Cre/loxP定位重组系统在植物雄不育和杂种优势中的利用研究

Cre—loxP是来源于噬菌体P1的一种位点特异性重组系统。目前，它已广泛应用于基因功能鉴定，动植物及微生物基因组的修饰以及基因的表达和调控。雄性不育系在杂种优势的利用中具有重要意义。本实验是以Cre—loxP系统调控细胞毒素基因barnase在植物雄性器官发育的特异时期在特异组织表达，从而达到控制植物育性的目的。1．在本实验中，通过PCR方法克隆了解淀粉芽孢杆菌Bacillus amyloliquefaciens的barnase基因，及其抑制因子barstar的DNA序列，同时，构建成功双元表达载体及基因枪转化载体，并已获得经分子险测呈阳性的小麦植株。2．osg6B是来源于水稻花药绒毡层的特异性的启动子，它在小孢子发育的四分体时期至单核花粉期在花药绒毡层中特异启动基因的表达，它具有较为严紧的时空特异性。以osg6B为启动子，构建了一组受控于Cre／loxP位点特异性重组系统的适用于基因枪法转化小麦的载体系统。其中，转不育基因(og6B—barnase)小麦植株生长正常，但开花后，花粉量明显减少，自交不能得到正常种子。进行花粉活力检测发现，转基因小麦大部分花粉表现为败育。花粉离体萌发试验表明，转不育基因小麦的花粉离体萌发率极低，几近败育。说明以osg6B驱动barnase在小麦花药中的表达可以导致雄性不育。3．以osg6B为启动子，构建了一组受控于Cre／loxP的用于控制双子叶植物育性的双元表达载体系统。以转cre基因的烟草作为受体，以含阻断序列的质粒农杆菌进行二次转化。二次转化株营养生长正常，但开花期花器官表现异常。具体表现为花瓣异常开裂或不开裂，花丝变短或粘连，花药提早萎蔫或异常开裂。推测这种现象的出现与启动子的组织特异性或barnase的渗露表达有关。4．二次转化株花粉染色因不同株系出现全部败育、部分不育和多数可育的情况。花粉离体萌发实验也得到了相似的结果。这说明通过筛选，利用Cre／loxP得到完全不育的植物株系是完全可行的。5．获得了9个barnase的突变体，初步推测第十四位氨基酸及第八十位氨基酸与核糖核酸酶活性有关。同时，发现barnase的第316位核苷酸序列为易突变位点，自然发生的突变多插入一个C，从而产生终止位点的改变。这或许会为今后barnase的克隆与利用提供一些参考。     

植被退化对土壤性质的影响及防治对策

植被和土壤在生态系统中相互影响、相互作用。由于植被老化、气候变化、载畜量过大、人类活动等因素的影响,破坏了植被的生存与发展,土壤性质也随之发生了变化并趋于恶化。分析了植被退化的概念、原因及其对土壤性质的影响,并提出了防治对策。     

A slowly progressive retinopathy in the Shetland Sheepdog

Objective To describe a slowly progressive retinopathy (SPR) in Shetland Sheepdogs. Animals Forty adult Shetlands Sheepdogs with ophthalmoscopic signs of SPR and six normal Shetland Sheepdogs were included in the study. Procedure Ophthalmic examination including slit‐lamp biomicroscopy and ophthalmoscopy was performed in all dogs. Electroretinograms and obstacle course‐test were performed in 13 affected and 6 normal dogs. The SPR dogs were subdivided into two groups according to their dark‐adapted b‐wave amplitudes. SPR1‐dogs had ophthalmoscopic signs of SPR, but normal dark‐adapted b‐wave amplitudes. Dogs with both ophthalmoscopic signs and subnormal, dark‐adapted b‐wave amplitudes were assigned to group SPR2. Eyes from two SPR2 dogs were obtained for microscopic examination. Results The ophthalmoscopic changes included bilateral, symmetrical, greyish discoloration in the peripheral tapetal fundus with normal or marginally attenuated vessels. Repeated examination showed that the ophthalmoscopic changes slowly spread across the central parts of the tapetal fundus, but did not progress to obvious neuroretinal thinning presenting as tapetal hyper‐reflectivity. The dogs did not appear seriously visually impaired. SPR2 showed significantly reduced b‐wave amplitudes throughout dark‐adaptation. Microscopy showed thinning of the outer nuclear layer and abnormal appearance of rod and cone outer segments. Testing for the progressive rod–cone degeneration ( prcd )‐mutation in three dogs with SPR was negative. Conclusion Slowly progressive retinopathy is a generalized rod–cone degeneration that on ophthalmoscopy looks similar to early stages of progressive retinal atrophy. The ophthalmoscopic findings are slowly progressive without tapetal hyper‐reflectivity. Visual impairment is not obvious and the electroretinogram is more subtly altered than in progressive retinal atrophy. The etiology remains unclear. SPR is not caused by the prcd‐mutation.     

粳型三系不育系混杂退化原因及对策的探讨

从理论与实践分析BT型、滇型粳稻不育系具有高度的遗传稳定性,发生混杂退化的主要原因在于:①繁殖过程中保持系的机械混杂;②杂交稻或恢复基因的迁移、扩散;③一般常规品种不育基因迁移及其连锁反应.针对BT型不育系混杂退化原因,提出“人工回交株系循环法“不育系原种繁种程序.     

水稻(Oryza sativa L.)正常和无花粉花药绒毡层解体过程孢粉素行为的比较观察(英文)

 利用派罗宁B-黄色组分诱导孢粉素产生荧光的特异性,观察了水稻M 101品种以及它的无花粉型胞核雄性不育(GMS)突变体Ⅰ-15系花药绒毡层解体过程中孢粉素的行为。M-101品种花药绒毡层的解体过程大体可分为两个阶段:在小孢子发育期,绒毡层主要分泌孢粉素;在花粉发育期,绒毡层主要降解非孢粉素物质。Ⅰ-15系花药绒毡层的解体过程虽然出现了一系列异常现象,但绒毡层解体的起始时间和阶段顺序性与M-101品种相似。对绒毡层解体过程孢粉素行为的有关事态及其与花粉发育的关系等问题进行了讨论。     

F1代西瓜亲本选择对防止退化的效应

1989～1993年对F1代西瓜亲本在相同条件下进行了连继世代选择和不加选择的加代繁殖两种处理，结果表明：F1代亲本的连续选择能不断提高株系的亲和力，产量比未加选择的增产25．88％。经选择后，母本x－4株系可溶性固形物从10．11％增至11．43％，增长13．06％．父本N－2株系的可溶性固形物含量从7．3％增至8．8％，增长21．92％。F1代炭疽病的发病率和病情指数从89．44％，33．23下降到61．17％和11．63。F1代西瓜亲本连续选择可增加商品率和品质，防止亲本连年自交衰竭和F1代种子退化，并可延长双亲和F1的使用年限，增加单位面积产量，增强抗病性。     

COMPARATIVE EVALUATION OF HYDROPIC DEGENERATION OF THE LIVER IN DAIRY CATTLE THROUGH BIOCHEMISTRY, ULTRASONOGRAPHY AND DIGITAL ANALYSIS

Ultrasonography of the liver of 181 Holstein-Friesian cows was performed and blood samples were collected for analysis. The hepatic ultrasonograms were evaluated and the echoes were analyzed digitally. After slaughter, liver specimens were taken and examined histopathologically. Of the 181 animals, 120 had a normal liver and 61 had hydropic degeneration of the liver, diagnosed through histopathologic examination. Diagnostic accuracy rates for hydropic degeneration were determined based on the following test positive conditions: a) for biochemical analysis—high levels of aspartate aminotransferase, alanine aminotransferase, total bilirubin and non-esterified fatty acids; b) for ultrasonography—presence of dark pattern and blurring of edges; and c) for digital analysis—low echo means at 1 cm and 3 cm from the hepatic surface. Digital analysis had the highest overall specificity, accuracy and positive predictive values for hydropic degeneration, followed by ultrasonography. The results suggest that ultrasonography and digital analysis of hepatic ultrasonograms can be used for diagnosis of hydropic degeneration of the liver in place of biochemical analysis.     

Photoreceptor cell dysplasia in two Tippler pigeons

Two 12-week-old Tippler pigeons were evaluated for ocular abnormalities associated with congenital blindness. The pigeons were emaciated and blind. Biomicroscopy and direct and indirect ophthalmoscopy findings of the Tippler pigeons were normal with the exception of partially dilated pupils at rest. Scotopic (blue stimuli) and photopic monocular electroretinograms were extinguished in the blind Tippler pigeons. Histological and electron microscopy studies revealed reduced numbers of rods and cones, and an absence of the double cone complex. The photoreceptor cells' outer segments were absent, and the inner segments were short and broad. The number of cell nuclei in the outer and inner nuclear layers was decreased, and the internal and external plexiform layers were reduced in width. Photoreceptor cell endfeet with developing synaptic ribbons were present in the external plexiform layer. Inflammatory cell and subretinal debris was not seen. The electroretinographic, histopathological, and ultrastructural findings of the blind Tippler pigeons support the diagnosis of a photoreceptor cell dysplasia.     

烟叶成熟过程相关基因变化及其叶绿素降解

烤烟叶片成熟过程与衰老是烟草发育的重要阶段,是一个复杂和高度调控的过程,涉及衰老相关基因的调控表达和叶绿体的分解,也是烟草质量和品质形成的关键时期,因此其成为烟草生产和研究的重要时段。介绍了烟叶衰老过程相关基因的表达变化和叶绿素分解的过程,为从分子水平定义烟叶的成熟度提供理论依据。     

生产用金针菇菌种退化的快速判断

取发生不同程度菌种退化的相同来源的金针菇(Flammulina velutipes)工厂化袋栽菌株和栽培菌包,观察其在PDA平板上的分纯情况和形态特征;采用LBL液体培养基检测不同菌株对培养基的脱色情况。实验结果表明:这些退化菌株经过分离可以得到不同菌落形态的分离菌株,而且对LBL液体培养基表现不同的脱色能力,退化严重的菌株脱色能力最差,说明LBL液体培养基的脱色反应结合菌种纯度的检测可以在一定程度上排除生产上的退化菌株。