构建基于CRISPR/Cas9技术敲除ALOX5的质粒

旨在利用CRISPR/Cas9技术构建敲除花生四烯5-脂氧合酶基因(Arachidonate 5-lipoxygenase gene,ALOX5)的重组质粒。设计合成3对靶向敲除ALOX5第六外显子的sgRNA,将其分别插入到CRISPR/Cas9质粒骨架pX458载体中,转化感受态大肠杆菌DH5α后挑取克隆,通过测序评估重组质粒是否构建成功。将构建好的重组质粒转染293T细胞,在荧光显微镜下观察转染效果,挑取转染成功的细胞,用试剂盒提取转染细胞基因组DNA,PCR扩增含敲除位点的DNA片段,用测序技术获得核苷酸序列,用DNAStar软件分析转染细胞中ALOX5基因敲除情况。测序结果表明2对双链sgRNA寡核苷酸已插入质粒,且序列正确,靶向ALOX5基因的重组质粒pX458-sgRNAs-ALOX5构建成功。其在293T细胞中的转染效率约为50%,用一代测序法未检测到sgRNAs的切割效果。初步表明利用CRISPR/Cas9技术成功构建靶向ALOX5基因的重组质粒pX458-sgRNAs-ALOX5。     

SD雌性大鼠性发育早期性器官等脏器和性激素的动态变化

目的探讨正常SD雌性大鼠性成熟前不同日龄段的脏器与促黄体生成素（LH）、促卵泡素（FSH）、雌二醇（E2）等性激素的变化及其关系。方法从生产群中取出60窝密度状态一致的SD大鼠,在不同日龄随机选取雌性大鼠,检测15、25、32、40日龄时大鼠体重、主要脏器指数,子宫、卵巢组织变化和15、25、32、40、60日龄大鼠血清LH、FSH、E2水平。结果记录了SD雌性大鼠性成熟前各脏器指数和卵巢、子宫组织变化,结果显示大鼠卵巢、子宫的增长速度大于体重的增长,而其他脏器增速大都小于体重的增长。本研究还记录了血清LH、FSH、E2水平在不同日龄段的变化规律,表明血清LH、E2浓度在32日龄时出现较为明显升高。结论不同日龄大鼠脏器指数的动态变化提示大鼠性器官在性发育早期得到机体的优先发育。血清LH、E2水平在32日龄时有了明显升高,提示性腺轴功能已经激活。60日龄大鼠血清性激素水平的波动类似于动情周期的规律性变化,推测大鼠在60日龄前即已进入性成熟,这些结果将为大鼠性发育的相关研究提供重要的参考数据。     

Effects of an exotic invasive macrophyte (tropical signalgrass) on native plant community composition,species richness and functional diversity

1. The issue of freshwater species being threatened by invasion has become central in conservation biology because inland waters exhibit the highest species richness per unit area, but apparently have the highest extinctions rates on the planet. 2. In this article, we evaluated the effects of an exotic, invasive aquatic grass (Urochloa subquadripara– tropical signalgrass) on the diversity and assemblage composition of native macrophytes in four Neotropical water bodies (two reservoirs and two lakes). Species cover was assessed in quadrats, and plant biomass was measured in further quadrats, located in sites where tropical signalgrass dominated (D quadrats) and sites where it was not dominant or entirely absent (ND quadrats). The effects of tropical signalgrass on macrophyte species richness, Shannon diversity and number of macrophyte life forms (a surrogate of functional richness) were assessed through regressions, and composition was assessed with a DCA. The effects of tropical signalgrass biomass on the likelihood of occurrence of specific macrophyte life forms were assessed through logistic regression. 3. Tropical signalgrass had a negative effect on macrophyte richness and Shannon and functional diversity, and also influenced assemblage composition. Emergent, rooted with floating stems and rooted submersed species were negatively affected by tropical signalgrass, while the occurrence of free‐floating species was positively affected. 4. Our results suggest that competition with emergent species and reduction of underwater radiation, which reduces the number of submersed species, counteract facilitation of free‐floating species, contributing to a decrease in plant diversity. In addition, homogenisation of plant assemblages shows that tropical signalgrass reduces the beta diversity in the macrophyte community. 5. Although our results were obtained at fine spatial scales, they are cause for concern because macrophytes are an important part of freshwater diversity.     

不同施肥处理对‘赤霞珠’葡萄根际土壤养分和真菌群落的影响

【背景】宁夏贺兰山东麓葡萄产区忽视有机肥的施用,果树枝条焚烧污染环境,造成土壤养分缺失,土壤质量下降。【目的】为解决长期施用化肥对土壤造成的一系列问题,通过大田试验研究施肥及喷施不同浓度菌剂对土壤理化性质、真菌群落组成及多样性的影响,为酿酒葡萄可持续健康发展提供科学依据。【方法】以‘赤霞珠’葡萄根际土壤为试验对象,采用Illumina MiSeq高通量测序技术,测定并分析根际土壤理化性质、真菌群落组成和多样性在7个处理[常规施肥(CK)、蚯蚓粪+腐熟枝条+100倍菌剂(T1)、蚯蚓粪+腐熟枝条+200倍菌剂(T2)、蚯蚓粪+腐熟枝条+300倍菌剂(T3)、蚯蚓粪+未腐熟枝条+100倍菌剂(A1)、蚯蚓粪+未腐熟枝条+200倍菌剂(A2)和蚯蚓粪+未腐熟枝条+300倍菌剂(A3)]的变化。【结果】相较于CK,葡萄根际土壤理化性质差异明显,施肥处理增加了土壤有机质含量,土壤pH含量无明显变化,改良了土壤结构,活化了土壤有效养分。相较于CK,各处理真菌分类操作单元(operational taxonomic unit, OTU)数均降低,A2处理根际土壤丰富度及多样性均显著增加。真菌群落组成...     

豆科黄华属植物种子表面特征的研究

在扫描电镜下观察了豆科黄华属Thermopsis 18种植物种子的表面纹饰,发现 T．alpina,T.bar- bata,T．inflata,T．lupinoides,T. licentiana,T.smithiana和T．turkestanica的种子表面为粗网状,T californica,T.divaricarpa,T. macrophylla,T．mollis的种子表面为细网状,T.gracilis,T.montana,T. fabacea的种子表面为相对平滑型纹饰,T．alterniflora的种子表面为不规则条形,T．chinensis的种子表 面为粘膜状,T.rhombifolia的种子表面为条形及 T．viuosa的种子表面为碎屑状纹饰。结果表明黄华属的种子表面特征对属下类群的划分有一定意义,对澄清某些混乱的种有一定价值。     

H-FABP基因的多态性和营养因素对猪肉质的影响

遗传和营养因素都能影响猪肉的品质。但是, 到目前为止同时研究遗传和营养因素对肉质影响的报道很少。在本研究中, 136头PIC5系杂交猪, 体重65 kg, 被随机分成4组, 各组分别给予不同日粮。在饲养35 d、体重大约90 kg时统一屠宰并且进行肉质测定、H-FABP基因分型及其与肉质性状的关联分析。结果表明: (1)所采用的3种日粮对肉色、屠宰后24 h的pH、肌内脂肪和肌肉蛋白含量有极显著的影响; (2)H-FABP基因型对肌内脂肪和肌肉蛋白含量存在极显著的影响; (3)H-FABP基因多态性和营养因素的交互作用对pH 和肌内脂肪含量均有显著的影响, 对照组的AA基因型具有最高pH值, 高维生素E组的AA基因型具有最高肌内脂肪值。实验结果提示在关于猪肉质的育种和生产过程中应该同时考虑营养因素和遗传因素。     

Diet-induced obesity in rats leads to a decrease in sperm motility

Background  

Obesity is rapidly becoming a worldwide epidemic that affects children and adults. Some studies have shown a relationship between obesity and infertility, but until now it remains controversial. Thus, the aim of the present study was to investigate the effect of high-fat diet-induced obesity on male reproductive parameters.     

Z:ZCLA长爪沙鼠微卫星标记的遗传多态性研究

通过对9个微卫星座位的扩增,研究了Z：ZCLA长爪沙鼠的遗传多态性。结果表明,Z：ZCLA长爪沙鼠在其中1个位点上只有一个等位基因,在其它位点上均有2～4个等位基因,平均等位基因数2．6个。平均杂合度0．4684,平均多态信息量0．4166,平均有效等位基因数2．1756。全群基因纯合度从0．1111～0．5555,平均0．3389,提示目前本群遗传多样性水平处于中度多态。     

丹参内生拮抗细菌DS-R5对丹参根际和根表土壤细菌群落结构的影响CSCD

目的 探究生防细菌DS-R5施入丹参植株后根际和根表土壤细菌群落组成及多样性变化。方法 向丹参植株根部施入生防细菌DS-R5,以未施用细菌为对照组,分别采集根际和根表土壤样品提取总DNA,扩增样品总DNA的V3-V4区,采用Illumina MiSeq测序平台对PCR扩增产物进行双端测序分析,利用生物信息学分析解析丹参植株根际土壤和根表土壤细菌群落结构组成及多样性。结果 菌株DS-R5处理后增加了根际土壤细菌群落的多样性和丰度,降低了根表土壤细菌群落的多样性和丰度;高通量测序得到的根际和根表土壤的有效序列数量和OTU数量相比对照组均有所下降,根际土壤处理样品中微生物种类最丰富,根表土壤处理样品中微生物种类最少,根际土壤处理样品与根际土壤对照物种种类更接近;在门水平上,根际土壤处理样品相比对照变形菌门丰度下降,酸杆菌门丰度升高,根表土壤处理样品相比对照变形菌门和酸杆菌门丰度均升高,放线菌门丰度降低;在属水平上,根际土壤处理样品中鞘氨醇单胞菌属、芽胞杆菌属、慢生根瘤菌属相比根际土壤对照占比均有升高,根表土壤处理样品相比对照黄杆菌属和伯克菌属丰度下降,而土壤中的优势菌属根瘤菌属和芽胞杆菌属丰度升高。结论 丹参植株施用生防细菌DS-R5后,改变了根际土壤和根表土壤中微生物群落结构和多样性。