisymbriifolium inoculation with verticillium pathogens) . The physical and chemical properties of SsFLS2 gene were analyzed by using bioinformatic analysis software, and real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expression of S. sisymbriifolium roots, stems and leaves, as well as the expression at different times after inoculation of verticillium pathogens.【Result】The results showed that SsFLS2 gene was 3655 bp in length, has a complete open reading frame (ORF), and encoded 1126 amino acids. The molecular formula of the encoded protein was C5596H8814N1494O1627S4, the theoretical molecular weight was 124.34 kD, the theoretical isoelectric point (pI) was 7.33, and the total mean hydrophilicity coefficient was 0.081. The secondary structure of SsFLS2 was mainly composed of α-helix (40.23%), random coil (42.81%), β-folding (3.73%), and extended chain (13.23%), and there was a transmembrane structure, localized on the cell membrane, in which the sites that can be phosphorylated and exceed the threshold line, a total of 151. The results of homologous sequence alignment and phylogenetic evolutionary analysis showed that the amino acid sequence of SsFLS2 protein was most closely related to the potato (Solanum tuberosum) homologous protein (XP 006358149.2) . RT-qPCR assay indicated that SsFLS2 gene was expressed in roots, stems, and leaves of S. sisymbriifolium, and that the relative expression level of SsFLS2 gene was highly significant higher in roots and leaves than in stems. The relative expression level of SsFLS2 gene was substantially and highly significantly higher at 24 h in both treatment and control groups than at other time points within 72 h after inoculation with verticillium pathogens.【Conclusion】In this study, SsFLS2 gene was successfully cloned from S. sisymbriifolium, and and its expression as well as the physicochemical properties of its encoded protein were analyzed. The results showed that SsFLS2 gene was closely correlated to the response of S. sisymbriifolium to verticillium wilt stress, which lays a theoretical foundation for further study on the role of SsFLS2 gene in verticillium wilt resistance in S. sisymbriifolium. [ABSTRACT FROM AUTHOR]

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract Background Cabbage (Brassica oleracea L. var. capitata) is an important crop within the Brassica oleracea species and is extensively cultivated worldwide. In recent years, outbreaks of downy mildew caused by Hyaloperonospora parasitica have resulted in substantial losses in cabbage production. Despite this, there have been limited studies on genes associated with resistance to downy mildew in cabbage. Results This study identified sister lines exhibiting significant differences in disease resistance and susceptibility. Using bulked segregant analysis followed by sequencing (BSA-seq) and linkage analysis, the cabbage resistance locus BoDMR2 was accurately mapped to an approximately 300 kb interval on chromosome 7. Among the candidate genes identified, several single nucleotide polymorphisms (SNPs) and a 3-bp insertion were found within the conserved domain of the Bo7g117810 gene, encoding a leucine-rich repeat domain protein, in susceptible genotypes. Additionally, real-time quantitative polymerase chain reaction (RT‒qPCR) analysis revealed that the expression level of Bo7g117810 in resistant specimens was 2.5-fold higher than that in susceptible specimens. An insertion‒deletion (InDel) marker was designed based on the identified insertion in susceptible materials, facilitating the identification and selection of downy mildew-resistant cabbage cultivars. Conclusions This study identifies Bo7g117810 as a potential candidate gene associated with adult-stage resistance to downy mildew in cabbage, supported by observed differences in gene sequence and expression levels. Furthermore, the development of an InDel marker I1-3, based on its mutation, provides valuable resources for breeding resistant cabbage cultivars.

High-yield strains of cold-active lipase are screened to provide production information for the industrial development of lipase. A cold-active lipase-producing strain was isolated from Xing'an larch forest soil in Mohe County using the Rhodamine B plate method. Its morphological, physiological, and biochemical characteristics were identified, and combined with the 16S rDNA gene sequence, the strain was determined to be Bacillus thuringiensis. The lipase gene Lip240 was cloned using the Bacillus thuringiensis genome as a template, and heterologous expression and enzymatic properties of Lip240 were analyzed. The results showed that the recombination lipase Lip240 had an optimal reaction temperature of 30 ℃, and could maintain more than 60% activity when treated at 4~30 ℃ for 6 hours, thus a cold-active lipase. Besides, its optimal pH was 8.0. Additionally, Ca2+, Mn2+, Fe2+, Fe3+, and Cr3+ had a certain enhancing effect on the activity of Lip240. Notably, Lip240 had a good organic solvent tolerance, while SDS could significantly (P

Abstract [Objective] The study aims to clone the fructokinase (FRK ) gene of red pitaya (Hylocereus polyrhizus), detect its expression and enzyme activity, and analyze its physiological function in soluble sugar accumulation during fruit development. [Methods] HpFRK1 gene was cloned from ‘Zihonglong’ fruits, and bioinformatics and subcellular localization analysis were carried out. Gene expression was detected by qRT-PCR. Recombinant protein was obtained by E.coli induction and enzyme activity was detected. [Results] The open reading frame (ORF) of HpFRK1 was 993 bp in length, encoding 330 amino acids with a phosphofructokinase type B (pfkB) domain. HpFRK1 had the closest genetic relationship with sugar beet BvFRK, both of which belonged to cytoplasm localized FRKs. HpFRK1 was expressed in different developmental stages of mature stems and fruits. The expression level of HpFRK1 was the highest at 20 days after flowering and the lowest at 30 days after flowering (fruit ripening), and was gradually decreased with fruit development. Subcellular localization assay showed that HpFRK1 was mainly localized in the nucleus and cytoplasm. HpFRK1 recombinant protein specifically catalyzed fructose phosphorylation (K m=11.01 mmol/L). [Conclusion] HpFRK1 specifically catalyzes fructose phosphorylation in the cytoplasm and negatively regulates fruit soluble sugar accumulation during the development of red-fleshed pitaya fruits

Abstract [Objective] As a peony breeding material, Paeonia delavayi has abundant flower color resources. Studying the regulatory effects of its MYB transcription factors on flower color will benefit molecular breeding of peony flower color. [Methods] Using the flowers of yellow and red P . delavayi as materials for hand sectioning. Based on the transcriptomic data of P . delavayi, a MYB transcription factor coding gene PdMYB57 was obtained through sequence alignment. Function of this gene was verified through gene cloning, phylogenetic tree construction, qRT-PCR, transient expression, and HPLC. [Results] PdMYB57 gene has an ORF of 798 bp, encoding an unstable hydrophilic protein of 265 amino acids. PdMYB57 was clustered with the Arabidopsis SG6 subfamily, as well as MYB transcription factors in P . qiui PqMYB113, P . suffruticosa PsMYB57/PsMYB58, and Vitis vinifera VvMYBA1/VvMYBA2. PdMYB57 had a R2R3 conserved domain [R/K] Px [P/A/R] xx [F/Y] motif. PdMYB57 gene was highly expressed in sepals of red P . delavayi and leaves of yellow P . delavayi. HPLC analysis showed that tobacco leaves transiently expressed PdMYB57 gene contained cyanidin-3-O-rutinoside (Cy3R). [Conclusion] PdMYB57 gene encodes an R2R3-MYB transcription factor. PdMYB57 gene expression is tissue-specific and promotes anthocyanin synthesis in plants.

Abstract [Objective] This study aims to explore the function of VuARI2 gene in response to low temperature stress, by cloning and expressing VuARI2 gene in Arabidopsis. [Methods] VuARI2 gene was cloned from the stem of bog bilberry by RACE and RT-PCR and conducted a comprehensive bioinformatics analysis. Expression of VuARI2 in different tissues after cold acclimation was analyzed by RT-qPCR. Furthermore, the transgenic Arabidopsis was analyzed for the expression of VuARI2, physiological indexes, and freezing resistance. [Results] VuARI2 was successfully cloned,with an open reading frame of 1 770 bp, encoding 589 amino acids. The multi-sequence alignment and phylogenetic tree indicated that ubiquitin ligase VuARI2 was closely related to the amino acid sequence of ARI2 proteins in wild camellia, coffee tree, grape, and riverside grape. RT-qPCR showed that the expression of VuARI2 in leaves and flower buds of bog bilberry was higher than that in roots and stems, and the expression of VuARI2 in stems and flower buds was induced under low temperature. After cold acclimation, expression levels of VuARI2 gene in transgenic plants were higher in wild type, and chlorophyll content was lower than the wild type. Proline content and soluble sugar content were higher than the wild type. Superoxide dismutase activity, peroxidase activity, and catalase activity were higher than the wild type, while relative conductivity and malondialdehyde content were lower than the wild type. After cold acclimation, the survival rate of transgenic plants was higher than the wild type,and the relative conductivity was lower than the wild type at freezing temperature,enhancing freezing resistance of plants. [Conclusions] VuARI2 gene is cloned from bog bilberry and expressed in Arabidopsis. Overexpress of VuARI2 gene enhances the cold resistance of plants.

To explore substitutes for natural tropomyosin as basic materials for the diagnosis and treatment of shrimp allergy, total RNA was extracted from Macrobrachium nipponense, and the full-length sequence of the tropomyosin gene was cloned by rapid amplification of cDNA ends (RACE). Specific primers were designed based on the cDNA sequence of tropomyosin and the expression plasmid pET-30a-TM was constructed. Finally, recombinant tropomyosin was expressed under the induction of isopropyl-β-D-thiogalactopyranoside (IPTG). The results showed that the full-length cDNA sequence of tropomyosin was 1 658 bp in length (GenBank accession number: OP974621). Its open reading frame (ORF) was 855 bp in length, encoding 284 amino acids, with a predicted protein isoelectric point of 4.70 and a predicted molecular mass of 32.8 kDa. The highest expression was obtained after 4 h of induction with a final IPTG concentration of 1 mmol/L at 37 ℃. The recombinant tropomyosin mainly existed as a soluble form in the supernatant of cell lysate with a molecular mass of approximately 38 kDa.

Abstract [Objective] WRKY is one of the plant specific transcription factor families, involved in various plant life activities. However, currently there are few reports related to the WRKY70 gene in strawberry. Analyzing the role of WRKY70 homologous genes in strawberry in response to stress will help accelerate the application of molecular breeding technology and cultivate new strawberry germplasm resources. [Methods] The FaWRKY70 gene was cloned from ‘Benihoppe’ strawberry fruits using homologous cloning method. Its conserved domain, physicochemical properties, protein structure, and evolutionary relationship were analyzed using bioinformatics. Expression pattern analysis was performed using qRT-PCR. [Results] The FaWRKY70 gene had a length of 1 020 bp and encodes 339 amino acids. Homologous gene alignment revealed that FaWRKY70 had a high degree of amino acid sequence similarity with species in the same family such as apple and peony. The homologous genes were mostly related to plant responses to biotic and abiotic stress, suggesting that FaWRKY70 may be involved in resistance to stress. FaWRKY70 was expressed in different organs of strawberry, with significant differences. The expression level was highest in flowers and lowest in fruits. Under salicylic acid treatment, the FaWRKY70 gene was quickly responded, reaching its highest expression level after 3 hours, and gradually decreased thereafter. Under the treatment with methyl jasmonate, the FaWRKY70 gene showed a slight degree of induction and an overall downward trend. [Conclusion] FaWRKY70 is involved in growth and hormone signal transduction through different response modes in strawberry.

【Objective】Trehalase (Tre) is a key enzyme in trehalose metabolism of Conopomorpha sinensis Bradley, which plays an important role in energy metabolism and growth development by specifically hydrolyzing trehalose into glucose. The study aims to clone two trehalase genes (CsTre1 and CsTre2) from Conopomorpha sinensis Bradley, to clarify its expression patterns in different developmental stages and tissues, and to analyze the molecular characteristics of the two genes and their enzyme proteins.【Method】Based on the transcriptome data of C. sinensis, the full-length cDNA sequences of CsTre1 and CsTre2 were cloned with the rapid amplification of cDNA ends (RACE)-PCR. Bioinformatics analysis was performed with software such as ORF Finder, ProtParam, SignalP 4.1, ProtScale, NetPhos2.0 Server and IQ TREE. The mRNA expression patterns of CsTre1 and CsTre2 in different developmental stages and tissues of C. sinensis were detected by using real-time quantitative PCR (RT-qPCR).【Result】The open reading frame of CsTre1 was 1 701 bp, encoding 566 amino acids, and the protein molecular weight was 64.53 kD. The open reading frame length of CsTre2 was 1 821 bp, encoding 606 amino acids, and the protein molecular weight was 69.08 kD. Signal peptide prediction analysis showed that both front-ends of CsTre1 and CsTre2 had a signal peptide, with positions of 1-16 and 1-17, respectively. The analysis on the secondary structure of the sequence showed that both CsTre1 and CsTre2 were mainly composed of α-helix and random coil, CsTre1 had 24 Sers, 15 Tyrs, and 10 Thrs that may serve as binding sites for protein kinases, while CsTre2 had 27 Sers, 10 Tyrs, and 13 Thrs that may serve as binding sites for protein kinases. The RT-qPCR results revealed that CsTre was expressed throughout all developmental stages of C. sinensis. In the expression pattern of adult stage, the expression level of CsTre1 was much higher than that of CsTre2, and the expression level of male adults of CsTre1 dropped sharply after the fourth day.【Conclusion】The study successfully cloned two trehalose genes of Conopomorpha sinensis Bradley. According to the results of analysis on their molecular characteristics and expression patterns, CsTre1 may be the main gene regulating trehalose metabolism in Conopomorpha sinensis Bradley. The research results provide important clues for elucidating the function of trehalase genes, which lay a solid foundation for the development pest control strategies.

[Objective] To explore the biosynthetic pathway of ergothioneine in Naematelia aurantialba and Stereum hirsutum. [Method] The ergothioneine synthase gene Egt1 and Egt2 of N. aurantialba and S. hirsutum were cloned by PCR amplification technology, respectively, and their functions were analyzed by bioinformatics software. High performance liquid chromatography（HPLC）was used to identify the intermediate products of hercynine, ergothioneine and their contents in two species. [Result] The complete DNA sequences of Egt1 and Egt2 genes of two species were successfully cloned. The analysis using bioinformatics software revealed that Egt1 of the two species contained functional binding domains such as EgtD and SAM-dependent methyltransferase. Egt2 contained the binding sites for pyridoxal phosphate （LPL）and cysteine desulfurase. Further analysis indicated that Egt1 and Egt2 shared similar functional domains and substrate binding sites with model fungi（Schizosaccharomyces pombe and Neurospora crassa）. This result showed that Egt1 and Egt2 may have similar gene functions as these model fungi. HPLC analysis revealed the presence of hercynine and ergothioneine in N. aurantialba blastospore（JEYB）, S. hirsutum fermentum broth（ShFJY）, S. hirsutum mycelium（ShJST）and N. aurantialba fruiting bodies（JEZST）. Additionally, the ergothioneine content in the JEZST was found to be the highest at 113.19 μg/g, which was 7.45 times, 26.14 times, and 27.74 times higher than that of the JEYB, ShFJY, and ShJST, respectively. [Conclusion] The Egt1 and Egt2 genes of N. aurantialba and S. hirsutum were identified for the first time. It is hypothesized that the biosynthetic pathway of N. aurantialba and S. hirsutum are involved the catalysis of histidine by the Egt1 enzyme, resulting in the formation of hercynine. Subsequently, the Egt1 enzyme catalyzes the conversion of hercynine into hercynylcysteine sulfoxide. Finally, the Egt2 enzyme catalyzes the transformation of hercynylcysteine sulfoxide into ergothioneine. [ABSTRACT FROM AUTHOR]

[Objective] Cytochrome P450 monooxygenase 98（CYP98）is a key rate-limiting enzyme in the phenylpropanoid pathway, and it is proposed to investigate whether it is involved in the biosynthesis of the phenylpropanoid pathway and in the anti-disease function in Hornworts, so as to provide a reference for the future study of the evolution of the early terrestrial plants and physiological mechanisms of adaptation to adversity stress. [Method] The full-length sequence of FfCYP98 cDNA was cloned from Folioceros fuciformis using RACE, analyzed for bioinformatics and subcellular localization, and functionally verified by constructing an overexpression vector to transform the Arabidopsis thaliana mutant FfCYP98-OE1. [Result] The FfCYP98 cDNA sequence, with an open reading frame of 1305 bp, is evolutionarily closest to the Bryophytes Anthoceros angustus and Physcomitrium patens CYP98, and subcellular localization results show that FfCYP98 is localized to both the nucleus and the cytoplasm. The overexpression of the FfCYP98 gene revealed that the expressions of total phenolics, total flavonoids, and C4H,4CL and C3H in the phenylpropanoid pathway was significantly higher in FfCYP98-OE1 plants than in A. thaliana WT plants. In addition, infestation of F. fuciformis and A. thaliana FfCYP98-OE1 plants with Botrytis cinerea revealed that the expression of FfCYP98 was significantly up-regulated, that FfCYP98-OE1 plants died significantly slower than WT plants, and that the expressions of total phenolics, total flavonoids, and four genes, C4H, HCT, C3H, and CAD in the phenylpropanoid pathway were significantly higher in A. thaliana FfCYP98-OE1 plants than in WT plants. [Conclusion] FfCYP98 may be involved in the biosynthesis of phenolics and flavonoids in the phenylpropanoid pathway by regulating the expressions of genes related to the synthesis of phenylpropanoid pathway, and may be related to disease resistance of plants. [ABSTRACT FROM AUTHOR]

[Objective] The heading date of rice（Oryza sativa）is an important agronomic trait and crucial for the regional adaptability and yields. Identification and functional analysis of heading date-associated genes may provide important gene resources for the breeding in rice. [Method] BSA-seq was performed to identify target gene of a yellowish and early heading mutant huangzao 1（hz1）. RT-qPCR was used to analyze the expression profile of target genes. The subcellular localization of HZ1 was determined by transient transformation between rice protoplasts and tobacco cells. Meanwhile, the heading dates, chlorophyll content and hydrogen peroxide content were measured to analyze the phenotype of hz1 mutant. [Result] Field phenotypic observations found that hz1 showed early heading phenotype. The heading date of hz1 mutant was the same under long-day（LD）and short-day（SD）conditions, which were 43 d and 26 d earlier than those of wild type（WT）respectively, indicating that hz1 was a photoperiod-insensitive mutant. Moreover, hz1 demonstrated a yellowish phenotype, with a decrease in chlorophyll content compared with WT. Genetic analysis revealed that hz1 was controlled by a recessive gene. BSA-seq（Bulk segregation analysis with whole-genome sequencing）of F2 pools indicated that the causal gene was linked with molecular markers on the chromosome 6 with 17.8 Mb region. Further analysis demonstrated that LOC_Os06g40080 site with a T-DNA insertion was completely linkage with hz1 mutant. LOC_Os06g40080 site was an identified SE5 gene encoding heme oxygenase 1（HO1）. Expression pattern analysis revealed that HZ1/SE5 was highly expressed in the leaves and dhad diurnal rhythm expression. Subcellular localization assay showed that HZ1/SE5 protein was localized in chloroplasts. Gene expression analysis showed that HZ1/SE5 regulated the expression of florigen genes Hd3a and RFT1 to modulate the heading date in rice. In addition, HZ1/SE5 can also regulate the expression of genes associated with chlorophyll synthesis pathway to modulate the change of chlorophyll levels. [Conclusion] hz1 mutant is a photoperiodic insensitive mutant due to the mutation of heme oxygenase gene SE5. HZ1/SE5 may regulate the expression of the florigen genes and chlorophyll synthesis-associated genes to influence the heading date and leaf yellowish in rice. [ABSTRACT FROM AUTHOR]

Various environmental stresses induce the production of reactive oxygen species (ROS), which have deleterious effects on plant cells. Glutathione (GSH) is an antioxidant used to counteract reactive oxygen species. Glutathione is produced by glutamylcysteine synthetase (GCS) and glutathione synthetase (GS). However, evidence for the GCS gene in sweetpotato remains scarce. In this study, the full-length cDNA sequence of IbGCS isolated from sweetpotato cultivar Xu18 was 1566 bp in length, which encodes 521 amino acids. The qRT-PCR analysis revealed a significantly higher expression of the IbGCS in sweetpotato flowers, and the gene was induced by salinity, abscisic acid (ABA), drought, extreme temperature and heavy metal stresses. The seed germination rate, root elongation and fresh weight were promoted in T3 Arabidopsis IbGCS-overexpressing lines (OEs) in contrast to wild type (WT) plants under mannitol and salt stresses. In addition, the soil drought and salt stress experiment results indicated that IbGCS overexpression in Arabidopsis reduced the malondialdehyde (MDA) content, enhanced the levels of GCS activity, GSH and AsA content, and antioxidant enzyme activity. In summary, overexpressing IbGCS in Arabidopsis showed improved salt and drought tolerance. Headings: •IbGCS gene was cloned from the sweetpotato cultivar Xu18. • The multiple abiotic stresses significantly increased the expression of IbGCS gene. • Overexpression of IbGCS significantly improved the salt and drought stress tolerance [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Northwest A & F University - Natural Science Edition is the property of Editorial Department of Journal of Northwest A&F University (Natural Science Edition) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Vibrio parahaemolyticus is a Gram-negative zoonotic pathogen, which can cause food-borne diseases such as human gastroenteritis. V. parahaemolyticus has many virulence factors and thermolabile hemolysin (TLH) is one of them. In this study, the tlh gene of V. parahaemolyticus was cloned and sequenced, and the bioinformatics analysis and structural and functional prediction of TLH protein were carried out. Results showed that the TLH consisted of 418 amino acids with a predicted molecular weight of 47.36 kDa. Several conserved domains and motifs were identified in TLH protein, including SGNH hydrolase domain and GDSL motif. Based on the homologous modeling and verification of the three-dimensional structure of TLH protein, the stability of TLH protein, the interaction ability with 4-nitrophenyl laurate and the influence of binding substrate on the structure compactness and residue flexibility were analyzed by molecular dynamics simulation. It was revealed that the periphery of the catalytic triad Ser153-His390-Asp393 is an important drug target active pocket with highly conserved residue sequences, and the residues showed flexibility difference after substrate binding. This study analyzed the properties and structure of V. parahaemolyticus TLH, which can provide theoretical support for ensuring the safety of aquatic products and improving the safety evaluation level of aquatic raw materials. [ABSTRACT FROM AUTHOR]

PhoR is a histidine kinase in a two-component regulatory system that regulates phosphorus metabolic pathways and undertakes the key mission of information transmission in pathogenic bacteria. The full-length phoR gene was successfully cloned from the Vibrio alginolyticus HY9901 strain. A comprehensive analysis of the cloned gene was conducted using bioinformatics. Sequence analysis revealed that the total length of the phoR gene (GenBank accession No.: KJ958404.1) is 1299 bp, with the coding region containing a total of 432 amino acid residues. The phylogenetic tree of PhoR revealed that it belongs to the same subclade as V. diabolicus. The SMART program was employed for the purpose of functional domain prediction, which revealed that PhoR possesses three major functional domains: PAS (amino acids 98 -166), His-KA (amino acids 205-272), and HATPase_c (amino acids 317429). [ABSTRACT FROM AUTHOR]

【Objective】The cytochrome P450 family is an important enzyme system involved in glucosinolate synthesis in cruciferous plants, where in the CYP83 subfamily plays a predominant role in core structure formation, this work aims to investigate the Chinese cabbage (Brassica rapa ssp. Pekinensis) CYP83B1 gene functional role.【Method】RT-PCR was used to clone the CYP83B1 gene. Bioinformatics analysis software was utilized to predict the encoded protein's physicochemical properties, homology of the encoded protein, and promoter cis-acting elements. The expression pattern of BrCYP83B1 was analyzed by RT-qPCR, and a plant overexpression vector was constructed for further experimentation.【Result】The cDNA length of BrCYP83B1 was 1 500 bp, encoding a total of 499 amino acids. The protein belonged to cytochrome P450 superfamily and predominantly located in the cytoplasm. Its secondary structure primarily comprised of α-helixes and irregular coil. Homologous comparison illustrated that BrCYP83B1 had close relationship with Brassica napus L and Brassica oleracea L. var. italica. The BrCYP83B1 promoter contained cis-acting elements that were involved in the response to salicylic acid（SA), abscisic acid（ABA), and methyl jasmonate（MeJA), suggesting that the expression of BrCYP83B1 gene may be regulated by hormones. The expression of BrCYP83B1was detected in various plant organs, including the roots, stems, leaves, flowers and fruits from RT-qPCR results. Notably, the highest expression was observed in the leaves. Moreover, BrCYP83B1 significantly presented induction upon treatment with MeJA, while its expression was repressed by SA. Additionally, ABA treatment initially up-regulated and subsequently down-regulated the gene.【Conclusion】BrCYP83B1 may be involved in the response regulation of Chinese cabbage to hormones. [ABSTRACT FROM AUTHOR]

[Objective] Trehalase (Tre) is a key enzyme in trehalose metabolism of Conopomorpha sinensis Bradley, which plays an important role in energy metabolism and growth development by specifically hydrolyzing trehalose into glucose. The study aims to clone two trehalase genes (CsTre1 and CsTre2) from Conopomorpha sinensis Bradley, to clarify its expression patterns in different developmental stages and tissues, and to analyze the molecular characteristics of the two genes and their enzyme proteins.[Method] Based on the transcriptome data of C. sinensis, the full-length cDNA sequences of CsTre1 and CsTre2 were cloned with the rapid amplification of cDNA ends (RACE)-PCR. Bioinformatics analysis was performed with software such as ORF Finder, ProtParam, SignalP 4.1, ProtScale, NetPhos2.0 Server and IQ TREE. The mRNA expression patterns of CsTre1 and CsTre2 in different developmental stages and tissues of C. sinensis were detected by using real-time quantitative PCR (RT-qPCR).[Result] The open reading frame of CsTre1 was 1 701 bp, encoding 566 amino acids, and the protein molecular weight was 64.53 kD. The open reading frame length of CsTre2 was 1 821 bp, encoding 606 amino acids, and the protein molecular weight was 69.08 kD. Signal peptide prediction analysis showed that both front-ends of CsTre1 and CsTre2 had a signal peptide, with positions of 1-16 and 1-17, respectively. The analysis on the secondary structure of the sequence showed that both CsTre1 and CsTre2 were mainly composed of α-helix and random coil, CsTre1 had 24 Sers, 15 Tyrs, and 10 Thrs that may serve as binding sites for protein kinases, while CsTre2 had 27 Sers, 10 Tyrs, and 13 Thrs that may serve as binding sites for protein kinases. The RT-qPCR results revealed that CsTre was expressed throughout all developmental stages of C. sinensis. In the expression pattern of adult stage, the expression level of CsTre1 was much higher than that of CsTre2, and the expression level of male adults of CsTre1 dropped sharply after the fourth day.[Conclusion] The study successfully cloned two trehalose genes of Conopomorpha sinensis Bradley. According to the results of analysis on their molecular characteristics and expression patterns, CsTre1 may be the main gene regulating trehalose metabolism in Conopomorpha sinensis Bradley. The research results provide important clues for elucidating the function of trehalase genes, which lay a solid foundation for the development pest control strategies. [ABSTRACT FROM AUTHOR]

Phenylalanine ammonia lyase (PAL) is a key enzyme regulating the biosynthesis of the compounds of the phenylpropanoid pathway. This study aimed to isolate and characterize PAL genes from Ferula pseudalliacea Rech.f. (Apiales: Apiaceae) to better understand the regulation of metabolite production. Three PAL gene isoforms (FpPAL1-3) were identified and cloned using the 3′-RACE technique and confirmed by sequencing. Bioinformatics analysis revealed important structural features, such as phosphorylation sites, physicochemical properties, and evolutionary relationships. Expression analysis by qPCR demonstrated the differential transcription profiles of each FpPAL isoform across roots, stems, leaves, flowers, and seeds. FpPAL1 showed the highest expression in stems, FpPAL2 in roots and flowers, and FpPAL3 in flowers. The presence of three isoforms of PAL in F. pseudalliacea, along with the diversity of PAL genes and their tissue-specific expression profiles, suggests that complex modes of regulation exist for phenylpropanoid biosynthesis in this important medicinal plant. The predicted interaction network revealed associations with key metabolic pathways, emphasizing the multifaceted roles of these PAL genes. In silico biochemical analyses revealed the hydrophilicity of the FpPAL isozyme; however, further analysis of substrate specificity and enzyme kinetics can clarify the specific role of each FpPAL isozyme. These comprehensive results increase the understanding of PAL genes in F. pseudalliacea, helping to characterize their contributions to secondary metabolite biosynthesis. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Dalian Ocean University is the property of Journal of Dalian Ocean University Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Transfection efficiency of the immortalized human breast epithelial cell line MCF-10A remains an issue that needs to be resolved. In this study, it was aimed to deliver a recombinant DNA (pCMV-Azu-GFP) to the MCF-10A cells by the magnetofection method using magnetic nanoparticles (MNPs) and a simple magnet to accelerate the DNA delivery. Surface positively modified silica-coated iron oxide MNPs (MSNP-NH2) were produced and characterized via TEM, FTIR, and DLS analyses. The recombinant DNA (rDNA) was obtained by the integration of codon-optimized azurin to produce a fusion protein. Then, rDNA cloned in Escherichia coli cells was validated by sequence analysis. The electrostatically conjugated rDNA on MSNP-NH2 with an enhancer polyethyleneimine (PEI) was studied by agarose gel electrophoresis and the optimum conditions were determined to apply to the cell. A dose-dependent statistical difference was observed on treated cells based on the MTS test. The expression of the fusion protein after magnetofection was determined using laser scanning confocal microscope imaging and western blot analysis. It was observed that the azurin gene could be transferred to MCF-10A cells by magnetofection. Thus, when the azurin gene is used as a breast cancer treatment agent, it can be expressed in healthy cells without toxic effects. [ABSTRACT FROM AUTHOR]

Copyright of Xinjiang Agricultural Sciences is the property of Xinjiang Agricultural Sciences Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The market demand for essential oil containing citral is increasing. Our research group identified a rare chemotype of Camphora officinarum whose leaves are high in citral content by examining over 1000 wild trees across the entire native distribution area of C. officinarum in China. Because C. officinarum is suitable for large-scale cultivation, it is therefore seen as a promising source of natural citral. However, the molecular mechanism of citral biosynthesis in C. officinarum is poorly understood. In this study, transcriptomic analyses of C. officinarum with different citral contents revealed a strong positive correlation between the expression of a putative geraniol synthase gene (CoGES) and citral content. The CoGES cDNA was cloned, and the CoGES protein shared high similarity with other monoterpene synthases. Enzymatic assays of CoGES with geranyl diphosphate (GPP) as substrate yielded geraniol as the single product, which is the precursor of citral. Further transient expression of CoGES in Nicotiana benthamiana resulted in a higher relative content of geranial and the appearance of a new substance, neral. These findings indicate that CoGES is a geraniol synthase-encoding gene, and the encoded protein can catalyze the transformation of GPP into geraniol, which is further converted into geranial and neral through an unknown mechanism in vivo. These findings expand our understanding of citral biosynthesis in Lauraceae plants. [ABSTRACT FROM AUTHOR]

Reducing plant height (PH) is one of the core contents of the "Green Revolution", which began in the 1960s in wheat. A number of 27 reduced-height (Rht) genes have been identified and a great number of quantitative trait loci (QTLs) for PH have been mapped on all 21 chromosomes. Nonetheless, only several genes regulated PH have been cloned. In this study, we found the interval of QTL QPh-1B included an EST-SSR marker swes1079. According to the sequence of swes1079, we cloned the TaOSCA1.4 gene. We developed a CAPS marker to analyze the variation across a natural population. The result showed that the PH was significantly different between the two haplotypes of TaOSCA1.4-1B under most of the 12 environments and the average values of irrigation and rainfed conditions. This result further demonstrated that TaOSCA1.4 was associated with PH. Then, we validated the TaOSCA1.4 via RNAi technology. The average PHs of the wild-type (WT), RNAi lines 1 (Ri-1) and 2 (Ri-2) were 94.6, 83.6 and 79.2 cm, respectively, with significant differences between the WT and Ri-1 and Ri-2. This result indicated that the TaOSCA1.4 gene controls PH. TaOSCA1.4 is a constitutively expressed gene and its protein localizes to the cell membrane. TaOSCA1.4 gene is a member of the OSCA gene family, which regulates intracellular Ca2+ concentration. We hypothesized that knock down mutants of TaOSCA1.4 gene reduced regulatory ability of Ca2+, thus reducing the PH. Furthermore, the cell lengths of the knock down mutants are not significantly different than that of WT. We speculate that TaOSCA1.4 gene is not directly associated with gibberellin (GA), which should be a novel mechanism for a wheat Rht gene. [ABSTRACT FROM AUTHOR]

[Objective]EARLY FLOWERING 3(ELF3), an important component of the core circadian oscillator, has been identified to function greatly in regulating circadian and flowering, and stress responses of plants. Up to now, however, there has been no report on banana ELF3. Our present study is aimed to explore the functions of ELF in banana responses to stresses.[Method]Four ELF3 genes(MaELF3- 1-MaELF3-4)identified from banana(Musa acuminata)genome were cloned using reverse transcription PCR and subjected to a series of bioinformatic analysis. Their expression patterns under treatments of high and low temperature, banana wilt disease pathogen Fusarium oxysporum f. sp. cubense Tropical Race 4(FocTR4), MeJA and ABA were analyzed using transcriptome data and quantitative real time PCR (RT-qPCR).[Result]The coding sequences of the four MaELF3s ranged from 2 058 bp to 2 301 bp, encoding proteins consisted of 685-766 amino acids. Except that MaELF3-4 had three exons, all the other three MaELF3s had four exons. Protein analysis revealed that all MaELF3s were nucleus-located unstable basic proteins; unlike the ELF3s from temperate plants, all ELF3s from banana and some other tropical plants did not contain the prion-like domain(PrD). Phylogenetic analysis results showed that MaELF3-2 and MaELF3-4 shared the closest relationship with Arabidopsis ELF3(At2g25930), while MaELF3-1 and MaELF3-3 was the closest to ELF3 from Ensete ventricosum and Musa balbisiana, respectively. The promoter regions of MaELF3s contained many light-, phytohormone(such as MeJA and ABA)- and stress(such as drought and low temperature)-responsive elements. Gene expression analysis revealed that, the expressions of all the four MaELF3s in the leaves were greatly influenced by JA and ABA treatments, and their expressions in the banana roots was significantly downregulated by FocTR4. Moreover, the expressions of all MaELF3s except MaELF3-4 were downregulated by both high and low temperature treatments.[Conclusion]The results obtained in this study indicate that MaELF3s participate in the banana responses to different stresses. [ABSTRACT FROM AUTHOR]

[Objectives]The paper aimed to provide a basis for studying the function of 14-3-3 protein and breeding of Jerusalem artichoke, we identified the 14-3-3 gene family of Jerusalem artichoke and analyzed their expression patterns in response to high temperature, cold, salt and drought stresses. [Methods]Cloning and bioinformatics analysis were used to study gene properties, and RNA-seq data analysis and RT-qPCR were used to study gene response patterns to abiotic stress. [Results]Ten members of the 14-3-3 gene family, namely HtGRF1-HtGRF10(GenBank accession No.OP132618-OP132627), were cloned from Jerusalem artichoke. According to the evolutionary relationship, these genes were divided into two subfamilies, HtGRF1-HtGRF7 belonged to the non-ε group, and HtGRF8-HtGRF10 belonged to the ε group, which usually formed homologous or heterodimers. The expression level of HtGRF2/3/6/9 in sprouts, roots, stems and leaves was higher than those of other genes. The expression level of HtGRF3/5/9 was higher in stolon and tuber formation stages than that in the tuber rapid intamescentia and maturity stages during the tuber development. Comprehensive analysis of HtGRF in response to abiotic stress in roots and leaves showed that HtGRF6 expression level decreased under high temperature, salt and drought stresses, while HtGRF2/3/7/8/9 expression level decreased under salt stress, but increased under drought stress. HtGRF1 expression level increased under cold stress, HtGRF4/5 expression level decreased under drought stress, and HtGRF10 expression level did not change significantly. [Conclusions]The 14-3-3 protein family in Jerusalem artichoke was highly conserved and encoded by multiple genes, and played an important role in the growth and development and in coping with complex environments. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Bioprocess Engineering is the property of Chinese Journal of Bioprocess Engineering Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The Cytochrome P450 (CYP450) family is the largest enzyme protein family in plants, distributed across various organs and involved in significant catalytic activities in primary and secondary metabolic processes. In this study, we cloned the PtCYP721A57 gene, characterized its open reading frame (ORF), and conducted comprehensive analyses including physicochemical properties, evolutionary relationships, subcellular localization, prokaryotic expression, and correlation between the relative expression of different parts and the content of tenuifolin, hormones, and abiotic stress response associated with the encoded protein. The ORF of PtCYP721A57 was 1,521 bp, with a secondary structure predominantly composed of α-helices and random coils. Subcellular localization experiments confirmed the presence of PtCYP721A57 in the endoplasmic reticulum. For prokaryotic expression, we constructed the recombinant plasmid pET28a-PtCYP721A57 using pET28a as the vector, which was then transformed into BL21(DE3). Induction with Isopropyl β-D-1-thiogalactopyranoside (IPTG) at temperatures of 16 and 25 °C and varying concentrations (0.1, 0.2, 0.5, 1, 2 mM) resulted in the formation of inclusion bodies, with higher expression observed at 25 °C. Our qPCR analyses revealed that PtCYP721A57 exhibited the highest expression in the cortex of Polygala tenuifolia, followed by roots and xylem, correlating with the observed tenuifolin content distribution. Induction with abscisic acid (ABA) and chitosan (CHT) initially decreased PtCYP721A57 expression followed by a subsequent increase, peaking at 48 h. Similarly, drought stress induced a gradual increase in PtCYP721A57 expression, also peaking at 48 h. NaCl treatment for 6 h significantly upregulated PtCYP721A57 expression. In conclusion, our study provides foundational insights into the PtCYP721A57 gene in Polygala tenuifolia, laying the groundwork for further exploration of its role in the biosynthesis pathway of triterpenoid saponins.

Abstract [Objective] Tartaric acid not only affects the taste of grape berries but also determines the color, texture, microbial stability, and aging potential of the wine. It is of great value to explore the transcription factors and mechanisms that regulate tartaric acid biosynthesis, regulate the accumulation of tartaric acid in grapes through biotechnology or to cultivate wine grape varieties with high tartaric acid accumulation. [Methods] In this study, using ‘Traminer’ and ‘Marselan’ wine grapes as experimental materials, we cloned the tartaric acid related transcription factor VvbHLH79 by PCR. We performed bioinformatic and subcellular localization analyses of VvbHLH79. Furthermore, we measured the tartaric acid content and VvbHLH79 expression levels in grape berries at different developmental stages using HPLC and qRTPCR techniques. We also constructed a plant expression vector and the expression and tartaric acid content of VvbHLH79 were detected after transient transformation of young grape berries using Agrobacteriummediated fruiting pedicle infection, in order to clarify the role of the VvbHLH79 gene in regulating the content of tartaric acid in grape berries. [Results] (1) A grapevine VvbHLH79 gene was obtained by RNA-seq, with a CDS of 831 bp, amino acids of 277, molecular weight of 66.22 kD, and isoelectric point of 5.15. It belonged to the class of unstable hydrophilic proteins without signal peptides and transmembrane structures. Positions 147-232 of amino acid sequence contained the bHLH-SF conserved domain, which belonged to the bHLH transcription factor family. The protein was localized in the cell nucleus. (2) The grapevine VvbHLH79-encoded protein had the highest sequence similarity with the bHLH89-encoded protein in riverbank grape (accession number: XP_034699340.1), with a sequence identity of 99.64%. (3) There was a gradual decrease of VvbHLH79 expression during the development of ‘Traminer’ grape fruits, decreasing 67.25% from veraison to harvest. The accumulation of tartaric acids occurred mainly before color change of the grapes, with a rapid decrease after color change. (4) Transient expression of VvbHLH79 in low tartaric acid grapes resulted in a increase in tartaric acids, whereas silencing of VvbHLH79 in high tartaric acid grapes resulted in a decrease in tartaric acids, confirming that the VvbHLH79 gene positively regulated tartaric acid content. [Conclusion] The VvbHLH79 transcription factor may play a role in the tartaric acid biosynthesis in grape berries.

The insulin-like growth factor (IGF) system plays a key regulatory role in biological processes, such as growth, reproduction, early development, cell differentiation, and proliferation, in fish and other vertebrates. The IGF family system consists of two IGFs (IGF1 and IGF2), two IGF receptors (IGF-1R and IGF-2R), and six IGF-binding proteins (IGF-binding proteins IGFBP1–6) in mammals. By binding to the cell surface-specific receptor (IGF-Rs), IGFs activate the activity of related protein kinases, initiate the signaling cascade in the cell, and participate in the regulation of various biological reaction processes. The research on fish IGFs mainly focuses on IGF1 and IGF2, and several studies have confirmed that they are widely involved in multiple biological processes, such as growth regulation, immune response, gonadal development, and maturation. IGF3 is a fish-specific sub-type; it was first detected in zebrafish (Danio rerio) and Nile tilapia (Oreochromis niloticus) in 2008 and plays an important role in gender differentiation in many reports. However, it is unclear whether it affects ovarian development.Turbot (Scophthalmus maximus) is an economically valuable flatfish species that is widely farmed in Europe and China. It cannot spawn eggs naturally in captivity and needs to be hand-stripped and artificially fertilized. The IGF system plays an important role during turbot ovary maturation. In this study, the full-length turbot IGF3 sequence was obtained via RACE cloning technology, its bioinformatics characteristics were analyzed, and the tertiary structure and docking situation were predicted. The relative expression levels during the whole tissue and ovarian development were detected using qRT-PCR. Hematoxylin-eosin staining was used to determine the gonadal development stage. The results showed that the full-length cDNA sequence of turbot IGF3 was 1 255 bp, encoding 259 amino acids, and displayed the highest homology with halibut (Hippoglossus stenolepis). A total of 33 amino acid signal peptides and 59 amino acid IGF-specific domains were found. It was a hydrophilic protein with a molecular mass of 26.39 kDa and a hydrophobic isoelectric point of 8.13. The turbot IGF3 is highly homologous with other fish. Turbot IGF3 is a hydrophobic protein, and its tertiary structure consists of three α spirals connected in series, closely integrated with IGF-1R and IGF-2R. igf3 is expressed differently between female and male turbot. In the liver, kidney, stomach, gonads, and brain, the male IGF3 expression was significantly higher than in females. In the heart, intestines, and pituitary gland, female IGF3 expression was significantly higher than males; however, the difference was not significant in the spleen. In male turbot, igf3 is mostly expressed in the brain, while in female turbot, igf3 is mostly expressed in the brain and intestine. During the five stages of development of turbot ovaries (previtellogenesis, early vitellogenesis, late vitellogenesis, migratory nucleus, and atresia), igf3 expression increased significantly in late vitellogenesis and maintained high expression in the migratory nucleus stage and decreased in the atresia stage. The results showed that turbot IGF3 has typical IGF family structure characteristics: local endocrine factors, widely distributed in tissues, significant sex dimorphism, and participates in regulating the growth and maturation of oocytes. Therefore, these findings lay a foundation for an in-depth research on the effects of IGF3 on ovarian development and oocyte maturation in turbot and provides crucial ideas for exploring the novel functions of IGF3 in fish.

[ Objective ] Chlorophyllide a oxygenase ( CAO ) is a key enzyme in the formation of chlorophyll b. Cloning and expression analysis of CAO genes may be helpful in exploring their function in regulating leaf color variation of Cymbidium hybrid. [ Methods ] ChCAO gene was cloned from leaves of Cymbidium hybrid by RACE and RT-PCR. ChCAO was analyzed for structure characteristics , physicochemical prop- erties , sequence alignment , and phylogenetic relationship. The expression of ChCAO in different tissues and leaves of K21-3 were analyzed by qRT-PCR. Meanwhile , ChCAO expression was silenced by VIGS technology. [ Results ] The results showed that ChCAO coding region was 1 608 bp , encoding 535 amino acids. ChCAO was closely related to Cymbidium sinense , which belonged to genus Cymbidium . qRT-PCR assay showed that the expression of ChCAO was the highest in leaves and the lowest in roots. The expres- sion of ChCAO in K21 leaves was significantly higher than that of K21-3. The VIGS silencing vector of ChCAO gene was constructed and transferred to tobacco. The viral vector was successfully invaded the to- bacco , and the old leaves of tobacco were turned yellow , while chlorophyll content and expression of ChCA O gene were reduced. [ Conclusion ] The ChCAO gene was obtained and its function was preliminarily characterized , which are helpful for the understanding of the formation of leaf art in Cymbidium hybrid.

Drought and land salinization are inhibiting factors for the sustainable development of forestry, and the plants will release volatile substances such as terpenoids during biological or non-biological stress. The 1-hydroxy-2-methyl-2-(E)-buteny-4-pyrophosphate reductase is the terminal active enzyme of the MEP pathway, which provides precursor terpenoids and has the main rate-limiting effect. In order to investigate whether HDR gene of Pinus massoniana is involved in stress response under drought and salt stress, the open reading frame of PmHDR gene was cloned, and bioinformation, tissue specific expression and preliminary function were analyzed. The results were as follows: (1) The coding region length of PmHDR gene is 1 458 bp, encoding 485 amino acids, and its encoded protein contains the core sequence of LytB/IspH gene superfamily and PLN02821 multifunctional structural domain, which belongs to the HDR family. (2) The PmHDR codon use preference was weak, with a preference for codons ending in A/U. Nicotiana tabacum, Arabidopsis thaliana and Saccharomyces cerevisiae were more suitable as its heterologous expression receptors. (3) Results of qRT-PCR showed that the PmHDR gene was most highly expressed in old needles of Pinus massoniana, followed by young needles, young stem and old stem and least expressed in root. (4) The gene expression vector pBI121-PmHDR was constructed and transformed into Arabidopsis thaliana. The transgenic A.thaliana showed greater resistance to drought and salt stress. These results indicate that PmHDR is involved in plant response and regulation to drought and salt stress, and provide some theoretical supports for Pinus massoniana stress-resistance breeding.

Sebastes schlegelii is a cold temperate ovoviviparous fish found near the seabed layer and is widely distributed in the Yellow Sea and Bohai Sea of China, as well as in the coasts of South Korea and Japan. It is characterized by its tender meat, delicious taste, rich nutrition, and high medicinal value, thus it is an economically important marine fish species in the northern coastal areas of China. In recent years, deep-water cage cultures have rapidly developed in China. Among the economically important fish species suitable for cage culture in northern China, S. schlegelii is the most important local breed with excellent quality and characteristics. However, with the expansion of the scale of the aquaculture industry, aquaculture diseases including those caused by Vibrio harveyi, Vibrio anguillarum, and Photobacterium damselae subsp. damselae, occur frequently. The authors isolated and identified a dominant pathogenic bacterium, the P5W of V. harveyi, from the lesions of diseased S. schlegelii and studied the response of S. schlegelii to V. harveyi infection. Multitissue transcriptome analysis screened the immune differential gene protein kinase B. Protein kinase B, also known as Akt protein, is a serine or threonine protein kinase, which mainly conducts signal transduction by phosphorylating other proteins. It plays an important role in various biological processes such as cell metabolism, transcriptional regulation, cell cycle regulation, immune defense, and embryonic development. Many studies have shown that during viral infection of cells, the Akt pathway is activated, thereby inhibiting viral proliferation. To clarify the sequence information, evolutionary characteristics, and biological functions of the Akt gene in S. schlegelii and to understand the immune response mechanism of S. schlegelii to bacterial invasion, the full-length cDNA of the SsAkt gene was obtained using molecular biology, and the sequence characteristics, tissue expression rules of the gene, and the response rules to Micrococcus luteus and V. anguillarum were studied. The results showed that the length of the open reading frame was 1 440 bp. The predicted relative molecular mass of the SsAkt protein was 55.80 kDa and the isoelectric point was 5.64. Sequence analysis revealed that the SsAkt protein contains three conserved domains: the PH domain (residues 6–109), serine/threonine protein kinase catalytic (S_TKc) domain (residues 148–405), and serine/threonine protein kinase (S_TK_X) domain (residues 406–475). The conserved amino acid site appears in the center of the S_TKc domain and at the end of the S_TK_ X domain: Thr305 and Ser472. Homologous alignment showed that the amino acid sequence of this gene was highly conserved among different species. Phylogenetic tree analysis showed that SsAkt is closely related to Akt in vertebrates, such as Sebastes umbrosus, Lates calcarifer, Larimichthys crocea, and Cyprinus carpio. The relative expression of SsAkt in different tissues of S. schlegelii was detected by real-time fluorescence quantitative PCR. The results showed that SsAkt was expressed in all tested healthy tissues of S. schlegelii, which indicated the relative expression of the SsAkt gene in S. schlegelii. This expression has distinct tissue specificity. The expression of SsAkt in the kidney is the highest, and in the blood and brain, it is also higher than that in other tissues. Therefore, it is speculated that higher expression of SsAkt in the kidney and brain is closely related to immune defense, neurodevelopment, and other processes. To study the role of SsAkt in the immune response of S. schlegelii, we designed a bacterial challenge experiment. The results showed that the transcription level of SsAkt changed significantly, and the relative expression of SsAkt in the three tissues increased after stimulation with M. luteus and V. anguillarum compared with the control group (PBS). After infection with M. luteus, its expression only increased sharply in the kidney, whereas its expression increased slowly in the blood and liver. In the experimental group infected with V. anguillarum, the expression increased significantly in the liver and kidney, and the two relative expression peaks in the kidney occurred at 12 h and 72 h after infection, respectively. The results revealed that the SsAkt gene may play an important role in the immune defense of S. schlegelii, which can further enrich the sequence information and evolutionary data of the Akt gene in marine fish and provide reference materials for an in-depth study of the biological function of this gene. In follow-up experiments, we will explore the specific mechanism of SsAkt in the innate immune defense of S. schlegelii to provide theoretical guidance for the study of the immune signaling pathway of S. schlegelii and to lay a theoretical foundation for revealing the role of SsAkt in other important biological functions in growth and development.

It is reported that ILF2 (interleukin enhancer binding factor 2) is involved in the immune response by regulating the transcription of interleukin 2 (IL2), which may play an important role in host-virus interactions. To investigate the role of ILF2 in WSSV infection in Penaeus japonicus, the ILF2 gene was characterized by bioinformatics analysis and RNAi. The sequence of 873 bp ORF region encoding 290 amino acids was obtained by PCR amplification in this study. The relative molecular weight of ILF2 protein is 32.29154 KDa. The bioinformatics analysis indicated that the ILF2 protein has an DZF domain and a RGG domain and contained a large number of alpha helices and irregular coils. The results of tissue-specific assays demonstrated that ILF2 was highly expressed in the hemolymph and gill of shrimp. After WSSV stimulation, qRT-PCR analysis showed that the expression of ILF2 was significantly increased in all tissues of shrimp. Using RNAi to knock down the expression of ILF2 in shrimp, the results showed that the copy number of WSSV in shrimp was significantly increased, suggesting that ILF2 could inhibit the replication of WSSV. The results showed that ILF2 had the potential to inhibit WSSV replication and to achieve antiviral properties, providing a theoretical basis for the control of WSSV in shrimp. [ABSTRACT FROM AUTHOR]

Insulin-like growth factors (IGFs) play central roles in the growth and development of vertebrates. Blotched snakehead (Channa maculata), an economically significant fish, exhibits obvious sexual dimorphism and achieves sexual maturity in one year. However, the role of IGFs in C. maculata remains unknown. Three IGF genes were identified in C. maculata, designated as CmIGF1-1, CmIGF1-2, and CmIGF2. The cDNA sequences of these genes are 1184, 655, and 695 bp, encoding putative proteins of 168, 131, and 215 amino acids, respectively, and all three proteins contain a conserved IGF domain. Quantitative real-time PCR (qPCR) revealed the predominant expression of CmIGFs in the liver of adult fish, with higher expression levels observed in males. Notably, CmIGF1-1, CmIGF1-2, and CmIGF2 displayed analogous expression profiles in the liver across various developmental stages, peaking at 365 days after hatching (dah). Subsequently, 600 individuals at 75 dah, at an early developmental stage, were randomly divided equally into six groups and reared in aerated 2 m × 2 m × 2 m cement ponds at 26.0 ± 1.0 °C. Following a one-week acclimatization period, fish without observed abnormalities were intraperitoneally injected with either 17α-ethynylestradiol (EE2) or 17α-methyltestosterone (MT) at a dose of 10 μg/g body weight. Three groups underwent short-term hormone treatment, and the remaining three groups underwent long-term hormone treatment, which included five injections at two-week intervals over ten weeks. The analysis of CmIGFs expression levels in the liver under different hormone treatments revealed that EE2 suppressed the expression of CmIGF1-1 and CmIGF1-2 while promoting CmIGF2 expression. In females, MT up-regulated the expression of CmIGF1-1 and CmIGF2 in a time-dependent manner, but consistently inhibited CmIGF2 expression. In males, MT promoted the expression of CmIGFs in a time-dependent manner, reaching peak levels for CmIGF1-1, CmIGF1-2, and CmIGF2 after 8, 10, and 2 weeks of injection, respectively. Additionally, CmIGF1 and CmIGF2 might exhibit a complementary relationship, with a compensatory increase in CmIGF2 expression in response to low CmIGF1 concentration. These findings highlight the potential key role of IGFs upon growth and their regulation by sex steroid hormones in C. maculata, providing a crucial foundation for future research aimed at elucidating the molecular mechanisms underlying the growth dimorphism between female and male blotched snakeheads. [ABSTRACT FROM AUTHOR]

【Objective】The BrMLP328 gene of Chinese cabbage（Brassica rapa subsp. pekinensis）was cloned, its expression pattern was analysed, and its function in regulating flowering was verified. This may lay the basis for further investigation into the mechanism of the role of this gene in regulating flower formation in Chinese cabbage.【Method】BrMLP328 was cloned by RT-PCR and analysed by bioinformatics. RT-qPCR was used to determine the relative expression of the BrMLP328. An overexpression vector was constructed and transformed into wild-type Arabidopsis thaliana by flower-dipping method, and the difference in flowering time between the T2 generation plants and the wild type was compared.【Result】The coding sequence（CDS）of BrMLP328 has a total length of 456 bp, encoding 151 amino acids. The relative molecular weight of the protein was 17 493.82 Da and was localized in the nucleus. The expression of BrMLP328 was the highest in the stems of Chinese cabbage, followed by roots and lowest in flower buds. The expression in the growing point of the stem tip showed an elevated level after vernalization, after which it declined rapidly at the stage of flower bud differentiation and was maintained at a very low level. Compared with the wild type, the flowering time of the BrMLP328 of T2 generation of transgenic plants delayed by 1.46-3.09 d.【Conclusion】BrMLP328 was cloned from Chinese cabbage. The expression of BrMLP328 was different in different tissues and different flowering stages, and the gene could delay flowering. [ABSTRACT FROM AUTHOR]

【Objective】MYB305 as a member of the MYB R2R3 subfamily, plays an important role in plant nectary development, nectar protein expression, starch accumulation and hydrolysis, and flavonoid synthesis. Therefore, it is of great significance to study the expression profile and function of MYB305 for the molecular mechanism study of Liriodendron tulipifera floral nectary.【Method】Having floral nectary of nectariferous plant L. tulipifera as materials, the anthocyanidin content in five stages of floral nectary was detected by spectrophotometry, LtMYB305 gene was cloned via RACE（rapid-amplification of cDNA ends）technology, and relative expressions among different tissues were analyzed by RT-qPCR assay. Then, subcellular localization of LtMYB305 protein was verified using tobacco leaves as materials, and the function of LtMYB305 gene was studied by genetic transformation approaches of wild type Arabidopsis thaliana（col）.【Result】The anthocyanidin in the nectaries of L. tulipifera was accumulated from the late expanding stage, increased sharply at the early flowering stage, and reached the highest of 27.14 μg/g at the withering stage, which was consistent with the process of nectar secretion and coloring. The full-length of LtMYB305 was 931 bp encoding 198 amino acids and its protein was hydrophilic and non-transmembrane. LtMYB305 was expressed highly only in the nectary of L. tulipifera at anthesis stage, and hardly in other tissues. LtMYB305 protein was localized in the nucleus. After the overexpression of LtMYB305, the nectary groove of the lateral nectary disappeared and deepened, the expressions of AtMYB30, AtPIN, AtSWEET9 genes related to flower nectary were up-regulated, and the expressions of AtCRC, AtBOP1/2, AtMYB21, AtSWEET3/4/7 genes were down-regulated.【Conclusion】 The LtMYB305 has typical characteristics of transcription factor and is involved to the nectary development in L. tulipifera. [ABSTRACT FROM AUTHOR]

【Objective】This study aims to clone the sterol glycosyltransferase gene AoSGT1 from Asparagus officinalis, to assess its catalytic properties, and to elucidate its role in the biosynthesis of asparagus saponins and their metabolic regulation.【Method】We designed specific primers based on asparagus transcriptome data to amplify the complete open reading frame（ORF）of AoSGT1. The gene was sequenced and analyzed through bioinformatics, and its expression profile was investigated via quantitative real-time PCR（RT-qPCR）. We also constructed a pGEX-4T-3-AoSGT1 expression vector, expressed it in Escherichia coli BL21（DE3）, and induced the recombinant protein production.【Result】AoSGT1 has a length of 1 800 base pairs, encoding 599 amino acids, with a relative molecular weight of 66.72 kD. It is a hydrophilic protein with no transmembrane domains or signal peptides. The phylogenetic analysis revealed a close homology between AoSGT1 and Dioscorea zingiberensis Dz3GT2, indicating their membership in the UGT80B1 subfamily. Multiple sequence alignment revealed that the protein sequence contained conserved domains“PSBD Box”and“PSPG Box”of sterol glycosyltransferase, indicating its potential glycosylation activity at the 3β-OH position of steroidal compounds. RT-qPCR results showed that AoSGT1 was highly expressed in the roots, while expression in stems and flowers was relatively low. Additionally, SDS-PAGE results revealed that the target protein was efficiently expressed in a soluble form within E. coli, matching the predicted size.【Conclusion】The AoSGT1 gene was successfully cloned and demonstrated to exhibit tissue-specific expression in asparagus, suggesting its potential involvement in the biosynthesis of steroidal saponins in asparagus. Additionally, heterologous expression of the target protein in E. coli was successfully achieved. [ABSTRACT FROM AUTHOR]