沙冬青AmCSDP基因特性及转基因烟草的抗寒性分析     

王红蕾  王艳萍  于婷乔  刘胜利  陈玉珍  卢存福 《园艺学报》2017,44(4):712-722

冷休克结构域蛋白(CSPs)是含有不同数量冷休克结构域(cold shock domain,CSD)的低温响应蛋白,在低温胁迫中发挥重要作用。以沙冬青(Ammopiptanthus mongolicus)为材料,克隆得到了冷休克结构域蛋白基因AmCSDP。生物信息学分析显示AmCSDP(Gen Bank登录号:KX756575)全长540bp,编码180个氨基酸,二级结构由8.9%的α–螺旋、47.8%的β–折叠和43.3%的无规则卷曲组成,其N端含有冷休克结构域。进化分析表明沙冬青与花生同源性最高。构建了AmCSDP真核表达载体Pcambia2300-35S-AmCSDP-OCS,经农杆菌介导转入本氏烟中。对转基因T1代和野生型烟草进行4℃低温胁迫处理,结果证实转基因本氏烟比野生型的耐冷能力强。  相似文献   

Role of TLR4 in process of colonic inflammation recovery induced by LPS     

LI Wang-lin  LIU Meng-ao  CAO Jie  YANG Ping  ZHENG Xiao-bin  DONG Bo-ye  LU Jia-bao 《园艺学报》2017,33(2):336-343

  相似文献   

Involvement of TLR4/NLRP3 inflammasome in contrast medium-induced inflammation and injury in renal tubular epithelial cells     

LIN Yan  LIN Jia-qiong  XIE Chu-li  GUAN Xiao-feng  TAN Xue-xian  HUANG Ze-na 《园艺学报》2017,33(12):2252-2258

AIM: To investigate whether Toll-like receptor 4 (TLR4) and Nod-like receptor protein 3 (NLRP3) inflammasome were involved in contrast medium (CM)-induced inflammation and injury in renal tubular epithelial cells. METHODS: Iopromide was used to injure NRK-52E cells in the study. The cell viability was measured by CCK-8 assay. The protein levels of TLR4, NLRP3, apoptosis-associated speckle-like protein (ASC), caspase-1 and cleaved caspase-3 were determined by Western blot. The releases of interleukin (IL)-1β and IL-18 were detected by ELISA. The apoptotic rate was evaluated by Hoechst staining, and mitochondrial membrane potential (MMP) was analyzed by JC-1 staining. siRNA was transfected into the NRK-52E cells to silence NLRP3 expression. RESULTS: CM decreased the viability of NRK-52E cells (P<0.05). CM also elevated the protein levels of cleaved caspase-3, TLR4, NLRP3, IL-1β and IL-18 (P<0.05). Silencing NLRP3 attenuated CM-induced releases of inflammatory cytokines. Moreover, treatment with TLR4 inhibitor TAK-242 or knockdown of NLRP3 by siRNA transfection both attenuated cell apoptosis and loss of MMP caused by CM. CONCLUSION: TLR4/NLRP3 inflammasome takes part in the pathogenesis of CM-induced acute kidney injury, and mediates CM-induced injury and inflammation in renal tubular epithelial cells.  相似文献   

Effect of xeroderma pigmentosum group D gene on proliferation of human umbilical arterial smooth muscle cells induced by Ox-LDL     

XIA Zi-rong  LI Qing  XIA Zhen  LI Ju-xiang  HONG Kui  WU Yan-qing  WU Qin-hua  CHENG Xiao-shu 《园艺学报》2017,33(12):2238-2244

AIM: To investigate the effects of xeroderma pigmentosum group D (XPD) gene on the proliferation of human umbilical arterial smooth muscle cells (HUASMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome. The cells were divided into blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL+pEGFP-N2 group and Ox-LDL+pEGFP-N2/XPD group. The proliferation rate of the cells was detected by MTT and EdU assays. The apoptotic rate and cell cycle distribution were analyzed by flow cytometry. The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with blank control group, the expression of XPD was increased in pEGFP-N2/XPD group (P<0.05). According to the results of MTT and EdU assays, the cell proliferation in pEGFP-N2/XPD group was reduced compared with blank control group (P<0.05). Compared with Ox-LDL group, the cell proliferation in Ox-LDL+pEGFP-N2/XPD group was significantly inhibited (P<0.05). According to the results of flow cytometry, the cell proportion of S phase decreased and the G₀/G₁-phase cell proportion increased significantly in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively (P<0.05). Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group, respectively (P<0.05). CONCLUSION: XPD inhibits the proliferation of HUASMCs and promotes their apoptosis, and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs. XPD may be the target for treatment of atherosclerosis.  相似文献   

Repressive effect of miRNA-363 via SOX4 on human osteosarcoma cell growth and apoptosis     

WAN Guang  YANG Hui  WU Da-peng  LIU Shen-peng  LIANG Qiu-dong 《园艺学报》2017,33(2):278-283

AIM: To detect the expression of miRNA-363 and SOX4 in osteosarcoma tissues and to investigate the effect of miRNA-363 on the viability and apoptosis of human osteosarcoma cell line MG-63.METHODS: Real-time PCR was used to detect the expression level and the relationship of miRNA-363 and SOX4 mRNA in the osteosarcoma tissues and the corresponding paratumorous tissues collected from 63 patients. The expression levels of miRNA-363 and SOX4 in osteosarcoma cell line MG-63 after transfected with miRNA-363 mimics were measured. The cell viability was measured by CCK-8 assay. Flow cytometry was used to monitor the changes of cell cycle and apoptosis. The changes of SOX4 and miRNA-363 expression levels in the MG-63 cells after transfection with SOX4 siRNA or pcDNA/SOX4 was detect by real-time PCR.RESULTS: The expressed level of miRNA-363 was lower, and the expression level of SOX4 was higher in the osteosarcoma tissues than those in the adjacent normal tissues. A significantly negative correlation between the expression levels of miRNA-363 and SOX4 was observed. The expression of miRNA-363 in the MG-63 cells after transfection with miRNA-363 mimics was significantly up-regulated, while the expression of SOX4 in the MG-63 cells was significantly down-regulated, with significant difference as compared with the cells transfected with miRNA-NC and control cells. The viability of MG-63 cells was inhibited, the cell cycle was arrested in G₀/G₁ phase, and the cell apoptosis was increased by transfection with miRNA-363 mimics. The relative protein expression levels of SOX4 in SOX4 siRNA group and pcNDA/SOX4 group were significantly different from those in negative control group, but the relative expression levels of miRNA-363 had no significant difference. Over-expression of SOX4 restored the viability of the MG-63 cells reduced by miR-363.CONCLUSION: The expression level of miRNA-363 is low in human osteosarcoma tissue. miRNA-363 may inhibits the viability of osteosarcoma cell line MG-63 and promotes cell apoptosis in vitro via inhibiting the SOX4 expression.  相似文献   

辣椒GRAS家族全基因组鉴定与表达分析   总被引：1，自引：0，他引：1  

张焕欣  董春娟  尚庆茂 《园艺学报》2017,44(12):2305-2317

基于辣椒全基因组数据信息,利用生物信息学方法对CaGRAS基因家族进行了系统鉴定和进化分析,并通过Real-time PCR方法检测了CaGRAS家族组织表达模式和对PEG6000、盐胁迫的应答。结果表明:在辣椒中存在54个CaGRAS基因,除11号染色体外其余染色体均有分布,外显子数1~3,等电点4.97~9.13;系统进化分析显示CaGRAS基因可分为8个进化群;CaGRAS基因具有不同的表达模式,在根、茎、叶片和茎尖中优势表达的基因分别有34、8、3和8个;多数CaGRAS基因能响应PEG6000和盐胁迫,其中CaGRAS11、CaGRAS30、CaGRAS40、CaGRAS44和CaGRAS50受到PEG6000和盐胁迫的强烈诱导。本研究为深入解析CaGRAS家族基因的功能奠定了一定基础。  相似文献   

干旱胁迫下‘宁玉’草莓生长发育及相关基因表达特点   总被引：1，自引：0，他引：1  

张超博  李傲  崔梦杰  刘众杰  陈立德  贾海峰  房经贵 《园艺学报》2017,44(12):2351-2360

为分析基因诊断草莓在干旱逆境条件下生长状态的可行性,以‘宁玉’草莓为试材,调查了不同干旱胁迫条件下的生长特点以及与生长素合成、脱落酸合成、花果发育、色素合成相关的10个基因的表达情况。研究结果表明:在不同干旱条件下草莓均可完成从营养生长到生殖生长的生命周期,所选基因也都行使并完成了其调控功能。与对照植株相比,在轻度和中度干旱胁迫(50%~65%土壤持水量)条件下,草莓植株的物候期较对照(80%土壤持水量)提早,而在严重干旱胁迫(35%土壤含水量时)条件下延缓,随着干旱胁迫条件的加重,草莓植株、叶片、果实明显变小,果实着色加深。每个基因在不同干旱条件下表达水平的变化趋势基本一致,且都与相应的物候期以及生长发育时间长短的变化情况相一致,但所选基因在几种干旱条件下其表达开始的早晚、表达水平的高低以及表达相应程度持续时间的长短存在不同。在轻度和中度干旱条件下,这些基因的表达时间提早,但持续时间较短,表达水平高于对照;而在重度干旱条件下,这些基因的表达开始的时间较对照延缓,表达水平较低但持续时间拉长。上述研究结果表明,利用基因的表达信息可以提前推断草莓植株在不同干旱条件下的生长发育状态,了解干旱胁迫对草莓生长发育的影响以及为是否需要提早采取管理措施等提供重要依据。  相似文献   

Effect of antisense locked nucleic acid blocking translation initiation region of c-myc exon 2 on apoptosis of hepatocellular carcinoma cells     

DENG Yi-bin  XIAO Shu-rong  XU Gui-dan  HUANG Zan-song 《园艺学报》2017,33(9):1602-1605

AIM: To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS: The anti-LNA that was complementary to the translation initiation region of c-myc exon 2 was designed, synthesized, and introduced into the HepG2 cells by cationic liposome-mediated transfection. The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot. The change of cell apoptosis was analyzed by flow cytometry, and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS: Five days after transfection, the mRNA level of c-Myc in anti-LNA group was 0.335±0.016, and the protein level was 0.448±0.037, significantly lower than those in control group (both P<0.05). The ratio of apoptotic cells in anti-LNA group was 32%±6%, which was higher than that in control group (P<0.05).CONCLUSION: Antisense locked nucleic acid targeting at the translation initiation region of c-myc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells.  相似文献   

番茄绿果与橙果间果实颜色及主要色素含量的遗传研究   总被引：1，自引：0，他引：1  

吴浪  赵菁菁  罗佳伟  王晨  李方曼  李云洲  张颜  梁燕 《中国蔬菜》2017,1(3):29-37

对番茄组合绿樱(绿果)×金珠1号(橙果)的6个世代遗传群体(P1、P2、F1、BC1、BC2和F2)进行果色性状、番茄红素含量、叶绿素含量和胡萝卜素含量等的遗传规律分析。结果表明:正反交F1的果色性状无明显差异,而色素含量存在显著差异;说明番茄果色性状受核基因控制,而色素含量遗传除受核基因控制外还可能存在胞质效应。采用多世代联合分析法的分析结果表明,番茄绿果与橙果间的果色性状符合2对加性主基因+加性-显性多基因(MX2-A-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为73.42%、78.25%和61.41%,多基因遗传率分别为22.87%、15.35%和34.94%,即果色性状遗传的主基因遗传力较强;叶绿素含量符合1对负向显性主基因+加性-显性多基因(MX1-AEND-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为0、1.73%和0.65%,多基因遗传率分别为45.47%、0和37.82%,即主基因遗传力在BC2群体中最高,多基因遗传力在BC1群体中最高;番茄红素含量与胡萝卜素含量均符合2对加性-显性-上位性主基因+加性-显性多基因(MX2-ADI-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为75.74%、1.79%、84.26%和61.53%、87.21%、81.05%,多基因遗传率分别为20.32%、74.12%、12.68%和0.68%、0、0,表明番茄红素含量和胡萝卜素含量的主基因遗传力较强。  相似文献   

甜瓜白粉病抗病基因的定位及候选基因分析     

李冰  赵玉龙  朱强龙  张志鹏  樊超  栾非时  高鹏 《中国蔬菜》2017,1(6):17-24

以甜瓜高抗白粉病自交系MR-1为母本,易感白粉病自交系Top Mark为父本,构建BC_1P_2和F_2群体,经13个国际通用的甜瓜白粉病生理小种鉴别寄主鉴定,2015年东北农业大学设施园艺工程中心的白粉病发病病菌为P.xanthii生理小种1,甜瓜MR-1对P.xanthii生理小种1的抗性由单显性基因控制。通过对266个F2分离群体的抗病性鉴定和CAPS标记分析,采用复合区间作图法对甜瓜抗白粉病性状进行QTL分析,最终构建了1张包含203个CAPS标记的甜瓜遗传连锁图谱,并将抗病基因PXR定位在第12号染色体上M12-GH和M12-TE 2个标记之间,该基因与两侧翼标记间的距离分别为0.63 c M和0.42 c M,两侧翼标记间在甜瓜参考基因组上对应的物理距离为303 kb,该区域内含有60个预测基因,其中10个基因含有非同义单碱基突变。10个基因荧光定量分析结果显示,在抗病与感病甜瓜表达量存在较大差异的4个基因MELO3C002434、MELO3C002437、MELO3C002441、MELO3C002457为甜瓜白粉病抗病候选基因。  相似文献