RESUMO
Calcium plays a critical role in regulating abiotic stress responses in plants. Calcineurin B-like (CBL) proteins are calcium sensors in calcium signaling pathways. However, the molecular mechanisms underlying calcium signaling remain to be elucidated. In this study, the CBL1 gene, which codes for the CBL protein, was isolated from the birch-leaf pear. One 2,969-bp sequence was cloned using PCR, and using the cloned 2,027-bp sequence was isolated from pear genomic DNA via genome walking. Sequencing analysis revealed that the 4,996-bp sequence was a PbCBL1 gene consisting of eight exons and seven introns, and the 2,027-bp sequence was identified as the promoter of the PbCBL1 gene, which contains the basic promoter elements TATA and CAAT boxes. In addition, some other cis-acting elements including heat, cold, drought, and hormone responsive elements were also present. To further investigate the activity of this promoter, the sequence was used to drive a GUS fusion gene into leaf discs of tobacco (Nicotiana benthamiana) with Agrobacterium-mediated transformation method. GUS gene expression could be regulated by the PbCBL1 promoter following induction by GA, ABA, SA, and MeJA. Furthermore, the results of real-time RT-qPCR indicate that the PbCBL1 gene can respond to changes in the intracellular calcium concentration, and that it can be induced by cold, heat, drought, and stress by several hormones including GA, ABA, SA, and MeJA. PbCBL1 gene may be involved in several signal transduction pathways, and play an important role in the condition of adversity stress in pear.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Clonagem Molecular , Proteínas de Plantas/genética , Regiões Promotoras Genéticas , Pyrus/genética , Região 5'-Flanqueadora , Sequência de Aminoácidos , Sequência de Bases , Expressão Gênica , Ordem dos Genes , Genes Reporter , Vetores Genéticos , Dados de Sequência Molecular , Motivos de Nucleotídeos , Proteínas Recombinantes de Fusão , Sequências Reguladoras de Ácido Nucleico , Reprodutibilidade dos Testes , Alinhamento de Sequência , Análise de Sequência de DNA , Transformação GenéticaRESUMO
Cotton (Gossypium spp) is one of the most economically important crops that provide the world's most widely used natural fiber. Diseases such as Fusarium wilt and particularly Verticillium wilt seriously affect cotton production, and thus breeding for disease resistance is one of the most important goals of cotton breeding programs. Currently, potential exists to improve disease resistance in cultivated cotton. Increasing the understanding of the distribution, structure, and organization of genes or quantitative trait loci for disease resistance will help the breeders improve crop yield even in the event of disease. To facilitate the mapping of disease-resistance quantitative trait loci to achieve disease-resistant molecular breeding in cotton, it is necessary to develop polymorphic molecular markers. The objective of this study was to develop simple sequence repeat markers based on cotton expressed sequence tags for disease resistance. The efficacy of these simple sequence repeat markers, their polymorphisms, and cross-species transferability were evaluated. Their value was further investigated based on genetic diversity and evolution analysis. In this study, the unique sequences used to develop markers were compared with the G. arboretum and G. raimondii genome sequences to investigate their position, homology, and collinearity between G. arboretum and G. raimondii.
Assuntos
Cromossomos de Plantas/química , Resistência à Doença/genética , Gossypium/genética , Doenças das Plantas/genética , Polimorfismo Genético/imunologia , Locos de Características Quantitativas , Sequência de Bases , Evolução Biológica , Mapeamento Cromossômico , Resistência à Doença/imunologia , Fusarium/patogenicidade , Fusarium/fisiologia , Marcadores Genéticos , Gossypium/classificação , Gossypium/imunologia , Gossypium/microbiologia , Repetições de Microssatélites , Dados de Sequência Molecular , Filogenia , Melhoramento Vegetal , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Verticillium/patogenicidade , Verticillium/fisiologiaRESUMO
Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease caused by non-excessive alcohol consumption and is the most common cause of elevated levels of serum liver enzymes. We examined changes in adiponectin (APN) and tumor necrosis factor-α (TNF-α) in type 2 diabetes mellitus (T2DM) complicated by NAFLD and their relationships with insulin resistance (IR). Forty-two T2DM, 39 NAFLD, and 45 T2DM complicated with NAFLD (complicated group) patients were enrolled in this study. Body mass index, fasting blood plasma glucose (FPG), fasting insulin, triglyceride (TG), alanine aminotransferase, gamma-glutamyl transpeptidase, APN, TNF-α, and homeostasis model of assessment (HOMA)-IR were determined. The degree of fatty liver was graded according to liver/spleen computed tomography ratio and intrahepatic vessel manifestations. Compared with the T2DM and NAFLD groups, fasting blood plasma glucose, alanine aminotransferase, gamma-glutamyl transpeptidase, TG, TNF-α, and HOMA-IR in the complicated group were significantly increased, while APN was significantly reduced. Body mass index in the complicated group was significantly higher than in the T2DM group. The complicated group was prone to severe fatty liver compared with the NAFLD group. APN was negatively correlated with body mass index, fasting blood plasma glucose, TG, TNF-α, and HOMA-IR. TNF-α was negatively correlated with APN, but positively correlated with FPG, fasting insulin, TG, and HOMA-IR. The complicated group had clear IR. A more severe degree of fatty liver was associated with higher HOMA-IR and TNF-α and lower APN. APN was an important factor for antagonizing inflammation and mitigating IR.