微卫星DNA技术用于中国对虾家系构建中的系谱认证

用6个微卫星标记对中国对虾（Fenneropenaeus chinensis）的5个家系进行系谱鉴别和遗传多样性研究。6个微卫星位点中有5个位点是多态的，并在所有家系中都显示了高度的遗传差异。5个家系中，5个多态微卫星位点共发现了30个等位基因，每个位点的等位基因数在5～8之间。实验中共发现了4个家系特异性等位基因：2^#家系及4^#家系各1个，5^#家系2个。根据已知亲本及子代基因型，可推断出5个家系中全部亲本的基因型，据此鉴别各家系。在EN0033位点，可将5#家系与其他4个家系相区别；在RS0859位点，可将3^#和4^#家系与其他3个家系相区分。因此，EN0033和RS0859标记可用于鉴别5^#、3^#和4^#家系的家系特异性标记。研究表明，用5个微卫星标记，且最少用2对微卫星标记即可鉴别5个中国对虾家系。     

小麦Wx基因突变体的建立及其淀粉特性的研究

以小麦品种藁城8901为母本,糯麦1号为父本,用“单粒传”法构建了一个含有8种Wx基因突变类型的重组自交系群体 (RIL) 。应用改良的SDS-PAGE方法对RIL-7的Wx基因突变类型进行检测,发现在228个株系中,正常类型的株系有34个,Wx-A1突变体为26个,Wx-B1突变体为32个,Wx-D1突变体30个,Wx-A1和Wx-B1位点同时突变的有28个,Wx-A1和Wx-D1位点同时突变的有20个,Wx-B1和Wx-D1位点同时突变的为28个,3个Wx基因位点均突变的为16个。卡方测验证明,3个Wx基因位点的突变符合孟德尔遗传规律,属于质量性状遗传。田间试验表明,8种类型之间在初花期、株高和穗粒数上无显著差异,但在穗长、每穗小穗数和千粒重上Wx-A1突变型显著低于其他7种类型,而7种类型间无显著差异。淀粉特性研究表明,不同Wx蛋白缺失显著影响直链淀粉含量、淀粉糊化特性和淀粉凝胶的质构剖面分析 (TPA) 特性。正常类型的直链淀粉含量最高(20.8%),糯麦的最低(1.1%)。糯麦淀粉的峰值黏度和稀澥值较高,但其低谷黏度、最终黏度和反弹值较低,与其他缺失类型间差异达5%显著水平。凝胶的TPA测试表明,随着直链淀粉含量的降低,凝胶的硬度、黏着性、弹性、胶着性和咀嚼性显著降低,而黏聚性和回复性显著升高。直链淀粉含量与糊化特性的低谷黏度、最终黏度、反弹值、峰值时间和糊化温度之间正相关达1%显著水平(r=0.892~0.965),与峰值黏度、稀澥值呈1%水平负相关( r=－0.892, r=－0.945);直链淀粉含量与凝胶TPA参数的黏聚性、回复性呈极显著负相关(r=－0.928, r=－0.829),与凝胶的硬度、黏着性、弹性、胶着性和咀嚼性呈极显著正相关( r=0.869~0.979)。     

Allelic Variation in Loci for Adaptive Response and Its Effect on Agronomical Traits in Chinese Wheat （Triticum aestivum L.）

Heading date was an important trait that decided the adaptation of wheat to environments. It was modiifed by genes involved in vernalization response, photoperiod response and development rate. In this study, four loci Xgwm261, Xgwm219, Xbarc23 and Ppd-D1 which were previously reported related to heading time were analyzed based on three groups of wheat including landraces （L）, varieties bred before 1983 （B82） and after 1983 （A83） collected from Chinese wheat growing areas. Generally, heading date of landrace was longer than that of varieties. Signiifcant differences in the heading time existed within the groups, which implied that diversiifcation selection was much helpful for adaptation in each wheat zone. Photoperiod insensitive allele Ppd-D1a was the ifrst choice for both landrace and modern varieties, which promoted the heading date about four days earlier than that of sensitive allele Ppd-D1b. The three SSR loci had different characters in the three groups. Predominant allele combination for each zone was predicted for wheat group L and A83, which made great contribution to advantageous traits. Xgwm219 was found to be signiifcantly associated with heading date in Yellow and Huai River Winter Wheat Zone （Zone II） and spike length in Middle and lower Yangtze Valley Winter Wheat Zone （Zone III）, which implied functional diversiifcation for adaption. Variation for earliness genes provided here will be helpful for whet breeding in future climatic change.     

甘肃黑猪H-FABP基因克隆、SNPs筛查及生物信息学分析

采用生物信息学方法从分子水平对甘肃黑猪H-FABP基因编码蛋白质的功能和性质进行分析。采用DNA池和直接测序技术对甘肃黑猪H-FABP基因进行SNPs快速筛查及其编码区蛋白质功能预测。结果表明,甘肃黑猪H-FABP基因编码区全长402 bp,编码133个氨基酸残基,共检测出4个SNPs,分别为C⁷³⁴T-Intron1、T¹⁵²C-Exon2、G³⁰A-Intron2、T¹²¹G-Intron2,其中T¹⁵²C-Exon2为错义突变,氨基酸由原来的异亮氨酸(Ile)转变为苏氨酸(Thr)。生物信息学预测发现,甘肃黑猪H-FABP蛋白是一种稳定的亲水蛋白,蛋白质二三级结构均以β-折叠为主的混合型蛋白。     

Bovine SLC11A1 3' UTR SSCP genotype evaluated by a macrophage in vitro killing assay employing a Brucella abortus strain

The 3' untranslated region (3' UTR) of the bovine natural resistance-associated macrophage gene (NRAMP1 or SLC11A1) was genotyped in Colombian Creole Blanco Orejinegro (BON) (Bos taurus) (n = 140) and Zebu Brahman (Bos indicus) (Z) (n = 20) cattle and their crosses (BON x Zebu Brahman [B x Z] [n = 10]; Zebu Brahman x BON [Z x B] [n = 10]), and in animals from a Holstein x BON (H x B) (n = 10) cross. Direct sequencing and single-strand conformation polymorphism analysis (SSCP) helped in detecting the polymorphic behaviour. The association between resistance to brucellosis infection and SSCP genotype was evaluated using a macrophage in vitro killing assay employing a virulent Brucella abortus strain. The 3' UTR (GT) repeated polymorphism was gentoyped and its association with resistance to brucellosis was evaluated. When all breeds were grouped, a high frequency in the homozygote GT(12) (AA genotype) (0.823) and a very low frequency in the homozygote GT(10) (BB genotype) (0.047) were detected. The BON (0.963), Z x B (0.60) and H x B (1.00) cattle showed high GT(12) allele frequencies, unlike that seen for the B x Z and Zebu cattle (0.3002 and 0.218, respectively). The GT(10) allele was only found in the Zebu cattle (0.391). A significant association (p < 0.001) was found between the B. abortus macrophage in vitro killing assay phenotypes and the bovine SLC11A1 3' UTR genotypes, which suggests that the A allele may be associated with resistance. Because only nine animals had the BB genotype, the results require some confirmation in more extensive populations.     

新疆小麦地方品种籽粒硬度及其Puroindoline 基因等位变异研究

[目的]籽粒硬度是重要的小麦品质性状,了解新生疆小麦地方品种籽粒硬度的变异类型和分布,为新疆小麦品质育种提供遗传信息.[方法]利用单籽粒硬度仪(SKCS)和PCR扩增、酶切及DNA测序技术,结合改良的Friabilin提取及电泳分析方法,对105份新疆小麦地方品种的籽粒硬度及Pina和Pint等位基因进行研究.[结果]新疆小麦地方品种以硬质类型为主,平均硬度值为63.5.新疆小麦地方品种Puroindoline基因共有9种突变型,分别是Pina-D1a/Pinb-D1a,Pina-D1a/Pinb-Dlb,Pina-D1a/Pint-D1p,Pina-D1k ()/,Pina-D1l/Pinb-D1a,Pina-D1r/Pinb-D1a,Pina-D1a/Pinb-D1ab,Pina-D1a/Pint-Dlac和Pina-D1b/Pinb-D1p,其中Pina-D1 r/Pinb-D1a为新发现的突变型.新疆小麦地方品种Puroindoline基因型以Pina-D1a/Pinb-D1p为主导类型,共有62份,占59.1;;其Pina-D1k()/和Pina-D1 a/Pint-D1ab两种突变型与其地理来源和生态型密切相关,主要分布在南疆冬小麦中.Pina-D1a/Pinb-D1a(野生型)的硬度值显著低于突变型,Pina-Dlk()/的硬度值最高,但Pins-D1k ()/ ,Pina-D1a/Pinb-D1p和Pina-D1a/Pinb-D1ab的硬度值差异不显著.[结论]新疆小麦地方品种籽粒硬度及其Puroindoline等位变异和分布信息能为新疆小麦品质改良和引种提供理论依据.     

Gene Dropping Analysis of Ancestral Contributions and Allele Survival in Japanese Thoroughbred Population

Genetic contributions of nine historically important ancestors and allelic diversity in the Japanese Thoroughbred population were examined by applying the gene dropping simulation to the foals produced from 1978 to 2005. Full pedigree records traced to ancestors (base animals) born around 1890 were used for the simulation. Alleles originated from some of the historically important ancestors were found to be at risk of future extinction, although their genetic contributions to the foal population have increased during the last three decades. The proportion of surviving alleles to the total alleles assigned to the base animals was 8.0% in the foal population in 2005, suggesting that a large part of genetic variability contained in the base animals is extinct in the current population.     

Effect of VRN‐1 and PPD‐D1 genes on heading time in European bread wheat cultivars

The variation of the vernalization (VRN‐1) and photoperiod (PPD‐1) genes offers opportunities to adjust heading time and to maximize yield in crop species. The effect of these genes on heading time was studied based on a set of 245 predominantly spring cultivars of bread wheat from the main eco‐geographical regions of Europe. The genotypes were screened using previously published diagnostic molecular markers for detecting the dominant or recessive alleles of the major VRN‐1 loci such as: VRN‐A1, VRN‐B1, VRN‐D1 as well as PPD‐D1. We found that 91% of spring wheat cultivars contain the photoperiod sensitive PPD‐D1b allele. Photoperiod insensitive PPD‐D1a allele has been found mainly in southern region of Europe. For this region the monogenic control of vernalization by VRN‐B1 or VRN‐D1 dominant alleles is common, whereas in the remaining part of Europe, the combination of photoperiod sensitive PPD‐D1b allele with dominant VRN‐A1, VRN‐B1 and recessive vrn‐D1 alleles represents the most frequent genotype. Also, we revealed a significantly later (5–8 days) heading of the monogenically dominant genotypes at VRN‐B1 as compared to the digenic VRN‐A1 VRN‐B1 genotypes.     

Improving Rice Blast Resistance by Mining Broad-Spectrum Resistance Genes at Pik Locus

Magnaporthe oryzae is known for its genetic diversity and pathogenic variability, leading to rapid breakdown of resistance in rice. Incorporating multiple broad-spectrum blast resistance genes into rice cultivars would extend disease resistance longevity. Effective resistance breeding in rice therefore requires continual enrichment of the reservoir of resistance genes and alleles. We conducted a large-scale screen of rice blast resistance in about 2 000 rice accessions. Among them, 247 accessions showed at least medium resistance to the natural infection of rice blast and 7 novel Pik alleles were identified from them. Variations in gene sequences were then correlated with the phenotypic trait to enable the identification of favorable alleles. Among the seven novel Pik alleles, the resistant rate of Pik-R0/ME/7017 donors was greater than 80%, and the disease score was less than 3. Through molecular marker-assisted backcross breeding, we successfully transferred the three Pik alleles, Pik-R0/ME/7017, into an elite cultivated line Kongyu 131 to obtain BC₃F₂ lines, which showed enhanced resistance to rice blast compared with the recurrent parent. Assessment of these near-isogenic lines in the greenhouse using 31 isolates of M. oryzae from Heilongjiang Province of China revealed that the resistant levels of the BC₃F₂ lines with Pik-R0/ME/7017 were significantly higher than those of the established cloned resistance genes Pik-m and Pi1. Exploring such alleles will enrich our gene library for resistance to rice blast.