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1.
【目的】克隆青花菜细胞色素家族P450CYP79F1,分析其序列特征,阐明其在不同发育时期器官中的表达情况及其与莱菔硫烷含量的关系,为进一步揭示莱菔硫烷合成机理提供科学依据,为青花菜品种改良和新品种选育奠定基础。【方法】通过5′和3′端RACE克隆技术获得青花菜CYP79F1全长序列,利用DNAMAN 6.0进行基因拼接、氨基酸序列分析和蛋白二级结构预测,结合在线分析程序和软件进行基因和编码蛋白生物信息学分析;利用荧光定量PCR技术研究CYP79F1在不同发育器官中的表达情况;结合HPLC方法测定各相应时期器官中莱菔硫烷含量,包括青花菜发育时期的根、茎、叶,成熟花球,抽薹期花蕾(顶端蕾、成熟蕾、开花前1 d的蕾和花)和种子;对CYP79F1表达量与和莱菔硫烷含量进行相关性分析。【结果】获得了CYP79F1全长序列,Gen Bank登录号为MG012890,该基因全长2 014 bp,包含一个1 620 bp的开放读码框(ORF),编码540个氨基酸;编码的蛋白与甘蓝、白菜、芥蓝和油菜等同源蛋白的相似性均在95%以上;生物信息学分析表明该基因编码的酶蛋白有2个信号肽和2个跨膜结构域,为亲水性稳定蛋白,Wolf Psort预测其亚细胞定位于细胞质中;CYP79F1在根和茎中的表达量较高,叶中表达量最低,在花器官发育时期,自顶端蕾到花发育过程中呈现逐渐降低的表达趋势,种子中该基因表达量与花处于同一水平。Pearson相关性分析显示,青花菜发育时期的根、茎、叶、成熟花球、抽薹期花蕾和种子中莱菔硫烷含量与CYP79F1表达量无显著相关关系,但抽薹期花蕾自顶端花蕾到花中莱菔硫烷生成量与CYP79F1表达量呈显著正相关关系(R=0.96,P0.05),CYP79F1在调控莱菔硫烷的生成方面起重要作用,尤其是在生殖器官花蕾的发育过程中,能够显著上调莱菔硫烷的生成量。【结论】CYP79F1在青花菜不同发育器官中对莱菔硫烷的生成发挥着重要调控作用,可能与青花菜不同器官中莱菔硫烷含量多样性密切相关。  相似文献   
2.
以西兰花为原料,采用高效液相色谱法(HPLC)测定西兰花的花、茎和叶3个不同部位的萝卜硫素含量,对比分析了微波干燥(微波功率900 W,时间10 min)和电热鼓风干燥(干燥温度90 ℃,时间2 h)两种脱水方式所得样品中萝卜硫素含量的差异,研究了微波干燥对西兰花中萝卜硫素含量的影响。结果表明:微波干燥西兰花不同部位的萝卜硫素含量由大至小顺序为茎、花、叶,分别为(152.50±0.27)、(82.75±1.28)和(23.72±0.18)mg/kg,西兰花茎中的萝卜硫素含量分别是花和叶中的2倍左右和7倍左右;电热鼓风干燥西兰花不同部位的萝卜硫素含量同样是茎中最高,为(156.32±1.37)mg/kg,花和叶中分别为(85.04±1.13)mg/kg和(53.26±2.78)mg/kg。微波干燥与电热鼓风干燥相比,对西兰花叶中的萝卜硫素含量影响较大,对茎和花的影响不大。西兰花叶子宜采用电热鼓风干燥,茎和花既可以采用微波干燥,也可以采用电热鼓风干燥。  相似文献   
3.
The study was aimed to explore the antioxident function of sulforaphane (SFN) on the leydig cells of cadmium (Cd) exposured mice.Cd and SFN were added to the cell culture medium of TM3 cell, half inhibitory concentration (IC50) of Cd and SFN safe dose range were determined.The tested model was set up,and the relative survival rate of TM3 cells, lactate dehydrogenase (LDH) activity and cell antioxidant levels were determined to study the antioxident function of SFN to cadmium exposured mice.The results showed that:①With the increase of Cd concentration,the relative survival rate of TM3 cells was decreased,and the IC50 of Cd was 51.4 μmol/L;Within a certain concentration range,SFN could increased the cells survive rate,but there was toxicity when the SFN concentration was more than the scope,and the greater the concentration,the greater the toxicity. At experimental condition,SFN safety concentration were 2.5,5 and 10 μmol/L.②Compared with the control group,the GSH content,the T-SOD and GSH-Px activities in Cd group were significantly or extremely significantly decreased (P < 0.05;P < 0.01),and the LDH activity,the MDA content were increased.Additionally,the LDH activity and MDA content of SFN groups were decreased,while others three indexes were increased. Compared with Cd group,the GSH content,the T-SOD and GSH-Px activities of Cd+SFN groups were significantly or extremely significantly increased (P < 0.05;P < 0.01),however,the LDH activity,MDA content were decreased. The result indicated that SFN had the antagonism effect on toxicity of Cd in TM3 cell.  相似文献   
4.
Recent evidence in in vitro and in vivo models suggests that sulforaphane (SFN), found in raw cruciferous vegetables, may have utility in chemoprevention, as an antineoplastic agent and as a free radical scavenger. The effects of SFN alone or with doxorubicin on cell viability were examined, as well as cell cycle kinetics, invasion capabilities and apoptosis in three canine osteosarcoma cell line (D17, OS 2.4 and HMPOS). Results showed that SFN could not induce cell death at potentially physiological concentrations (<50 μM), but significantly diminished cell invasion and downregulation of focal adhesion kinase (FAK) signaling. Modest cell cycle changes were observed in each cell line. When doxorubicin was used in conjunction with SFN, there was a protective effect to doxorubicin‐induced cytotoxicity in D17 and OS 2.4 cells. Further studies examining SFN as a supplement are warranted, particularly in light of pro‐proliferative and cytoprotective properties in canine osteosarcoma.  相似文献   
5.
The study was aimed to explore the protective effect of sulforaphane (SFN) on the reproductive function of male mice with cadmium poisoning.40 healthy clean grade male Kunming mice were randomly divided into four groups:control group (H2O),cadmium chloride group (2.3 mg/kg CdCl2),sulforaphane group (10 mg/kg SFN),sulforaphane + cadmium chloride group (10 mg/kg SFN+2.3 mg/kg CdCl2),and continuous administration for 10 d,all mice were executed by dislocated cervical vertebra at 2 d after the last administration,and then the pathologic changes of testicular tissues,organ coefficient of testicle and epididymis,sperm quality and concentration of testosterone were tested.Additionally,the contents of GSH and MDA,and the activities of T-SOD in testis were also detected at the same time. Compared with the control group,pathology damages were observed in cadmium chloride group,organ coefficient of testis and epididymis,sperm quality and levels of testosterone extremely significantly decreased (P<0.01),the activities of T-SOD and GSH content were extremely significantly decreased (P<0.01),and the concentration of MDA was extremely significantly enhanced (P<0.01).Compared with the control group,the activity of T-SOD and concentration of GSH in sulforaphane group were significantly increased (P<0.05),and the concentration of MDA was not significant different between the control group and sulforaphane group (P>0.05).While compared with the cadmium chloride group,the sperm motility rate and sperm total count in sulforaphane and cadmium chloride group were extremely significantly increased (P<0.01),the organ coefficient of testicle and epididymis was increased significantly (P<0.05),the concentration of GSH and activity of T-SOD in testicular tissue were extremely significantly increased (P<0.01),and the concentration of MDA was extremely significantly decreased (P<0.01).The results indicated that sulforaphane had the protection effect on reproduction function of male mice with cadmium poisoning.  相似文献   
6.
萝卜硫素调控机制的研究进展   总被引:1,自引:0,他引:1  
姚丹燕  吴秋云  李倩  黄科 《园艺学报》2014,41(5):1020-1026
 综述了萝卜硫素(sulforaphane,1–异硫氰酸–4–甲磺酰基丁烷)的合成机理,以及影响 萝卜硫素含量的内源和外源因素,为开展萝卜硫素合成的调控机制研究提供线索。  相似文献   
7.
高萝卜硫素青花菜新品种‘福青1号’   总被引:1,自引:0,他引:1  
黄科  吴秋云  李宾  郑金贵 《园艺学报》2008,35(12):1854-1854
 青花菜新品种‘福青1号’为中晚熟一代杂种,株高60 cm,开展度65 cm,叶片15片,叶面蜡粉中等,叶色绿,花球绿,花蕾中细,定植后80 d开始收获。萝卜硫素含量高达984.47 μg·g-1,是对照品种的9.45倍。  相似文献   
8.
甘蓝和青花菜不同器官中莱菔硫烷含量差异研究   总被引:1,自引:0,他引:1  
十字花科蔬菜含有抗癌活性成分莱菔硫烷,为了筛选富含莱菔硫烷的甘蓝和青花菜材料,本研究以48份甘蓝和29份青花菜为试材,对其不同器官中莱菔硫烷含量进行HPLC分析。结果表明,不同甘蓝和青花菜基因型间莱菔硫烷含量差异显著,青花菜同一基因型不同器官中莱菔硫烷含量差异较大。甘蓝叶球中莱菔硫烷含量变化范围为4.28~57.18 mg·kg~(-1)DW,并平均含量为25.72 mg·kg~(-1)DW,筛选到莱菔硫烷含量较高的甘蓝自交系2份和杂交F1材料1份。青花菜中莱菔硫烷变化范围为:花球:3.58~1 137.47 mg·kg~(-1)DW;幼茎:13.79~259.58 mg·kg~(-1)DW;叶片:0~297.05 mg·kg~(-1)DW。花球、幼茎和叶片中莱菔硫烷平均含量分别为301.23、84.25和47.96 mg·kg~(-1)DW,初步筛选到莱菔硫烷含量较高的青花菜自交系3份。本研究结果为甘蓝类蔬菜新品种选育与改良提供了依据和材料。  相似文献   
9.
为探究西兰花叶中生物活性成分的具体含量,以西兰花叶为原材料,对西兰花叶中萝卜硫素、总酚、总黄酮、总蛋白质的含量和组成进行了测定与分析。结果表明,西兰花废弃叶中萝卜硫素含量为92.50 μg·g-1 DW,总酚含量为3.15 mg·g-1 DW,总黄酮含量为2.48 mg·g-1 DW,总蛋白质含量为89.38 mg·g-1 DW,蛋白质组成中核酮糖-1,5-二磷酸羧化酶/加氧酶(Rubisco)所占比例最大。本研究明确了西兰花叶片中4种生物活性成分的具体含量,为进一步开发利用西兰花废弃物提供了一定的数据支持。  相似文献   
10.
以中甘21(ZG21)、06-115两个甘蓝品种为试验材料,研究叶面喷施6种氨基酸对甘蓝硫代葡萄糖苷组分及其含量的影响。结果表明,不同种类氨基酸处理甘蓝硫代葡萄糖苷组分相同,均含有9种硫苷。氨基酸对甘蓝硫苷含量有一定的影响,半胱氨酸和甲硫氨酸显著提高06-115硫苷的总含量、脂肪族硫苷的含量和4-甲基硫氧丁基硫苷含量,其中半胱氨酸处理含量最高,与对照相比分别提高63.0%、69.6%和69.1%;半胱氨酸处理提高了3-吲哚基甲基硫苷含量,达到725.6 μmol/kg, FW;半胱氨酸处理也显著提高了中甘21硫苷的总含量、脂肪族硫苷的含量、吲哚族硫苷含量和4-甲基硫氧丁基硫苷含量,与对照相比分别提高了27.2%、39.6%、15.3%和26.2%。半胱氨酸、苯丙氨酸、甘氨酸和谷氨酸处理都提高了3-吲哚基甲基硫苷的含量,其中半胱氨酸处理最高,达到292.7 μmol/kg, FW,其次是谷氨酸和甘氨酸处理。  相似文献   
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