万寿菊(Tagetes erecta L.)中含有类胡萝卜素,不仅可用于天然色素提取,还是研究类胡萝卜素合成代谢的植物材料.番茄红素β-环化酶(lycopene beta cyclase,LCYB)是类胡萝卜素合成代谢途径中的一个关键酶,文章通过对万寿菊中TeLCYB基因克隆、植物表达载体构建、并利用农杆菌介导法进行番茄遗传转化,获取TeLCYB基因转化番茄植株.分子鉴定和表型分析结果表明,万寿菊TeLCYB基因在番茄中表达,并改变转基因番茄果实颜色.高效液相色谱测定显示,TeLCYB基因的表达显著增加了番茄果实中β-胡萝卜素的量.该文研究结果表明TeLCYB基因具有功能活性,可用于改良植物果实的色素营养品质.

酵母双杂交技术作为检测蛋白质之间相互作用的有效手段,不仅能验证与已知蛋白质的相互作用,还可从文库中筛选与特定蛋白互作的未知蛋白.文章为研究番茄抗病相关蛋白Pti(Pto interaction protein)的生物学功能,筛选与其互作的蛋白,以未处理野生型番茄叶片、真空渗透法接种丁香假单胞杆菌番茄致病变种DC 3000(Pseudomonas syringae pv.tomato DC3000,Pst DC3000)的感病株系prf3、抗性株系PtoR的植株叶片提取RNA,构建酵母双杂交文库.经检测,文库容量为1.25×107 CFU,阳性率大于95％,插入片段平均长度约1200 bp.通过构建Pti5重组诱饵载体并转化酵母菌,筛选与其相互作用的蛋白质.初步获得的Pti5互作的靶标基因,可用于进一步研究番茄抗病基因Pti的作用机制.

水稻在生长过程中会受到各种逆境胁迫的影响,特别是环境污染引起的土壤金属离子胁迫.P1B-ATPase又称为重金属ATP酶(heavy metal transporting ATPase,HMA),作为P型ATP酶的一个亚族,目前已被证明参与植物体内金属离子转运.水稻基因组中有多个基因被推断编码该类蛋白,但其功能尚未完全解析.文章通过对水稻第二染色体HMA基因(OsHMA,基因登录号AK107997)进行生物信息学分析,发现该基因编码蛋白含有金属结合结构域;表达模式分析结果表明:OsHMA在水稻幼苗、成体根中表达量最高;在铜离子诱导下表达水平上调,提示OsHMA可能在水稻铜离子转运中发挥作用.构建OsHMA基因RN A干涉载体,以用于后续分析其在水稻抗土壤金属离子胁迫中的功能.

番茄Pto基因编码丝氨酸/苏氨酸蛋白激酶,是抵御丁香假单胞菌(Pseudomonas syringae)的重要抗性蛋白,但对其抗病分子途径尚未完全阐明.Pti6是可以与Pto互作的蛋白之一,被认为是Pto介导抗病途径的下游基因.文章通过农杆菌介导方法进行番茄遗传转化,对Pti6过表达转基因植株进行分析.结果 表明获得了Pti6过量表达的转基因阳性植株;对过表达番茄果实的过氧化氢酶(Catalase,CAT)、过氧化物酶(Peroxidase,POD)活性测定表明,其酶活比野生型有显著提高,而丙二醛(Malondialdehyde,MDA)的量有所降低,表明其在番茄抗病中发挥了作用.Pti6过表达株系的获得为对其功能的深入研究打下了基础.

Pseudomonas syringae pv. tomato (Pst) is a pathogenic microorganism that causes bacterial speck disease and affects tomato yield and quality. Pto is a disease resistant gene for plant to recognize and defense against Pst. Pto interacts with Pti (Pto interacting) proteins, which include three transcription factors, Pti4, Pti5, Pti6, and they were thought to be downstream of Pto-mediated pathway to promote the expression of disease-related genes. In the present work, the overexpression plants of Pti4, Pti5 or Pti6 were obtained by Agrobacterium-mediated transformation in tomato. The Pti4/5/6-overexpressed lines indicated enhanced expression of pathogenesis related genes and resistance to pathogenic bacteria Pst DC3000. Meanwhile, the transgenic plants showed that Pti4/5/6 function in ripening but performed no obvious adverse influence on flowering time, seed-setting rate, weight and soluble solids content of fruits. Furthermore, Pti-overexpressed fruits exhibited increased enzymatic activities of phenylalnine ammonialyase, catalase, peroxidase and decreased content of malondialdehyde. Additionally, cell-free and in vivo ubiquitination assay indicated that Pti4, Pti5 and Pti6 degraded by 26S proteasome which suggested that these Pti transcription regulators' functions could be regulated by ubiquitinmediated post translational regulation in tomato.

根据万寿菊转录组测序分析结果克隆PDS基因(TePDS1),并构建植物表达载体,利用农杆菌介导法将TePDS1进行番茄遗传转化.试验结果显示,TePDS1已整合于番茄基因组中,获得了表达万寿菊PDS基因的番茄植株,且转化番茄果实中的番茄红素含量明显高于野生型,表明TePDS1基因具有功能活性,可用于提高植物果实中色素含量.

以番茄的根、叶、花及不同发育时期的果实提取RNA,构建酵母双杂交文库.经检测,文库容量为1.1×107 CFU,阳性率大于95％,插入片段平均长度大于1000 bp,可用于互作蛋白的筛选.依据番茄抗病途径相关基因Pti4编码序列设计引物,构建重组诱饵载体并转化酵母菌,筛选与Pti4相互作用的蛋白质.经初步检测获得6个可能与Pti4互作的转录因子,为进一步研究番茄Pti基因的作用机制提供了基础.

拟南芥抗坏血酸过氧化物酶2(A rabidopsis thaliana ascorbate peroxidase 2,A tA PX2)是A PX基因家族成员,在拟南芥逆境胁迫应答中起着重要作用.文章通过A tA PX2基因过表达载体构建、农杆菌介导遗传转化,获得A tA PX2基因水稻过表达植株.对过量表达A tA PX2的转基因水稻阳性植株的分析结果显示,其抗过氧化氢能力提高,对高温、盐胁迫耐受能力显著增强,田间植株秕谷率也明显低于野生型.研究结果表明,拟南芥抗坏血酸过氧化物酶基因A tA PX2在水稻中的过量表达,可通过提高清除活性氧能力而用于增强水稻对多种非生物胁迫逆境的耐受能力.

丁香假单胞菌所导致的细菌性斑点病是影响番茄产量和品质的重要病害,而番茄色氨酸苏氨酸激酶(Pto)是植物识别、防御这一病原菌的重要抗性蛋白.文章通过克隆番茄Pti(Pto interaction protein)基因Pti4、Pti5和Pti6,分别构建植物表达载体,利用农杆菌介导方法在烟草叶片中进行瞬时表达和western blot-ting检测,并进一步利用免疫共沉淀技术,在植物体系中检测了它们与番茄Pto蛋白的相互作用.实验结果表明,利用所构建Pti4、Pti5和Pti6基因植物表达载体,在烟草中获得了预期大小Pti4、Pti5和Pti 6蛋白,并在植物体系中验证了它们与Pto蛋白的相互作用.该工作为深入研究Pti基因在番茄分子免疫途径的功能奠定了基础.

Marigold(Tagetes erecta)is one of important plant resources for natural xanthophylls extraction.China has become the world’s leading producer of xanthophyll from marigold.In this paper,eight marigold cultivars which were renowned at home and abroad were cultivated on a small scale in Hefei,Anhui Province for preliminary screening to be suitable for local scale planting by agronomic traits and xanthophyll content.The results indicated that all tested marigold cultivars exhibited good growth adaptability.High performance liquid chromatography determination showed that xanthophyll content of orange flowers was higher than that of yellow flowers.Comprehensive analysis of agronomic traits and pigment content suggests that the domestic cultivar"JUWANG"is suitable for local scale planting and breeding.This study can provide the basis for further investigation of pigment synthesis regulation of marigold and its industrialization development in Anhui Province.

This study focused on the analysis of total phenolics and antioxidative activities in speciality tea including YongXiHuoqing,TaQuanYunWu,JingTingLvXue and XiKouYunJian of Xuancheng city by Folin-Ciocalteu colorimetric method,DPPH and ABTS methods.Moreover,the difference of the total phenols and antioxidative activities for JingTingLvXue at different picking times were investigated and the effects of processing on tea polyphenols were discussed using HPLC-MS.The results showed that YongXiHuoQing contained much more tea polyphenols (161 mg catechin/g dry weight),XiKouYunJian just 127 mg catechin/g dry weight.YongXiHuoQing showed the highest antioxidant activity by ABTS and DPPH methods.Tea polyphenols content and antioxidant activities were increased when picked after Qingming festival.The content of tea polyphenols was increased and caffeine was decreased after tea producing process.