利用CRISPR/Ca9技术靶向编辑芥蓝BoaZDS

以芥蓝(Brassicaoleraceavar.alboglabra)为材料,以ζ–胡萝卜素脱氢酶(ζ-Carotene desaturase,ZDS)基因为目标基因,建立其CRISPR/Cas9基因组编辑体系。在BoaZDS的编码区近5′端选择靶位点,构建了CRISPR/Cas9表达载体,通过农杆菌介导的遗传转化方法获得了19个芥蓝转基因阳性植株,Sanger测序分析发现其中13株成功突变,CRISPR/Cas9载体在芥蓝上的突变效率为68.42%,且所有突变植株均表现出明显的白化表型。     

Effects of ZIP14 on biological behaviors of hepatocellular carcinoma cell line BEL-7404

AIM: To study the expression of zinc transporter ZRT/IRT-like protein 14 (ZIP14) in the hepatocellular carcinoma (HCC) tissues, and to investigate the effects of ZIP14 over-expression on the biological behaviors of HCC cells. METHODS: The expression of ZIP14 at mRNA and protein levels in the HCC tissues and adjacent non-tumor tissues were detected by real-time PCR and immunohistochemical staining, respectively. The lentivirus expression system containing GV365-ZIP14 was constructed, and was used to infect the HCC cell line BEL-7404, which had relatively poor expression of ZIP14. The expression of ZIP14 at mRNA and protein levels in the transfected cells were detected by real-time PCR and Western blot, respectively. Under the conditions of zinc sulfate stimulation at different concentrations, the cell viability, the cell cycle, and the cell migration and invasion abilities were detected by MTT assay, DNA ploid detection, and Transwell assay, respectively. RESULTS: The mRNA expression level and the strong-positive rate of protein expression of ZIP14 in the HCC tissues were significantly lower than those in the adjacent non-tumor liver tissues (P<0.01). The expression of ZIP14 at mRNA and protein levels in the BEL7404 cells was significantly enhanced by infection of GV365-ZIP14 expression lentivirus. Compared with negative control group (transfected with negative control lentivirus), the cell viability, migration and invasion in ZIP14 over-expression group (transfected with GV365-ZIP14 expression lentivirus) were significantly reduced, and the percentage of the cells in G₂/M phase was significantly increased, all of which were more obvious with the elevation of zinc concentration in the culture medium. CONCLUSION: ZIP14 is low expressed in the HCC tissues. The ZIP14 over-expression has inhibitory effects on the viability, migration and invasion of HCC cells, and blocks the cell cycle in G₂/M phase, which might be closely related to the elevation of zinc concentration in cytoplasma of HCC cells due to enchanced zinc transport by ZIP14.     

miR-204 inhibits proliferation of Hodgkin lymphoma cells by targeting Sirt1/p53 signaling pathway

AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53.     

不同嫁接方法对番茄嫁接工效和嫁接苗种苗质量的影响

为获得一种适应番茄集约化嫁接育苗的高效嫁接方法,以中研红2号为接穗和砧木,同时播种,比较了劈接、套管嫁接和贴接3种嫁接方法对番茄嫁接工效和嫁接苗质量的影响。结果表明,贴接法的嫁接速率和嫁接成活率显著高于劈接法和套管嫁接法;3种嫁接方法的嫁接苗形态指标无显著差异;嫁接后22 d,贴接法嫁接苗相对叶绿素和可溶性糖含量显著高于劈接法和套管嫁接法。综合来看,砧木和接穗苗龄相同时,番茄贴接法的嫁接工效最高,嫁接苗质量最好,可在生产中推广应用。     

辣椒花药开放式培养及不同基因型对花药培养的影响

为了创新花药培养方法,并用于筛选出再生频率较高的辣椒品种,通过改进花蕾灭菌和花药接种方式,以及向NTH培养基中添加抑菌剂,建立了1种辣椒花药开放式培养的方法,具体内容为:对剥去3/4花萼的花蕾灭菌时更易杀死花蕾表面细菌和内生菌;徒手挤压花药降低了对花药的伤害,且花蕾接种速度为镊子夹取处理的2.18倍;添加25 mg/L的氨苄青霉素和25 mg/L的两性霉素B可使花药染菌率降低至1.42%;简化了培养环节,可用于大规模胚状体诱导和双单倍体植株的获得。同时,比较了23个辣椒品种的花药培养效果,筛选到了3个再生频率高的辣椒品种,分别为博辣红牛、H15和辣八螺丝王。     

施肥对雨养区马铃薯干物质累积及肥效的影响

针对宁夏雨养农业区,选择具有代表性的海原县树台乡马铃薯种植基地,设置氮磷钾肥不同施肥处理,于2016~2017年开展了马铃薯田间试验,探讨雨养区施肥对马铃薯干物质累积及肥效的影响。试验结果表明,不同施肥处理对马铃薯干物质积累、产量及水肥利用效率影响不同,其中氮磷钾肥配施处理马铃薯干物质快速积累期增长速度最快,施用氮、磷、钾在淀粉积累期马铃薯块茎累积量分别提高了36.4%、40.5%、18.5%。两年期间,氮磷钾肥配施处理的水分利用效率最大,2016、2017年较无肥区分别增长35.3%、51.0%,平均增长43.2%。在氮磷钾配施条件下,氮、磷、钾肥增产量分别是5 496.6 kg/hm^2、4 739.4 kg/hm^2、3 044.1 kg/hm2,其增产率分别为31.1%、30.6%、17.1%。每千克N、P2O5、K2O分别增产马铃薯30.5 kg、52.7 kg、67.7 kg。氮、磷、钾肥利用率分别为38.3%、18.2%、53.9%,其中钾肥利用率较高。     

不同LED光质对紫甘蓝幼苗生长的影响

以紫甘蓝品种‘紫辉甘蓝’为试材,在南方塑料大棚内密闭式光照植物培养架中,采用新型LED光源研究了不同光质对紫甘蓝幼苗生长的影响,试验共设置6个处理:R(红)、B(蓝)、8R2B(红光∶蓝光=8∶2)、5R5B(红光∶蓝光=5∶5)、2R8B(红光∶蓝光=2∶8)和荧光灯(对照CK)。结果表明:(1)在8R2B处理下,紫甘蓝幼苗地下部鲜重、地上部鲜重、单株鲜重、株高、根体积、根总表面积和根投影面积均最大;(2)与CK相比,不同光质处理均提高了幼苗叶片叶绿素a、叶绿素b和类胡萝卜素的含量,以2R8B处理最高;(3)与CK相比,除R处理的幼苗可溶性蛋白质含量降低外,其它处理均有助于幼苗可溶性蛋白质含量的积累,以8R2B处理最高;(4)可溶性糖含量以R处理最高;(5)与CK相比,各光质处理均显著降低了幼苗MDA含量,以B处理效果最显著,8R2B和5R5B次之;(6)各光质处理对紫甘蓝幼苗叶片脯氨酸含量的影响与CK相比无显著性差异。     

漳平水仙茶饼多酚的纯化及其体外活性研究

为了研究大孔树脂对水仙茶饼多酚的纯化效果,试验以水仙茶饼多酚粗提液为原料,比较了9种不同型号大孔树脂对水仙茶饼粗多酚的静态吸附和解吸性能,再以吸附率和解吸率为指标通过静态、动态吸附解吸试验。考察了各因素对LX-28树脂纯化水仙饼茶粗多酚的影响,最后探讨了纯化前后的水仙茶饼多酚的体外抗氧化和抑菌效果。结果表明:最佳纯化树脂LX-28,最佳吸附和解吸条件为1.5 mg/mL的质量浓度的水仙茶饼多酚粗提取液(pH4.0~5.0)以2.0 mL/min的流速上样95 mL,吸附平衡后去离子水冲洗至无色,再用70 mL,65%体积分数乙醇溶液(pH6.0)为洗脱剂,以3.0 mL/min流速洗脱,在此纯化条件下LX-28树脂对水仙茶饼多酚的吸附率和解吸率分别为(93.587±0.379)%和(95.330±1.282)%,树脂可重复使用4次,纯度提高了约3.04倍;对DPPH·和ABTS+·自由基清除率的IC50值分别从纯化前的1.279、3.682 mg/mL降低到0.295、1.525 mg/mL;对供试的大肠杆菌、金黄色葡萄球菌、米根霉和紫红曲霉均有不同程度的抑制作用,纯化后的水仙茶饼多酚的抑菌效果优于纯化前。     

白茶寿眉预防小鼠肥胖作用研究及安全性评价

取1、3、7年寿眉(白茶)制备白茶提取物(White tea extracts,WTEs)。在对雄性C57BL/6J小鼠喂饲高脂饲料的同时给予WTEs干预,诱导建立预肥胖模型。观察小鼠体质量增量、摄食量、脂肪和肝脏组织病理学变化,统计脂肪湿重,测定血脂及血转氨酶水平。q PCR测定小鼠肝脏中脂代谢相关基因表达。综合评价WTEs预防小鼠肥胖功效及其安全性,并对其机制进行探究。结果显示,WTEs在无肝毒性和生长抑制作用条件下,能抑制脂肪细胞分化和脂肪的过度积累,还能下调SREBP-1c、FAS和ACC1的转录水平,抑制肝脏脂肪酸合成和脂肪过度积累,有效预防肥胖和脂肪肝的发生。高剂量下,WTEs预防肥胖功效随白茶年份的增加而显著降低。     

绿茶、EGCG预防及缓解香烟烟雾诱导COPD的研究进展

慢性阻塞性肺疾病是由吸烟引发的最常见肺部疾病之一。香烟烟雾诱导的氧化应激、炎症反应及蛋白酶/抗蛋白酶失衡与慢性阻塞性肺疾病的发生、恶化密切关联。饮用绿茶、摄入EGCG能够清除人体内活性氧自由基,预防及缓解其带来的一系列呼吸道疾病。本文总结了近10年来众多学者对绿茶、EGCG作用于慢性阻塞性肺疾病的研究文献,以期探讨绿茶、EGCG预防及缓解慢性阻塞性肺疾病的机理,提高人们对绿茶、EGCG预防及缓解慢性阻塞性肺疾病的认识。