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Background: The 12-oxo-phytodienoic acid reductase (OPR) enzyme is crucial for the synthesis of jasmonates (JAs), and is involved in the plant stress response. However, the OPR gene family in sweetpotato, an important horticultural crop, remains unidentified. Results: In this study, we employed bioinformatics techniques to identify nine IbOPR genes. Phylogenetic analysis revealed that these genes could be divided into Group I and Group II. Synteny analysis indicated that IbOPR evolution was driven by tandem duplication, whole-genome duplication (WGD), and segmental duplication events. The promoter sequences of IbOPRs were found to be associated with stress and hormonal responses. Additionally, we successfully cloned four IbOPRs from "Haida HD7791" and "Haida HD7798" using homologous cloning technology. These sequences were 1203 bp, 1200 bp, 1134 bp, and 1137 bp in length and encoded 400, 399, 377, and 378 amino acids, respectively. The protein sequence similarity between the salt-tolerant variety "Haida HD7791" and the salt-sensitive variety "Haida HD7798" was determined to be 96.75% for IbOPR2, 99.75% for IbOPR3, 92.06% for IbOPR6, and 98.68% for IbOPR7. Phylogenetic analysis categorized IbOPR2 and IbOPR3 proteins into Group II, while IbOPR6 and IbOPR7 proteins belonged to Group I. Subcellular localization experiments showed IbOPR2 protein present in the peroxisome, while IbOPR3, IbOPR6, and IbOPR7 proteins were found in the cytoplasm and nucleus. Salt stress induction experiments demonstrated that IbOPR2, IbOPR3, and IbOPR7 were significantly upregulated only in 'Haida HD7791' after 6 h. In contrast, IbOPR6 was induced in 'Haida HD7798' at 6 h but inhibited in 'Haida HD7791' at later time points (12, 24, 48, and 72 h), highlighting functional differences in salt stress responses. Conclusions: Our findings suggest that IbOPR2 may play a crucial role in sweetpotato's response to salt stress by participating in JAs synthesis. These results provide a foundation for future functional analyses of OPR genes in sweetpotato. [ABSTRACT FROM AUTHOR]

The full-length cDNA sequence of Penaeus monodon glucose transporter-2 (PmGLUT2) was cloned in this study using the RACE method. Quantitative real-time PCR was used to analyze the differential expression of PmGLUT2 during the development of P. monodon larvae in different tissues and under low salinity stress. The PmGLUT2 cDNA exhibited a total length of 2018 base pairs, with 94 base pairs located in the 5' untranslated region (UTR) and 352 base pairs in the 3' UTR. Additionally, the sequence contained 29 base poly (A) tails and 1572 base pairs within the open reading frame (ORF), capable of encoding 523 amino acids. Through a comparative analysis of the amino acid sequences of PmGLUT2 and LvGLUT2, it was determined that PmGLUT2 had 94.77% homology with Tret1 gene of Litopenaeus vannamei, and 60.54% homology with GLUT2 gene of L. vannamei. The results indicated that there was a fluctuation in PmGLUT2 expression levels from zygote to postlarva development, with initial reduction followed by an increase, although the difference was not statistically significant. According to the molting analysis, the highest expression level of PmGLUT2 was observed in the hepatopancreas during the premolt period, while the gill and gut exhibited peak expression levels during the intermolt period. PmGLUT2 was found to be most abundant in lymph tissue, followed by gill tissue, and least abundant in ootheca, according to the tissue expression analyses. Following 96 hours of acute salt stress, there was a notable inhibition in the expression of PmGLUT2 in the hepatopancreas and gills. Additionally, the expression level in the gills at 96 hours was significantly lower compared to the baseline level at 0 hours. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Gene cloning is an important step in investigating gene structure and function. To verify gene sequence, Sanger sequencing is used, which may produce several overlapping sequencing files that need to be merged before alignment to the target gene sequence is performed. Previously, we reported the Python program to Merge Sanger sequences (), which ran in command line and relied heavily on EMBOSS suite. In this updated version of the program, we have made several remarkable improvements. It provides a graphical user interface (GUI) written with tkinter, which is convenient and stable. It does not require users to rename the input sequences before performing merging. With regard to the implementation, the updated version utilizes Python function (Align.PairwiseAligner) to align adjacent sequences, which is more flexible (can adjust program parameter i.e., the number of first-time consecutive matching bases). The new version of the program makes merging Sanger sequences much more convenient and facilitates gene study. [ABSTRACT FROM AUTHOR]

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 FROM AUTHOR]

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]

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.)

[Objectives] This study was conducted to understand the structure and function of MsrA protein. [Methods] With Vibrio alginolyticus HY9901 as the object of study, primers were designed to amplify the full-length gene of msrA, and its bioinformatics analysis was carried out. [Results] The full length of msrA gene was 639 bp, encoding 212 amino acids, and its theoretical molecular weight was about 23 729.60 Da. The protein had a stable structure, and it was hydropho-bic overall. The structure of signal peptides at the N terminal of the amino acid sequence was predicted, and it was found that there was no signal peptide cleavage site and no transmembrane region. The amino acid sequence of MsrA contained multiple signal binding sites. Protein subcellular localization showed that MsrA protein was most likely located in the cytoplasm. Homology analysis showed that MsrA of V. alginolyticus had high homology with other Vibrio species, and the highest homology with V. alginolyticus. In the prediction of functional domains, MsrA had the function of methionine sulfoxide reduction. In secondary structure prediction, MsrA contained random coils at a proportion of 46.70%, which was the highest. The similarity between the tertiary structure model of MsrA and template Q87SW6. 1. A was 89. 15%. PTM analysis showed that MsrA protein had many PTM modification sites such as phosphorylation and glycosylation sites. [Conclusions] This study provides some reference value for further study on the role of MsrA in bacterial antioxidant stress. [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.)

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]

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]

The WRKY transcription factor family plays a key role in plant growth and development, hormone signaling, and resistance to environmental stress. In this study, we investigated the gene sequence, subcellular localization, and response pattern of a member of the WRKY transcription factor family to reveal its protein structure and involvement in the resistance signaling pathway.The BsWRKY51 gene was cloned by RT-PCR, and the structural characteristics of its encoded protein WRKY51 were analyzed by bioinformatics. The vector was next transiently transformed into tobacco to analyze the subcellular localization, and real-time fluorescence quantitative PCR was performed to analyze the changes in the expression pattern of BsWRKY51. The BsWRKY51 gene has a coding sequence (CDS) length of 987 bp.The respective unstable hydrophilic protein BsWRKY51 is localized in the nucleus. It most closely related to the WRKY protein of Dendrobium catenatum in the Orchidaceae family. Fluorescence quantitative PCR results showed that the BsWRKY51 expression in the leaves was significantly higher than that in the roots, stems, and pseudobulbs of Bletilla striata seedlings. Under the conditions of salt and drought stress, the BsWRKY51 expression gradual increased and then a slightly decreased, and under salicylic acid (SA) treatment, the expression of BsWRKY51 showed an overall decreasing trend.The BsWRKY51 gene of Bletilla striata may play an important regulatory role in its salt and drought stress responses. Our present findings provide the foundation for elucidating the mechanisms of salt and drought tolerance in Bletilla striata and for breeding new varieties. [ABSTRACT FROM AUTHOR]

MYB (myoblast) protein comes in large quantities and a wide variety of types and plays a role in most eukaryotes in the form of transcription factors (TFs). One of its important functions is to regulate plant responses to various stresses. However, the role of MYB TFs in regulating stress tolerance in strawberries is not yet well understood. Therefore, in order to investigate the response of MYB family members to abiotic stress in strawberries, a new MYB TF gene was cloned from Fragaria vesca (a diploid strawberry) and named FvMYB108 based on its structural characteristics and evolutionary relationships. After a bioinformatics analysis, it was determined that the gene belongs to the R2R3-MYB subfamily, and its conserved domain, phylogenetic relationships, predicted protein structure and physicochemical properties, subcellular localization, etc. were analyzed. After qPCR analysis of the expression level of FvMYB108 in organs, such as the roots, stems, and leaves of strawberries, it was found that this gene is more easily expressed in young leaves and roots. After multiple stress treatments, it was found that the target gene in young leaves and roots is more sensitive to low temperatures and salt stimulation. After these two stress treatments, various physiological and biochemical indicators related to stress in transgenic Arabidopsis showed corresponding changes, indicating that FvMYB108 may be involved in regulating the plant's ability to cope with cold and high-salt stress. Further research has found that the overexpression of this gene can upregulate the expression of AtCBF1, AtCOR47, AtERD10, and AtDREB1A related to low-temperature stress, as well as AtCCA1, AtRD29a, AtP5CS1, and AtSnRK2.4 related to salt stress, enhancing the ability of overexpressed plants to cope with stress. [ABSTRACT FROM AUTHOR]

Leaf morphology is a crucial aspect of plant architecture, yet the molecular mechanisms underlying leaf development remain incompletely understood. In this study, a narrow leaf mutant, m625, was identified in rice (Oryza sativa L.), exhibiting pleiotropic developmental defects. Pigment measurement revealed reduced levels of photochromic pigments in m625. Cytological analysis demonstrated that the m625 gene affected vascular patterns and cell division. Specifically, the narrowing of the leaf was attributed to a decrease in small vein number, shorter vein spacing, and an abnormal V-shaped arrangement of bulliform cells, while the thickening was caused by longer leaf veins, thicker mesophyll cells, and an increased number of parenchyma cell layers. The dwarf stature and thickened internode were primarily due to shortened internodes and an increase in cell layers, respectively. Positional cloning and complementation assays indicated that the m625 gene is a novel allele of NAL1. In the m625 mutant, a nucleotide deletion at position 1103 in the coding sequence of NAL1 led to premature termination of protein translation. Further RNA-Seq and qRT-PCR analyses revealed that the m625 gene significantly impacted regulatory pathways related to IAA and ABA signal transduction, photosynthesis, and lignin biosynthesis. Moreover, the m625 mutant displayed thinner sclerenchyma and cell walls in both the leaf and stem, particularly showing reduced lignified cell walls in the midrib of the leaf. In conclusion, our study suggests that NAL1, in addition to its known roles in IAA transport and leaf photosynthesis, may also participate in ABA signal transduction, as well as regulate secondary cell wall formation and sclerenchyma thickness through lignification. [ABSTRACT FROM AUTHOR]

The formation of fruit color in pepper is closely related to the processes of carotenoid metabolism. In this study, red wild-type pepper XHB, SP01, PC01 and their corresponding mutants H0809 (orange), SP02 (yellow), and PC02 (orange) were used as research materials. The Ggps, Psy, Lcyb, Crtz, Zep, and Ccs genes involved in carotenoid biosynthesis were cloned, and bioinformatics and expression analyses were carried out. The results showed that the full lengths of the six genes were 1110 bp, 2844 bp, 1497 bp, 2025 bp, 510 bp, and 1497 bp, and they encoded 369, 419, 498, 315, 169, and 498 amino acids, respectively. Except for the full-length Ccs gene, which could not be amplified in the yellow mutant SP02 and the orange mutant PC02, the complete full-length sequences of the other genes could be amplified in different materials, indicating that the formation of fruit color in the SP02 and PC02 mutants could be closely related to the deletion or mutation of the Ccs gene. The analytical results of real-time quantitative reverse transcription PCR (qRT-PCR) showed that the Ggps, Psy, Lcyb, Crtz, and Zep genes were expressed at different developmental stages of three pairs of mature-fruit-colored materials, but their patterns of expression were not consistent. The orange mutant H0809 could be amplified to the full Ccs gene sequence, but its expression was maintained at a lower level. It showed a significant difference in expression compared with the wild-type XHB, indicating that the formation of orange mutant H0809 fruit color could be closely related to the different regulatory pattern of Ccs expression. The results provide a theoretical basis for in-depth understanding of the molecular regulatory mechanism of the formation of color in pepper fruit. [ABSTRACT FROM AUTHOR]

Members of the light-harvesting chlorophyll a/b binding protein (CAB) gene family play an important role in plant leaf yellowing variation. In this study, the CAB family members were identified from tea plant 'Tieguanyin' genomic data. The bioinformatics and expression patterns were analyzed. Furthermore, the expression patterns of the CABs gene were analyzed by gene cloning and qRT-PCR in tea cultivars with different leaf colors. The key CAB genes related to tea yellowing were screened by correlation analysis of leaf color parameters and chlorophyll SPAD values. The results show that 25 members of the CAB gene family were identified, their amino acid length ranged from 167-337 and the protein molecular weight ranged from 18.5-37.1 kDa. Most CAB members were stable and hydrophobic proteins, and distributed in chloroplast by the subcellular localization prediction. According to the evolutionary relationship, CAB family members are divided into 13 subfamilies, and the Lhcb1 subfamily has the most members. Cis-acting element analysis of promoter shows that CAB family members have a lot of light-responsive elements, as well as other elements related to growth and development, hormone response, and adversity stress. The members of Lhcb1 subfamily were cloned from tea plants, CAB1, CAB6, and CAB7 genes were screened by sequence alignment. The expression analysis shows that CAB1, CAB6, and CAB7 genes had tissue expression characteristics with higher expression levels in buds, leaves and fruits, and could respond to various stresses. Finally, the qRT-PCR indicates that the expressions of CAB1, CAB6, and CAB7 genes were consistent in the yellow and green leaves. Compared with green leaves, the expression of CAB genes in yellow leaves were significantly down-regulated. The correlation analysis of gene expressions and related leaf color parameters shows that the gene expressions of CAB1, CAB6, and CAB7 were significantly correlated with leaf color parameters a, b, L, and chlorophyll SPAD values (P<0.01). Among them, the expression of CAB1 shows the highest correlation coefficient. The subcellular localization analysis shows that CAB1 was distributed in the nucleus, cytoplasm, and chloroplasts. The studies analyzed the basic characteristics of CAB family members in tea plants and the key genes related to tea color variation were identified, which provided a theoretical basis for the molecular regulation mechanism of tea color variation. [ABSTRACT FROM AUTHOR]

MYB transcription factors (TFs) have been shown to play a key role in plant growth and development and are in response to various types of biotic and abiotic stress. Here, we clarified the structure, expression patterns, and function of a MYB TF, SlMYB86-like (Solyc06g071690) in tomato using an inbred tomato line exhibiting high resistance to bacterial wilt (Hm 2-2 (R)) and one susceptible line (BY 1-2 (S)). The full-length cDNA sequence of this gene was 1226 bp, and the open reading frame was 966 bp, which encoded 321 amino acids; its relative molecular weight was 37.05055 kDa; its theoretical isoelectric point was 7.22; it was a hydrophilic nonsecreted protein; and it had no transmembrane structures. The protein also contains a highly conserved MYB DNA-binding domain and was predicted to be localized to the nucleus. Phylogenetic analysis revealed that SlMYB86-like is closely related to SpMYB86-like in Solanum pennellii and clustered with other members of the family Solanaceae. Quantitative real-time PCR (qRT-PCR) analysis revealed that the expression of the SlMYB86-like gene was tissue specific and could be induced by Ralstonia solanacearum, salicylic acid, and jasmonic acid. The results of virus-induced gene silencing (VIGS) revealed that SlMYB86-like silencing decreased the resistance of tomato plants to bacterial wilt, suggesting that it positively regulates the resistance of tomatoes to bacterial wilt. Overall, these findings indicate that SlMYB86-like plays a key role in regulating the resistance of tomatoes to bacterial wilt. [ABSTRACT FROM AUTHOR]

[Objective] This study cloned Uncaria rhynchophylla horse money nucleoside acid methyltransferase (loganic acid methyltransferase, LAMT) gene and promoter, analyzed UrLAMT gene expression and its biological and abiotic stress response, and conducted bioinformatics analysis on UrLAMT and its promoter to improve further study of UrLAMT transcriptional regulation. [Method] Primers were designed based on the transcriptome data of Uncaria rhynchophylla, UrLAMT sequences were cloned by PCR from cDNA, and bioinformatics analysis was performed. Quantitative real-time fluorescence PCR (qRT-PCR) was used to analyze its expression in different tissues (root, stem, leaf, flower, fruit and hook) and its response to exogenous methyl jasmonate (MeJA) and shading/relighting stress. The upstream promoter sequence of UrLAMT was cloned by FPNI-PCR and its activity was verified. The regulatory relationship between the corresponding transcription factors and the UrLAMT promoter was analyzed by yeast ingle hybridization test. [Result] The sequence length of UrLAMT was 1 137 bp, encoding 378 amino cids. The relative molecular weight of UrLAMT protein was 42.64 ku, and the theoretical isoelectric point was 5.76. It was a hydrophilic protein, without signal peptide and transmembrane structure, and it was located in cells. The expression of UrLAMT gene was the highest in leaves, followed by roots. UrLAMT was responsive to MeJA and light. It was closely related to the evolution of Ophiorrhiza pumila and Catharanthus roseus. The sequence length of UrLAMT promoter was 1141 bp, containing multiple photo responsive elements in addition to core response elements. UrLAMT promoter interacted with photo-regulated transcription factor HY5 in yeast. [Conclusion] The UrLAMT gene and its promoter sequence were obtained. The expression level of UrLAMT gene was the highest leaves, which was possibly induced by light. [ABSTRACT FROM AUTHOR]

[Objective] The function of AcSCL4 in flowering was preliminarily studied to provide basis or understanding molecular regulation mechanism of onion flowering regulation. [Method] AcSCL4 was dentified by qRT-PCR and bioinformatics analysis based on the transcriptome data of a long-day onion inred line SB2. The AcSCL4 overexpression vector pB221-3300-AcSCL4 was constructed and transformed o Arabidopsis by Agrobacterium tumefaciens. Homozygous Arabidopsis transgenic lines (T3) were used or further phenotypic characterization and genes expression identification. [Result] The cDNA of AcSCL4 gene was 1 515 bp and encoded 504 amino acids. AcSCL4 had conserved GRAS domain at the C terminus and was highly variable at the N terminus. AcSCL4 was phylogenetically closer to the SCL4 of Asparagus fficinalis and Gossypium hirsutum. Overexpression of AcSCL4 delayed the flowering time of wild-type Arabidopsis. The expressions of CO and FT were significantly decreased in AcSCL4 overexpression trans-genic lines. [Conclusion] AcSCL4 suppressed flowering by regulating CO and FT expressions. [ABSTRACT FROM AUTHOR]

Basic helix-loop-helix (bHLH) proteins are important in abiotic stress control. Here, a specific bHLH transcription factor gene, CabHLH18, from a strong waterlogging-tolerant pepper cultivar, 'ZHC2', was successfully cloned. The CabHLH18 gene presented a coding sequence length of 1,056 bp, encoding 352 amino acids, and the protein was the closest to Capsicum annuum XM016694561.2 protein. The CabHLH18 protein was located in the nucleus. The transformation of the CabHLH18 overexpression vector into the plumules of hot peppers, 'DFZJ' and 'ZHC1', exhibited 21.37% and 22.20% efficiency, respectively. The root length, plant height, and fresh weight of the 'DFZJ' overexpression lines were greater than those of wild-type (WT) plants under waterlogging conditions. Compared with the WT plants, the overexpression lines generally showed greater contents of water, the amino acid, proline, soluble sugar, root viability, and superoxide dismutase activity, but lower malondialdehyde content under waterlogging conditions. Plant fresh weight, amino acids, proline, and soluble sugar levels of the overexpression lines were 39.17%, 45.03%, 60.67%, and 120.18% greater, respectively, compared with the WT plants at 24 h after waterlogging stress. Therefore, the CabHLH18 gene could be implicated in conferring waterlogging tolerance in hot peppers and holds promise for enhancing their overall waterlogging tolerance. [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 structure and function of yak heart fatty acid binding protein 3 (FABP3) gene were analyzed and its expression levels in adult and fetal yak were detected to provide references for xploring biological functions of the gene in yak breeding. [Method] The FABP3 gene was amplified by PCR from heart tissue of Meiren yak, and bioinformatics analysis was carried out after the CDS region was obtained. The expression levels of FABP3 in heart, liver, spleen, lung, kidney, and muscle tissues of adult and fetal yak were detected by real-time quantitative PCR (RT-qPCR). [Result] The length of the coding egion of FABP3 was 402 bp, encoding 133 amino acids. The advanced structure of the protein included mainly β-turn and extended chain. There was no transmembrane region or signal peptide on yak FABP3 protein, indicating it was a stable protein with certain hydrophilicity. Homology analysis of the nucleotide and amino acid sequences of the CDS region showed that the FABP3 gene was conserved in cattle. Accordng to the phylogenetic tree, yak had the closest genetic relationship with wild yak and zebu, and the farhest genetic relationship with chicken. RT-qPCR results showed that the FABP3 gene was expressed in heart, liver, spleen, lung, kidney, and muscle tissues of adult and fetal yak. The expression in liver, spleen, kidney, and muscle of fetal cattle was significantly or extremely significantly higher than that of adult yak, and the expression level in lung of adult yak was significantly higher than that of fetal yak. [Conclusion] The FABP3 gene of yak was cloned, and its tissue expression pattern was explored. This study provided asic data for further study on the role of FABP3 in fat deposition. [ABSTRACT FROM AUTHOR]

The purpose of this study was to examine STC-1's structure, function, and differential expression in large and miniature pigs. We cloned the Hezuo pig's coding sequence, compared its homology, and used bioinformatics to assess the structure. RT-qPCR and Western blot were used to detect the expression in ten tissues of Hezuo pig and Landrace pig. The results showed that Hezuo pig was most closely related to Capra hircus and most distantly related to Danio rerio. The protein STC-1 has a signal peptide and its secondary structure is dominated by the alpha helix. The mRNA expression in the spleen, duodenum, jejunum, and stomach of Hezuo pigs was higher than that of Landrace pigs. And except for heart and duodenum, expression of the protein in Hezuo pig was higher than in another. In conclusion, STC-1 is highly conserved among different breeds of pigs, and the expression and distribution of its mRNA and protein are different in large and miniature pigs. This work can lay the foundation for future study into the mechanism of action of STC-1 in Hezuo pigs and the enhancement of breeding in miniature pigs. [ABSTRACT FROM AUTHOR]

To further reveal the molecular mechanisms underlying the formation of continuous cropping disorders in patchouli, this study analyzed the function of calcium dependent protein kinase (CDPK) genes at the molecular level in patchouli continuous cropping disorders. The findings unveiled the presence of 45 PcCDPK genes within the complete patchouli genome dataset. These genes exhibited a range of molecular weights from 50.78 to 78.96 kDa and aliphatic index values spanning from 74.42 to 88.49, and are shown to be hydrophilic proteins. The evolution of 45 PcCDPK members was divided into 4 subfamilies, with a total of 65 pairs of collinear genes. Each PcCDPK contains a STKc-CAMK domain and four EF-hand structures exhibiting a certain degree of conservatism during evolution. Transcriptome data further supported the significance of PcCDPK25 and PcCDPK38 genes, showing substantial upregulation, which was corroborated by qRT-PCR results. The 1629 bp and 1716 bp CDS sequences were obtained by cloning the PcCDPK25 and PcCDPK38 genes, respectively, and subcellular localization showed that both proteins were localized on the plasma membrane. This comprehensive study provides molecular-level confirmation of the pivotal roles played by CDPK genes in the emergence of continuous cropping challenges in patchouli plants, establishing a crucial foundation for a deeper comprehension of the molecular mechanisms underpinning these obstacles. [ABSTRACT FROM AUTHOR]

Aquaculture species are often exposed to acute stressors such as low water levels and handling during routine aquaculture procedures. This might result in oxidative stress by the increased reactive oxygen species (ROS)' production (e.g., superoxide anion). The harmful effects of ROS are eliminated by a defense system, referred antioxidant defense system (ADS). sod1 is the first gene involved in the ADS. Therefore, we cloned and characterized the open reading frame of the sod1 in brown trout. Then, we determined the effects of low water level and handling stress on sod1 mRNA expression in the liver and gills at 0 min, 1 and 2 h. The total RNA isolated was used to synthesize cDNA for RT-qPCR analysis. Phylogenetic tree, identity/similarity percentages, genomic organization, and conserved gene synteny analyses were applied to characterize Sod1/sod1. While low water level stress upregulated sod1 expression in the liver compared to the control group, no significant differences were observed in the gills between experimental groups. However, brown trout differently responded to handling stress at different time intervals in both tissues. Transcriptional differences were also noted between the sexes. This study contributes to the current understanding of the molecular mechanism between oxidative stress and ADS. [ABSTRACT FROM AUTHOR]

【Objective】CMB1 gene plays an important role in the regulation of inflorescence structure and fruit ripening in plants, but the function and regulatory mechanism of CMB1 gene in pepper have been rarely reported. Therefore, the role of transcription factors related to the regulation of carotenoid synthesis and CMB1 gene in pepper was studied so as to provide a reference for pepper breeding.【Method】The coding sequences of CMB1 were cloned by the homology sequence method, and the expression patterns of CMB1 in fruits of red and yellow peppers with wrinkled skin at different development stages (green ripening stage, color change stage, mature stage) were analyzed by real-time quantitative PCR. 【Result】The ORF length of CMB1 gene was 732 bp in both red pepper and yellow pepper, encoding 243 amino acid residues. Sequence alignment showed that the mutation occurred at the 11th site of CMB1 base sequence, G mutated to T, resulting in amino acid sequence mutated from G (Glycine) to V (Valine). Bioinformatics analysis revealed that the CMB1- encoded protein of wrinkled skin red pepper had a molecular mass of 20 923.21, a theoretical isoelectric point of 5.59, an instability coefficient of 57.28, and a total mean hydrophilicity (gravimetric) index of -1.001. The CMB1-encoded protein of wrinkled skin yellow pepper had a molecular mass of 27 958.47, a theoretical isoelectric point of 7.10, an instability coefficient of 53.54, and a total mean hydrophilicity (gravimetric) index of -0.809. Both CMB1 of wrinkled skin red and yellow peppers were unstable hydrophobic proteins, both contained 38 phosphorylation sites, both were devoid of transmembrane structures and signal peptides, and both had typical MADS-box structural domains and K-box structural domains. Evolutionary tree analysis revealed that pepper CMB1 was most closely related to Solanaceae and most distantly related to Leguminosae. The results of fluorescence quantitative PCR showed that the expression of CMB1 increased sequentially in the three stages of green ripening, color change, and maturity of pepper fruits, and the expression of CMB1 in the three stages was significantly higher in wrinkled skin red pepper than in wrinkled skin yellow pepper.【Conclusion】Pepper CMB1 is a member of MADS-box, and its expression in pepper fruit is similar to that of carotenoid in pepper, suggesting that CMB1 is mainly involved in regulating the synthesis of carotenoid like capsanthin. [ABSTRACT FROM AUTHOR]

Flower organ development is one of the most important processes in plant life. However, onion CMS (cytoplasmic male sterility) shows an abnormal development of floral organs. The regulation of MADS-box transcription factors is important for flower development. To further understand the role of MADS-box transcription factors in the regulation of cytoplasmic male sterility onions. We cloned the full-length cDNA of five MADS-box transcription factors from the flowers of onion using RACE (rapid amplification of cDNA ends) technology. We used bioinformatics methods for sequence analysis and phylogenetic analysis. Real-time quantitative PCR was used to detect the expression patterns of these genes in different onion organs. The relative expression levels of five flower development genes were compared in CMS onions and wild onions. The results showed that the full-length cDNA sequences of the cloned MADS-box genes AcFUL, AcDEF, AcPI, AcAG, and AcSEP3 belonged to A, B, C, and E MADS-box genes, respectively. A phylogenetic tree construction analysis was performed on its sequence. Analysis of MADS-box gene expression in wild onion and CMS onion showed that the formation of CMS onion was caused by down-regulation of AcDEF, AcPI, and AcAG gene expression, up-regulation of AcSEP3 gene expression, and no correlation with AcFUL gene expression. This work laid the foundation for further study of the molecular mechanism of onion flower development and the molecular mechanism of CMS onion male sterility. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences 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.)

In this study, PsMYB2 was successfully cloned using cDNA from Potentilla sericea as a template. It was used to construct the plant overexpression vector pBI121-PsMYB2-GFP, which was transferred into Arabidopsis thaliana wild-type plants. We studied the gene function using real-time quantitative PCR and performed a preliminary characterization and analysis of the function of PsMYB2 under abiotic stresses. This study showed that under cadmium stress, the gene expression of PsMYB2 gene in roots, stems, and leaves was up to 3-6 times higher than the control. The germination rate of transgenic A. thaliana T3 generation seeds reached more than 95%. The O2·-, H2O2, and malondialdehyde contents of the transgenic P. sericea plant lines were increased but lower than those of the wild-type strain. The superoxide dismutase, peroxidase, and hydrogen peroxidase activities were increased in both wild-type and transgenic strains, and the transgenic strains showed higher enzyme activities than the wild-type. We concluded that PsMYB2 could improve plant resistance to cadmium, which provides a theoretical basis for using transgenic plants to remediate cadmium-contaminated soil and for sustainable land use. [ABSTRACT FROM AUTHOR]

Cumulus granulosa cells (CGCs), an important type of ovarian somatic cells, carries out various functions related to oogenesis, follicular development, and steroidogenesis. Studying the biological mechanisms involved in the development and function of CGCs makes a great contribution to understanding the reproductive regulation in female animals. Stanniocalcin-1 (STC1) is an important Ca2+-regulated glycoprotein hormone that exhibits high expression levels in ovaries. In this study, we cloned the coding sequence of the yak STC1 , predicted the structure of STC1 protein, detected the expression and localization of STC1 in yak ovaries, and analyzed the functions of STC1 in yak CGCs. The CDS (coding sequence) region of yak STC1 gene was found to be 744 bp and encoded 247 amino acids. Homology comparison revealed that STC1 protein was highly conserved among mammals. The STC1 mRNA displayed dynamic expression profiles in different stages of yak ovaries, and the highest expression was found in the follicular phase. Regarding localization, STC1 protein was widely distributed in various kinds of yak ovarian cells, including oocytes, mural granulosa cells, CGCs, and thecal cells. Repressing the expression of STC1 resulted in defective proliferation and survival of yak CGCs. In addition, knockdown the expression of STC1 repressed the secretion of progesterone and promoted the secretion of estrogen. Overexpression of STC1 partially rescued the proliferation of CGCs and resulted in opposite effects on the secretion of progesterone and estrogen. Several apoptosis and steroidogenesis-related genes, including BAX , BCL2 , HSD3B1 , HSD17B1, CYP11A1 and CYP17A1 showed altered expressions after repressing or increasing the expression of STC1 in yak CGCs. To the best of our knowledge, this study is the first to focus on the role of STC1 in yak CGCs, and the outcomes offer fresh insights into the mechanism governing yak reproduction. • Cloning the coding sequence of the yak STC1 gene and predicting the structure of STC1 protein. • Identifying the expression and localization of STC1 protein in yak ovaries. • STC1 is essential for the survival and steroidogenesis in yak cumulus granulosa cells. [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.)

Bioluminescence is interesting, among other reasons, for the various technological applications that have been derived from it. Among these applications, developing visualization techniques to record the expression of one or more genes simultaneously in real-time are particularly useful. With this in mind, this study aimed to generate a recombinant Pyrocystis lunula luciferase protein (Luci D2-3 partial CDS). As the main results, i) a fragment of 1467 bp of the luciferase (LCFb) mRNA of the dinoflagellate P. lunula, containing part of domain 2 and all of the domain 3, was cloned in the pET28a vector; ii) the constructed vector was used to transform Escherichia coli to express the recombinant protein and subsequently purify it through an affinity chromatography procedure using a His-Tag; and iii) the purified protein (~50 kDa) was further analyzed by mass spectrometry to confirm its identity. Despite being unable to perform activity tests with the luciferin substrate, the evidence from previous studies indicates that the recombinant protein obtained in this case is enzymatically active. Due to the limited number of currently available luciferases, synthesizing this recombinant protein represents a useful tool, especially in designing expression assays coupled to multiple reporter genes, thus expanding the palette of proteins available for developing this type of biotechnological advances. [ABSTRACT FROM AUTHOR]

【Objective】Fresh-cut taro is prone to turning yellow and brown during storage, even stored at low temperature, which greatly diminishes its commercial values. In order to identify the important functional genes involved in the browning process of fresh-cut taro, the allene oxide synthase (AOS) gene AOS1 was cloned from fresh-cut taros. The correlation between the expression of AOS1 and the browning development of fresh-cut taro was analyzed, with a view to offering references for the study of taro browning mechanism.【Method】With cDNA of fresh-cut taro as a template, the coding sequence (CDS) of AOS1 gene was cloned by the RT-PCR method. The expression patterns of AOS1 during the browning of fresh-cut taro were analyzed by using qRT-PCR, the impact of exogenous browning inhibitors on AOS1 expression was examined by using transcriptome sequencing data, and the promoter subsequence was analyzed.【Result】Fresh-cut taros showed obvious yellowing and browning symptoms on the cut surface after 24 hours of storage at 20 ℃, while the obvious browning symptoms were occurred after 12 d of cold storage at 4 ℃. These results suggested that storage temperature was a crucial factor influencing the browning process of fresh-cut taros, with low temperature storage effectively inhibiting the browning. The CDS of taro AOS1 gene had a total length of 1 560 bp and encoded 519 amino acid residues. The molecular weight of the protein encoded by AOS1 gene was 57.63 kD, with an isoelectric point of 8.68, and located in chloroplast. The similarity of AOS1 protein sequences across different plant species was relatively low, with notable differences observed between taro, tomato, and Arabidopsis. However, high sequence homology was found among AOS1 proteins in Araceae family plants. The promoter region of taro AOS1 contained various plant hormone and stress response elements. The expression of AOS1 significantly increased after fresh-cut processing, indicating that the cutting treatment induced AOS1 expression. Moreover, as the browning of freshcut taro progressed, the expression of AOS1 gradually increased. Applications of exogenous glycolic acid and vanillic acid significantly decreased the expression of AOS1 in fresh-cut taros, showing its correlation with the browning process.【Conclusion】 A significant positive correlation was observed between the browning of fresh-cut taros and the expression of AOS1 during cold storage. Treatments such as cutting and peeling may accelerate the browning process by inducing AOS1 expression. [ABSTRACT FROM AUTHOR]

In this study, the complete cDNA sequence of argininosuccinate synthetase was cloned in the saltwater clam Meretrix meretrix using the rapid amplification of cDNA ends (RACE) approach. The complete MmASS cDNA measures 1588 bp in length with an open reading frame encoding 415 amino acids. The molecular weight of MmASS was 46.81 kD and the theoretical isoelectric point was 5.51. The deduced MmASS protein comprises six conserved sequences, containing the substrate-binding residues for ATP, aspartate and citrulline. The results of the multiple amino acid alignment and the phylogenetic analysis showed that MmASS exhibited high amino acid sequence identity with other species. MmASS also had a close phylogenetic relationship with Sinonovacula constricta. Overall, similar functions might exist in M. meretrix and other species. Predicted subcellular localization suggested that MmASS is a cytosolic enzyme. The expression levels of MmASS were distributed in all of the examined tissues. MmASS was most highly expressed in the gill (P<0.05), followed by the hepatopancreas. These results indicated that MmASS was involved in various physiological activities of different tissues and may play an important role in the immune mechanisms. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences 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.)

Phenylalanine ammonia-lyase (PAL) is a key enzyme in the metabolism of phenylpropane that regulates the biosynthesis of flavonoids in raspberry. To elucidate the function and regulation mechanism of the RiPAL1 gene in raspberry, total RNA was extracted from raspberry fruits to synthesize first-strand cDNA. The gene was successfully cloned, and related bioinformatics analyses were carried out. Our results showed that the coding sequence (CDS) of the RiPAL1 gene was 2 133 bp in length, encoding 710 amino acids, with a molecular weight of 77.5 ku and an isoelectric point of 6.21. The maximum hydrophobicity of RiPAL1 protein was 4.50, its minimum hydrophobicity was -4.50, and its average hydrophobicity index was -0.49. Based on these, it was determined that it is a hydrophilic protein with no transmembrane structure and no signal peptide. The RiPAL1 protein contained 4 N-glycosylation sites and 67 specific kinase sites. Its secondary structure was mainly composed of α-helices, while its tertiary structure was a homotetramer. Multiple sequence alignment showed that the RiPAL1 protein has a typical conserved domain of phenylalanine ammonia-lyase. Phylogenetic analysis indicated that the RiPAL1 protein is closely related to that of strawberry. This study provides a reference for further research on the molecular mechanism of the RiPAL1 gene in the biosynthesis of flavonoids in raspberry fruit. [ABSTRACT FROM AUTHOR]

bHLH（basic helix-loop-helix）transcription factors play crucial roles in the biosynthesis of flavonoids in plants. In order to explore the function and molecular regulation mechanism of bHLH transcription factor in flavonoid biosynthesis in tartary buckwheat, RTPCR was used to clone the FtbHLH3, one bHLH transcription factor. Then the bioinformatics subcellular localization, transcriptional activation activity, gene expression, gene co-expression, and the correlation between gene expression level and total flavonoid content of it were conducted. As results, the CDS length of FtbHLH3 was total 783 bp and it encoded total 260 amino acids. Conserved domain and phylogenetic analysis showed that FtbHLH3 was a member of non-IIIf subfamily of bHLH transcription factor family. Subcellular localization and transcriptional activation assay suggested that FtbHLH3 was located in the nucleus and had no transcriptional activation activity. Gene co-expression analysis revealed that FtbHLH3 was co-expressed with the Arabidopsis TT8 homologous gene FtTT8 and 12 structural genes of flavonoid biosynthesis in the tartary buckwheat. The correlation analysis between gene expression level and total flavonoids content in different tissues indicated that there was a highly positive correlation between the expressions of FtbHLH3 and the contents of total flavonoids. All these results suggest that FtbHLH3 is a non-IIIf subfamily bHLH transcription factor that was located in the nucleus and has no transcriptional activation activity. It may regulate the flavonoids biosynthesis in tartary buckwheat through interacting with other transcription factors. [ABSTRACT FROM AUTHOR]

In order to improve the activity and stability of β-mannanase, an endophytic Bacillus subtilis YZ-21 β-mannanase gene ManA was cloned and expressed by gene cloning technique. The enzyme ac‐ tivity and stability were determined by 3,5- dinitrosalicylic acid (DNS) method. The results showed that ManA sequence length of the sequence was 1 104 bp, and the homology with β-mannosidase from the same source was 97.56%. The ManA conserved domain of β-mannanase gene was consistent with the typical characteristics of GH26 glycoside hydrolase family. ManA was connected to the pET-32a expression vector, the recombinant expression plasmid pET-ManA was constructed, and was introduced into Escherichia coli BL21(DE), and the high expression of the enzyme was obtained by induction. The expres‐ sion products were analyzed by SDS-PAGE with a band of about 56.07 ku. The enzyme activity was 372.86 U/mL, and the optimum reaction temperature was 40 ℃, the optimum pH was 7.0, and the optimum reaction substrate was guar gum, whose enzyme activity could reach 392.54 U/mL. After treatment at 55 ℃ for 120 min, the enzyme activity remained 69.8%, and after treatment at pH 4.0- 9.0 for 30 min, the enzyme activity remained more than 60%, and metal ions Ca2+, Co2+, Ba2+ had obvious activation effect on the recombinant enzyme. In conclusion, β-mannanase gene cloned from endophytic Bacillus subtilis YZ-21 can be efficiently expressed in Escherichia coli. [ABSTRACT FROM AUTHOR]

SUMMARY: Rust, caused by the fungus Puccinia helianthi Schwein., is one of the most devastating diseases of sunflower (Helianthus annuus L.), affecting global production. The rust R gene R11 in sunflower line HA‐R9 shows broad‐spectrum resistance to P. helianthi virulent races and was previously mapped to an interval on sunflower chromosome 13 encompassing three candidate genes annotated in the XRQr1.0 reference genome assembly. In the current study, we combined ethyl methane sulfonate (EMS) mutagenesis with targeted region capture and PacBio long‐read sequencing to clone the R11 gene. Sequencing of a 60‐kb region spanning the R11 locus from the R11‐HA‐R9 rust‐resistant line and three EMS‐induced susceptible mutants facilitated the identification of R11 and definition of induced mutations. The R11 gene is predicted to have a single 3996‐bp open reading frame and encodes a protein of 1331 amino acids with CC‐NBS‐LRR domains typical of genes conferring plant resistance to biotrophic pathogens. Point mutations identified in the R11 rust‐susceptible mutants resulted in premature stop codons, consistent with loss of function leading to rust susceptibility. Additional functional studies using comparative RNA sequencing of the resistant line R11‐HA‐R9 and R11‐susceptible mutants revealed substantial differences in gene expression patterns associated with R11‐mediated resistance at 7 days post‐inoculation with rust, and uncovered the potential roles of terpenoid biosynthesis and metabolism in sunflower rust resistance. Significance Statement: The rust R gene R11 in sunflower line HA‐R9 shows broad‐spectrum resistance to Puccinia helianthi virulent races. We combined EMS (ethyl methane sulfonate) mutagenesis with targeted region capture and PacBio long‐read sequencing to clone the R11 gene. The R11 gene is predicted to have a single 3996‐bp open reading frame that encodes a protein of 1331 amino acids with CC‐NBS‐LRR domains typical of genes conferring plant resistance to biotrophic pathogens. [ABSTRACT FROM AUTHOR]

Background: Potentilla sericea is a heavy metal hyperaccumulator landscaping plant. MYB transcription factors play an important role in regulating plant stress response to adversity. However, there are few studies on MYB transcription factors in stress tolerance in Potentilla sericea. In this study, the PsMYB62 gene was successfully cloned from Potentilla sericea. Methods: Bioinformatic analysis and real-time quantitative PCR (qPCR) methods were used to evaluate this gene. The transgenic A. thaliana were obtained by flower dipping and the gene function was identified by determining physiological indicators under cadmium stress. Results: The open reading frame of PsMYB62 is 942 bp, which encodes 313 amino acids (aa) and belongs to the R2R3 MYB transcription factor. The plant overexpression vector PBI121-PsMYB62-GFP was constructed and successfully transferred into A. thaliana. The relative expression level of PsMYB62 was significantly increased by CdCl2, NaCl, ABA, and mannitol treatments. The germination rate of transgenic seeds was higher than those of wild type (WT) and empty vector (EV) under different concentrations of cadmium treatment. Upon treatment with 100 µmol·L-1 of CdCl2·2.5H2O, the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in the transgenic plants were significantly higher than those in the WT and EV. The contents of H2O2, O2·- and malondialdehyde (MDA) in transgenic lines were increased, but lower than those in WT and EV. The expression levels of AtGSH, AtPCS, and AtNAS4 that were related to the regulation of cadmium were increased, but the expression levels of transgenic lines were higher than those of WT and EV. Conclusion: The above results showed that PsMYB62 could be induced by cadmium and could improve the cadmium resistance of plants. [ABSTRACT FROM AUTHOR]

Based on the whole genome data of tea plant（Camellia sinensis）, members of tea plant TATA element modulatory factor （CsTMF）gene family were screened and identified. The full-length coding sequence（CDS）of CsTMFs in the tea plant leaves was further cloned. The conserved domain, gene structure, physical and chemical properties, protein secondary structure and phylogenetic relationship of CsTMFs were analyzed by bioinformatics method. Tissue-specific expression was analyzed based on transcriptome data, and qPCR was used to detect the expressions of CsTMFs in the tea plant leaves under cold stress and drought stress. The result showed that there were two genes of CsTMF gene family in the tea plant. The lengths of CDS region in two CsTMF genes were 2 934 bp and 2 904 bp, respectively. There were complete conserved domains, TMF-DNA-bd and TMF-TATA-bd in both two CsTMF genes. CsTMF1 and CsTMF2 had tissue expression specificity. CsTMF1 was highly expressed in the flowers and stems, while CsTMF2 was highly expressed in the mature leaves and stems. The results of qPCR showed that CsTMF1 was induced by cold stress, while CsTMF2 was inhibited by cold stress and drought stress. [ABSTRACT FROM AUTHOR]

Copyright of Bulletin of Botanical Research is the property of Bulletin of Botanical Research 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.)

Recently, phosphatidylglycerol (PG) focused on its important role in chloroplast photosynthesis, mitochondrial function of the sperm, an inhibitory effect on SARS-CoV-2 ability to infect naïve cells, and reducing lung inflammation caused by coronavirus disease 2019. To develop an enzymatic PG determination method as the high-throughput analysis of PG, a PG-specific phospholipase C (PG-PLC) was found in the culture supernatant of Amycolatopsis sp. NT115. PG-PLC (54 kDa by SDS-PAGE ) achieved the maximal activity at pH 6.0 and 55°C and was inhibited by detergents, such as Briji35, Tween 80, and sodium cholate, but not by EDTA and metal ions, except for Zn2+. The open reading frame of the PG-PLC gene consisted of 1620 bp encoding 515-amino-acid residues containing the preceding 25-amino-acid residues (Tat signal peptide sequence). The putative amino acid sequence of PG-PLC was highly similar to those of metallophosphoesterases; however, its substrate specificity was completely different from those of known PLCs. [ABSTRACT FROM AUTHOR]

To investigate whether the expression of the chloroplast gene Iborf56 in sweet potato is induced by salt stress and its effect on growth and development, and provide a reference for sweet potato breeding, in this study, the sweet potato variety, Chestnut Fragrance, was used as the research material to clone the Iborf56 gene. The protein sequence features, subcellular localization prediction, phylogenetic analysis, regulatory element analysis, expression patterns under different tissues and salt stress were analyzed using bioinformatics methods, and qPCR was used to validate the expression level of Iborf56. The results showed that Iborf56 had a 99.28% similarity with known sequences from NCBI and an open reading frame length of 929 bp, encoding 60 amino acids with 8 phosphorylation sites. The Iborf56 protein lacked signal peptides and transmembrane regions and did not possess the characteristics of secreted proteins. It was mainly composed of α-helices and its subcellular localization was in the chloroplast stroma. Phylogenetic tree analysis showed that Iborf56 had the closest genetic distance to Ipomoea trifida and Ipomoea triloba. Promoter element analysis revealed that the promoter region of this gene contained functional elements such as light response, stress response, and hormone response. Transcriptome and qRT-PCR quantitative analysis showed that the expression level of Iborf56 was the highest in sweet potato leaves and increased significantly with growth time, inferring that this gene may be involved in the growth and development of sweet potatoes. In addition, the expression level of the Iborf56 gene showed an upward trend first and then a downward trend with increasing salt stress time, and the critical value was reached after 12 h under salt stress, indicating that it may be involved in the salt stress response of sweet potatoes. [ABSTRACT FROM AUTHOR]

Aquaporins are highly conserved water channel proteins involved in the transport of solutes across membranes. The few aquaporins identified to date play major roles in excretion in insects. Excretion is a major source of indoor allergens from the house dust mite species, but, to date, aquaporins have not been well studied in these species. We used RNA sequencing to study the transcriptome of the house dust mite Dermatophagoides pteronyssinus to identify putative aquaporin family members. RNA was isolated from more than 2000 mites for 5΄-RACE and 3΄-RACE. Putative aquaporins were identified through BLAST alignments, and primers were designed to amplify the five identified sequences. These cDNA fragments were then inserted into bacterial plasmids for molecular cloning and sequencing of DerpAQP1, DerpAQP2, DerpAQP3, DerpAQP4, and DerpAQP5 (GenBank accessions nos. KY305293, KY305294, KY305295, KY305296, and KY305298, respectively). Phylogenetic analysis of cloned sequences revealed the clustering of these putative aquaporins into two groups with human aquaporins (hAQPs): DerpAQP3 was clustered with hAQP8, hAQP11, and hAQP12; and DerpAQP1, DerpAQP2, DerpAQP4, DerpAQP5 were clustered with hAQP3, hAQP7 hAQP9, and hAQP10. This study provides a foundation for future studies of the importance of these proteins to dust mite functions and allergenicity. [ABSTRACT FROM AUTHOR]

Simple Summary: There are many existing studies related to goat adipose deposition. However, no studies have reported the role of the CXCL17 gene on adipose deposition in goats before. The aim of this study was to explore the effect of CXCL17 on goat subcutaneous preadipocytes proliferation and provide potential targets for goat-directed molecular breeding. The results showed that the exogenous expression of CXCL17 at different doses had different regulatory effects on the proliferation of subcutaneous preadipocytes of goats. The presence or absence of subcutaneous adipose accumulation will affect the energy storage, insulation resistance and metabolism of animals. Proliferation and differentiation of preadipocytes play a significant role in lipid deposition. The objective of this study was to clone the goat CXCL17 gene and investigate its potential functions on goat subcutaneous preadipocytes' proliferation by gaining or losing function in vitro. The goat CXCL17 gene was cloned by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and bioinformatics analysis was performed. The expression of the CXCL17 gene in the different goat tissues and adipocytes at different differentiation stages was detected by real-time fluorescence quantitative PCR (qPCR). The results showed that the cloned sequence of goat CXCL17 gene is 728 bp and the CDS region is 357 bp, encoding 118 amino acids. CXCL17 protein is located in nucleus, cytoplasm, mitochondria and extracellular matrix. Tissue-expression profiles revealed that CXCL17 expressed in all of the examined tissues. In visceral tissues, the highest expression level was found in lung (p < 0.01); in muscle tissues, the highest CXCL17 expression level was found in the longissimus dorsi (p < 0.01) and in adipose tissues, the highest expression level was found in subcutaneous adipose (p <0.01). Compared with those cells before differentiation, CXCL17 expression levels upregulated at 48 h (p < 0.01), 72 h (p < 0.01), 120 h (p < 0.01) and downregulated at 96 h (p < 0.01). Furthermore, the results of crystal violet staining and semi-quantitative assay showed that transfection with 1 μg CXCL17 expression plasmid reduced the cell numbers in vitro. Meanwhile, the expression of CCND1 was significantly decreased. A similar consequence happened after interfering with CXCL17 expression. However, plasmid transfected with 2 μg pEGFPN1-CXCL17 increased the number of cells in vitro. These results suggest that CXCL17 is involved in the proliferation of goat subcutaneous preadipocytes. [ABSTRACT FROM AUTHOR]