野蚕基因克隆研究进展

野蚕是重要的吐丝类鳞翅目昆虫,在不同的功能和抗性上表现优势。本文概述了野桑蚕、柞蚕、蓖麻蚕、柳蚕及其他野蚕基因的克隆研究进展,阐明了部分野蚕基因的特性及表达情况,比对野蚕和家蚕的氨基酸序列,分析亲缘关系,以便开展目标明确的分子育种,缩短育种进程,并且对野蚕基因克隆研究发展前景进行分析和展望,作为进一步研究的参考。     

提高斑翅山鹑人工孵化率的措施

斑翅山鹑俗称"沙半鸡",是我国北方传统4大狩猎珍禽之一,被人们视为野味珍品。斑翅山鹑的肉质细嫩、味道鲜美,有生精、滋补壮阳、敛伤生肌的功效。近年来,随着我国人民生活水平的不断提高,斑翅山鹑的需求量日益增大,斑翅山鹑的人工养殖业也随之逐步发展起来。相比其他动物养殖,斑翅山     

野桑蚕抗性基因P450 cDNA片段的克隆

本实验在对野桑蚕添食NaF的基础上,提取其总RNA,利用特异性引物,通过砌RT-PCR方法,获得了野蚕P450基因的cDNA,经克隆和序列测定,得到了野蚕P450基因片段的序列,经与家蚕P450基因比较分析,两者的同源性达99％。     

重要外文学术期刊发表蚕学论文简介

<正>1 ZHOU Q Z,FU P,LI S S,ZHANG C J,YU Q Y,QIU C Z,ZHANG H B,ZHANG Z.A comparison of co-expression networks in silk gland reveals the causes of silk yield increase during silkworm domestication. Front Genet,2020,11:225.DOI:10.3389/fgene.2020.00225题目家蚕与野蚕丝腺共表达网络的比较分析揭示了家蚕驯化过程中丝产量增加的原因摘要长期驯化和选择性育种使家蚕的产丝量是其野生祖先野桑蚕产丝量的几倍。然而,人们对驯化过程中蚕丝产量增加的分子机制知之甚少。根据家蚕和野蚕丝腺功能分化的动态模式,重庆大学ZHOU等发现在丝腺发育的早期和中期,家蚕中的表达上调基因主要涉及与细胞分裂和生长相关的DNA整合、核酸结合和转运蛋白活性,这导致家蚕的后部丝腺(PSG)细胞(丝蛋白合成"工厂")显著多于野蚕。在丝腺发育的后期,家蚕中的表达上调基因在蛋白质加工和核糖体途径富集,表明在家蚕驯化过程中蛋白质合成效率得到了提高。虽然家蚕丝腺中的丝素蛋白合成增加,但丝胶蛋白的产量也同时减少。这反映了家蚕和野蚕处于不同的选择压力下。重要的是,发现家蚕丝蛋白基因共表达的网络比野蚕的大。此外,在家蚕中与丝蛋白基因共表达的基因比在野蚕中更多地受到人工选择。研究结果表明,在家蚕驯化过程中,蚕丝产量的增加与生物系统的改善有关,该系统不仅包括蛋白质合成"工厂"(PSG细胞)的扩大,还包括丝蛋白基因和丝生产相关基因(如生物能量、运输和核糖体途径基因)的高表达。     

基于微卫星标记的中国斑翅山鹑遗传多样性及遗传结构分析

斑翅山鹑(Perdix dauuricae)是具有很高经济价值的猎用禽.为了获得其群体遗传多样性及遗传变异信息,为我国斑翅山鹑进一步有效保护和合理利用提供科学依据,本研究共采用了8个微卫星标记位点对分布于我国北方的斑翅山鹑23个地理种群285个个体进行了群体遗传多样性及遗传结构分析.结果表明,我国斑翅山鹑为遗传多样性较丰富的群体,且每一群体均表现出较高的遗传多样性,其中柴达木盆地地区的种群拥有最高的遗传多样性.遗传分化分析显示组间及组内种群间均表现为遗传分化显著,遗传分化系数(FST)值表明大部分种群间表现出显著的遗传分化.系统树构建和贝叶斯聚类分析结果均显示斑翅山鹑23个地理种群被分化为明显的两个组群.另外,通过BOTTLENECK对各地理种群的分析显示斑翅山鹁种群曾经历过近期瓶颈效应.本研究结果表明,柴达木盆地地区种群、华北平原地区种群、静宁种群、张家川种群以及六盘山地区种群应予以相应关注,以保证有足够的种群大小来保持这些种群的遗传多样性.     

蓖麻蚕线粒体基因组中nd1及其侧翼tRNA基因的克隆与结构分析

按KoichiroTamura等 (1988)的方法制备出蓖麻蚕线粒体DNA ,用EcoRⅠ和XbaⅠ双酶解后 ,以pSK(+)(Ampr)为载体进行克隆 ,获得 1个 3 3kb的克隆片段。测序结果表明 ,所测序列包括部分 16SrRNA、完整的tRNA Leu ,nd1和tRNA Ser基因。其中 16SrRNA(3’端部分 )、nd1基因与家蚕具有很高的同源性。同时以nd1基因为依据构建了 17种动物的分子进化树 ,显示出蓖麻蚕与家蚕、野蚕有最近的演化关系。根据tRNA Leu和tRNA Ser基因的一级结构 ,推定了可能的二级结构 ,并与家蚕进行了比较。     

家蚕抗血液型脓病新品种"锦绣1号"通过国家畜禽遗传资源委员会审定

"十二五"期间,湖南省蚕桑科学研究所在国家蚕桑产业技术体系建设项目的支持下,与体系岗位科学家司马杨虎教授开展了长期的产学研协作,利用国家桑蚕品种改良中心长沙分中心丰富的种质资源,发明了普斑品种限性转育新方法(一种家蚕普斑品种的斑纹限性转育方法ZL201510236166 .8) ,挖掘了深色花翅基因资源(家蚕遗传育种中深色蛾翅基因的利用方法ZL201510044662 .3).     

野蚕染色体的观察分析

<正> 野蚕(Bombyx mandarina L.)染色体的观察,对研究家蚕(Bombyx mori L.)的起源、进化及家蚕品种的培育具有重要的参考价值。日本野蚕染色体已报道为n=27;中国野蚕染色体根据曾锦标报道是n=28。但中国野蚕染色体的观察均取材于长江下游(镇江、杭州、上海等地),而黄河中游野蚕的染色体数目为多少?未见报道。本文首次对黄河中游的野蚕染色体进行了观察分析,并与家蚕染色体进行了初步比较。     

中国野蚕染色体结构及其变异的研究

采用细胞生物学的电离辐射方法研究了中国野蚕染色体形态和结构变异。结果表明 :中国野蚕 2 8条染色体粗线期大小在 1～ 3 μm之间 ,而且变异范围小 ,在生物界属于小染色体类。在电离辐射作用下 ,野蚕染色体结构异常的表现主要为易位、缺失、倒位等 ,其中易位的类型为相互易位和简单易位 ;缺失的类型为中间缺失和顶端缺失。本文从染色体变异角度对蚕的起源与分化作了探讨     

野蚕卵和野蚕黑卵蜂寄生卵的空间分布型及其应用

用7种方法、11个聚集度指标,测定分析了越冬代和发生代野蚕卵块、卵粒、野蚕黑卵蜂寄生卵块、卵粒的空间分布型。结果,越冬代和发生代野蚕卵块均符合所拟合的Poisson分布,Neyman A型分市及负二项分布;聚集度指标表明:越冬代野蚕卵块、卵粒、被寄生卵块、卵粒呈聚集分布,发生代则以均匀分布为主。同时还考查了卵块在桑树各部位的分布,并确定了资料代换公式和最适抽样数,制定了发生代野蚕卵块的序贯抽样表。     

Single nucleotide polymorphism in avian uncoupling protein gene is associated with thermoregulation in chicks

Avian uncoupling protein (av-UCP) is a key protein for thermoregulation in poultry. A single nucleotide polymorphism (SNP) in the av-UCP gene has been reported in chickens. The purpose of the current study was to clarify the association between this av-UCP gene mutation and thermoregulation in chickens. Wild and mutant type chicks for the av-UCP gene SNP (g. 1270 of the av-UCP gene exon 3 with C to T substitution and amino acid substitution) were exposed to high ambient temperature. Rectal temperature, radiation temperature on the body surface, and the expression of heat dissipation behavior (wing drooping and panting) during heat exposure were measured. In addition, oxygen consumption rate in the thermoneutral zone in wild and mutant type chicks was measured. Changes in wing temperature during heat exposure in wild-type chicks were lower than those in mutants. The latency of continuous wing drooping during heat exposure in wild-type chicks was shorter than in mutant chicks. It was also found that the SNP in the av-UCP gene caused reduced oxygen consumption. These results suggest that the av-UCP gene mutation affects thermoregulation, especially heat production, in chickens.     

不同遗传背景猪群体黏病毒耐药蛋白1基因多态性分析

为探讨不同遗传背景猪黏病毒耐药蛋白1（myxovirus resistance,Mx1）基因的遗传多态性,本试验采用RT-PCR方法扩增克隆出猪Mx1基因并进行生物信息学分析,采用高分辨率熔解曲线法（HRM）对民猪、长白山野猪与民猪杂交猪（简称野杂猪）和大白猪3个群体Mx1基因外显子2区多态性进行分析。结果显示,从民猪及野杂猪中成功克隆出4条Mx1基因mRNA序列。该序列含有1个1 992 bp的完整开放阅读框（ORF）,编码663个氨基酸。比较发现4条Mx1基因序列中共含有30个SNPs位点,其中17个SNPs位点引起了氨基酸序列的变异。功能结构域分析发现,大部分氨基酸突变位点位于Mx1基因功能结构域两侧。HRM多态性分析显示,Mx1基因外显子2区DD基因型作为优势基因型在3个群体中具有最高的基因型频率,其中大白猪中DD基因型频率为100%,野杂猪为62.5%,民猪为56.2%。基因多态性分析还发现该区域存在更多的核苷酸及氨基酸突变位点,表明民猪和野杂猪Mx1基因存在更丰富的基因多态性。     

The Polymorphism Analysis of Mx1 gene in Pig Populations from Different Genetic Background

In order to explore the polymorphisms of myxovirus resistance (Mx1) gene in pig populations from different genetic background,we cloned Mx1 gene and tested the polymorphisms on exon 2 of Mx1 gene in Min pig, crossbred from wild boars in Changbai Mountain with Min pig (crossbred wild boars) and Large White pig population using high resolution melting (HRM). The results showed that four Mx1 mRNAs were cloned from Min pig and crossbred wild boars, and they had whole ORF which was 1 992 bp in length and coded 663 amino acids. In the four Mx1 mRNA, there were 30 SNPs in nucleotide and 17 mutations in amino acid. The location of SNPs displayed that almost mutations located on two sides of the function domains in Mx1 protein. The polymorphism data in HRM test showed that the DD genotype was the dominant genotype which had the highest frequency in three populations. The DD genotype frequency was 100% in Large White pig population, 62.5% in crossbred wild boars, and 56.2% in Min pig.Some novel mutations in both nucleotide and amino acid on exon 2 region were explored in polymorphism test. In conclusion, there was more polymorphism of Mx1 gene in Min pig and crossbred wild boars.     

家蚕蛾黄翅的遗传分析与黄血基因的多效作用

在保存家蚕突变基因资源过程中调查发现：黄血蚕蛾翅在羽化之初呈鲜黄色,白血蚕均不然。通过与白血蚕（非黄翅）和黄血抑制基因（I）进行杂交试验,分析黄翅的遗传规律,结合普查家蚕突变系统蛾翅色性状,证明黄翅性状与幼虫黄脚一样,为黄血基因支配。进而记录了因黄血基因的多效作用在卵、幼虫、茧、成虫各阶段表现的形态特征。     

桑突变体Cty-Ym叶片蛋白质的双向电泳及质谱分析

为探讨桑树叶色突变体CtyYm变异的分子机理,采用蛋白质组学分析技术,从突变体和对照桑中提取并分离叶片蛋白质,获得了IEF/SDSPAGE双向电泳图谱,在CBB染色条件下,检测到比较清晰的蛋白质点184个;经PDQuest软件分析,发现两者蛋白质点的相对含量有36处存在显著差异(±2倍),尤其是突变体中蛋白质点SSP2801的相对含量仅为对照桑的258%,经胶内酶切、LCESIQTOFMS分析、肽质量和碎片离子检索分析,鉴定该蛋白组分为RuBisCO大亚基,推测突变体CtyYm叶色黄化可能与RuBisCO大亚基含量减少有关。     

湖北有机猕猴桃主要虫害的生活习性及其OPM技术

猕猴桃(Actinidia deliciosa)病虫害防治是制约有机猕猴桃发展的主要问题之一。本文通过对湖北猕猴桃害虫的调查研究,发现湖北省猕猴桃主要害虫有十多种,主要包括：金龟甲、隆背花薪甲、介壳虫、斑衣腊蝉、叶蝉、茶翅蝽、鸟嘴壶夜蛾、蝙蝠蛾等;文章分别对其生活习性及其OPM技术进行了介绍。     

家蚕BmAWD基因的表达和蛋白纯化

家蚕BmAWD与家蚕翅膀发育密切相关,控制其翅膀发育的完整性及可用性。在对本实验室构建的家蚕蛹期cDNA文库测序中,我们筛选到-条编码AWD蛋白的cDNA序列,将其命名为BmAWD基因。通过生物信息学分析,发现该基因序列全长为689bp,ORF框为465bD,编码154个氨基酸残基,含有-个NDPK保守结构域。通过PCR扩增获得该基因ORF框并克隆到表达载体pGEX-5X-1上,得到重组表达质粒pGEX-5X-1-AWD,将该重组子转化大肠杆菌BL21,经PCR、酶切鉴定证明重组正确,在37℃下以IPTG诱导表达后收集菌体并裂解,SDS-PAGE分析发现在43kD左右处有一特异性蛋白条带,与预期值相符,但目的蛋白主要以沉淀即包涵体形式存在,25℃下诱导表达,SDS—PAGE分析表明目的蛋白主要存在于上清中。采用亲和层析法纯化融合蛋白GST-AWD,目的蛋白通过与层析柱上的GST磁珠结合而被纯化分离。     

Isolation and characterisation of Rhodococcus equi from submaxillary lymph nodes of wild boars (Sus scrofa)

Rhodococcus equi has been isolated from the submaxillary lymph nodes of domesticated pigs, but little is known about the presence of R. equi in wild boars. The aim of the study was the evaluation of the incidence of R. equi in wild boars and the characterisation of them. Of 482 submaxillary lymph nodes of wild boars shot in 39 settlements throughout Hungary, R. equi was isolated from 60 specimens, and plasmid types of 82 isolates were examined. The isolates were tested for the presence of 15-17-kDa (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes by polymerase chain reaction (PCR). Plasmid DNAs were isolated and analysed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. None of the 82 isolates contained vapA gene but 21 isolates (25.6%) were positive for vapB gene showing 827bp product of the expected size in the PCR amplification. Sixty-one strains (74.4%) did not contain plasmid. The 21 isolates of intermediate virulence contained virulence plasmids that were identified as types 1 (1 isolate), 5 (16 isolates), 21 (1 isolate), and three new distinct plasmid variants (1-1-1 isolate), respectively. On the basis of restriction digestion patterns of plasmid DNAs, we tentatively designated the new variants as types 25-27, respectively. The prevalence of R. equi strains of intermediate virulence among the isolates originated from the submaxillary lymph nodes of wild boars (25.6%) is very similar to those of domestic pigs (26.8%) in Hungary, and plasmid type 5 is the predominating one in both groups. This is the first report of isolation of VapB-positive R. equi from wild boars in the world.     

Orthopedic conditions that affect the avian pelvic limb.

Birds are often presented to the veterinarian with injury or disease that affects their ability to use one or both legs. Although in wild birds traumatic injuries usually involve the wing, captive birds especially falconer's birds, are usually presented with injuries to their legs. This article focuses on conditions affecting practice: birds of prey, and injured wild birds. The birds are presented with lameness or loss of use of the leg. The conditions described can be initiated by injury or by disease affecting an area other than the leg, such as the kidney or spinal cord, or they can be toxic or metabolic.     

Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil

The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15–17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.