镧对镉胁迫羽衣甘蓝的调节效应

[目的]为花卉植物用于cd污染土壤栽培提供依据。[方法]采用水培试验研究了镧（工丑）对羽衣甘蓝受到镉（Cd）胁迫的调节效应。[结果]低浓度的cd对羽衣甘蓝的出芽率、株高、根系、地上和地下鲜重与千重等指标均有一定的抑制作用，高浓度的cd对其有明显的抑制作用，而一定浓度的La可以缓解这种抑制作用。[结论]5mg／LLa缓解高浓度cd对羽衣甘蓝种子出芽以及生长等各项指标的抑制作用效果最好。     

Estimating theoretical radiation‐use efficiency for kale crops

Establishing the radiation‐use efficiency (RUE) of forage brassica crops will aid our understanding of their photosynthetic performance. The concept of RUE has been developed for cereals and legumes, but there is limited information for forage brassica crops. Three experiments defining the influence of different sowing dates on ‘Gruner’ kale (Brassica oleracea acephala L.) dry matter production were conducted at Hastings (Hawkes Bay) and Lincoln (Canterbury) in New Zealand between 2002 and 2009. These trials were also evaluated for radiation interception and RUE. Delayed sowing increased RUE in two out of three experiments across sites: from 1·93 g MJ^?1 photosynthetically active radiation (PAR) for December‐sown crops to 2·72 g MJ^?1 PAR (P < 0·001) for January‐sown crops at Hastings and from 1·50 for September‐sown crops to 2·00 g MJ^?1 PAR (P < 0·001) for November‐sown crops at Lincoln. The different sowing dates and years of experimentation provided a range of mean temperatures (from 13 to 16°C) during the vegetative period. Across years and sowing dates, RUE was strongly correlated with mean temperature (R² = 0·81) and sowing date (R² = 0·64), but weakly correlated with season length (R² = 0·11) and dry matter (R² = 0·002). There was also a strong correlation (R² = 0·83) between sowing date and mean temperature. The increase in RUE with delayed sowing was therefore mainly attributed to increased mean temperatures. Radiation‐use efficiency increased at about 0·41 g MJ^?1 for each 1°C increase from 13 to 16°C.     

RATIO OF CALCIUM TO MAGNESIUM INFLUENCES BIOMASS,ELEMENTAL ACCUMULATIONS,AND PIGMENT CONCENTRATIONS IN KALE

Producers use elemental ratios, such as calcium (Ca): magnesium (Mg), in fertility programs to ensure sufficient nutrient uptake. Kale (Brassica oleracea L. var. acephala D.C.) accumulates high levels of carotenoids which can be beneficial for human health. Objectives were to determine the influence of Ca:Mg fertilization on 1) biomass, 2) essential nutrients, and 3) carotenoids in kale leaf tissues. ‘Redbor’ kale was greenhouse-grown in solution culture. Ca:Mg ratio treatments were 9:1, 6:1, 3:1, 1:3, 1:6, and 1:9. Ca:Mg ratio significantly affected biomass, nutrient accumulation, and carotenoids. Plant biomass decreased linearly (P ≤ 0.001) and β-carotene, lutein, neoxanthin, and antheraxanthin all increased, then decreased quadratically (P ≤ 0.001) as the ratio of Ca:Mg changed from 9:1 to 1:9. Ca:Mg ratio also affected leaf tissue Ca, Mg, potassium (K), sulfur (S), boron (B), manganese (Mn), molybdenum (Mo) and zinc (Zn). Results indicate that producers wishing to maximize elemental uptake and carotenoid content of kale need to consider the ratio of Ca:Mg in their fertility programs.     

Identification and quantification of anthocyanins in different coloured cultivars of ornamental kale (Brassica oleracea L. var. acephala DC)

Anthocyanins are responsible for the colour of many fruits, vegetables, flowers, and coloured-leaved trees. Ornamental kale (Brassica oleracea L. var. acephala DC) is widely cultivated for its colourful inner leaves. To investigate the relationship between the degree of colouration and anthocyanin distribution, content, and composition in ornamental kale, the authors studied the pigment characteristics of five cultivars with different coloured leaves (white, pink, red, purple, and purple-black). Microscopy observation, spectrophotometer, and high-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis of fresh inner leaves revealed that pink, red, and purple colourations were associated with high levels of anthocyanin, while purple-black was the result of the combination of anthocyanins and chlorophyll. In the coloured cultivars, anthocyanins were abundant mainly in the first and second cell layers below the epidermis in both the hypocotyls and inner leaves. No anthocyanin was found in the white-leaved phenotype cultivar. Anthocyanin content increased as leaf colour deepened from pink, red, to purple cultivars, which had little chlorophyll and carotenoid. The authors identified eight anthocyanins in the four coloured cultivars, including one non-acylated, four monoacylated, and three diacylated cyanidin glycosides. Cyanidin-3-(sinapoyl)(feruloyl)-diglucoside-5-glucoside was the most abundant anthocyanin in the four coloured cultivars followed by cyanidin-3-(sinapoyl)-diglucoside-5-glucoside. The analysis of anthocyanin accumulation characterisation provides important information on evaluating colouration patterns in coloured plants, and will be helpful for breeding desired leaf colours in ornamental kale.     

羽衣甘蓝抗寒生理特性研究

观赏羽衣甘蓝作为秋冬季节重要的园林绿化植物,抗寒性直接影响其推广和应用。为了阐明羽衣甘蓝响应低温的生理特性,揭示抗寒机理,以北京市农林科学院蔬菜研究中心生物技术课题组选配的优良F1杂种作为研究试材,分析不同低温处理下叶片电导率变化和半致死温度,以及渗透调节物质含量和抗氧化酶活性的变化。结果显示,羽衣甘蓝叶片电导率在-5℃以下迅速升高,-10℃左右达到最大值;半致死温度为-10.3~-13.7℃,红色类型低于白色类型,羽叶和圆叶低于皱叶类型。上述研究结果与观赏羽衣甘蓝田间抗寒表现一致。低温处理下渗透调节物质含量和抗氧化酶活性响应特性表明,不同指标呈现不同的变化趋势,除了可溶性糖在-10℃含量最高外,可溶性蛋白和游离脯氨酸含量,以及超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均在-5℃或-8℃达到最大值,并随温度进一步降低而下降。主成分分析结果表明半致死温度、可溶性糖和游离脯氨酸含量是导致羽衣甘蓝抗寒性差异的主要因素。研究结果为建立羽衣甘蓝抗寒性评价体系,进一步解析抗冻机理,培育抗寒新品种奠定理论基础。     

羽衣甘蓝ARC1 蛋白的原核表达、纯化及泛素连接酶活性分析

 ARC1 是植物特有的一类含有U-box/ARM 结构域的蛋白,在芸薹属植物自交不亲和 （self-incompatibility,SI）信号转导中起着正向调控因子的作用。将羽衣甘蓝（Brassica oleracea var. acephala）BoARC1 编码区的序列连接到原核表达载体pET-14b 上,通过酶切鉴定和测序分析,构建pET-14b- BoARC1 表达质粒;将获得的阳性表达质粒转化到大肠杆菌表达菌株BL21（DE3）pLysS 中,利用IPTG 进行诱导表达。SDS-PAGE 结果显示,在分子量69 kD 处有BoARC1 蛋白特异性地诱导表达;利用Ni2+-NTA 树脂通过亲和层析的方法获得BoARC1 融合蛋白。在泛素激活酶（E1）、泛素结合酶UBC7（E2）和泛素 体外泛素化反应后,通过免疫印迹的方法检测,显示出BoARC1 融合蛋白能够将底物进行多泛素化修饰; 当U-box 中保守位点第323 位Pro 突变为Ala 或其他泛素化组分缺少时,底物不能被泛素化修饰。     

羽衣甘蓝ARC1的基因分离、表达及与SRK相互作用的分析

 以羽衣甘蓝（Brassica oleracea var. acephala）自交不亲和系（S_13-b S_13-b）为试验材料,利用RT-PCR的方法分离其ARC1基因cDNA序列（命名为BoARC1,GenBank登录号为EU344909）。BoARC1 cDNA序列中含有1个1 992 bp的开放读码框（ORF）,编码1个含有663个氨基酸的蛋白质。羽衣甘蓝BoARC1与油菜BnARC1的氨基酸序列具有94%的一致性;Southern杂交结果显示BoARC1在基因组中只存在单一拷贝;Northern杂交结果显示BoARC1特异性地在柱头组织中表达;在酵母双杂交试验分析中,BoARC1能够与SRK₃、SRK _13-b和SRK₉₁₀的胞内激酶域发生相互作用。     

七份观赏羽衣甘蓝品种遗传相似性的分析

本文选取7份白色观赏羽衣甘蓝材料,分别进行9种农艺性状观测数据的聚类分析和基因组DNA的SRAP标记多态性结果的聚类分析.农艺性状的聚类分析结果显示分为两大类群,第一类群为皱叶类型,包括两个亚群,其中名古屋、帝王为一个亚群,骑士为另一亚群.SRAP标记聚类分析结果完全验证了这个结果;第二类群为圆叶类型,也包括两个亚群,其中白舞、白鹰、火烈鸟为一个亚群,乐园为另一个亚群.而SRAP标记聚类分析结果显示第二类群也包括两个亚群,但乐园、火烈鸟、白舞为一个亚群,白鹰为另一个亚群,这与农艺性状聚类有点不同.综合结果表明,7份白色羽衣甘蓝材料间的遗传背景存在不同的差异;SRAP标记分析的聚类结果与形态特征分类具有较高的相关.本研究确立了可以利用于羽衣甘蓝基因组分析的SRAP标记引物序列;证明了SRAP标记分析的聚类结果,能够较准确地从分子水平上对材料进行分类,可以作为羽衣甘蓝杂交育种中亲本亲缘关系远近的判断标准之一.     

羽衣甘蓝ARC1的抗体制备及蛋白表达分析

ARC1是芸薹属植物自交不亲和(Self-incompatibility,SI)信号复合体下游传递因子,特异性地介导SI信号传递。利用原核表达系统对羽衣甘蓝(Brassica oleracea var.acephala)ARC1的全长蛋白(1~663 aa,BoARC1-F)、UND结构域蛋白(1~279 aa,BoARC1-N)和ARM结构域蛋白(361~663aa,BoARC1-C)进行重组表达。SDS-PAGE结果显示这3种蛋白分别在分子量68、33和38 kD处特异性地诱导表达。利用Ni~(2+)-NTA树脂亲和层析技术获得重组蛋白。将BoARC1-C蛋白免疫小鼠并通过细胞融合技术制备单克隆抗体,获得2株单克隆抗体。交叉反应试验结果显示制备的单克隆抗体具有ARC1识别特异性。通过免疫印迹技术检测BoARC1在不同组织和不同发育阶段柱头中的表达,结果显示BoARC1特异性地在羽衣甘蓝柱头中表达,而且在开花阶段的柱头表达量最高。     

羽衣甘蓝花粉SCR13-b基因的分离及其功能分析

 以羽衣甘蓝(Brassica oleracea var. acephala) S_13-bS_13-b自交不亲和系为试验材料, 分离了控制花粉自交不亲和基因———S_13-b单倍型富含半胱氨酸基因(SCR_13-b) , GenBank收录号为EF577028。SCR_13-b cDNA序列含有237 bp的开放读码框(ORF) , 编码一个含78个氨基酸的蛋白。SCR_13-b gDNA含有一个758 bp的内含子, 内含子5′供体位点和3′受体位点边界序列符合GU2AG规则; DNA blot分析表明SCR_13-b在基因组中只有单一拷贝; 授粉生物活性分析试验显示原核表达SCR132b融合蛋白能够启动SI反应, 抑制杂交亲和花粉的萌发和生长。