母猪诱导发情技术

本文就母猪各种诱导发情方法的原理及应用现状进行了概述。认为诱导排卵技术是解决母猪生理性乏情的根本途径。     

β- 氨基丁酸诱导甜( 辣) 椒抗疫病作用的研究

 报道了用β- 氨基丁酸( DL-β-aminon-butyric acid, BABA) 喷雾处理辣椒叶片和茎后的诱导抗疫病作用。研究证明: 高浓度BABA ( 1 000 g/mL) 对离体辣椒疫霉病菌无抗菌活性, 用其喷雾处理辣椒的茎叶所诱导的抗疫病作用可完全控制其危害; 用BABA 诱导处理后3 d 接种辣椒疫霉病菌, 辣椒植株开始表达出较高的诱导抗性, 这种抗病作用可持续20 d 以上, 并表现出与数量抗病性相似的特性。     

β-氨基丁酸、茉莉酸及其甲酯诱导辣椒抗TMV作用的研究

β -氨基丁酸 (BABA)、茉莉酸 (JA)及其甲酯 (MeJA)喷雾处理辣椒叶片和茎秆后可诱导辣椒获得对TMV的抗性。研究结果表明 :3种诱抗剂对不同辣椒品种所表现的诱抗效果没有显著差异 ;诱抗效果受诱抗剂的使用浓度、作物的生育期和TMV的接种浓度所影响 ;最高的诱抗效果可达到 5 0 %以上 ;BABA处理后 5～ 6d ,JA与MeJA处理后 6～ 10d接种TMV ,辣椒植株开始表达出较高的诱导抗性 ,这种抗性可持续 15d以上 ,并表现出与数量抗病性相似的特性。     

sikFBA4基因启动子的克隆及低温表达模式分析

天山雪莲生长于常年积雪覆盖高海拔处的高山区域,具有很强的极端低温生境适应能力,属于研究植物低温适应的良好模式植物。前期的研究表明,sikFBA4基因可以显著提高番茄的抗寒能力。为进一步研究sikFBA4基因的耐寒响应模式,以天山雪莲为材料,采用实时荧光定量PCR分析sikFBA4在低温胁迫下的表达模式;利用高效热不对称PCR法(Hi-Tail PCR)克隆sikFBA4启动子序列并进行生物信息学分析。为分析PsikFBA4序列克隆的完整性及转录表达特性,将PsikFBA4与GUS基因融合在烟草中进行瞬时表达。结果表明,sikFBA4在低温胁迫条件下的表达发生瞬时显著上调,并在1 h达到峰值,而后表达下调。通过启动子克隆分析,在PsikFBA4序列-1648 bp位置处具有冷响应元件LTRE;进一步的GUS染色和酶活力测定结果表明,低温能够显著提高GUS基因的表达活性,说明sikFBA4属于低温诱导型基因。SikFBA4基因启动子的克隆、序列分析以及表达分析,为进一步探究雪莲果糖-1,6-二磷酸醛缩酶(sikFBAs)基因的表达与调控机制奠定了基础。     

菊花ClERF1基因的克隆与表达分析

以甘菊叶片为试验材料,采用RT-PCR和Tail-PCR以及生物信息学的方法,研究了甘菊ClERF1及其启动子序列特征、表达模式,以及甘菊ClERF1在低温胁迫下的响应。以期为进一步研究甘菊ClERF1功能提供参考依据。结果表明:ClERF1基因序列全长1 242 bp,最大开放阅读框(ORF)618 bp,可编码206个氨基酸。经理化性质分析,ClERF1分子量23 631.35 Da,理论等电点5.19,不稳定系数53.84,脂肪指数62.04,平均亲水性-0.786,亚细胞定位在细胞核内。系统进化树表明,ClERF1与拟南芥At3g23240亲缘关系最近,属于ERF亚家族。qRT-PCR分析表明ClERF1在叶片中表达最高,其次在茎段中。该研究克隆了ClERF1启动子序列1 534 bp,主要包括低温反应和乙烯响应元件、生长素和茉莉酸甲酯反应元件等。甘菊低温处理幼苗ClERF1表达量显著高于未低温处理的幼苗,其中5℃时ClERF1相对表达量最高。     

Vitamin C enhanced myocardial differentiation of dedifferentiated fat cells

AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.     

荔枝FKBP16-2基因启动子的克隆与瞬时表达分析

前期研究中发现了一个果肉低表达而叶片中高表达的荔枝基因FKBP16-2。本文克隆了该基因1 578 bp的启动子片段并对其功能进行了初步分析,结果表明:荔枝FKBP16-2基因启动子序列中含有大量的TATA-box和CAAT-box保守元件,以及TCA-element,ARE,HSE,GCN4_motif,O2-site等各种转录调控相关的顺式作用元件。该启动子能驱动GUS基因在荔枝的花、叶、根、果皮以及种子中表达而在果肉中不表达,表达具有组织特异性。     

农业技术模式转换与对策探讨——以河南省欠发达平原农业区为例

面对当前农村劳动力转移给农业发展带来的挑战和机遇,转换农业技术模式尤为重要.采用理论分析和调查访谈相结合的方法,得出河南欠发达平原农业区农业技术模式转换必须结合本地实际,遵循诱致性轨迹,注重转换过渡模式,即以农民社会资本为纽带进行土地流转,形成土地适度渐进规模经营,以村集体、宗族等为核心发展种子、生物科技、中小型机械设...     

鸡脂肪组织TCF21基因启动子区DNA甲基化与其表达的关系

旨在研究鸡脂肪组织中TCF21基因启动子区DNA甲基化水平与其表达的关系。以东北农业大学高、低腹脂双向选择品系（简称高、低脂系）第24世代7周龄肉鸡为试验材料，利用RT-qPCR检测高、低脂系肉鸡腹部脂肪组织中TCF21基因的mRNA表达水平；利用生物信息学和双荧光素酶报告系统分析TCF21基因启动子的结构与功能；利用Sequenom MassARRAY飞行质谱检测高、低脂系肉鸡腹部脂肪组织中TCF21基因启动子区CpG位点的甲基化水平；利用CpG甲基转移酶处理TCF21启动子报告基因质粒，分析DNA甲基化对TCF21基因启动子活性的影响。结果显示，高脂系肉鸡腹部脂肪组织中TCF21基因的mRNA表达水平极显著高于低脂系（P<0.001）；TCF21基因的启动子区存在40个CpG位点，且在启动子的近端和远端均有分布，但不存在CpG岛；将TCF21基因的启动子划分为5个功能区域，分别为R1区域（-2 000~-1 500 bp）、R2区域（-1 500~-1 000 bp）、R3区域（-1 000~-500 bp）、R4区域（-500~-200 bp）和Core区域（-200~-100 bp）；高脂系R2、R3和R2+R3区域的DNA甲基化水平显著或极显著高于低脂系（P<0.05或P<0.001）；R2、R3、R2+R3区域的DNA甲基化水平与TCF21基因mRNA表达水平呈显著正相关（R2区域：r=0.438，P<0.05；R3区域：r=0.371，P<0.05；R2+R3区域：r=0.489，P<0.05）；R2区域的DNA甲基化显著抑制其转录活性（P<0.05）。综上所述，TCF21基因在高、低脂系肉鸡腹部脂肪组织中的表达水平主要与其启动子R2区域的DNA甲基化水平有关。     

Polymorphism of P53‐Ets/AP1 transactivation region of MDM2 oncogene and its immunohistochemical analysis in canine tumours

Mouse Double Minute‐2 (MDM2) is an ubiquitin ligase which is overexpressed or its promoter polymorphism has been reported in different tumours. The objective of this study was to examine the MDM2 protein expression and its promoter polymorphism in some canine tumours. Twenty specimens were collected from 20 dogs with 15 mammary gland carcinomas, 3 lymphomas, 1 transmissible venereal tumour and 1 trichoblastoma. Samples were analysed immunohistochemically using human antibody against MDM2 protein. PCR and DNA sequencing were carried out to identify MDM2 promoter polymorphism. MDM2 gene was expressed in 13 of 20 samples including 11 mammary carcinomas, 1 lymphoma and 1 trichoblastoma. We found 94% homology between canine and human sequences. Four mutations including G169C, A177G, G291T and A177G were identified in different types of breast carcinomas. An extra p53 response element was found in a mixed mammary carcinoma.