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تطوير إقتصاد دائري في الصين
هذا العمل الجماعي المشترك، كما يشير عنوانه : (التاريخ المديني الحضري للصين) هو كتاب حضاري بامتياز. كتاب يستعرض جوانب أساسية من حضارة الشعب الصيني القديمة-المتجددة الراسخة، الضاربة جذورها عميقا في تربة التاريخ البشري الشامل. يعد الكتاب سفر حضاري قيم وبحث متصل رصين حول التطور التاريخي العمراني في المدن الصينية وهو إذ يتناول بشكل أساسي، فن العمارة والبناء والتصميم وتخطيط المدن والشوارع والأحياء والأسواق والمقار الحكومية والحدائق والمعابد والساحات ومسالك الملاحة النهرية والري، فإنه لا يغفل في الوقت نفسه عن ما يتصل به هذا الموضوع، من عشرات المواضيع الحضارية والثقافية والمعيشية الأخرى، مثل : العبادات والاعتقادات والتقاليد والعادات والآداب والطقوس والحرف والفنون والنظم الاجتماعية والصراع السياسي والغزو الخارجي والتنظيم الحكومي والقبلي والاجتماعي والطبقي والعسكري والإداري والإنمائي والعمراني والإسكاني والتعليمي والتجاري والأدبي والفني والموسيقي والفولكلوري. إنه كتاب جميل يقدم العديد من اللوحات الرشيقة الرحبة المتقابلة المتلاحقة المتكاملة التي ترحل بنا بين القصور والأنهار والأسوار والأحياء والمعاهد والمعابد وهياكل عبادة الأجداد ومواقع التنقيب والساحات والحدائق والميادين ودواوين الشعر وسجلات التاريخ ولا يخلو كل ذلك من جولات من البحث المقارن في غير أمر من هذه الأمور ولن نطيل عليكم في الإضاءة على مواضيع هذا الكتاب لأكثر من ذلك، آملين لكم سلاسة القراءة ومتعة الاكتشاف.
Book
Conciliatory Anti-Allergic Decoction Attenuates Pyroptosis in RSV-Infected Asthmatic Mice and Lipopolysaccharide (LPS)-Induced 16HBE Cells by Inhibiting TLR3/NLRP3/NF-κB/IRF3 Signaling Pathway
Respiratory syncytial virus (RSV) infection can deteriorate asthma by inducing persistent airway inflammation. Increasing evidence elucidated that pyroptosis plays a pivotal role in asthma. Conciliatory anti-allergic decoction (CAD) exhibits an anti-inflammatory effect in ovalbumin (OVA)-induced asthma; however, the effects and mechanisms of CAD in RSV-infected asthmatic mice have not yet been elucidated. The RSV-infected asthmatic mice model and lipopolysaccharide (LPS)-induced 16HBE cell pyroptosis model were established, respectively. Pulmonary function, ELISA, and histopathologic analysis were performed to assess the airway inflammation and remodeling in mice with CAD treatment. Furthermore, ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry (UPLC-Q-TOF/MS) was conducted to identify the chemical compounds of high-dose CAD (30 g/kg). Cell viability and apoptosis of 16HBE cells were assessed by CCK-8 and flow cytometry assays, respectively. Finally, the expression levels of apoptosis-, pyroptosis-, and TLR3/NLRP3/NF-κB/IRF3 signaling-related genes were measured with qRT-PCR or western blotting, respectively. Pulmonary function tests showed that CAD significantly ameliorated respiratory dysfunction, airway hyperresponsiveness, inflammation cell recruitment in BALF, pulmonary inflammation, collagen deposition, and cell death in lung tissues. CAD significantly decreased the content of TNF-α, IL-13, IL-4, IL-1β and IL-5 in the bronchoalveolar lavage fluid (BALF), IL-17, IL-6, and OVA-specific IgE in serum and increased serum IFN-γ in asthma mice. The results of UPLC-Q-TOF/MS showed that high-dose CAD had 88 kinds of chemical components. In vitro, CAD-contained serum significantly suppressed LPS-induced 16HBE cell apoptosis. Additionally, CAD and CAD-contained serum attenuated the up-regulated expressions of Bax, Cleaved caspase-3, NLRP3, ASC, Cleaved caspase-1, GSDMD-N, IL-18, IL-1β, TLR3, p-P65, p-IκBα, and IRF3 but increased Bcl-1 and GSDMD levels in the asthma mice and LPS-induced 16HBE cells, respectively. These results illustrated that CAD may have a potential role in improving airway inflammation and pyroptosis through inhibition of the TLR3/NLRP3/NF-κB/IRF3 signaling pathway.
Journal Article
Investigation of the role of AcTPR2 in kiwifruit and its response to Botrytis cinerea infection
Background
Elucidation of the regulatory mechanism of kiwifruit response to gray mold disease caused by
Botrytis cinerea
can provide the basis for its molecular breeding to impart resistance against this disease. In this study, ‘Hongyang’ kiwifruit served as the experimental material; the TOPLESS/TOPLESS-RELATED (TPL/TPR) co-repressor gene
AcTPR2
was cloned into a pTRV2 vector (
AcTPR2
-TRV) and the virus-induced gene silencing technique was used to establish the functions of the
AcTPR2
gene in kiwifruit resistance to
Botrytis cinerea
.
Results
Virus-induced silencing of
AcTPR2
enhanced the susceptibility of kiwifruit to
Botrytis cinerea
. Defensive enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and phenylalanine ammonia-lyase (PAL) and endogenous phytohormones such as indole acetic acid (IAA), gibberellin (GA
3
), abscisic acid (ABA), and salicylic acid (SA) were detected. Kiwifruit activated these enzymes and endogenous phytohormones in response to pathogen-induced stress and injury. The expression levels of the IAA signaling genes—
AcNIT
,
AcARF1
, and
AcARF2
—were higher in the
AcTPR2
-TRV treatment group than in the control. The IAA levels were higher and the rot phenotype was more severe in
AcTPR2
-TRV kiwifruits than that in the control. These results suggested that
AcTPR2
downregulation promotes expression of IAA and IAA signaling genes and accelerates postharvest kiwifruit senescence. Further,
Botrytis cinerea
dramatically upregulated
AcTPR2,
indicating that
AcTPR2
augments kiwifruit defense against pathogens by downregulating the IAA and IAA signaling genes.
Conclusions
The results of the present study could help clarify the regulatory mechanisms of disease resistance in kiwifruit and furnish genetic resources for molecular breeding of kiwifruit disease resistance.
Journal Article
Expression of the SPL-like gene LaSPL9 in Japanese larch (Larix leptolepis) is regulated by miR156 during somatic embryogenesis
Key messageThe Larix leptolepis SPL-like gene LaSPL9 was identified. Additionally, premiR156b and LaSPL9 expression patterns during somatic embryogenesis and the effects of gibberellin and salicylic acid on their transcription revealed the post-transcriptional regulatory functions of the miR156–LaSPL9 module.A full-length cDNA sequence of a SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE (SPL) gene was isolated from Japanese larch (Larix leptolepis) and designated as LaSPL9. The cDNA comprised 2735 bp, including a 1779-bp open reading frame encoding a protein (592 amino acids) with conserved SBP domains. The LaSPL9 promoter region included several gibberellin (GA)- and salicylic acid (SA)-inducible elements. Further analyses revealed that LaSPL9 mRNA is targeted by miR156. An examination of the LaSPL9-GFP fusion protein in tobacco leaf epidermal cells indicated that LaSPL9 is a nuclear protein. Additionally, LaSPL9 expression was induced by GA and the miR156b precursor (premiR156b) transcript accumulation was inhibited by SA. Moreover, abscisic acid (ABA) was identified as the main factor down-regulating LaSPL9 expression during the early stage of somatic embryogenesis (SE), which can induce premiR156b expression. Furthermore, LaSPL9 transcription peaked at 10 days, implying it might encode an important regulator of early embryonic pattern formation. The LaSPL9 transcript level subsequently decreased gradually and was lowest at 42 days, which is consistent with the gradual increase in miR156 expression during SE. The miR156–LaSPL9 module might regulate SE in L. leptolepis by responding to ABA. Overall, our results provide evidence that miR156–LaSPL9 is important for the crosstalk between GA and SA pathways during SE.
Journal Article
Screening and Functional Evaluation of Four Larix kaempferi Promoters
Promoters are powerful tools for breeding new varieties using transgenic technology. However, the low and unstable expression of target genes is still a limiting factor in Larix kaempferi (Lamb.) Carr (Japanese larch) genetic transformation. In this study, we analyzed L. kaempferi transcriptome data, screened out highly expressed genes, cloned their promoters, and constructed plant expression vectors containing the β-glucuronidase (GUS) reporter gene driven by these promoters. Recombinant vectors were introduced into the L. kaempferi embryogenic callus by means of the Agrobacterium-mediated transient or stable genetic transformation method, and the promoter activity was then determined by measuring GUS expression and its enzyme activity in the transformed materials. Four highly expressed genes were identified: L. kaempferi Zhang Chen Yi-1 (LaZCY-1), Zhang Chen Yi-2 (LaZCY-2), Translationally Controlled Tumor Protein (LaTCTP), and ubiquitin (LaUBQ). The 2000 bp fragments upstream of ATG in these sequences were cloned as promoters and named pLaZCY-1, pLaZCY-2, pLaTCTP, and pLaUBQ. Semi-quantitative and quantitative RT-PCR analyses of transient genetic transformation materials showed that all four promoters could drive GUS expression, indicating that they have promoter activities. Semi-quantitative and quantitative RT-PCR analyses and the histochemical staining of stable genetic transformation materials showed that the pLaUBQ promoter had higher activity than the other three L. kaempferi promoters and the CaMV35S promoter. Thus, the pLaUBQ promoter was suggested to be used in larch genetic transformation.
Journal Article
Transcriptome-wide analysis to dissect the transcription factors orchestrating the phase change from vegetative to reproductive development in Larix kaempferi
The timing of phase change from vegetative to reproductive development during aging of forest trees is important for wood and seed production. In a previous study, we investigated the effects of aging on wood formation by measuring the transcriptomic changes in the uppermost main stems of 1-, 2-, 5-, 10-, 25-, and 50-year-old Larix kaempferi. Based on the published transcriptome data, here we investigated the transcriptomic differences between the juvenile vegetative (1- and 2-year-old) and adult reproductive (25- and 50-year-old) phases to determine the molecular mechanisms underlying the phase change. In total, 12,789 transcripts were identified as differentially expressed genes, including 573 transcription factors. Further analysis showed that 27 transcription factors belonging to 8 families were common to all four comparisons between old and young life stage categories: (I) 25 vs 1 year old; (II) 50 vs 1 year old; (III) 25 vs 2 years old; and (IV) 50 vs 2 years old. The analysis of their expression patterns in six age categories showed that members of the AP2 and Dof families were expressed highly in 1- and 2-year-old trees, weakly in 25- and 50-year-old trees, while members of the C3H, G2-like, GRAS, MYB-related, and MADS families had the opposite patterns. Notably, one member of the MADS family was only detected in 25- and 50-year-old trees. These results suggest that the phase change might (1) occur in the early stage of the L. kaempferi lifetime and (2) be controlled by a complex regulatory network of different transcription factors, some of which are known to play roles in the phase change in model plants. These findings not only provide molecular markers to distinguish different stages of tree growth and development and potential targets for genetic manipulation to improve the reproductive traits of trees, but also improve our understanding of the phase change with aging in trees.
Journal Article
Identification of Target Gene and Interacting Protein of Two LaSCL6 Alternative Splicing Variants Provides Novel Insights into Larch Somatic Embryogenesis
Somatic embryogenesis is valuable for clonal propagation and genetic improvement, and it also serves as an ideal system for studying plant development mechanisms. In Larix kaempferi, microRNA171 and its target gene L. kaempferi SCARECROW-LIKE6 (LaSCL6), which has two alternative splicing variants, can regulate somatic embryogenesis; however, the underlying molecular mechanism is still unknown. In this study, we overexpressed these two LaSCL6 variants in Oryza sativa and Arabidopsis thaliana and then used the RNA-Seq method to screen genes from O. sativa and A. thaliana, whose expression patterns are related to those of LaSCL6 variants. The screened genes were then used to search L. kaempferi proteins to identify the candidate target genes of LaSCL6. After yeast one-hybrid and dual- luciferase transcriptional activity assays, cytochrome P450, family 89, subfamily A, polypeptide 5 (CYP89A5), and wall-associated receptor kinase-like 20 (WAKL20) were confirmed to be the target genes of LaSCL6-var1; in addition, WAKL20 and UDP-glycosyltransferase 85A3 (UGT85A3) were confirmed to be the target genes of LaSCL6-var2. Moreover, APETALA2-like protein 2, a transcription factor from the AP2/ERF family, was shown to interact with LaSCL6-var1 and LaSCL6-var2. Taken together, our results suggest a regulatory network of miR171-LaSCL6. The findings presented here not only provide novel insights into the regulation of the miR171-LaSCL6 module but also explain the mechanism underlying larch somatic embryogenesis and other biological processes.
Journal Article
Overexpression of Larch SCL6 Inhibits Transitions from Vegetative Meristem to Inflorescence and Flower Meristem in Arabidopsis thaliana (L.) Heynh
SCARECROW-LIKE6 (SCL6) plays a role in the formation and maintenance of the meristem. In Larix kaempferi (Lamb.) Carr., an important afforestation tree species in China, SCL6 (LaSCL6) has two alternative splicing variants—LaSCL6-var1 and LaSCL6-var2—which are regulated by microRNA171. However, their roles are still unclear. In this study, LaSCL6-var1 and LaSCL6-var2 were transformed into the Arabidopsis thaliana (L.) Heynh. genome, and the phenotypic characteristics of transgenic A. thaliana, including the germination percentage, root length, bolting time, flower and silique formation times, inflorescence axis length, and branch and silique numbers, were analyzed to reveal their functions. It was found that LaSCL6-var1 and LaSCL6-var2 overexpression shortened the root length by 41% and 31%, respectively, and increased the inflorescence axis length. Compared with the wild type, the bolting time in transgenic plants was delayed by approximately 2–3 days, the first flower and silique formation times were delayed by approximately 3–4 days, and the last flower and silique formation times were delayed by about 5 days. Overall, the life cycle in transgenic plants was prolonged by approximately 5 days. These results show that LaSCL6 overexpression inhibited the transitions from the vegetative meristem to inflorescence meristem and from the flower meristem to meristem arrest in A. thaliana, revealing the roles of LaSCL6-var1 and LaSCL6-var2 in the fate transition and maintenance of the meristem.
Journal Article
Regulation of LaSCL6 expression by genomic structure, alternative splicing, and microRNA in Larix kaempferi
SCR-LIKE-6 (SCL6), a member of the GRAS transcription factor family, plays important roles in many aspects of plant growth and development. In a previous study, we showed that the post-transcriptional regulation of Larix (La)SCL6 by the microRNA miR171 functions in regulating the mode of cell division and the maintenance of embryogenic potential during somatic embryogenesis in larch. To investigate its expression pattern during tree aging, which has been studied by comparative transcriptomic analysis, we re-examined the annotation and expression of every transcript and found one that was longer; we annotated it as SCL6, indicating that it might be a variant of LaSCL6. To verify this, we cloned the DNA and cDNA sequences of this transcript and found that alternative splicing indeed occurred in its expression. Moreover, both variants were detectable in embryogenic cultures and several organs. Notably, the DNA sequence of LaSCL6 contained simple sequence repeats and a single-nucleotide polymorphism which might result in the expression of two variants with a change in their tertiary structures. Regulation of LaSCL6 by miR171 was also found. Taken together, the expression of LaSCL6 is controlled at several levels, and this study not only provides further information about the expression of LaSCL6 but also offers a means to study the regulation of gene expression.
Journal Article
Transcriptional and post-transcriptional regulation of the miR171-LaSCL6 module during somatic embryogenesis in Larix kaempferi
Key messageExpression analysis of Larix kaempferi mature miR171s and their primary transcripts and target gene LaSCL6 during somatic embryogenesis revealed the transcriptional and post-transcriptional regulation of the miR171-LaSCL6 module.Somatic embryogenesis provides a useful experimental system for studying the regulatory mechanisms of plant development. The level and activity of microRNA171 (miR171) fluctuate during somatic embryogenesis in Larix kaempferi, but the underlying mechanisms are still unclear. Here, in L. kaempferi we identified five members of the miR171 family, which cleave LaSCL6 mRNA at different sites. In addition, we improved the method of measuring miRNA activity in a more direct way. Furthermore, we measured the expression patterns of mature miR171s and their primary transcripts during somatic embryogenesis in L. kaempferi and found that their patterns differed, indicating that the transcription of MIR171 genes and the subsequent cleavage of their intermediate products are regulated. Taken together, our findings not only offer a means to study the regulation of miRNA activity, but also provide further insight into the regulation of L. kaempferi somatic embryogenesis by miR171-LaSCL6.
Journal Article