芪合酶基因Fm-STS在何首乌毛状根中的过量表达及dsRNA干扰

目的:建立一套探究植物基因功能的方法体系,验证由芪合酶基因保守序列通过RACE扩增技术在何首乌中得到的基因Fm -STS的功能.方法:由含CaMV 35S启动子驱动的gfp基因的植物转基因表达基础质粒pBIN-35S-GFP构建过表达质粒pBIN-35S-STS-GFP(阳性)和双链RNA干扰重组质粒pBIN-35S-正向-反向-GFP(阴性),并同空白表达质粒pBIN-35S-GFP(空白)均导入野生型发根农杆菌ATCC15834中,转化何首乌外植体,诱导生成毛状根并培养,对毛状根进行高效液相色谱分析以及实时荧光定量检测.结果:在过表达组、空白组和干扰组中毛状根中发根农杆菌Ri质粒中的rolB基因和外源基因gfp均有表达,高效液相色谱法分析芪合物二苯乙烯苷含量依次为4.67mg/g、2.18mg/g和0.65 mg/g,在mRNA水平上测试荧光定量检测基因Fm-STS表达量:RNAi组是空白组的1/433.53,过表达组是空白组的2.41倍.结论:结果表明过量表达与双链RNA干扰相结合在植物基因功能研究中有良好的应用,何首乌中芪合酶Fm-STS是二苯乙烯苷主要的合成酶.     

超富集植物短毛蓼对锰的富集特征

通过野外调查和营养液培养试验,研究了锰在短毛蓼体内的富集特征和对其生长的影响.在锰含量高达2.5×105mg/kg的锰矿废弃地上短毛蓼生长良好,叶锰含量高达1.66×104mg/kg.营养液培养条件下,随着生长介质中Mn浓度的升高,短毛蓼根、茎、叶中的Mn含量逐渐增加,当锰供应水平为1.000mmol/L时,叶锰含量超过10000mg/kg;当锰供应水平为20 000mmol/L时,短毛蓼仍能生长,根、茎和叶3部分的锰含量均达到最大值,分别为9923,18112mg/kg和55750mg/kg.在所有锰供应水平下,短毛蓼茎和叶中的锰含量都比根部的高.结果表明,短毛蓼是一种锰超富集植物,这一发现为锰污染土壤的植物修复和探讨锰在植物体内的超富集机理提供了一种新的种质资源.     

Nitrogen fertilizers promote plant growth and assist in manganese (Mn) accumulation by Polygonum pubescens Blume cultured in Mn tailings soil

Abstract

This study examined how different nitrogen (N) forms and application levels promote plant growth and assist in manganese (Mn) remediation of Polygonum pubescens Blume (P. pubescens) cultured in soil with a high Mn level. The effects of ammonium chloride (a) and urea (u), at three application levels (10, 20, and 30?mg L^?1 N) and control (no N addition, CK) on the growth, Mn accumulation, and enzymatic anti-oxidative defenses of P. pubescens were examined. In general, both ammonium-N and urea-N promoted the plant mass and height of P. pubescens. The total Mn amount of roots, stems, and leaves in N treatments were higher (p?<?0.05) than that of CK. The ammonium-N treatments showed greater plant biomass and Mn accumulation compared to the urea-N ones. In general, the accumulations of Mn, Cr, Zn, and Cu were significantly lower (p?<?0.05) in the N fertilizer treatment than those in the control; while the accumulations of Pb were higher (p?<?0.05) in P. pubescens across all N fertilizer treatments than those in the control. The N addition decreased the contents of O₂^? and H₂O₂ in the leaves of P. pubescens, while increasing the activities of enzymatic anti-oxidative defenses.     

气象因子和矿质元素对虎杖根茎白藜芦醇含量的影响

对全年各月虎杖材料中白藜芦醇含量,及其Ca、Cu、Fe、K、Mg、Na、Zn 8种矿质元素积累进行检测,结合气象因子动态变化,采用主成分分析法,研究了不同季节虎杖白藜芦醇含量动态变化与其矿质元素、气象因子动态变化的关系．结果表明,Cu与Fe,Mg与K、Zn,Ca与月日照时数,K与Zn相关系数分别为0．812、0．871、0．793、0．602、0．729,呈极显著正相关;Cu、Mn与月平均气温,Fe与K相关系数分别为-0．738、-0．712、-0．766,呈极显著负相关;Cu与白藜芦醇含量呈显著负相关．找出了影响虎杖白藜芦醇含量动态变化的5个主成分,其中“酶促反应促进性矿质营养因子”,“高温、强照射气象因子”的贡献率分别为35．539％、33．358％,合计为68．897％,二者对不同季节虎杖白藜芦醇的变异贡献率最大．同时进行白藜芦醇含量的动态变化与主成分的多元相关、回归分析,建立了回归方程。     

Indigo and indirubin derivatives from indoles in Polygonum tinctorium tissue cultures

Substituted indoles (5-bromo-, 5-fluoro-, 5-chloro-, 5-hydroxy-, 4-methyl-, 5-methyl-, 6-methyl-, and 7-methylindole) were fed at 1 mM for 10 to 21 days into the tissue cultures of Polygonum tinctorium including cell suspension, root and shoot cultures. They were converted into the corresponding indirubin and indigo derivatives with the exception of 5-hydroxyindole. The yield ranged between 0.7–1.1 mg/g dried weight for indigos and 0.007–0.024 mg/g dried weight for indirubins. Presence of the corresponding indican derivatives in the shoot culture suggested that the pigment production proceeded through the indican formation. © Rapid Science Ltd. 1998     

Indigo production in hairy root cultures of Polygonum tinctorium Lour.

The transformed root culture of Polygonum tinctorium Lour. was established by infecting leaf explants with Agrobacterium rhizogenes A4. These cultures were examined for their growth and indigo content under various culture conditions. Among the four different culture media tested, SH medium showed the highest yield for root growth (28 mg dry wt/30 ml) and indigo production (152 g/dry wt). In SH medium, 30 g sucrose l^–1, 2500 mg KNO₃ l^–1, 300 mg NH₄H₂PO₄ l^–1 were the best conditions for indigo production at pH 5.7. The production of indigo in hairy roots slightly increased with the addition of 200 mg chitosan l^–1 (186 g/dry wt) and 20 U pectinase l^–1 (181 g/dry wt).     

何首乌中新的二苯乙烯甙

用反相层析法从何首乌（Polygonum multiflorum Thunb.)根茎的水提物只发得2个新的二苯乙烯甙（1,2）及9个已知化合物：没食子酸（3）,2,6－二羟基－苯甲酸（4）,吲哚－3－（L－）α氨基－α－羟基－丙酸）甲酯（5）1,2－二羟基丙烷－1－（4－羟基－苯基）（6）,大黄素（7）,大黄素－8－β－D－葡萄糖甙（8）,（＋）－lyoniresinol-3α－O－β－D－葡萄糖甙（9）,2,3,4、,5－四羟基反式二苯乙烯－2－O－β－D－吡喃葡萄糖甙（10）和2,3,4',5－四羟基反式二苯乙烯－2,3,－二－O－β－D－吡喃葡萄糖甙（11）。亲化合物结构通过理化性质与波谱分析特别是2D NMR得以确定。化合物2表现出很强的DNA裂解活性,化合物1,2与10具有很强的抗脂质过氧化活性。     

锰胁迫对杠板归细胞超微结构的影响

锰是植物生长必需的微量元素,然而锰含量过高将影响酶活性、造成植物毒害,不同植物的锰耐性差异很大。杠板归(Polygonum perfoliatum L.)是一种生长在锰尾矿废弃地的耐性植物。通过温室培养,应用透射电子显微镜-能谱联用仪研究不同锰处理条件下(5,1000,10 000μmol/L)杠板归根、茎和叶细胞超微结构的变化和锰在叶细胞内可能的存在形式,结果表明:(1)生长介质锰含量为5μmol/L时,杠板归细胞超微结构未见异常;即使锰处理浓度为1000μmol/L时,杠板归根、茎和叶细胞结构依然完整,细胞器清晰可见,未见明显损伤;(2)当锰处理浓度为10000μmol/L时,杠板归细胞器未见缺失现象,但根细胞内线粒体数量减少,茎细胞叶绿体开始受损,叶细胞叶绿体膜结构出现破损,基粒片层结构减少,嗜锇颗粒数量明显减少;(3)与对照相比,1000μmol/L或10000μmol/L锰处理30 d,植物细胞内出现黑色团聚物。10000μmol/L锰处理条件下,杠板归叶细胞内和细胞间隙出现类似针状物质,这可能是杠板归体内锰积累和分布的形态之一。研究成果有助于阐明植物的锰耐性机制,为锰耐性植物筛选和废弃锰尾矿库生态重建提供科学参考。     

何首乌愈伤组织的诱导

研究了外植体、光暗条件、植物激素等因子对何首乌愈伤组织诱导的影响 ,以及 6 -BA和何首乌组织提取液对愈伤组织增殖的影响。结果表明 :茎段的愈伤组织诱导优于带侧芽茎段和叶片 ;光照有利于愈伤组织的诱导 ;4因子 4水平的正交实验表明 ,对何首乌茎段愈伤组织诱导的影响 2 ,4 -D >6 -BA >IBA >IAA ,最佳激素搭配是 :MS +2 ,4 -D 2mg L +6 -BA 1mg L +IBA1mg L或MS +2 ,4 -D 2mg L +6 -BA 2mg L +IAA 1mg L ;MS培养基中附加 6 -BA或组织提取液均促进愈伤组织的增殖。