转基因水稻对稻瘟病的抗性研究

 采用苗期初筛、复筛、抗谱测定和田间自然诱发试验等不同鉴定方法,对经分子检测证明已整合有碱性几丁质酶基因和β-1,3-葡聚精酶基因的22个转化系的转基因水稻植株进行稻瘟病抗性鉴定研究,筛选出对稻瘟病的抗性比原种对照七丝软占有明显提高的一系列转基因水稻品系,其中表现高抗的有来自F4-9转化株系的7个品系。高抗材料的R7代品系,经室内抗谱测定及田间病圃试验结果,仍然表现高抗稻瘟病。本研究通过转基因技术,成功地将优质感病品种改良成高抗品系,研究结果证明了利用基因工程手段培育抗病水稻新品种是一个非常有希望的育种途径。     

植物苯丙氨酸解氨酶基因的表达调控与研究展望

苯丙氨酸解氨酶(phenylalanineammonia-lyase,PAL,EC4.3.1.5)是催化苯丙烷代谢途径第一步反应的酶,也是这个途径的关键酶,对植物有非常重要的生理意义。根据有关文献综述了植物PAL的分布与定位、酶学性质,总结了生长发育、钝化因子与调节因子、末端产物等内部因素及光、温、机械损伤与生长调节剂等外部因素对PAL的调控作用,得出外部因子是在转录水平上对酶活性实施调控的结论,并运用酶学和分子生物学方面的知识阐述了其调控机理。还着重阐述了PAL酶在果树上的研究现状与进展,并对其今后的研究及在果树上的应用进行了展望。     

Familial Congenital Methemoglobinemia in Pomeranian Dogs Caused by a Missense Variant in the NADH‐Cytochrome B5 Reductase Gene

Background

In veterinary medicine, congenital methemoglobinemia associated with nicotinamide adenine dinucleotide (NADH)‐cytochrome b5 reductase (b5R) deficiency is rare. It has been reported in several breeds of dogs, but little information is available about its etiology.

Objectives

To analyze the NADH‐cytochrome b5 reductase gene, CYB5R3, in a Pomeranian dog family with methemoglobinemia suspected to be caused by congenital b5R deficiency.

Animals

Three Pomeranian dogs from a family with methemoglobinemia were analyzed. Five healthy beagles and 5 nonrelated Pomeranian dogs without methemoglobinemia were used as controls.

Methods

Methemoglobin concentration, b5R activity, and reduced glutathione (GSH) concentration were measured, and a turbidity index was used to evaluate Heinz body formation. The CYB5R3 genes of the affected dog and healthy dogs were analyzed by direct sequencing.

Results

Methemoglobin concentrations in erythrocytes of the affected dogs were remarkably higher than those of the control dogs. The b5R activity of the affected dogs was notably lower than that of the control dogs. DNA sequencing indicated that this Pomeranian family carried a CYB5R3 gene missense variant (ATC→CTC at codon 194) that resulted in the replacement of isoleucine (Ile) by leucine (Leu).

Conclusions and Clinical Importance

This dog family had familial congenital methemoglobinemia caused by b5R deficiency, which resulted from a nonsynonymous variant in the CYB5R3 gene. This variation (c.580A>C) led to an amino acid substitution (p.Ile194Leu), and Ile194 was located in the proximal region of the NADH‐binding motif. Our data suggested that this variant in the canine CYB5R3 gene would affect function of the b5R in erythrocytes.     

患红体病凡纳滨对虾肝胰腺NAGase基本性质的改变研究

[目的]为虾养殖过程中红体病的诊断和控制提供重要的理论依据。[方法]分别测定健康和患红体病的凡纳滨对虾（Litopenaeus vanname）的肝胰腺N-乙酰-β-D-氨基葡萄糖苷酶（EC3.2.1.52,NAGase）的比活力及其性质差异。[结果]2种来源NAGase的比活力、基本酶学性质等均存在差异。对虾患红体病后,肝胰腺NAGase的比活力和最适温度下降,但催化反应动力学常数Km、Vm值和活化能上升。[结论]该研究表明NAGase与对虾患病可能存在着密切关系。     

日粮中添加β-胡萝卜素对牦牛皮下脂肪颜色及组织中维生素A含量的影响

试验旨在探究日粮中添加不同水平β-胡萝卜素（β-C）对牦牛皮下脂肪颜色及组织维生素A含量的影响。试验选取12头2~3岁牦牛为研究对象，随机分为4个组，每组3个重复。各处理组日粮分别在基础日粮中添加0（对照）、720（低剂量）、1 440（中剂量）、1 620 mg/d（高剂量）β-C，饲养90 d后，分别测定牦牛脂肪、肌肉色度、背膘厚度及不同组织中β-C和代谢产物维生素A的含量，结果表明：日粮中添加不同水平β-C，牦牛背部、胸部脂肪及半腱肌的黄度值（b^*）显著高于对照组（P<0.05），亮度值（L^*）和红度值（a^*）没有显著变化（P>0.05）；牦牛背膘厚度在中剂量组达到最大值；对照组牦牛血清中β-C及维生素A含量显著低于各试验组（P<0.05），且维生素A含量随β-C添加量的上升呈上升趋势（P<0.05）；牦牛皮下脂肪、腹腔脂肪、心脏、肝脏、脾脏、肺脏、肾脏及空肠等组织器官中β-C含量在中剂量组均达到最大值，除肝脏中维生素A含量较高外，其他组织中维生素A含量在各处理组间差异不显著（P<0.05）。综上所述，日粮添加β-C影响牦牛皮下脂肪颜色，且在中剂量组牦牛皮下脂肪、心脏、肝脏、脾脏及空肠等组织器官中β-C含量达到最大值，故推荐其最佳摄入量为1 440 mg/d；在日粮中添加β-C影响牦牛肝脏和血清中维生素A含量，对其余组织器官中维生素A含量没有影响。     

白羊草R2R3-MYB转录因子的挖掘及对干旱胁迫反应的研究

本文以白羊草（Bothriochloa ischaemum）为材料，利用其目前已有的转录组序列，通过搜索、比对、功能域多态性分析、进化树构建和基因表达分析等手段，对白羊草R2R3-MYB型转录因子进行挖掘和干旱胁迫下的表达模式分析。结果从白羊草挖掘出43个R2R3-MYB转录因子，它们具有高度保守的[W]-X（19）-[W]-X（19）-[W]-X（n）-[W]-X（18）-[W]结构，主要参与生长发育、次生代谢和逆境响应等生物过程。通过进一步的分析，本研究筛选出8个可能参与干旱胁迫响应的R2R3-MYB转录因子。这为白羊草R2R3-MYB基因的功能研究和开发利用奠定了基础。     

普通烟草β-胡萝卜素羟化酶2基因克隆与生物信息学分析

β-胡萝卜素羟化酶(beta-carotene hydroxylase,BCH)是植物类胡萝卜素合成代谢中的关键限速酶,为获得烟草β-胡萝卜素羟化酶2(NtBCH2)基因序列,采用同源克隆法从烟草中分离NtBCH2的基因组DNA序列,基因登录号为JX101477。结果表明:1)同源克隆法从烟草中克隆出NtBCH2基因,NtBCH2基因组DNA全长2131bp,cDNA为1 083bp,含7个外显子和6个内含子。2)NtBCH2蛋白有309个氨基酸,分子量为34.79kD,具有7个糖基化位点,11个磷酸化位点,4个跨膜域。3)系统发育树与BLAST分析表明,NtBCH2是番茄的SlBCH和辣椒CaBCH2的同源基因;GENEVESTIGATOR数据库芯片结果表明,NtBCH2在幼嫩的茎和子叶中表达量最高。     

Study on Developmental Expression of ApoCⅡ Gene mRNA in Liver Tissues of Pigs

The aim of this study was to investigate the developmental patterns of ApoCⅡ gene mRNA in liver in Mashen and Large White pigs, and study the relationship between the expression level of ApoCⅡ and the lipid metabolism in pigs.The mRNA relative expressions of ApoCⅡ gene in liver at seven stages of 1,30,60,90,120,150,and 180-day old in Mashen and Large White pigs were determined by quantitative Real-time PCR.The results showed that the developmental trend of ApoCⅡ mRNA expression in liver between Mashen and Large White pigs was different.The ApoCⅡ mRNA abundance was decreased from birth to 60-day old,then increased at 90-day old,and decreased again after that in Mashen pig.However,the relative expression amount in Large White pig was gradually decreased from birth to 150-day old and increased again at 180-day old.Except for the ApoCⅡ mRNA expression amount at 1-day old,the differences of the expression amount at other stages in Mashen and Large White pigs were significant or extremely significant (P<0.05 or P<0.01).The ApoCⅡ mRNA expression in liver was affected by age and breed,and could play an important role in lipid metabolism in pigs.     

重组猪抑制素蛋白质中毒素残留检测及其理化稳定性

为探索纯化的重组猪抑制素蛋白质毒素残留情况以及重组猪抑制素蛋白质稳定性,本研究利用鲎试剂、气相色谱和SDS-PAGE方法对重组猪抑制素蛋白质中的细菌内毒素、β-巯基乙醇的残留量及其蛋白质在不同温度、极端p H值和反复冻融条件下的稳定性进行检测。结果显示,经过3次等电点沉淀洗涤后,重组猪抑制素蛋白质中细菌内毒素和β-巯基乙醇的残留量大幅减少。通过在不同温度下的处理发现,重组猪抑制素蛋白质在4℃环境下较稳定,可以长期保存;但在37℃时,稳定性快速下降,随着温度上升而降解加速。通过极端p H值和反复冻融试验发现,重组猪抑制素蛋白质对极端p H值和反复冻融具有较好的耐受性。     

人参中人参皂苷Ro和姜状三七皂苷R_1的含量测定

目的测定不同种类人参,人参不同部位中,红参加工过程中人参皂苷Ro(简称Ro)和姜状三七皂苷R1(简称R1)的含量含量。方法:采用HPLC的方法,对样品预处理及色谱条件进行优化。结果:鲜参中Ro和R1含量比红参中含量高;不同部位中Ro和R1的含量差异为:芦头参须主根;红参加工过程中的蒸制使Ro含量升高,R1含量降低。结论本文所建立的人参中人参皂苷Ro和姜状三七皂苷R1的含量测定方法准确、稳定可靠。