鹅HMGR基因内含子5的SNP及其与鹅重要经济性状的关联分析

本文屠宰测定了116只皖西白鹅胴体性状、羽绒性状和肉质性状，克隆了鹅3－羟基－3－甲基戊二酸单酰辅酶A还原酶（HMGR）基因内含子5，采用PCR-SSCP检测其SNP，并分析了SNP与鹅重要经济性状之间的关联性。结果显示，鹅HMGR基因内含子5长704bp，与鸡的同源性44.0%；PCR-SSCP检测发现，内含子5存在一对等位基因A和B，A和B的基因频率分别为0.7716和0.2284。分析SNP基因型与鹅重要经济性状的关联性，结果表明，AA型可以显著提高鹅的胸肌重和腺胃重（P<0.05），增加千朵重（P<0.01），和降低绒系水率（P<0.05）；AB型则具有显著降低胸肌纤维直径（P<0.01）和提高胸肌45pH的作用（P<0.05）。     

奶牛IGFBP-3基因部分序列PCR-SSCP多态性与产奶量和生长性能的相关性

采用PCR-SSCP技术,对荷斯坦奶牛及荷斯坦公牛冻精杂交改良的黄牛后代(回交二代)共105头奶牛胰岛素样生长因子结合蛋白3(IGFBP-3)基因内含子2部分序列和外显子3全部序列进行了多态性研究,分析了该基因与奶牛生产性状的相关性.结果表明:IGFBP-3基因第2内含子共存在2个等位基因3种基因型,A、B基因频率为O.7667和O.2333,AA、AB和BB基因型频率分别为0.6000、O.3333和0.0667.该群体在这一位点上处于Hardy-Weinberg平衡状态,遗传杂合度(H(e))和多态信息含量(PIC)分别为O.3578和为O.2937.测序结果显示,与普通黄牛该基因序列(AF305712)相比,B等位基因在第8 515碱基处发生了G→A的转换,而A等位基因在此处与AF305712序列相同.此外,A、B等位基因在第8 634碱基位置(属第3外显子区)均为c,而普通黄牛在此处为T碱基,该突变为同义突变.最小二乘法分析表明,AA基因型305 d产奶量和12月龄体重极显著高于AB和BB基因型(P<0.01);AA型12月龄体长和体高均极显著高于AB型(P<0.01),而BB型与AB型和AA型差异不显著;不同基因型的胸围和腹嗣差异不显著.     

五指山猪GH、IGFBP3基因SNPs检测及与生长性状的关联分析

以五指山猪为试验材料,采用PCR-SSCP和测序相结合技术,对GH基因和IGFBP3基因进行多态性检测,并分析其与生长性状（体质量,体高,体长和胸围）的相关性。结果表明,GH基因第2外显子存在一处G→A转换,属沉默突变,该位点与生长性状关联分析显示,BB基因型的体重显著高于AA基因型和AB基因型;IGFBP3基因第2外显子存在一处G→C颠换,属沉默突变,该位点与生长性状关联分析显示,AA基因型的体高显著高于BB基因型;IGFBP3基因第4内含子存在一处碱基C缺失,该位点与生长性状关联分析显示,AA基因型体重显著高于AB基因型和BB基因型。研究将为五指山猪生长发育规律、系统选育及矮小机制等方面研究提供了遗传学依据。     

贵州本地山羊STAT5A基因多态性及其与生长性状关联分析

本研究通过探讨STAT5A基因SNP与山羊生长性状关联性,旨在为山羊选种选育提供更好的科学依据。实验以贵州白山羊和贵州黑山羊为研究对象,采用DNA池法及PCR-SSCP技术检测STAT5A基因单核苷酸多态性。结果发现,在贵州白山羊和贵州黑山羊STAT5A基因内含子10均检测到1个SNP位点G127A,表现为3种基因型,分别命名为GG、GA和AA。基因型与生长性状关联分析显示,贵州黑山羊基因型GA个体的胸围和体重指标显著高于基因型GG个体（p〈0.05）,而其余3个指标均差异不显著（p〉0.05）;贵州白山羊基因型GA个体的体斜长、胸围和管围指标显著高于基因型GG个体（p〈0.05）,而其余指标均差异不显著（p〉0.05）。研究结果提示：STAT5A基因可能是影响山羊体重、体斜长、管围和胸围的主效基因或与主效基因连锁,G127A位点可望作为提高山羊个体生长性状的分子遗传标记。     

基于DNA池重测序技术研究细毛羊LAMB1基因的SNPs分析

层粘连蛋白β1 (laminin-beta-1,LAMB1)作为层粘连蛋白家族成员之一,在生物过程中发挥极为重要的作用.为了分析细毛羊(Ovis aries)毛性状相关候选基因LAMB1的遗传效应,本研究通过DNA池重测序技术筛选了LAMB1基因外显子区的5个单核苷酸多态性(single nucleotide polymorphisms,SNPs),对其使用聚合酶链反应-单链构象多态性(single strand conformation polymorphism analysis of polymerase chain reaction products,PCR-SSCP)技术分型,在此基础上,利用SAS 8.1的GLM程序分析其对新疆巩乃斯种羊场418只细毛羊毛性状的遗传效应,利用HaploView 4.2软件进行连锁不平衡(linkagedisequilibrium,LD)分析.结果显示,该细毛羊群体遗传变异处于中等水平,LAMB1的5个SNPs发生错义突变.SNP2中AA和GA基因型个体鉴定时体重、剪毛后体重极显著高于GG基因型个体(P＜0.O1);SNP4中CC、TT和TC基因型个体间的自然长度差异极显著(P＜0.01),CC基因型个体的剪毛后体重极显著高于TT和TC基因型的个体(P＜0.01);而其他SNPs各基因型个体间的部分毛性状有显著差异(P＜0.05).SNP1和SNP3、SNP2和SNP3处于连锁不平衡状态,LAMB1基因可作为具有潜在应用价值的细毛羊毛性状分子标记之一.本研究揭示了L4MB1基因的分子遗传特征及其与细毛羊群体的遗传关系,为高效选育细毛羊经济性状及对其种质资源的保护与利用提供了分子学基础.     

郏县红牛跨膜蛋白18基因(TMEM18)SNPs检测及其与生长性状的关联分析

跨膜蛋白18基因(Transmembrane protein 18,TMEM18)在神经系统表达.全基因组关联分析(GWAS)表明,该基因的突变会影响人类肥胖和Body Mass Index(BMI)值变化.本研究采用DNA池测序技术检测了郏县红牛(Bos taurus) TMEM18基因5个外显子和部分内含子区单核苷酸多态(single nucleotide polymorphism,SNP)位点,通过PCR-RFLP技术对所发现的SNP位点进行分型验证,然后分析不同基因型与郏县红牛生长性状的关联性.结果表明,在郏县红牛TMEM18基因上共发现了3处新的SNPs:NW_003099175.1∶g.2303G＞A、g.3835 G＞A和g.3865 A＞G(aa.Gly＞Ser).关联分析结果表明,郏县红牛TMEM18基因g.2303G＞A位点AA型个体体高、体长、胸围和体重显著大于GG型(P＜0.05),郏县红牛TMEM18基因g.3835 G＞A位点AG型个体胸围和体重显著大于AA型(P＜0.05).研究结果提示,这两个位点可作为郏县红牛品种体尺和体重生长性状选择的分子育种候选标记.     

催乳素(PEL)和生长激素(GH)基因对高邮鸭产蛋性能的遗传效应分析

分别采用PCR-RFLP和PCR-SSCP方法检测催乳素(PRL)和生长激素(GH)基因的多态性,并分析了PRL和GH基因与高邮鸭产蛋性能的关系.结果表明:①PRL基因内含子1经酶切得出3种基因型AA/AB/BB,等位基因A/B频率分别为0.226/0.774;经测序发现PRL基因内含子1在1 326 bp处发生T→C突变.GH基因外显子4经SSCP分析显示3种基凶型CC/CD/DD,等位基因C/D基因频率分别为0.222/0.778;经测序发现GH基因外显子4在3 701 bp处发生C→T突变.②PTL内含子1多态与高邮鸭的30周龄蛋重存在极显著关联(P<0.01);PRL多态与双黄蛋率显著关联(P<0.05);GH基因多态与最长连产天数和双黄率间相关性达显著性水平(P<0.05).     

清远麻鸡生长激素基因(GH)的SNPs检测及其对生长及繁殖性状的遗传效应

为了研究生长激素基因(GH)对清远麻鸡部分生长及繁殖性状的遗传效应,基于聚合酶链反应-连接酶检测反应(PCR-LDR)基因多态性并行检测系统,对清远麻鸡(Gallus gallus)生长激素(CH)基因内含子1的G662A位点和内含子4的T3094C,C3199T位点进行检测,并分析其对部分生长及繁殖性状的遗传效应.检测结果显示,清远麻鸡在3个位点上均产生了3种基因型,LDR分型结果同直接测序结果一致.G662A位点对生长及繁殖性状均不存在显着的效应;T3094C位点上,TT型个体的开产日龄显着晚于CC和CT型个体,其18周龄体重又显著高于CC和CT型个体;C3199T位点上,不同基因型个体仅在50周平均蛋重上存在显着差异,TT型个体的50周平均蛋重显着高于CC和CT型个体.3个位点的合并基因型对开产蛋重、50周平均蛋重以及生长性状均有显着的影响,且对生长性能的遗传效应更加明显,合并基因型AACTCC可以作为筛选清远麻鸡高的体重的分子标记.     

猪促甲状腺激素β基因(TSH-β)多态分布及与经济性状的关联分析

为研究猪促甲状腺激素口基因(TSH-β)的多态分布情况及与生长肉质性状的关联性,本研究对TSH-β基因全序列进行再测序,发现3个单核苷酸突变座位:位于5'非转录区多态座位A-175G、位于编码区非同义突变座位G+52A,以及位于内含子的突变座位G+570A.采用PCR-RFLP和四引物ARMS-PCR方法对皮特兰、长白、杜洛克、约克夏、金华猪、岔路黑猪和嘉兴黑猪七个纯种猪群以及金皮资源家系F2代群体共计548头个体进行多态分型,并在F2代群体中分析了TSH-β多态座位与生长和肉质性状的关联性.结果表明,猪TSH-β基因具有较高的遗传多样性(Shannon多样性指数总体均值为0.48),且存在相当程度的品种间遗传分化(Fst=0.5389).其中,座位A-175G在4个国外品种和岔路黑猪中的优势等位基因为A,而在金华猪和嘉兴黑猪中的优势等位基因为G,关联分析显示AA型个体在胴体长、日增重、后腿重和后腿肌肉重方面显著高于GG型个体(P<0.05):座位G+52A的多态性主要存在于金华猪中,而在其他猪种中表现为低杂合度或完全纯合于等位基因G,且与肌内脂肪含量有显著关联;座位G+570A的多态性仅存在于我国的嘉兴黑猪和金华猪中,其他猪种表现为完全纯合于等位基因G,关联分析结果表明GG型个体在胴体长和后腿肌肉重方面显著高于AA型个体.研究结果提示,TSH-β基因可考虑为猪4号染色体相关QTL的功能候选基因.     

奶牛IGFBP-3基因PCR-SSCP多态性与产奶量和生长性能相关性

采用PCR-SSCP技术,对荷斯坦奶牛及荷斯坦公牛冻精杂交改良的黄牛后代(回交二代)共105头奶牛IGFBP-3基因第3外显子进行了多态性研究, 分析了该基因与奶牛生产性状的相关性。结果表明：IGFBP-3基因第3外显子共存在2个等位基因3种基因型,A、B基因频率及AA、AB、BB基因型的频率分别为0.7667 、0.2333和0.6000、0.3333、0.0667。该群体在这一位点上处于Hardy-Weinberg平衡状态,遗传杂合度（He）和多态信息含量(PIC)分别为0.3578和为0.2937。测序结果显示: 与AF305712相比,B等位基因在第8515碱基处发生了G-A的转换,而A等位基因在此处与AF305712序列相同;此外,A、B等位基因在AF305712的第8634碱基处均发生了T-C的无义突变。最小二乘法分析表明：AA基因型305天产奶量、12月龄体重极显著高于AB和BB基因型( P < 0.01);AA型12月龄体长和体高均极显著高于AB型（p<0.01）,而BB型与AB型和AA型差异不显著;不同基因型的胸围、腹围差异不显著。     

The flavonol quercetin-3-glucoside inhibits cyanidin-3-glucoside absorption in vitro

At present, little is known about the mechanisms responsible for intestinal absorption of anthocyanins (ACNs). For example, it has not yet been established if ACNs are absorbed through an active transport mechanism, such as the sodium-dependent glucose transporter (SGLT1), or by passive diffusion. Previously, we found that the absorption of ACNs differs between regions of the digestive tract and is maximal in the jejunum, suggesting that an active transport mechanism is involved. In the present study, we examined the effect of d-glucose (main substrate of SGLT1), phloridzin (inhibitor of SGLT1), and quercetin-3-glucose (Q3G, a flavonol) on the absorption of cyanidin-3-glucoside (C3G; approximately 5 micromol/L) by mouse jejunum mounted in Ussing chambers. We found that the presence of either D-glucose (10, 20, and 40 mmol/L) or phloridzin (50, 100, and 200 micromol/L) resulted in a small but insignificant inhibition of C3G disappearance from the mucosal solution (decrease of disappearance with glucose, 33%; with phloridzin, 18%; NS). However, when the flavonol Q3G (50 micromol/L) was added to the mucosal solution together with the C3G, the disappearance of C3G was significantly decreased (74%; p < 0.001), and Q3G disappeared instead. In addition, we found phloretin and quercetin, the aglycones of phloridzin and Q3G, respectively, present in the mucosal solution and tissue extracts, indicating hydrolysis of these compounds by the enterocytes of the jejunum. In contrast, the aglycone cyanidin was not detected at all. Our results show that in the mouse small intestine, ACN absorption is not solely dependent on the activity of the SGLT1 transporter, as d-glucose and phloridzin had only a slight effect on uptake. Q3G, however, clearly inhibited C3G disappearance. These results suggest that there might be a competitive inhibition between C3G and Q3G absorption. It is possible that an absorption mechanism other than the SGLT1 is involved, which has a structural preference toward flavonols.     

Intracellular mediators of procyanidin-induced lipolysis in 3T3-L1 adipocytes

We have previously reported that grape seed procyanidins stimulate long-term lipolysis on 3T3-L1 fully differentiated adipocytes. To unravel the molecular mechanism by which procyanidins exert this effect, we checked the involvement of two main cellular targets in adipose cells: protein kinase A (PKA) and peroxisome proliferator-activated receptor-gamma (PPAR-gamma). Procyanidin treatment increased intracellular cAMP levels in 3T3-L1 adipocytes, and their lipolytic effect was inhibited by simultaneous treatment with H89, a PKA specific inhibitor. BRL49653, a very highly specific ligand of PPAR-gamma, totally abolished the lipolytic effect of procyanidins. Simultaneous to this long-term lipolytic effect, the mRNA levels of some differentiation adipocyte markers decreased, although there were no changes in the triglyceride content of the cells. BRL49653 did not antagonize the decrements of differentiation markers. These results support a mediation of PPAR-gamma and PKA on the lipolytic effects of procyanidins on 3T3-L1 adipocytes.     

Increase in lipid content in potato tubers modified by 14-3-3 gene overexpression

Recently, transgenic potato plants were created with overexpression of the 14-3-3 protein derived from Cucurbita pepo. Detailed analysis of those plants suggested that the function of the isolated 14-3-3 isoform is in the control of carbohydrate and lipid metabolism in the plants. 14-3-3 protein overexpression gave rise to an increase in soluble sugar and catecholamine contents in both leaves and tubers. It is proposed that 14-3-3 protein affects carbohydrate metabolism in potato plants via regulation of catecholamine synthesis. Furthermore, genetically modified potato tubers with 14-3-3 protein overexpression showed changes in lipid content and composition. The transgenic potato tubers contained 69% more total fat compared to the wild-type plant. Separation of tuber lipids into polar and nonpolar fractions revealed that the transgenic potato tubers contained almost 3 times more nonpolar lipids than the control. Analysis of fatty acid composition, conducted by the means of gas chromatography, showed that linoleic acid was the main fatty acid present in the tubers of both modified and control potato plants. In the nonpolar fraction of the fat of the transgenic tubers the unsaturated fatty acids exhibited a higher participation in the sum of all fatty acids.     

“3S”技术在水土保持“三区”划分中的应用

为准确、科学划分河南省水土流失重点防治区 ,河南省依据遥感TM影像 ,运用“3S”技术 ,修改、完成了全省 66个山区县的水土流失防治区通告图的电子版制作和图集编印工作。介绍了应用“3S”技术划分“三区”的操作过程     

GIS、RS与3DS MAX在园林三维景观设计中的应用

目前应用计算机进行园林景观设计的方法可以分成两大类:一类应用地理信息系统(G IS)技术,另一类应用传统的图形图像处理软件。前者制约了园林景观的建模和效果展示,后者却忽略了作为园林景观设计基础的地形研究与应用。为了克服上述不足,提出"G[地理信息系统(G IS)]R[遥感(RS)]M[3DSM AX]三维园林"综合方法来进行园林景观设计:先用G IS生成园林设计区域数字高程模型,用RS创建3D数字地形模型,再把3D数字地形模型导入3DS M AX,制作基于三维地形的园林景观模型。该方法应用于广西都乔山森林公园设计,其设计质量和效率明显提高。     

Cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside as primary phenolic antioxidants in black raspberry

Anthocyanin constituents in black raspberries (Rubus occidentalis L.) were investigated by HPLC-DAD, and their involvement as potent, significant antioxidants in black raspberries was demonstrated by three common antioxidant assays (FRAP, DPPH, ABTS) in this study. Five anthocyanins were present in black raspberries: cyanidin 3-sambubioside, cyanidin 3-glucoside, cyanidin 3-xylosylrutinoside, cyanidin 3-rutinoside, and pelargonidin 3-rutinoside. Their identities and structures, with particular emphasis on cyanidin 3-xylosylrutinoside, were confirmed by NMR spectroscopy. Two of these anthocyanins, cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside, predominated, comprising 24-40 and 49-58%, respectively, of the total anthocyanins in black raspberries. On the basis of both potency and concentration, cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside were found to be the significant contributors to the antioxidant systems of black raspberries. These findings indicate that these two anthocyanin compounds may function as the primary phenolic antioxidants in black raspberries. These two compounds exhibit potential biological activities that may be exploited in conjunction with other naturally occurring bioactive compounds in black raspberry fruit-based products used in clinical trials for the treatment of various types of cancer.     

Beta-conglycinin embeds active peptides that inhibit lipid accumulation in 3T3-L1 adipocytes in vitro

Obesity is a worldwide health concern because it is a well-recognized predictor of premature mortality. The objective was to identify soybean varieties that have improved potential to inhibit fat accumulation in adipocytes by testing the effects of soy hydrolysates having a range of protein subunit compositions on lipid accumulation and adiponectin expression in 3T3-L1 adipocytes. The results showed that differences in the protein distribution of 15 soy genotypes led to different potentials for the reduction of fat accumulation. The inhibition of lipid accumulation of soy alcalase hydrolysates in 3T3-L1 adipocytes ranged from 29 to 46%. Soy hydrolysates made from genotypes with 45.3 +/- 3.3% of total protein as beta-conglycinin, on average, showed significantly higher inhibition of lipid accumulation compared to those with 24.7 +/- 1.5% of extracted total protein as beta-conglycinin. Moreover, after in vitro simulated digestion with pepsin-pancreatin of the soy alcalase hydrolysates, 86% of the original activity remained. Adiponectin expression was induced in 3T3-L1 adipocytes treated with 15 soy hydrolysates up to 2.49- and 2.63-fold for high and low molecular weight adiponectin, respectively. The inhibition of lipid accumulation calculated from a partial least squares (PLS) analysis model correlated well with experimental data (R(2) = 0.91). In conclusion, it was feasible to differentiate soy varieties on the basis of the potential of their proteins to reduce fat accumulation using a statistical model and a cell-based assay in vitro. Furthermore, beta-conglycinin embeds more peptides than glycinin subunits that inhibit lipid accumulation and induce adiponectin in 3T3-L1 adipocytes. Therefore, soy ingredients containing beta-conglycinin may be important food components for the control of lipid accumulation in adipose tissue.     

Enzymatic synthesis of [3'-O-methyl-(3)H]malvidin-3-glucoside from petunidin-3-glucoside

Malvidin-3-glucoside has been labeled by enzymatic synthesis in a single-step experiment. Catechol-O-methyl transferase catalyzed the B-ring methylation of petunidin-3-glucoside, and S-Adenosyl-L-[methyl-(3)H] methionine was the methyl donor. Solid phase extraction and preparative high-performance liquid chromatography were necessary to separate [3'-O-methyl-(3)H]malvidin-3-glucoside from an isomer and the starting material. The specific activity was 2.2 Ci mmol(-1), and the yield of incorporation was 1.1%. A possible application of the labeled material is the study of anthocyanin reactions in complex mixtures such as red wine where products are difficult to isolate and analyze.     

Comparative toxicity of Al3+, Yb3+, and La3+ to root-tip cells differing in tolerance to high Al3+ in terms of ionic potentials of dehydrated trivalent cations

Abstract

Green manure legumes are often used to compare biomass production as well as nitrogen-fixing capacity. Mineral deficiency often limits the symbiotic nitrogen fixation of many legumes, thus limiting their productivity despite their high yielding potential (O’Hara et al. 1988; Flis et al. 1993). Leguminous species require large amounts of P for growth, nodulation, and nitrogen fixation. Consequently, they are often unable to grow in acid soils with low available P. The low P availability in tropical acid soils often arises from fixation of P by Al and Fe in soil. Generally, Al and Fe-phosphates are relatively unavailable to plants (McLachlan 1976; Ae et al. 1990).     

NH3 Volatilization from Urea-NBPT in Eucalyptus

High losses of nitrogen (N) by volatilization of ammonia from urea applied in Eucalyptus are expected due to the influence of plant residues on the soil surface. The study evaluated the N losses by volatilization of ammonia from urea coated with Thiophosphate N-(n-butil) triamide (NBPT) applied in soil with eucalyptus residues in surface under moisture treatments: fertilization in dry soil without irrigation; fertilization in dry soil with posterior irrigation depth (3 mm); fertilization in moist soil without irrigation and fertilization in moist soil with irrigation depth (3 mm). NBPT is a potential inhibitor of urease. Urea with NBPT shows lower losses by volatilization of ammonia when it is applied in dry soil; however in soil conditions of high moisture the losses as well as inhibitor effect of the NBPT are lower. The inhibitor effect of NBPT is reduced over time when it is subjected to moisture conditions.