饲料中添加不同水平醋糟对獭兔肉品质的影响研究

为研究醋糟作为粗纤维饲料对獭兔肉品质的影响,选择90日龄健康的生长獭兔180只,随机分成6组,分别饲喂6种不同比例醋糟替代谷草与小米壳(0%、7%、14%、21%、28%和35%)的饲粮。结果表明:各组氨基酸总量、鲜味氨基酸含量差异不显著(P>0.05);在人体必需氨基酸含量方面,各试验组都有一定程度的提高,其中35%组最高(P>0.05);兔肉中蛋白、灰分与水分含量各组差异不显著(P>0.05)。脂肪含量35%组与对照组接近,与21%组差异显著(P<0.05);各组滴水损失、熟肉率及剪切力差异不显著(P>0.05);肉色方面,35%组与对照组最低;在pH值方面,对照组与各试验组之间差异不显著(P>0.05)。因此,从兔肉的营养价值和饲料成本等综合考虑,在獭兔日粮中添加35%的醋糟对肉品质效果较好。     

猪红细胞生成素受体基因突变与血常规性状关联分析

本研究旨在探索红细胞生成素受体基因(EPOR)影响红细胞相关性状的分子机理.对EPOR基因第1内含子和第4内含子突变位点g.705G＞T和g.2373C＞T,以飞行时间质谱进行个体基因型分型,在构建的大白猪×民猪F2代资源群体内开展其与6种血常规性状(红细胞压积、血红蛋白、平均红细胞血红蛋白量、平均红细胞体积、红细胞计数及红细胞分布宽度)的关联分析.结果表明,第1内含子突变位点g.705G＞T未发现与血常规性状显著关联;第4内含子突变位点g.2373C＞T群体内只发现2种基因型,其中CT基因型个体的红细胞压积显著高于CC基因型个体(P＜0.05),但并未发现与其他性状显著关联.结果显示,EPOR基因突变可显著影响红细胞压积,说明该基因可作为影响红细胞压积的主效候选基因开展深入研究.     

口蹄疫病毒Asial/JS/China/2005株基因组全长感染性克隆的构建

利用长距离RT-PCR技术扩增病毒基因组3'端覆盖口蹄疫病毒Asial/JS/China/2005株近全长的3个片段(共约7.5 kb),并利用单一酶切位点将其分别克隆到pBlueseriptSK+载体上.利用融合PCR扩增到基因组5'端含有15个C碱基的基因片段(约700 bp),并将其连接至pGEM-T载体.最后将这4个片段的阳性克隆装配至剔除T7启动子的低拷贝载体pcDNA3.1/Zeo(+)中构建该病毒株的全长cDNA克隆.以构建的FMDV Asial/JS/China/2005株全长cDNA为模板,使用TTRNA聚合酶在体外转录得到病毒RNA,通过脂质体将其导入BHK细胞获得拯救病毒.对收获的病毒分别用RT-PCR、间接免疫荧光、电子显微镜观察和乳鼠致病性分析结果证实,通过体外转录获得了具有感染性的口蹄疫病毒.该株感染性克隆的构建为深入研究口蹄疫病毒的致病机制及研制新型疫苗等奠定了基础.     

粉纹夜蛾(Trichoplusia ni)培养细胞对家蚕微孢子虫(Nosema bombycis)的吞噬过程观察

观察微孢子虫在非天然宿主细胞系的生物行为过程,有助于探究微孢子虫的侵染特异性机制。选用鳞翅目昆虫粉纹夜蛾(Trichoplusia ni,Tn)细胞系为材料,接种家蚕微孢子虫(Nosema bombycis,Nb)孢子后观察其吞噬过程。结果发现:经氢氧化钾发芽预处理的Nb孢子虽然可以在Tn培养细胞液中发芽,但未见发芽孢子的孢原质进入Tn培养细胞;在接种Nb孢子浓度为2.0×105mL-1以上时,观察到孢子通过内吞作用进入了Tn培养细胞,但未见孢子的增殖;随着时间的延长和进入Tn培养细胞的Nb孢子的增加,可引起Tn培养细胞的超微结构变化,并导致细胞的裂解。以上结果说明,虽未见Nb以发芽形式感染Tn培养细胞,但Nb能够被吞噬进入Tn培养细胞并破坏其结构。     

用卡片PCR法检测卡耶他环孢子虫

使用FTA卡片法提取卡耶他环孢子虫DNA,利用特异性引物组F1E/R2B、CC719/CRP999进行巢式PCR,得到1条298bp的特异条带。并以柔嫩艾美尔球虫、贝氏隐孢子虫等作为对照,结果显示,除卡耶他环孢子虫外,均无特异性条带。用卡耶他环孢子虫卵囊进行巢式PCR敏感性试验,结果显示敏感度最高达1.0×100个/mL。     

Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis

AIM: To analyze the feasibility of PCR-high-resolution melting curve analysis (HRM) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR-restriction fragment length polymorphism (RFLP), the present research used the method of PCR-HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group (n=100) and liver cirrhosis group (n=100). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR-HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index (FLI), fasting insulin (FINS) and insulin resistance index estimated by homeostatic model assessment (HOMA-IR) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index (ISI) and soluble leptin receptor (sOB-R) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA-IR (r=0.45, r=0.53, P<0.05), and negatively with ISI (r=-0.34, P<0.05). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI-IR, leptin, sOB-R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB-R levels (P<0.05). CONCLUSION: PCR-HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

壳聚糖涂膜对常温贮藏兔眼蓝莓果实品质的影响

以“圆蓝”兔眼蓝莓为试材,研究了0．5％壳聚糖（0．4％高分子壳聚糖＋0．1％低分子壳聚糖）涂膜对常温贮藏蓝莓果实品质的影响。结果表明,在20℃常温条件下贮藏20d的过程中,壳聚糖涂膜可以有效减缓蓝莓果实感官品质及果实硬度的下降速度,降低腐烂率和失重率,保持较稳定的TSS和有机酸含量;抑制PPO活性的快速升高和MDA的大量积累：有效延长常温条件下的贮藏期,有利于保持采后蓝莓的品质。但对采后蓝莓果实中的花色苷含量无明显影响．对类黄酮的增加有一定的抑0作用。     

室内模拟条件下玉米秸秆的分解特征及物质组成变化

以玉米秸秆为材料,研究了在室内模拟的条件下不同处理的玉米秸秆的腐解率、粗纤维以及全钾含量的变化。结果表明,随着腐解时间的延长,各处理的腐解率不断增大,在其他条件一致的前提下,添加微生物的处理平均腐解率 大于未添加微生物的处理,秸秆长度为1 cm的处理平均腐解率大于秸秆长度为3 cm的处理;粗纤维含量的变化总体趋势是下降,其他条件一致的前提下,秸秆长度为1 cm的处理粗纤维含量小于秸秆长度为3 cm的处理,在添加物微生物的处理中秸秆长度为1 cm且C/N=25的处理粗纤维平均含量最小,其含量为31.57%;其他条件一致的前提下,秸秆长度为1 cm的处理全钾含量大于秸秆长度为3 cm的处理。     

水生蔬菜设施高效栽培技术及其应用

水生蔬菜是我国蔬菜生产中的一大特色,其品种之多,分布之广,生产面积之大,在世界各国中均居首位。我国栽培的水生蔬菜主要有莲藕、水芹、茭白、慈姑、荸荠、芡实、莼菜、菱、蒲菜、水蕹菜、水芋和豆瓣菜等10多种。水生蔬菜的生长环境与旱生蔬菜有显著的差异。主要表现为要求气候比较温暖,地势比较低洼,雨量