西林水牛群体的RAPD遗传多样性分析

【目的】了解西林水牛群体的遗传变异和遗传结构,为其辅助标记育种、遗传资源保护及利用提供参考依据。【方法】从广西西林县的8个乡（镇）采集184份西林水牛血样,采用优化后的RAPD技术检测其多态性DNA,统计多态性条带数,并应用Popgene32软件对西林水牛群体的有效等位基因数、Nei氏基因多样性指数及Shannon多样性指数进行分析。【结果】从18条RAPD引物中筛选出8条扩增产物稳定、条带清晰可辨的引物,扩增出的DNA片段分子量为100-1000 bp;从184个样本中共扩增出4968个多态性条带,平均每条引物可扩增出621个多态性条带,多态性频率达60.0%-100.0%,平均为89.7%。西林水牛群体的平均有效等位基因数为1.6745,平均Nei氏基因多样度指数为0.3583,平均Shannon多样性指数为0.5510。【结论】西林水牛群体的遗传变异、遗传分化及遗传多样性均较高,但选育程度较低,育种潜力较大,有待进一步保种和开发利用。     

A gamete abortion locus detected by segregation distortion of isozyme locus Est-9 in wide crosses of rice (Oryza sativa L.)

Summary A locus for male gamete abortion in hybrids for Japonica and Indica rice was identified with the aid of marker genes Rc and Est-9 on chromosome 7. In an Indica-Japonica cross, AKAMAI 1/IR50, the Indica allele Est-9 ² was transmitted via the male gamete with a ratio of 0.29 instead of the normal 0.5, whereas no segregation distortion was observed for the Rc locus. The recombination value (p) for Est-9 and Rc was estimated to be 0.38 by a least square method after adjusting Mendelian segregation ratios with the male transmission ratios of 0.29 (Tr) for Est-9 ² and 0.71 (1-Tr) for Est-9 ¹. The recombination value (q) for the new locus for male gamete abortion, ga-11, and Est-9 was estimated to be 0.23 by using 56 F₃ lines from F₂ plants which were heterozygous for the Est-9 locus. No linkage for Rc and ga-11 was found. Therefore, the two markers and ga-11 were located in the order of ga-11-Est-9-Rc. Using the estimated recombination value (q), the male transmission rate (k) of ga-11 ^a was estimated to be 0.11 with the F₂ data and-0.07 with the F₃ line data. Thus, it was apparent that male gametes possessing ga-11 ^a were frequently aborted in the Indica-Japonica hybrid.     

大豆四向重组自交群体单株产量QTL单标记分析

以(垦丰14×垦丰15)×(黑农48×垦丰19)衍生的含160个株系的大豆四向重组自交系群体为试验材料,应用154个SSR标记鉴定个体基因型,利用单标记分析方法,对2013和2014年在哈尔滨和克山两地4个环境下的单株产量进行QTL定位分析。结果显示:共检测到46个与单株产量相关的QTL位点,主要分布在A1、A2、C1、C2、D2、D1b、L、K、B2、N、E、J、F、G和H连锁群上,遗传率为0.15%~9.37%。遗传率较高的标记位点有BARCSOYSSR_02_0607、BARCSOYSSR_03_1620、BARCSOYSSR_19-0451、Sat_153、Sat_367、Satt229和Satt529,优异等位基因型为BARCSOYSSR_02_0607(Q1Q1)、BARCSOYSSR_03_1620(Q2Q2)、BARCSOYSSR_09_0183(Q1Q1)、Satt229(Q2Q2)、Sat_367(Q3Q3)、Satt338(Q1Q1)、Satt229(Q1Q1)和Satt668(Q1Q1)。在检测的标记位点中,BARCSOYSSR_08_0966、Sat_36、Sat_153、BARCSOYSSR_02_0607和Satt529在两个环境中重复检测,表明这5个QTL可用于分子设计育种改良单株产量。     

Grain Quality as Affected by Down-regulation of Expression of Different ALK Alleles in indica Rice (Oryza sativa L.)

【Objective】Gelatinization temperature (GT) is one of the key physicochemical properties in rice quality, which is mainly regulated by ALK (SSII-3) gene. In general, there are two ALK alleles among indica cultivars. To detect their functional differentiation in indica rice,【Method】Zhenshan 97B (a high GT variety carrying ALKc allele) and Longtefu B (a low GT variety carrying ALKb allele), were used as receptors for the generation of transgenic rice with down-regulation of ALK expression by RNA interference (RNAi).【Result】Down-regulation of ALK gene significantly decreased the GT of the transgenic lines. Due to the difference of original GT between the two receptors, the GT of transgenic rice lines derived from Zhenshan 97B (a high GT variety) decreased significantly, but it is slightly decreased in transgenic plants derived from Longtefu B (a low GT variety). The differential scanning calorimetry (DSC) results showed that the initial temperature of RNAi transgenic rice was significantly lower than the corresponding control and the transgenic lines were gelatinized in advance. The peak value of GT(Tp) in RNAi rice grains was significantly lower than that of the control under Zhenshan 97B background. However, Tp of RNAi rice grains under Longtefu B background was significantly lower than the control to a lesser extent. Also, down-regulation of ALK expression had a significant effect on rice physical-chemical characteristics. An increase of apparent amylose content in RNAi transgenic plants was detected due to the decreased expression of ALK gene. Besides, the pasting properties showed that down-regulation of ALK gene had obvious effects on peak viscosity and breakdown value, improving the taste of the transgenic rice. The gel consistency was significantly different among Zhenshan 97B RNAi lines and their parents, but no difference was found in Longtefu-derived transgenic lines.【Conclusion】RNA interference to ALK allele expression had a significant effect on rice quality, especially the gelatinization characters. Down-regulated expression level of ALKc allele would cause larger variation of physical-chemical characteristics between transgenic rice and their parent than that of ALKb allele.     

A rapid real-time PCR assay for detecting columnar growth in apples (Malus × domestica)

Columnar growth in apples can have considerable positive consequences for orchard maintenance, but the lack of columnar cultivars producing fruits with desired market quality prevents their broader utilization in orchards. New crossings focused on their breeding are therefore conducted throughout the world every year. Unfortunately, the reliable discrimination between columnar growth and normal phenotype is difficult until seedlings are 2–3 years of age. However, molecular marker–assisted selection (MAS) can be used in seedlings several weeks old providing immense help in breeding programmes focused on columnar apple production. Here, we describe a rapid and reliable method for the detection of both wild-type (WT) and columnar (Co) alleles originating from ‘McIntosh Wijcik’ in a single reaction using a real-time PCR allelic discrimination approach. Our assay was tested in 130 genotypes (80 normal and 50 columnar habit) from various crossings and their parents, and in all cases, genotype corresponded to phenotype. The assay is thus suitable for laboratories interested in high-throughput MAS for columnar tree habit breeding.     

Genome‐wide association study for plasma very low‐density lipoprotein concentration in chicken

The plasma very low‐density lipoprotein (VLDL) concentration is an effective blood biochemical indicator that could be used to select lean chicken lines. In the current study, we used Genome‐wide association study (GWAS) method to detect SNPs with significant effects on plasma VLDL concentration. As a result, 38 SNPs significantly associated with plasma VLDL concentration were identified using at least one of the three mixed linear model (MLM) packages, including GRAMMAR, EMMAX and GEMMA. Nearly, all these SNPs with significant effects on plasma VLDL concentration (except Gga_rs16160897) have significantly different allele frequencies between lean and fat lines. The 1‐Mb regions surrounding these 38 SNPs were extracted, and twelve important regions were obtained after combining the overlaps. A total of 122 genes in these twelve important regions were detected. Among these genes, LRRK2, ABCD2, TLR4, E2F1, SUGP1, NCAN, KLF2 and RAB8A were identified as important genes for plasma VLDL concentration based on their basic functions. The results of this study may supply useful information to select lean chicken lines.     

等位基因特异性表达形成机制及其应用的研究进展

基因作为遗传信息的介质,决定了生物的性状表现。基因序列变异的发生是生物体适应环境变化的必然趋势。在二倍体生物中等位基因是一对控制着相对性状的基因,因此,等位基因特异性表达效应根据细胞类型和发育阶段的不同而变化,表现出不同个体之间的差异。等位基因差异表达对基因的表达起到重要的调控作用,并最终可能与生物的表型多态性联系起来。等位基因特异性表达分析已经在动物研究中得到广泛应用,有助于提高鉴定调控遗传变异的能力。对等位基因特异性表达的形成机制及其应用研究进展进行了综述,以期为进一步揭示等位基因特异性表达的遗传学机制,以及加快其在动物生产中的应用提供理论依据。     

中国荷斯坦牛TLR2基因SNPs的快速筛查及等位基因频率的估算

为了快速筛查Toll样受体2基因的SNPs,采用DNA池和直接测序法确定了荷斯坦牛TLR2基因的多个SNP突变方式,估算了各SNP的等位基因频率,并利用生物信息学方法预测突变对TLR2 mRNA和蛋白的影响。结果显示,在荷斯坦牛TLR2 基因内共发现6个核苷酸突变位点,分别为T602A、G10900C、G11047C、A11076T、C12077T、T10634G,其中T10634G为错义突变,导致第64位氨基酸由原来的谷氨酸(Gly)替换为天冬氨酸(Asp)。研究结果可为中国荷斯坦牛的抗病育种提供实验依据。     

甘蓝型油菜粒色遗传的复等位模型假说

通过分析GH06×94005和GH06×A800两组合各世代粒色分离情况,提出复等位遗传模型假说。据此模型,较好地解释了两组合各世代粒色显隐性遗传关系。有关复等位遗传模型假说的应用范围也做了初步讨论。     

金黄地鼠（Mesocricetus auratus）朊蛋白基因的克隆及其原核表达

采用RT-PCR技术从金黄地鼠(Mesocricetus auratus)脑组织获得朊蛋白基因(Prion protein nucleic acid,PRNP)开放阅读框(Open reading frame,ORF),截去其N端信号肽(66 bp)和C端GPI锚定位点(69 bp)形成朊蛋白编码区PRNPx;将编码区重组于融合表达质粒Pet-DsbA中,并在大肠杆菌BL21(DE3)plysS中经IPTG诱导表达,并经Western-blot验证。结果表明,金黄地鼠PRNP基因ORF区全长为765 bp,编码254个氨基酸的前体蛋白,核苷酸序列与已发表金黄地鼠序列(M14054)同源性为99.87%,其第116个氨基酸发生了同义突变由GCT变为GCC;朊蛋白在大肠杆菌得到高效表达,产物是相对分子质量为47 000的融合蛋白。