沉淀法制备的紫杉醇白蛋白微粒包封率测定

采用沉淀法制备紫杉醇白蛋白微粒,采用高效液相色谱法(HPLC)测定紫杉醇质量浓度,以二氯甲烷为有机相,采用萃取法分离白蛋白微粒与游离药物,测定其包封率。结果表明:紫杉醇在10~100 mg·L-1范围内线性良好,平均回收率可达到97.6%~101.0%;有机溶剂萃取法测定紫杉醇白蛋白微粒包封率的平均回收率,达94.80%~101.67%。这种方法测定紫杉醇白蛋白微粒回收率准确可靠,适用于沉淀法制备的白蛋白微粒的包封率测定。     

马铃薯AP2/ERF转录因子的克隆及植物表达载体构建

AP2/ERF基因家族转录因子普遍存在于植物中,参与植物体内的各种生物学过程,包括植物的生长发育、生物和非生物胁迫响应等。前期转录组测序的结果发现马铃薯‘加湘1号’一个AP2/ERF家族基因(PGSC0003DMG400012154)在接种晚疫病菌(Phytophthora infestans)24 h后被显著激活。从接种P.infestans 24 h的‘加湘1号’的总RNA中通过RT-PCR获得了该基因CDS序列为885 bp,BLAST分析显示该基因编码一个295个氨基酸残基的蛋白,并且含有1个AP2/ERF结构域,是AP2/ERF转录因子家族中的ERF亚家族的一员。本研究将该基因与XcmⅠ酶切的表达载体pCXSN连接,转化大肠杆菌,通过测序挑选插入正确克隆酶切验证,并成功转化农杆菌。本研究结果为进一步研究该基因的功能提供了帮助。     

川西北地区上二叠统吴家坪组凝灰岩分布地震预测

通过对川西北地区的老井进行复查,发现多口井在上二叠统吴家坪组存在大套的凝灰岩,对上述井的地质、测井、地震资料综合分析发现,凝灰岩的岩性以凝灰质泥岩、凝灰质粉砂岩、凝灰质灰岩、沉凝灰岩、凝灰岩、凝灰质白云岩为主,次生溶蚀孔隙发育;凝灰岩在测井曲线和地震剖面上均有明显的响应特征:在测井曲线上表现为明显的低纵波速度、高自然伽马特征,凝灰岩底界在地震剖面上均表现为波峰反射特征.为此,采用模型正演分析技术及波阻抗反演技术,首次对四川盆地川西北地区晚二叠世吴家坪组凝灰岩的分布进行了地震预测:双鱼石构造-剑阁区块的北部地区凝灰岩分布最厚,南部减薄;双鱼石构造东部凝灰岩分布较厚,西部减薄,直至不发育.研究结果表明,晚二叠世吴家坪组在川西北地区沉积了一套凝灰岩,从西北至东南方向减薄;由于凝灰岩物性好,孔隙度高,可成为四川盆地火山岩气藏勘探全新的后备领域,为川西北地区下一步新的勘探领域开发提供了重要研究基础.     

养分管理对安溪茶园土壤肥力及茶叶品质的影响

安溪是铁观音的主产区,茶园养分管理滞后已成为该地区茶叶生产的瓶颈,本研究旨在为该地区茶园的养分管理提供科学依据。试验于2015年在安溪县采集了50个茶园的茶青与土壤样品,测定了土壤5项主要肥力指标和茶青中9种主要次生代谢物含量;并根据茶农氮肥用量调研数据,初步将茶园养分管理划分为:少量、中量和过度型等三种方式。限制性主坐标轴分析发现,此分类方式可解释34.4%(P0.001)茶园土壤肥力参数的总体差异,说明此分类方式能反映出不同茶园养分管理的总体水平;并且,养分管理对表层土壤有效磷的影响最为显著。进一步分析表明,养分管理对茶青综合品质的影响可解释品质总差异的7.48%(P0.001);大部分茶青次生代谢物在中量型管理下含量最高。说明养分管理影响安溪茶园土壤的肥力状况,施肥量过高或过低均不利于高品质茶叶的生产,该地区的建议施氮量约为200～400 kg·hm~(–2)·a~(–1)。     

贪食迈阿密虫(M.avidus)对4种海水养殖鱼类组织匀浆液的趋化性

通过对贪食迈阿密虫(M. avidus)对4种养殖鱼类组织匀浆液的趋化特性进行研究,以探讨贪食迈阿虫对养殖鱼类的侵染特点。试验结果显示,贪食迈阿密虫对大黄鱼、大菱鲆、黄姑鱼、鲈鱼的眼、脑、脑脊液匀浆液有强烈的趋化效果(P<0.01),对皮肤黏液、鳃、肝、肌肉、肾组织匀浆液的趋化性较弱(P>0.05);对不同鱼类组织匀浆液对贪食迈阿密虫的趋化诱导差异比较结果推测,贪食迈阿密虫对宿主具有一定的选择性。本研究有助于深化贪食迈阿密虫致病性的认识,并为海水养殖鱼类盾纤虫病的预防提供参考资料。     

Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.     

工程设施技术在稻渔共作模式中的发展

稻渔共作模式是一种继承传统农业中遵循自然协调、绿色生态的农业发展理念并以现代农业技术为指导的生态循环农业模式，该模式具有稳粮、促渔、增收、提质、环境友好和可持续发展等多种生态系统功能，但缺乏稻田生态理论研究及新的生态管理技术，且基础设施不足，与种养技术匹配设施缺乏。工程设施技术属于稻渔共作模式配套技术，决定稻渔共作模式的发展潜力。目前工程设备设施在稻渔共作模式中的应用普及程度低，尚未开发出与稻渔共作模式特点相匹配的专用设备。结合稻渔共作模式的发展历史和水平，对配套工程设施技术的发展进行分析。      

Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino

Single‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.     

一个潜在催化莱鲍迪D苷合成的新型糖基转移酶

尿苷二磷酸糖基转移酶(uridine diphosphate glycosyltransferases,UGTs)催化糖基转移反应,与植物次生代谢密切相关。本研究根据甜叶菊(Stevia rebaudiana)转录组数据库,克隆到一个催化莱鲍迪D苷(rebaudioside D,RD)合成的新型糖基转移酶候选基因,对其开展生物信息学分析。结果表明,该基因开放阅读框长1380 bp,编码459个氨基酸,等电点(pI)预测为5.54,理论分子量约49.66 kD,系统发育分析表明该基因与向日葵中的UGT89A2同源,故将其命名为SrUGT89A2。构建pET28a-SrUGT89A2原核表达载体,并在大肠杆菌(BL21(DE3))中诱导表达得到重组蛋白,HPLC检测表明粗酶液能催化甜叶菊提取液形成一个新的色谱峰,该峰保留时间与莱鲍迪D苷一致。经进一步纯化UGT89A2蛋白,添加不同甜菊糖苷标准品为催化底物,但未鉴定出该蛋白催化的具体糖苷。该潜在催化甜菊糖RD苷合成的新型糖基转移酶基因SrUGT89A2的发现,为RD苷的生物合成和甜菊糖苷的生物途径研究提供新的理论依据。     

山羊肺淋巴系的研究

24例山羊肺经肺实质和肺胸膜下注射30％普鲁士蓝氯仿溶液，剖查其器官内淋巴管及淋巴流向。结果表明，山羊肺的器官内淋巴管有浅淋巴管和深淋巴管2种。左肺尖叶淋巴管注入左支气管肺淋巴结和气管支气管左淋巴结，左肺心叶淋巴管注入左支气管肺淋巴结、气管支气管左淋巴结和气管支气管中淋巴结，左肺膈叶淋巴管注入气管支气管左淋巴结、气管支气管中淋巴结和纵隔后淋巴结；右肺尖叶淋巴管注入右支气管肺淋巴结和气管支气管前淋巴结，右肺心叶淋巴管注入右支气管肺淋巴结和气管支气管右淋巴结，右肺膈叶淋巴管注入气管支气管右淋巴结、气管支气管中淋巴结和纵隔后淋巴结，右肺副叶淋巴管注入右支气管肺淋巴结、气管支气管中淋巴结和纵隔后淋巴结。