聚多曲霉菌原生质体和液泡的制备及优化

原生质体的大量制备是研究原生质体转化、诱变、融合等技术的关键,液泡的制备对研究液泡中分解、转运有机物的特性具有重要意义,但目前分离技术还不够成熟.本研究从聚多曲霉菌菌丝中分离原生质体和液泡并对分离条件进行优化,以聚多曲霉菌DJ515-2菌丝为材料,探索不同因素对原生质体和液泡制备分离效果的影响.结果表明,聚多曲霉菌DJ515-2在菌龄42 h,以3％纤维素酶、1％蜗牛酶和3％的溶壁酶组成复合酶液,25℃下酶解4 h,原生质体达到最大产量,为5.167×105个/mL.同时,在此基础上裂解液泡的最优条件为pH7.5、1.0mol/L KCl、0.020％Triton X-10,其产量达2.63×105个/mL,产率为原生质体的50.8％,相比采用多元碱化合物诱导真菌原生质体裂解释放液泡的产率增加了 40％～45％.本研究为真菌和植物的各种原生质体技术及基于亚细胞层面的研究提供了材料基础.     

缅甸稻飞虱的发生和防控情况(英文)

稻飞虱是缅甸水稻种植区常见且分布广泛的一类害虫,可造成农作物不同程度的减产.稻飞虱的爆发与高产品种的大规模推广种植具有一致性,同时其种群变化也与天气条件有关.近年来,缅甸中部部分省份稻飞虱大量孳生,在一定程度上,这与氮肥施用水平有关.雨季稻飞虱种群增加,7月和8月为高峰期.在缅甸,主要通过培育抗虫、抗旱、抗逆等水稻品种来防控稻飞虱.同时用诱虫灯进行早期入侵的虫源的监测,必要时,采用化学杀虫剂防治.缅甸部分农场还采用了病虫害综合管理系统(IPM),以建立健康、安全、可调节的水稻生态系统及可持续的病虫害管理.     

Evolution and diversity of axon guidance Robo receptor family genes

The diversity of axon guidance (AG) receptors reflects gains in complexity of the animal nervous system during evolution. Members of the Roundabout (Robo) family of receptors interact with Slit proteins and play important roles in many developmental processes, including AG and neural crest cell migration. There are four members of the Robo gene family. However, the evolutionary history of Robo family genes remain obscure. We analyzed the distribution of Robo family members in metazoan species ranging in complexity from hydras to humans. We undertook a phylogenetic analysis in metazoans, synteny analysis, and ancestral chromosome mapping in vertebrates, and detected selection pressure and functional divergence among four mammalian Robo paralogs. Based on our analysis, we proposed that the ancestral Robo gene could have undergone a tandem duplication in the vertebrate ancestor; then one round of whole genome duplication events occurred before the divergence of ancestral lamprey and gnathostome, generating four paralogs in early vertebrates. Robo4 paralog underwent segmental loss in the following evolutionary process. Our results showed that Robo3 paralog is under more powerful purifying selection pressure compared with other three paralogs, which could correlate with its unique expression pattern and function. Furthermore, we found four sites under positive selection pressure on the Ig1‐2 domains of Robo4 that might interfere with its binding to Slits ligand. Diverge analysis at the amino acid level showed that Robo4 paralog have relatively greater functional diversifications than other Robo paralogs. This coincides with the fact that Robo4 predominantly functions in vascular endothelial cells but not the nervous system.     

中国人胎盘组织ABCB1基因多态性与P-gp表达水平的相关性

为了探讨中国人群胎盘组织中ABCB1基因单核苷酸多态性与其mRNA及蛋白表达水平的相关性,为妊娠期安全用药提供理论依据,本研究选取2016年6月至2018年5月于昆明医科大学第一附属医院正常分娩的200例中国汉族妇女的胎盘组织为研究对象,剪取母体侧胎盘组织,用于DNA、RNA和蛋白质的提取.通过测序确定ABCB1基因rs1045642、rs1128503及rs2032582多态性位点的基因型;采用实时荧光定量PCR和Western blot法测定组织中P-gp的mRNA和蛋白表达水平;对ABCB1基因多态性与P-gp的mRNA及蛋白表达水平的相关性进行分析.研究发现,对于rs1045642、rs1128503及rs2032582多态性位点,各基因型之间P-gp的mRNA及蛋白表达水平差异均无统计学意义(F=0.813,0.219,P＞0.05;F=0.132,0.086,P＞0.05;F=1.617,0.794,P＞0.05).本研究表明ABCB1 rs1045642、rs1128503及rs2032582基因多态性与中国汉族人群胎盘组织P-gp mRNA及蛋白表达水平可能无关.     

Congestive heart failure in COX2 deficient rats

Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit prostaglandin (PG) formation by targeting cyclooxygenase (COX) 1 and 2.Long-term use of NSAIDs that selectively inhibit COX2 increases the risk for thrombotic events,cardiac failure,and hypertension.However,the underlying mechanisms remain unclear.In this study,COX1- and COX2-deficient rats were created via Cas9/RNA-mediated gene targeting.DNA genotyping and Western blot analysis confirmed successful generation of COX1~(-/-)and COX2~(-/-)rats.Adult COX1~(-/-)rats grew normally,while more than 70%of COX2~(-/-)rats after wean died within 2 months.Echocardiography showed markedly reduced left ventricular ejection fraction and fractional shortening in adult COX2~(-/-)rats compared to those in wildtype (WT) controls.Histological analysis revealed accumulation of inflammatory cells and severe interstitial and perivascular fibrosis in COX2~(-/-)cardiac tissues.Moreover,cardiac ATP and acetyl-Co A production was dramatically decreased in COX2~(-/-)rats.Consistently,the expression of genes related to mitochondrial oxidation,such as those that encode for subunits of pyruvate dehydrogenase complex and acyl Co A dehydrogenases,were downregulated,while glycolytic hexokinase 1 (HK1) was upregulated in COX2~(-/-)heart tissues.These observations indicate that COX2-deficient rats developed spontaneously heart failure,likely as a result of dysregulated cardiac energy metabolism.     

基于CRISPR/Cas系统的单碱基编辑技术研究进展*

基于CRISPR/Cas系统出现的单碱基编辑技术可以实现高效且简便的单个碱基的替换编辑,其原理是将胞嘧啶脱氨酶(cytosine deaminase)或腺苷脱氨酶(adenosine deaminase)与Cas9n(D10A)形成融合蛋白,通过CRISPR/Cas精准识别和定位DNA上的靶位点后,利用胞嘧啶脱氨酶或腺苷脱氨酶将靶点距离sgRNA位点基序(protospacer adjacent motif,PAM)序列端的4~7位的单个碱基发生单碱基转换或颠换。对基于CRISPR/Cas系统的单碱基编辑技术发现的历史、组成和分类、工作原理进行了概述,并总结了该系统最新进展及应用。     

甘肃省大豆品种生育期组的划分及地理分布研究

甘肃省地域广大,地形复杂,气候多样,大豆品种资源丰富,但甘肃各地大豆品种生育期生态类型并不明确。本试验以25份MG 0~Ⅴ的北美大豆生育期组标准品种为对照,将从甘肃各地收集到的151个大豆品种在甘肃境内5个生态区进行生育期性状鉴定和生育期组划分。结果表明,甘肃地区种植的大豆品种的生育期组在MG 00和MGⅥ之间。在所鉴定的品种中,属MGⅣ的比例(44.37%)最高,其次为MGⅤ(29.14%)、MGⅢ(11.26%)、MGⅡ(4.64%)、MG 0(3.31%)、MGⅥ(3.31%)、MG 00(2.65%)和MGⅠ(1.32%)。归属于MGⅢ的品种主要分布在河西灌区,占该区域参试品种总数的47.62%;归属于MGⅣ的品种主要分布在中部地区和陇东旱塬区,占中部地区参试品种总数的56.67%,占陇东旱塬区的63.83%;归属于MGⅤ的品种主要分布在陇南地区,占该区域参试品种总数的61.40%。根据甘肃不同地区可成熟的最晚熟大豆品种的生育期组与当地经度、无霜期建立回归方程,推断甘肃省大豆各生育期组的地理分布区间,提出了基于生育期组的甘肃省大豆品种引种方案,以期为甘肃地区大豆育种和品种布局提供科学依据。     

一种六妹羊肚菌的新型柄腐病害

近年来,四川等地种植的六妹羊肚菌先后爆发柄腐病害,造成严重经济损失。本研究对该病害发生情况进行了调查,从四川、河南、甘肃、贵州4个省收集的12份六妹羊肚菌柄腐病子实体,分离纯化得到37株真菌和3株细菌。根据柯赫氏法则,对所有分离纯化得到的疑似病原菌进行了回接试验,最终确定GJB-3菌株为柄腐病的病原菌。经ITS、RPB2和EF1α序列分析,结合菌落形态和显微特征鉴定柄腐病的病原菌为Fusarium nematophilum。本研究为后期羊肚菌生产病害防控提供了有效的基础数据。