家蚕AFLP连锁框架图谱的构建

利用改进的AFLP分子标记方法,对家蚕Bombyx mori回交一代BC1群体进行连锁图谱的构建。经17组AFLP引物的选择性扩增,共得到430个多态位点,卡方检验后有253个为有效位点。利用Mapmaker/Exp (Version 3.0b)软件作连锁分析,其中163个标记分属28个连锁群,连锁群标记数变化范围是2～28个,平均每个连锁群标记数为5.8个,该图覆盖的基因组长度为2.998.9cM(图距单位),连锁群长度变化范围为4.5～652.8 cM,连锁群的平均长度为107.1 cM,平均图距为4.5～36.7 cM。     

改性聚二甲基硅氧烷 (iPDMS) 蛋白芯片在肿瘤相关抗原自身抗体筛查中的应用

肿瘤标记的快速筛查是临床早期诊断的难题。利用化学发光蛋白质芯片技术,对低丰度的肿瘤相关抗原的自身抗体进行高灵敏度的筛选,是一种有益的尝试。本研究首先将带烯烃末端的、引发聚合反应的表面引发剂加入到常规聚二甲基硅氧烷材料中,再通过热交联反应固定到聚二甲基硅氧烷的三维结构中,形成改性聚二甲基硅氧烷 (iPDMS)。为了使iPDMS材料具有抗蛋白质非特异性吸附的特性,在活性引发位点处通过表面引发的原子转移自由基聚合反应合成poly(OEGMA) 高分子刷。最后将20种肿瘤相关的抗原利用高通量喷点打印技术打印到芯片的特定区域,并组装成iPDMS芯片的48孔检测微孔板。对临床上常见的8种肿瘤患者血清进行分析,发现VEGFR1和VEGF121自身抗体对常见的8种肿瘤具有检测价值,有望成为肿瘤快速筛查的检测指标。     

红鲫C1HD近交系RAPD标记的建立

目的建立红鲫C1HD近交系的RAPD标记。方法从80条随机引物中筛选出20条扩增效果和多态性较好的引物,对8尾红鲫C1HD近交系和8尾普通红鲫基因组DNA进行RAPD扩增。结果S333引物扩增出一条特异性条带,大小约为2.1 kb。结论S333引物扩增出的特异性条带可以作为区分普通红鲫与红鲫C1HD近交系的分子遗传标记。     

Development of nuclear microsatellite markers for the threatened wetland plant Geranium soboliferum var. kiusianum (Geraniaceae)

Nuclear microsatellite markers were developed for the threatened plant Geranium soboliferum var. kiusianum, which has decreased its population size as a result of loss of its wetland habitat in Kyushu, Japan. Utilizing RNA‐seq data obtained by next‐generation sequencing techniques, 10 polymorphic microsatellite markers with 3–16 alleles in a nuclear genome were developed and characterized. Two to 15 alleles were observed in G. soboliferum. These markers will be used to investigate the genetic circumstance of remnant populations of G. soboliferum var. kiusianum and their phylogenetic relationship with G. soboliferum.     

芽孢杆菌TS02特异RAPD标记的SCAR标记转化和验证

利用自主分离的蜡状芽孢杆菌菌株TS02,采用RAPD 方法对TS02及其同源性相近的5株芽孢杆菌(地衣芽孢杆菌、枯草芽孢杆菌、凝结芽孢杆菌、巨大芽孢杆菌、短小芽孢杆菌)进行了RAPD条带特异性分析,从TS02基因组中筛选获得了一个533 bp的特异RAPD标记TSR1.TSR1克隆、测序后,根据其序列设计出一对特异引物P1/P2进行扩增,结果只在TS02中扩增得到目的片段,而其余对照菌株扩增为阴性,从而证明试验得到了在种水平上对该菌种进行准确鉴定的特异SCAR标记.     

The high level of wide-compatibility of variety ‘Dular’ has a complex genetic basis

Wide-compatibility varieties (WCVs) are a special class of rice germplasm that is able to produce fertile hybrids when crossed to both indica and japonica rice varieties. WCVs may differ greatly in their spectrum and level of compatibility. The objective of this study was to determine the genetic basis of wide-compatibility conferred by ‘Dular’, a landrace variety from India that has demonstrated a high level of wide-compatibility in previous studies with a broad range of indica and japonica varieties. A three-way cross (‘Balilla/Dular//Nanjing 11’) was made and the resulting F₁ population evaluated in the field for spikelet fertility. A total of 235 plants from this population was assayed individually for restriction fragment length polymorphisms (RFLPs) at 159 marker loci covering the entire rice genome at regular intervals. Quantitative trait locus (QTL) analysis identified 5 loci, located on chromosomes 1, 3, 5, 6 and 8, as having significant effects on hybrid fertility, which jointly explained 55.5% of the fertility variation in this population. The QTL on chromosome 5 ( f5) showed the largest effect on hybrid fertility, followed by those on chromosomes 6 ( f6), 3 ( f3) and 1 ( f1), with the one on chromosome 8 ( f8) having the smallest effect. Genotypes each composed of an allele from ‘Dular’ and an allele from ‘Nanjing 11’ at four ( f3, f5, f6 and f8) of the five QTLs contributed to the increase of fertility in the population. In contrast, the genotype composed of alleles from ‘Balilla’ and ‘Nanjing 11’ at the fifth locus ( f1) was in the direction of increasing fertility. Analysis of variance using marker genotypes at the five QTLs as the groups detected two interactions involving four of the five loci, a 2-locus interaction between f5 and f8 and a 3-locus interaction among f3, f5 and f6. The level of hybrid fertility is the result of complex interactions among these loci. The implication of the present findings in the utilization of the wide-compatibility of ‘Dular’ in rice breeding programs is also discussed. Received: 21 October 1997 / Accepted: 30 December 1997     

AFLP在分子生物学研究中的应用

扩增片段长度多态性（AFLP）可靠性强,多态检出率高,因而被认为是最有效的DNA指纹分析技术。AFLP已广泛应用于分类学、病理学、种群遗传学、DNA指纹分析的研究和建立数量性状基因图谱,成为最主要的遗传标记。介绍了AFLP的原理、影响因素及其在分子生物学研究中的应用。     

玉米转基因方法研究进展

玉米是重要的粮食作物,玉米的遗传转化取得了重大成就,而玉米的遗传转化又是玉米遗传改良的有效手段。总结了玉米转化过程中受体的选择标准,比较了各种转化方式,如：农杆菌介导法、基因枪法、花粉管通道法,并探讨了如何有效地选择转化后的阳性事件。     

苗期标记性状在我国作物育种及种子纯度鉴定上的研究应用进展

利用苗期标记性状进行作物杂交新品种培育和种子纯度鉴定的技术方法,具有直观准确、简便快速、成本低廉的特点,优越于异地种植、同工酶电泳和DNA分子标记的鉴定方法.本文综述了这种方法应用的技术方法、材料来源、研究内容、应用的类型和成果,指出了存在的问题,对其应用前景进行了展望.     

合浦珠母贝三个养殖群体微卫星遗传多样性

以广东徐闻金碧公司养殖场、广西北海营盘镇养殖场和南海水产研究所海南实验基地3个合浦珠母贝养殖群体为对象,利用8个微卫星位点M1、M2、M3、M4、M5、M6、M7、M8的引物进行了遗传多样性分析.结果表明:8个微卫星标记位点在3个养殖群体中共检测到58个等位基因,观测等位基因数为2～9个,平均有效等位基因数3.72～5.06,平均观察杂合度0.41～0.56,平均期望杂合度0.67～0.75,3个群体的平均多态信息含量PIC值为0.62～0.70,全部为高度多态(PIC≥0.5),表明这几个合浦珠母贝养殖群体目前仍具有较高的遗传多样性,遗传信息丰富,遗传变异大,可以作为良好的育种材料;在这3个养殖群体中,南海水产研究所海南实验基地的养殖群体的遗传多样性最高,广西北海营盘镇养殖群体遗传多样性最低,这一结果可以为今后选择育种、种质保护提供可资借鉴的资料.

Abstract：

By using eight mierosatellite loci (M1, M2, M3, M4, M5, M6, M7 and MS), the genetic diversity of three Pinctada fucata populations from the pearl farms in Xuwen of Guang-dong and Beihai of Guangxi, and from the experimental base of South China Sea Fisheries Re-search Institute in Hainan was studied. A total of fifty eight alleles of these eight microsatellite lo-ci were detected, among which, the observed allele number was 2-9, average effective allele number was 3.72-5.06, average observed population heterozygosity was 0. 41-0. 56, and aver-age observed expected heterozygosity was 0. 67-0. 75. All the three populations had a polymor-phie information content (PIC) of 0. 62-0.70, suggesting their high polymorphism (PIC > 0. 5). Among the three populations, the cultured population from the experimental base of South China Sea Fisheries Institute had the highest polymorphism, and that from Beihai of Guangxi had the lowest one. These results provided useful information for the selective breeding and germplast conservation of P. Fucata.