Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

论高校图书馆随书光盘的数字化管理

本文讨论了随书光盘的管理和利用的特点,结合多媒体阅览服务的实践,给出了随书光盘数字化管理服务中的思路和具体作法.     

~(86)R_b与K的作物吸收及其在土壤中移动的相关性

实验结果表明:西红柿、红麻、高梁和玉米作物之间对R_b与K吸收的相关系数为r=0.9842,各生育期对R_b与K吸收及分布的相关系数均在0.9以上,因此可以用R_b做K的示踪剂研究这些作物对钾的吸收。R_b与K在土壤中移动、分布的相关性受质地、水分状况影响较大,用R_b做示踪剂研究K在土壤中的移动要慎重。     

对高校图书馆随书光盘管理的探讨

本文从我馆对随书光盘管理模式经历的3大阶段进行探讨,建议从4大方面做好随书光盘的管理与应用:一是严格把守光盘入库的质量关,二是大力提升馆员的技术素质,三是培养读者对光盘的正确使用习惯,四是采用适合各馆的随书光盘下载管理系统.     

CD14沉默的猪小肠上皮细胞系的建立及其对大肠杆菌黏附能力的变化分析

白细胞分化抗原14(cluster of differentiation antigen 14,CD14)在先天性免疫和适应性免疫反应中都发挥重要调控作用,本研究旨在构建猪(Sus scrofa)CD14基因的短发夹RNA (short hairpin RNA,shRNA)干扰载体,包装成慢病毒,转染猪小肠上皮细胞,在细胞水平上进行相关功能探讨.本研究参照CD14基因(GenBank登录号:EF626695.1)全长编码区序列,设计4个靶向猪CD14基因的shRNA干扰序列,将其克隆至慢病毒表达载体质粒中,分别命名为pGLV3-CD14-1、pGLV3-CD14-2、pGLV3-CD14-3和pGLV3-CD14-4,以及阴性对照pGLV3-CD14-NC.包装成功后转染猪小肠上皮细胞系IPEC-J2,利用qRT-PCR检测其干扰效率.选择干扰效率最高的慢病毒进行药筛,获得CD14基因稳定沉默的猪小肠上皮细胞系.大肠杆菌(Escherichia coli)黏附实验检测大肠杆菌F18ab和F18ac对CD14基因沉默小肠上皮细胞黏附能力的变化.结果表明,本研究成功构建4个shRNA表达载体,包装成的慢病毒均能降低猪CD14mRNA表达水平,其中pGLV3-CD14-3慢病毒的干扰效果最好,其干扰效率达到94.6％;CD14基因沉默后F18ab对小肠上皮细胞的黏附能力显著增强,而F18ac虽然也有上调,但并未产生显著性差异.研究结果表明,CD14基因沉默后的小肠上皮细胞对大肠杆菌F18ab的黏附能力显著增强,CD14基因可能在小肠上皮细胞抵御大肠杆菌F18ab感染过程中发挥了重要的调控作用.慢病毒介导的CD14沉默的猪小肠上皮细胞系的建立,不仅为在细胞水平研究CD14基因功能提供了有效模型,也为进一步探究其所在的Toll样受体/白介素-1受体(toll-like receptors/interleukin-1 receptor,TLRs/IL-1 R)信号通路在猪肠道病原微生物引起的免疫反应和抵御病原入侵的调控机制奠定了基础.     

草鱼_中华鳖淋巴细胞扫描电镜结构_省略_绵羊红细胞受体_CD_2_的检测

草鱼淋巴细胞表面扫描电镜结构特征为有孔穴且较为光滑、具短小细锥状突起;中华鳖淋巴细胞、胸腺细胞表面扫描电镜结构特征为凹凸不平、较为光滑;它们均以表面凹凸不平特征的占大多数。E花环试验的光镜和扫描电镜观察显示：草鱼淋巴细胞E花环形成不明显,而中华鳖淋巴细胞、胸腺细胞的成花率分别为25％－34％和36％－47％,能形成花环的淋巴细胞、胸腺细胞以表面结构特征凹凸不平（类似哺乳动物T淋巴细胞）的为主。草鱼、中华鳖血液淋巴细胞和中华鳖胸腺细胞与抗人CD2单克隆抗体交叉反应的免疫组化检测显示中华鳖血液淋巴细胞和胸腺细胞的CD2阳性率分别为24％和33％。草鱼血液淋巴细胞CD2阳性反应不明显。     

西番莲果皮黄酮微胶囊的制备及其对运动耐力的影响

为提高西番莲果皮黄酮的稳定性,并考察其对机体运动的影响。以羟丙基-β-环糊精为壁材、西番莲果皮黄酮为芯材,采用喷雾干燥法制备西番莲果皮黄酮微胶囊,通过单因素与响应面试验确定该微胶囊的最佳制备工艺条件,同时采用动物实验研究其抗运动疲劳活性。结果表明,西番莲果皮黄酮微胶囊的最佳制备工艺条件为：芯壁比为1∶10（g/mL）,包埋温度49 ℃,搅拌时间53 min,进风干燥温度163 ℃。该条件下制得的西番莲果皮黄酮微胶囊外观略呈球形,大小基本均一,表面完整且光滑,包埋率达84.7%,平均粒径为7.52 μm,含水量为3.72%,堆密度为0.51 g/mL,休止角为35.12°。在相同储藏条件下,西番莲果皮黄酮微胶囊中黄酮类化合物的保留率高于其纯化物,表明微胶囊化有助于提高西番莲果皮黄酮的稳定性。通过不同组别小鼠的运动耐力考察指标结果对比分析发现,西番莲果皮黄酮微胶囊可明显延长小鼠的力竭游泳时间,减少运动后血清中乳酸与尿素氮的含量,从而帮助机体进一步提高运动耐力。该研究可为西番莲果皮的利用提供新的思路。     

猪瘟病毒Shimen株p80基因的克隆及其真核表达质粒的构建

根据已发表的猪瘟病毒(classicalswinefevervirus,CSFV)Alfort株和Brescia株的全基因组序列,设计2对引物P1/P2和B1/B2,在B1和B2的5''端分别加上XhoI和ApaI位点,以CSFVShimen株细胞毒为材料一步法提取总RNA,并以此为模板采用反转录PCR(RT-PCR)和套式PCR(nPCR),成功地扩增到约2.0kb的片段,将此PCR产物回收后与pMD18-T连接、转化,获得重组质粒,经PCR扩增,限制性酶切(BamhI/HindШ)和序列部分测定鉴定为阳性重组质粒P80-T。将P80-T分别经XhoI和ApaI酶切消化、回收后,与经XhoI/ApaI酶解的真核表达载体PEGPF-C1连接、转化,获得重组质粒,经PCR,XhoI和ApaI限制性酶切和序列测定鉴定为真核表达质粒P80-P,目的基因的插入位置、方向和读码框完全正确。为下一步在哺乳动物细胞中表达猪瘟病毒p80蛋白奠定了基础。     

Study on Antimicrobial and Antiviral Activities of Lysozyme From Marine Strain S-12-86 In Vitro

In this study, the in vitro antimicrobial and antiviral activities of the lysozyme from marine strain S-12-86 （LS） were investigated, The antimicrobial activity of LS was tested by minimum inhibition concentration （MIC） and minimum bactericidal concentration （MBC） method. The inhibiting effects of LS on pseudo rabies virus （PRV） in swine kidney cells （PK-15 ceils） were judged by cytopathogenic effect test （CPE）, The results showed LS had a broad antimicrobial spectrum against several standard strains including gram-positive bacteria, gram-negative bacteria, fungi, etc, The MIC of LS was 0.25-4.00 mg mL^-1 and its MBC was 0.25-8.00 mg mL^-1, respectively, Observation under the transmission electron microscope revealed that the cell wall of Candida albicans was distorted seriously, and the cytoplasm with many cavities was asymmetrical after being hydrolyzed by LS, The median cytotoxicity concentration （TC50） of LS was 100.0 μg mL^-1, the median effective concentration （EC50） was 0.46 μg mL^-1, and the selectivity index （TI = TC50/EC50） was 217. LS could inhibit PRV in PK-15 cells when it was added to cell culture medium at 0, 2, 4, 6, and 8 h after PK-15 cells had been infected by PRV. From the results, we concluded that LS had broad antimicrobial spectrum and good inhibiting effects on PRV,     

杂交稻新组合抗优80对褐稻虱抗性的初步研究

通过对褐稻虱苗期抗性测定,杂交稻新组合抗优80对褐稻虱表现抗性,抗性级别为1.0级,对褐稻虱存活率、取食量、种群建立等有很强的抑制作用;田间系统调查,抗优80上的虫量仅为感性品种汕优63上的1/7～1/20。因此,在褐稻虱中等或偏重发生年,抗优80上的虫量达不到防治指标,不需进行防治。