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Flavonoid extracts from chrysanthemum with appropriate anthocyanins turn blue when exposed to iron ions

Horticultural Plant Journal 2024年第3期10卷 837-852页

作者： Yanfei Li Jiaying Wang Chenfei Lu Zhongman Wang Chengyan Deng Kang Gao Jingjing Li Zhijun Fang Hao Liu Yan Hong silan dai Beijing Key Laboratory of Ornamental Plants Germplasm Innovation&Molecular Breeding National Engineering Research Center for FloricultureBeijingLaboratory of Urban and Rural Ecological EnvironmentKey Laboratory for Genetics and Breeding of Forest Trees and Ornamental Plants of Ministry of EducationSchool of Landscape ArchitectureBeijing Forestry UniversityBeijing 100083China Beijing Greenland Maintenance and Management Service Center Beijing 102211China Beijing North Asia Landscape Co. Beijing 102211China

Although some species that accumulate only cyanidin(Cy)in nature can produce blue flowers through iron ions,there has been no evidence of blue chrysanthemums being generated in this manner.This study revealed that flavonoid extracts from the ray florets of the chrysanthemum cultivar‘Wandai Fengguang’turned blue when exposed to Fe^(3+).Samples that could turn blue were labeled as CB(Cy-determined blue flowers),while samples that did not turn blue were labeled as CN(Cy-determined non-blue flowers).After a series of experiments,a stable screening system was established using flavonoid extracts containing NaAc buffer at pH 5.5 and a total anthocyanin concentration(TAC)of 30 μmol·L^(-1),and the addition of Fe^(3+)from 0 to 0.25 μmol·L^(-1)allowed for the selection of five CB samples from 39 chrysanthemum cultivars.All five CB samples exhibited flower color phenotypes that belonged to Cluster-I with redness(a*)values ranging from 29.03 to 45.99,yellowness(b*)values from-11.31 to 3.77,and brightness(L*)values from 29.07 to 45.99.Additionally,the ratio of TAC to total luteolin concentration(TLC)was found to be a critical factor for distinguishing between CB and CN samples.To realize the desired blue hue in the flavonoid extracts with the participation of Fe^(3+),a TAC to TLC ratio of 2.25 and above is required.Moreover,the protoplasts and ray florets of CB samples that turned blue with the involvement of Fe^(2+)showed great potential for cultivating blue chrysanthemums through ferric-anthocyanin chelate.Overall,this study reveals that blue flowers can be cultivated through the increase in the iron ion concentration,combined with the accumulation of Cy.

关键词： Chrysanthemum×morifolium Cyanidin Copigment Ferric-anthocyanin chelate Blue flower

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Genotype Screening of Recipient Resources with High Regeneration and Transformation Efficiency in Chrysanthemum

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Phyton-International Journal of Experimental Botany 2022年第4期91卷 869-888页

作者： Yajun Li Yumeng Cui Bingjie Wang Yue Li Mengmeng Zhang silan dai He Huang Beijing Advanced Innovation Center for Tree Breeding by Molecular Design Beijing Key Laboratory of Ornamental Plants Germplasm Innovation&Molecular BreedingNational Engineering Research Center for FloricultureBeijing Laboratory of Urban and Rural Ecological EnvironmentKey Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of EducationMinistrySchool of Landscape ArchitectureBeijing Forestry UniversityBeijing100083China Beijing Botanical Garden Beijing100093China

Genetic transformation is one of the key steps in the molecular breeding of chrysanthemum,which relies on an optimal regeneration and transformation system.However,the regeneration system of different chrysanthemum cultivars varies,and the regeneration time of most cultivars is long.To screen cultivars with highly efficient regeneration,leaves and shoot tip thin cell layers(tTCL)from eight chrysanthemum cultivars with different flower colors and flower types were cultured on Murashige and Skoog media(MS)supplemented with 1.0-5.0 mg L^(−1)6-benzylaminopurine(6-BA)and 0.1-1.0 mg L^(−1)α-naphthaleneacetic(NAA).The results showed that the most efficient regeneration media were MS+6-BA 1.0 mg L^(−1)+NAA 0.5 mg L^(−1)for leaf explants and MS+6-BA 5.0 mg L^(−1)+NAA 0.1 mg L^(−1)for tTCL explants.Subsequently,another 13 chrysanthemum cultivars were screened by using the media,and finally,three cultivars with high regeneration efficiency were obtained from 21 cultivars.Among these,C1 had the highest regeneration efficiency:the regeneration rate of leaf explants reached 80.0%after 42 days of culture,and the regeneration rate of tTCL explants reached 100%after 31 days of culture.Furthermore,we also established the transformation system for C1 as follows:preculturing for one day,infecting with Agrobacterium suspension(OD_(600)=0.6)for 10 min,and cultivating in the regeneration medium with 350 mg L^(−1)carbenicillin and 10 mg L^(−1)kanamycin,thus ultimately achieving a transformation rate of 4.0%.In this study,a new chrysanthemum cultivar with an efficient regeneration and transformation system was screened,which is beneficial to enrich the flower color of chrysanthemum transgenic plant recipients and to the functional research of flower color or type-related genes.

关键词： Chrysanthemum tissue culture regeneration system genetic transformation cultivars screening

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瓜叶菊花色性状形成机理的研究进展

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植物遗传资源学报 2024年第5期25卷 695-703页

作者：崔宇萌黄河戴思兰北京林业大学园林学院/花卉种质创新与分子育种北京市重点实验室/国家花卉工程技术研究中心北京100083

瓜叶菊是拥有多种花色品种资源的观赏植物,同时具有蓝色、斑色等观赏植物中稀缺的花色表型,不同花色品种具有多种花青素代谢途径,解析其花色性状形成的分子机制能够为观赏植物的分子育种提供理论依据,其中鉴定的关键基因能够为蓝色花新... 详细信息

瓜叶菊是拥有多种花色品种资源的观赏植物,同时具有蓝色、斑色等观赏植物中稀缺的花色表型,不同花色品种具有多种花青素代谢途径,解析其花色性状形成的分子机制能够为观赏植物的分子育种提供理论依据,其中鉴定的关键基因能够为蓝色花新品种培育提供宝贵的基因资源。在对瓜叶菊多年研究的基础上,本文从瓜叶菊特殊的花青素结构、花青素生物合成调控途径及其花色研究的主要技术手段方面,综述了近20年来瓜叶菊花色研究的进展,具体内容包括:(1)瓜叶菊不同色系呈色的色素基础,尤其是蓝色花品种中特殊的多聚酰化色素结构;(2)多聚酰化修饰相关糖基化、酰基化修饰的瓜叶菊花青素代谢途径的基因,以及调控花色、花斑形成的MYB、MADS-box等转录因子的功能;(3)瓜叶菊中进行花色研究相关的高效遗传转化体系和病毒介导的基因沉默(VIGS,virus-induced gene silencing)体系,及其在花色研究中的研究进展。本文旨在为后续瓜叶菊及其他花卉花色研究和分子育种提供参考。

关键词：瓜叶菊花色花青素生物合成及调控

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矢车菊不同颜色花瓣酵母cDNA文库的构建

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北京林业大学学报 2024年第3期46卷 115-122页

作者：邓成燕王佳颖戴思兰临沂大学农林科学学院山东临沂276000 北京林业大学园林学院北京100083

【目的】矢车菊花瓣的蓝色呈色和品种间花色变异的分子调控机制尚不明晰。本研究采用Gateway技术构建了矢车菊6个不同花色品种花瓣的酵母cDNA文库,以期进一步通过酵母单杂交或双杂交技术筛选参与调控花瓣呈色的关键互作蛋白。【方法】... 详细信息

【目的】矢车菊花瓣的蓝色呈色和品种间花色变异的分子调控机制尚不明晰。本研究采用Gateway技术构建了矢车菊6个不同花色品种花瓣的酵母cDNA文库,以期进一步通过酵母单杂交或双杂交技术筛选参与调控花瓣呈色的关键互作蛋白。【方法】本研究以白色、粉色、红色、蓝色、紫色和墨色矢车菊花瓣为材料,提取总RNA后分离和纯化mRNA,合成双链cDNA后依次进行BP重组反应和LR重组反应,分别获得初级和次级文库。最后将次级文库质粒转化酵母Y187,获得矢车菊不同颜色花瓣的酵母cDNA文库。【结果】质量鉴定结果显示:初级文库的库容量为1.3×10^(7) CFU,重组率为100%,且插入片段长度均在1000 bp以上;次级文库的库容量为1.6×10^(7) CFU,重组率为100%,且插入片段长度均在1000 bp以上。酵母文库的滴度为3.5×10^(7) CFU/mL,随机挑选的24个单克隆经PCR检测后均扩增出明亮条带,重组率为100%,插入片段长度均大于1000 bp。【结论】本研究构建的矢车菊不同颜色花瓣酵母cDNA文库的质量较高,能满足酵母文库筛选的试验要求,为后续探究矢车菊花瓣的蓝色呈色和品种间花色变异的分子调控机制提供了材料基础。

关键词：矢车菊花色酵母 cDNA文库 Gateway技术

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Tobacco Rattle Virus-induced Phytoene Desaturase (PDS) Silencing in Centaurea cyanus

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Horticultural Plant Journal 2021年第2期7卷 159-166页

作者： Chengyan Deng Fan Zhang Jiaying Wang Yanfei Li He Huang silan dai Beijing Key Laboratory of Ornamental Plants Germplasm Innovation&Molecular Breeding National Engineering Research Center for FloricultureBeijing Laboratory of Urban and Rural Ecological Environment and College of Landscape ArchitectureBeijing Advanced Innovation Center for Tree Breeding by Molecular DesignBeijing Forestry UniversityBeijing 100083China

Virus-induced gene silencing(VIGS)is a genetic tool used to assess gene function.Tobacco rattle virus(TRV)is a VIGS vector commonly used to induce endogenous gene silencing in plants.However,there is no VIGS system established for Centaurea spp.We evaluated the effectiveness of a TRV-based VIGS system using phytoene desaturase(PDS)as a reporter gene in Centaurea cyanus.Three methods including pressure-,vacuum-and apical meristem-infiltrationwere tested to infect C.cyanus seedlings.Photobleached leaveswere only obtained using apicalmeristem-infiltration after a 14 d treatment.The CcPDS transcripts in photobleached leaves were significantly reduced compared with that in green leaves treated with empty TRV.Four C.cyanus cultivars were tested to detect their VIGS responses,and‘Dwarf Tom Pouce Blue’was the most sensitive.The agro-infiltration condition was optimized by screening for the optimal seedling stage as well as the optimum Agrobacterium density for efficient silencing.Seedlings with four true leaves and infiltration with an Agrobacterium density of OD_(600)0.5 were optimal conditions to obtain more photobleached leaves and more intense photobleached phenotype.The results demonstrated the feasibility of TRV-based VIGS for functional analysis of genes in C.cyanus.

关键词： Centaurea cyanus VIGS tobacco rattle virus PDS seedling photobleached

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High-density genetic map construction and identification of loci controlling flower-type traits in Chrysanthemum(Chrysanthemum×morifolium Ramat.)

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Horticulture Research 2020年第1期7卷 1469-1481页

作者： Xuebin Song Yuhui Xu Kang Gao Guangxun Fan Fan Zhang Chengyan Deng silan dai He Huang Huaigen Xin Yingying Li Beijing Advanced Innovation Center for Tree Breeding by Molecular Design Beijing Key Laboratory of Ornamental Plants Germplasm Innovation&Molecular BreedingNational Engineering Research Center for FloricultureBeijing Laboratory of Urban and Rural Ecological EnvironmentKey Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of the Ministry of EducationBeijing Forestry UniversitySchool of Landscape ArchitectureBeijing Forestry University35 East Qinghua RoadBeijing 100083China College of Landscape Architecture and Forestry Qingdao Agricultural UniversityQingdao 266109 ShandongChina Biomarker Technologies Co. LTDBeijing 101300China LC Science Co. LTD.Hangzhou 310018China

Flower type is an important and extremely complicated trait of chrysanthemum.The corolla tube merged degree(CTMD)and the relative number of ray florets(RNRF)are the two key factors affecting chrysanthemum flower type.However,few reports have clarified the inheritance of these two complex traits,which limits directed breeding for flower-type improvement.In this study,305 F1 hybrids were obtained from two parents with obvious differences in CTMD and RNRF performance.Using specific-locus amplified fragment sequencing(SLAF-seq)technology,we constructed a high-density genetic linkage map with an average map distance of 0.76 cM.Three major QTLs controlling CTMD and four major QTLs underlying RNRF were repeatedly detected in the 2 years.Moreover,the synteny between the genetic map and other Compositae species was investigated,and weak collinearity was observed.In QTL regions with a high degree of genomic collinearity,eight annotated genes were probed in the Helianthus annuus L.and Lactuca sativa L.var.ramosa Hort.genomes.Furthermore,20 and 11 unigenes were identified via BLAST searches between the SNP markers of the QTL regions and the C.vestitum and C.lavandulifolium transcriptomes,respectively.These results lay a foundation for molecular marker-assisted breeding and candidate gene exploration in chrysanthemum without a reference assembly.

关键词： breeding controlling traits

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早花露地小菊杂交后代株系的综合评价与筛选

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中国农业科学 2024年第1期57卷 173-189页

作者：宋想王钟曼张秋玲魏媛媛赵小刚刘波戴思兰北京林业大学园林学院/北京市林木分子设计育种高精尖创新中心/花卉种质创新与分子育种北京市重点实验室/城乡生态环境北京实验室/林木花卉遗传育种教育部重点实验室/国家花卉工程技术研究中心

【目的】建立早花露地小菊综合评价体系，筛选出花期为9月中旬至10月中旬、花色接近纯正红色或黄色、花型和株型等综合表现优异的株系，丰富华北地区早秋节庆用花。为进一步培育早花露地小菊新品种提供育种材料和选择方法。【方法】以... 详细信息

【目的】建立早花露地小菊综合评价体系，筛选出花期为9月中旬至10月中旬、花色接近纯正红色或黄色、花型和株型等综合表现优异的株系，丰富华北地区早秋节庆用花。为进一步培育早花露地小菊新品种提供育种材料和选择方法。【方法】以露地小菊3个早花亲本和1个重瓣性高且花色鲜艳亲本及其25个经田间初选的杂交后代无性株系为供试材料，划分花色、花期、花型、株型和叶型5个层次，从观测的30个原始性状中筛选出18个关键性状指标作为评价因子，利用层次分析法及K-Means聚类分析法建立综合评价体系，开展杂种株系的综合筛选。【结果】以早花、花型圆整丰满、红黄两种花色稳定为育种目标确定了层次分析法各性状的权重值，其中初花期起始时间（0.1817）权重值最大，其次是花色稳定性（0.1695）、盛花期单株着花数量（0.1383）、开花持续时间（0.1301），再次是花色适宜度（0.0565）、盛花期着花整齐度（0.0565）、花序直径（0.0539）、舌状花数量（0.0574）。通过K-Means聚类分析法将29份材料划分为3个等级，Ⅰ级10个（34.38%），Ⅱ级11个（37.93%），Ⅲ级8个（27.59%）；杂交子代性状分离广泛，花色、花型和株型均出现了超亲性状，但花期仍以2个早花亲本初花期最早，符合前人研究认为花期为多基因控制的数量性状。因此，早花露地小菊育种仍需筛选更多亲本，增加可早秋使用的露地小菊资源。【结论】本评价方法综合各类评价体系优势，以数量化结果充分表征田间观测的性状特征，以育种目标为导向进行层次分析赋值评价，为综合评价和筛选早花露地小菊优良品种提供了量化方法。建立的评价体系可以很好地对早花露地小菊进行评价与筛选，为后期早花露地小菊新品种的选育提供了一种可靠方法，有助于提高后代优良种质筛选效率。最终筛选出10个观赏性状优良的早花露地小菊株系，可作为新品种或新品系应用，也可作为育种材料。

关键词：露地小菊杂交后代综合评价层次分析法 K-Means聚类分析早花性状

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Application of Real-time Fluorescent Quantitative PCR in Studies on Plants

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Agricultural Biotechnology 2012年第1期1卷 1-7页

作者： Yueping MA silan dai Yanrong MA College of Sciences Northeastern University College of Landscape Architecture Beijing Forestry University College of Agriculture Ningxia University

Real-Lime fluorescent quantitative PCR is a method for quantitative analysis of gene expression developed in recent years, which has been widely used in various fields such as basic scientific research, clinical diagnosis, disease study, drug research and development since its appearance. It starts relatively late in study on plants, but has already been used for analysis of gene expression in plants and gene identification of exogenous genes. The principles or advantages and dis- advantages of real-time fluorescent quantitative PCR, or its potential problems and condition optimizations in tests were introduced in this study, and then the appli- cation and prospect of real-time fluorescent quantitative PCR in study on plants were also been discussed.

关键词： Real-time fluorescent quantitative PCR （FQ-PCR） Plant C, ene expression

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毛华菊3种瓣型株系再生体系的建立

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植物学报 2024年第2期59卷 245-256页

作者：武晓云廖敏凌李雪茹舒梓淳辛佳潼张伯晗戴思兰北京林业大学园林学院城乡生态环境北京实验室花卉种质创新与分子育种北京市重点实验室国家花卉工程技术研究中心北京100083

毛华菊(Chrysanthemum vestitum)是栽培菊花(C.×morifolium)的近缘六倍体野生种之一,其自然群体中的舌状花与栽培菊花一样具有典型的平瓣型、管瓣型和混合瓣型变异,是研究菊属植物瓣型变异的理想材料。其舌状花发育过程受生长素和花器... 详细信息

毛华菊(Chrysanthemum vestitum)是栽培菊花(C.×morifolium)的近缘六倍体野生种之一,其自然群体中的舌状花与栽培菊花一样具有典型的平瓣型、管瓣型和混合瓣型变异,是研究菊属植物瓣型变异的理想材料。其舌状花发育过程受生长素和花器官发育关键差异基因的影响,但目前缺乏稳定高效的不同瓣型毛华菊株系的再生体系,制约了毛华菊瓣型相关基因的研究。利用在河南省伏牛山收集的3种瓣型毛华菊株系,以叶片和茎间薄层为外植体建立再生体系。结果表明,以平瓣型叶片为外植体,其愈伤组织诱导和不定芽分化最佳培养基为MS+1 mg·L^(-1)NAA+2mg·L^(-1)6-BA,接种20天愈伤组织诱导率为100%,不定芽分化率达100%;最佳生根培养基为1/2MS+0.2 mg·L^(-1)NAA,生根率达100%。平瓣型毛华菊的叶片最佳再生体系也适用于管瓣型和混合瓣型株系,不定芽分化率分别为83.46%和91.67%,生根率均为100%。移栽后对开花植株进行观察,发现利用叶片再生体系获得的3种不同瓣型再生植株花型稳定,为后续利用不同瓣型株系解析舌状花形态变异机理提供了技术方法。

关键词：毛华菊不同瓣型叶片再生体系茎间薄层

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植物类胡萝卜素代谢及调控研究进展

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园艺学报 2022年第12期49卷 2559-2578页

作者：陆晨飞高月霞黄河戴思兰北京林业大学园林学院北京市林木分子设计育种高精尖创新中心花卉种质创新与分子育种北京市重点实验室国家花卉工程技术研究中心城乡生态环境北京实验室林木花卉遗传育种教育部重点实验室北京100083

本文概述了植物类胡萝卜素的代谢途径及关键的代谢流控制节点,重点总结了光合器官和类胡萝卜素存储器官中色素代谢的转录调控及转录后、翻译后、表观遗传等调控机制。

关键词：光合器官存储器官类胡萝卜素代谢调控

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