绵羊BMP15基因B2突变可视化检测技术的建立

骨形态发生蛋白15（bone morphogenetic protein 15,BMP15）基因突变对绵羊排卵率、产仔数以及繁殖力有很大影响,本研究旨在建立快速可视化检测BMP15基因B2突变的技术。将改良的扩增阻滞突变系统（amplification refractory mutation system,ARMS）与核酸染料SYBR Green Ⅰ结合,针对BMP15基因B2突变设计ARMS特异性引物,使引物下游3'端的最后一个碱基为A,并在下游引物3'端的第3位碱基处设计额外的错配,以提高引物的特异性;经ARMS PCR扩增后,向产物中加入SYBR Green Ⅰ,通过肉眼观察PCR管内的颜色变化来检测BMP15基因的B2突变。经测序发现所采集的样本均为野生型,因此,利用重叠延伸PCR构建BMP15基因B2突变型模板,测序结果显示BMP15基因B2突变型模板构建成功。ARMS PCR结果显示,ARMS特异性引物能够只扩增突变型模板,而不会扩增野生型模板,且扩增效果良好。ARMS PCR扩增后加入SYBR Green Ⅰ发现已知野生型模板与阴性对照一致,呈SYBR Green Ⅰ原始颜色橙黄色,而已知突变型模板呈亮绿色,且色差明显,肉眼可辨。用建立的可视化检测BMP15基因B2突变的技术检测50个小尾寒羊基因组DNA （随机向其中20个样本中加入构建的BMP15基因B2突变型模板）,将可视化检测的突变型样本编号与混合样本时记录的编号对比,结果表明该技术能够准确检测绵羊BMP15基因的B2突变,且准确率高达100%。将构建的BMP15基因B2突变型的模板进行梯度稀释,对本试验可视化检测体系的灵敏度进行检测,发现ARMS可视化检测技术的灵敏度达到0.006 ng/μL。因此,本研究建立的技术能够可视化地检测绵羊BMP15基因B2突变,且准确率高,有望为高繁殖力绵羊的选育提供技术支持。     

紫花苜蓿根腐新病原-厚垣镰孢菌生物学特性研究

针对紫花苜蓿根腐致病菌厚垣镰孢菌(Fusarium chlamydosporum)在不同温度、光照、p H和多种碳氮源条件下的生物学特性,采用菌饼法进行了测定,并对不同碳氮源下菌落产色素情况进行了连续观察。结果表明:该病原菌在5~40℃均可生长,最适温度为25℃;当p H 4.0~12.0时均能生长,最适宜p H为7.0;全黑暗条件有利于菌丝生长,菌丝生长最适碳源、氮源分别为山梨醇和蛋白胨,碳源氯醛糖及除蛋白胨以外8种氮源均不利于菌丝的生长。含不同碳氮源培养基可诱导菌丝产生不同色素,且颜色变化差异大,在不同的生长阶段,色素颜色也会发生变化。     

黑豆色素的研究进展

本文综述了近年来黑豆色素的研究进展.提出了综合开发利用的前景.     

大果沙棘杂交后代果实主要成分遗传变异倾向研究

本文以黑龙江省绥棱浆果研究所选育的"楚伊×蒙优1"、"楚伊×蒙优2"、"楚伊×蒙优3"、"楚伊×蒙优4"、"HS-21×蒙优1"、"HS-22×阿列伊"、"绥1×蒙优7"和"HS-20×阿列伊"8个大果沙棘杂交组合为研究对象,对杂交后代果实中糖、酸和黄色素成分的遗传变异倾向进行了初步研究.结果表明:以蒙古沙棘系列雄株为父本的杂交组合后代果实中糖含量呈低亲变异的趋向,以阿列伊为父本的杂交组合后代果实中糖含量在一定程度上呈高亲变异倾向.阿列伊作为父本优于蒙古沙棘系列雄株."高糖"母本楚伊杂交组合的F1代株系果实中糖含量均呈现出衰退趋势.8个沙棘杂交组合后代果实中酸含量呈现一定程度的高亲变异."HS-21×蒙优1"杂交组合后代果实中酸含量变异广泛.8个沙棘杂交组合后代果实中黄色素含量一定程度上亦呈高亲变异倾向.在"楚伊×蒙优4"杂交组合后代中进行选择,获得超亲株系的几率最大.     

蓝靛果忍冬色素提取工艺的研究

以蓝靛果忍冬果实为原料,进行了天然色素提取工艺研究。结果表明:蓝靛果忍冬色素最佳的提取工艺条件为:提取剂采用无水乙醇效果较好,用量为35mL;提取pH 3、最适合的提取温度为80℃、60min为最佳的提取时间。     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

“荧光”蚕品种雌雄荧光色素的定性及相对量分析

用高效液相色谱分析和纸上色层分析法 ,对家蚕“荧光”品种雌雄幼虫和茧层的荧光色素进行了定性和相对量分析。结果表明 :构成雌雄茧层荧光色素的种类无显著差异 ,雌雄茧荧光色不同的原因是组成茧荧光色的各种荧光色素相对量不同 ;茧层中的一种Rf值为 0 77的黄荧光色素 ,在熟蚕丝腺中没有检出 ,说明营茧过程中有新的荧光物质形成或有较大的变化 ;5龄幼虫不同发育时期 ,雌雄各荧光色素的相对量不同 ,以荧光黄色素为例 ,雄性随日龄的增加其相对量不断增加 ,而雌性则以 5龄 96h为最高 ,其后逐渐下降 ;雌性幼虫肠壁组织中的Rf值为0 19和 0 5 9的两种明黄色荧光色素 ,雄性幼虫和雌性幼虫的其它组织中没有检出 ,说明雌雄幼虫在色素代谢上也存在差异     

Genomic and Pathogenic Studies on a Glycoprotein E Variant Field Isolate of Bovine Herpesvirus 1

Glycoprotein E-negative (gE–) laboratory strains of bovine herpesvirus 1 (BHV-1) were recently introduced as novel marker vaccines, allowing serological discrimination between vaccinated and naturally infected animals on the basis of lack or presence of antibodies against gE epitopes. The applicability of this approach is based on the genetic stability of the gE. However, mutant field variants of BHV-1 with a variable response in anti-gE ELISA have been isolated. The molecular characterization of a gE variant field isolate (Salwa strain) is presented here. By comparing the gE nucleotide and amino acid sequences of the Salwa strain with those of the wild strain Jura, ten mutated bases were found in the gE strain of Salwa, six of which alter the amino acid sequence, leading to changes in five amino acids. Both strains caused respiratory disease in experimentally infected calves, but Salwa generated slightly milder signs. Both viruses were excreted in nasal and ocular discharges, and were reactivated by dexamethasone treatment. In conclusion, the rather close similarities observed in the gE gene structure and pathogenicity features of the gE mutant and of the wild strain of BHV-1 confirm the genetic stability of gE. The findings indicate that the Salwa isolate is virulent, but less virulent than wild strains. Our data support the use of gE-negative marker vaccines in eradication programmes.     

产单核细胞李氏杆菌突变株的感染动力学研究

以BALB/c小鼠为实验动物模型,对actA基因和plcB基因双缺失的产单核细胞李氏杆菌（LM）yzuLM1-2菌株进行感染动力学研究。分别用减毒突变株yzuLM1-2和野生型菌株yzuLM4给BALB/c小鼠静脉注射或灌服,并对脏器中LM的分布动态进行测定。结果显示,在显著高于野生型菌株感染剂量的条件下注射或灌服接种后,减毒突变株组小鼠肝和脾中的LM数量迅速下降,被清除的速度显著快于野生型菌株组。溶血素（LLO）抗体的测定结果显示,减毒突变株菌能和野生型菌同样激发较高水平的抗体。小鼠免疫保护试验结果显示,减毒突变株免疫组对同种血清型LM的攻击具有较好的免疫保护力,可达100%。结果表明,yzuLM1-2突变株侵袭力下降,致病力降低,能激发较高水平的抗体产生,可以作为预防动物李氏杆菌病的候选疫苗菌株,并为用作运送外源保护性抗原的载体提供了材料。     

毛樱桃红色素的提取及应用研究

以东北毛樱桃为原料,用酸性食用乙醇提取制得樱桃红色素,并对该色素的应用范围进行了研究,结果表明,该色素在酸性条件下对光、热和常用食品添加剂较稳定,是一种价廉易得、安全可靠、使用方便的天然植物色素.