Sex-linked SSR markers in hemp

Hemp is a dioecious plant with sex chromosomes X and Y, the male sex being heterogametic. The quality of the fibre depends on the sex type. The sex chromosomes can be characterized by molecular markers. In this report, sex‐linked simple sequence repeat (SSR) markers are described. One SSR marker was polymorphic in both the populations derived from single crosses, two other markers in but one of the two populations. Three alleles were detected for two SSR markers indicating polymorphism not only between X and Y, but also between different X chromosomes. In addition, several sex‐linked RAPD markers were detected in one population. Recombination within the sex chromosomes was observed for nearly all markers.     

微型冷库复合加热循环除霜系统的研制与试验

针对冷冻冷藏库传统除霜方法高能耗、冷库温湿度波动大等缺陷,通过设计系统除霜结构,完善控制策略,研发了一种智能除霜方法。复合加热循环除霜采用时间—压差联合智能控制策略界定最佳除霜点,配备排管辅助制冷技术,基于欧姆龙PLC高精度控制系统使各设备协同完成除霜过程。对复合加热循环除霜系统进行试验研究,对比分析不同除霜方法的工作性能及能耗情况。结果显示:相比于传统电加热除霜,复合加热循环除霜过程冷库温度波动减小3.8℃,相对湿度波动降低21.1百分点,除霜时间缩短14 min,系统除霜节能率达34.5%。     

表达H5亚型禽流感病毒HA基因的重组马立克氏病病毒的构建

采用聚合酶链反应或反转录聚合酶链反应扩增出H5亚型禽流感病毒(AIV)的HA基因、网状内皮增生症病毒的长末端重复序列(LTR)、马立克氏病病毒(MDV)Rispens CVI988毒株基因组的sorf 1和sorf 2序列、两端带loxp位点的lac/smGFP标志基因,构建含这些基因的转移载体质粒pMHA;以MDV Rispens CVI988毒株的基因组DNA和PMHA质粒DNA共转染鸡胚成纤维细胞(CEF),采用同源重组方法将LTR、lac/smGFP和HA基因插入到MDV基因组,获得重组病毒rMDV-HA/GFP;以cre介导的同源重组去除lac/smGFP标志基因,再转染CEF,获得仅带LTR启动子和HA基因的重组MDV疫苗毒株rMDV-HA.rMDV-HA仍保留了MDV RispensCVI988疫苗毒株的复制特点,并能稳定表达AIV的HA.     

哺乳动物卵母细胞非整倍体产生机制的研究进展

雌性生殖细胞进行减数分裂时易发生染色体分离错误而产生非整倍体卵母细胞,其受精后会产生非整倍体胚胎,导致出生缺陷或胚胎致死,是影响哺乳动物繁殖的重要因素。卵母细胞在第一次减数分裂前期发生同源染色体联会,此时DNA双链断裂引发重组。重组时缺乏交叉、重组事件数量的减少及交叉靠近端粒或着丝粒导致染色体发生同向分离或不分离,从而产生非整倍体卵母细胞。减数分裂期间,当染色体的端粒共向于同一极或没有完全附着在纺锤体微管上时,纺锤体组装检查点（spindle assembly checkpoint,SAC）被激活,E3泛素连接酶APC/Cyclome （APC/C）沉默,保护分离酶抑制蛋白（securin）和细胞周期蛋白B （cyclin B）不被降解,从而抑制分离酶和染色体的分离。直到所有染色体与纺锤体实现稳定的双极定向并正确排列到赤道板上,SAC关闭,染色体正确分离。卵母细胞中SAC蛋白缺失,导致SAC不能有效地监测端粒在纺锤体上的正确附着,发生染色体分离错误,从而产生非整倍体卵母细胞。因此,通过现代分子技术手段解析非整倍体卵母细胞所涉及的机制是保护哺乳动物生育的重要目标。作者主要介绍了卵母细胞减数分裂的特点,详细阐述了卵母细胞非整倍体发生的染色体分离错误的分子机制,以期为开发卵母细胞非整倍体的治疗手段提供参考。     

Genetic Diversity and Global Distribution of Citrus tristeza virus （CTV） Strains

Citrus tristeza virus （CTV）, the most devastating viral pathogen in citrus, causes tremendous economic losses to citrus industry worldwide. The CTV isolates exhibit variable pathogenicities on their ho...     

人IL-1Ra基因的克隆及原核表达的研究

IL-1Ra是第一个被发现的天然存在的细胞因子拮抗剂,结合于IL-1受体后不引起IL-1效应。文章通过RT-PCR从人的胎盘组织中克隆了人IL-1Ra基因,使其在大肠杆菌DH5α中重组表达。结果表明,克隆到了460bp左右的目的基因,并在大肠杆菌中成功表达了18ku左右的目的蛋白,目的蛋白在大肠杆菌中表达量占总蛋白20%左右,为IL-1Ra的进一步研究及应用奠定了基础。     

猪源隐孢子虫卵囊的分离及基因型鉴定

应用RT-PCR方法对南京、上海和合肥猪源隐孢子虫卵囊SSU rRNA部分序列进行扩增,产物测序后提交GenBank,收录号为DQ855266、DQ855267;用BLAST和DNAStar软件与GenBank参考序列进行比较,分析其同源性,绘制系统发育进化树,结合卵囊形态学观察和对小鼠、大鼠、兔、山羊和鸡的传染性试验确定隐孢子虫种类或基因型。结果表明,3地区猪源隐孢子虫分离株与微小隐孢子虫(C.parvum)同源性达94%~100%,与C.parvummouse型有99.8%~100%的同源性,并处于进化树的同一分支。因此,3地区猪源隐孢子虫是C.parvummouse型,提示猪和鼠之间存在交叉传播的可能。     

布鲁氏菌M5-90△WboA基因缺失株的构建及免疫效果的初步评价

为获得毒力较弱并能区分自然感染和疫苗免疫的布鲁氏菌候选疫苗株,本研究用PCR方法扩增WboA基因的上下游同源臂序列,构建重组质粒pGEM-7zf-△WboA-Sac,电转化布鲁氏菌M5-90感受态细胞,筛选布鲁氏菌疫苗株M5-90的WboA基因缺失株,并对获得的M5-90△WboA遗传稳定性、毒力、免疫保护性、抗体水平等指标进行检测.实验结果表明M5-90△WboA株的毒力比M5-90株明显减弱,差异极显著(p＜0.01),体液免疫和细胞免疫结果表明M5-90△WboA株与亲本M5-90株相比差异不显著(p＜0.05),M5-90△WboA株和亲本株的保护率分别为10％和20％,表明M5-90△WboA株与M5-90株具有相似的保护性.凝集试验和western blot试验显示M5-90△WboA株免疫小鼠的血清反应结果为阴性.本研究构建的布鲁氏菌基因缺失株M5-90△WboA具有较好的遗传稳定性,毒力比亲本株更弱,免疫保护性与亲本株相当,并能以血清学检测方法区分野毒株感染和缺失疫苗株免疫的动物.     

Genomic structure of androgenic progeny of pentaploid hybrids, Festuca arundinacea × Lolium multiflorum

To assess the usefulness of the doubled haploid (DH) method in the breeding of forage grasses, a sample of anther-derived progeny of pentaploid F₁ hybrids of Festuca arundinacea × Lolium multifiorum was karyotyped using genomic in situ hybridization (GISH). The technique allowed scoring of the total number of chromosomes, the number of chromosomes contributed by each parent, and the number and positions of the Festuca-Lolium translocation breakpoints. Among 27 plants analysed, 13 belonged to three clones, effectively reducing the number of different progeny karyotyped to 17. These included 10 haploids, five doubled haploids and two plants for which the origins could not be explained. In all plants analysed, a mixture of chromosomes of both parents was present, including an average of 1.88 intergeneric translocations per plant. The translocation breakpoints were distributed along almost the entire length of the chromosome arms. Chromosome variation among androgenic progeny appeared much wider than that in the conventional backcross but low vigour and high mortality suggest that this additional variation may be difficult to exploit directly in breeding. However, a change in the pattern of recombination makes the entire genome accessible to manipulation.     

Development and Identification of Triticum aestivum L.-Thinopyrum bessarabicum L(o)ve Chromosome Translocations

With ass7istance of chromosome C-banding and genomic in situ hybridization(GISH)combined with meiotic analysis,five germplasms with homozygous wheat-Th. Bessarabicum chromosome translocations were developed and identified among BC1F5 progenies of the cross between T. Aestivum cv. Chinese Spring and Chinese Spring-Th. Bessarabicum amphiploid. These lines included Tj01 and Tj02(2n=44)containing a pair of wheat-Th. Bessarabicum translocation chromosomes besides a pair of added Th. Bessarabicum chromosomes,Tj03(2n=44)with a pair of added interspecific translocation chromosomes,Tj04(2n=44)containing a pair of interspecific translocation chromosomes besides an added pair of Th. Bessarabicum chromosome arms and Tj05(2n=46)containing a pair of interspecific translocation chromosomes besides two pairs of added intact alien chromosomes. The breakpoints of all the translocations were found to be not around centromere. Meanwhile,all the lines showed normal plant growth,development and fertility,while the translocation chromosomes transmitted regularly. The obtained translocations might be of use for transferring elite genes from Th. Bessarabicum into wheat.