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| 1 | 外源褪黑素处理对月季幼苗盐胁迫的缓解效应显示文摘【目的】研究外源褪黑素(MT)处理对盐胁迫下中国古老月季‘月月粉’Rosa chinensis‘Old Blush’的缓解效应及其机制,为解决月季栽培中的盐害问题提供理论依据。【方法】以90 d扦插苗为材料,分别通过根部浇灌0、5、10和20μmol·L^−1褪黑素进行预处理5 d后,再对根部浇灌200 mmol·L^−1氯化钠溶液进行盐胁迫处理20 d,以浇灌清水的植株为空白对照,研究了外源褪黑素处理对月季盐胁迫的缓解效应。【结果】与空白对照相比,不同浓度外源褪黑素处理可显著降低盐胁迫下月季叶片的相对电导率和丙二醛质量摩尔浓度(P<0.05),同时可显著提高可溶性蛋白质、抗坏血酸和脯氨酸的质量分数(P<0.05),特别是显著提高了超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)的活性,从而增强了活性氧的清除能力。【结论】外源褪黑素处理可增强月季‘月月粉’的抗氧化酶活性以及渗透调节物质的质量分数,从而抑制活性氧的积累,防止膜脂过氧化,提高耐盐性。 | 刘政 胡孙田 沈晓飞 朱培 褚旭东 李卓娅 罗平 | 2020 | 浙江农林大学学报2020，37，5： | 10 |
| 2 | TRV介导的葡萄叶片VvANR基因瞬时表达分析显示文摘为建立葡萄叶片TRV-VIGS系统,分析验证VvANR基因功能,本研究以烟草脆裂病毒(Tobacco rattle virus,TRV)介导基因沉默表达载体pTRV2-ANR,采用真空侵染和主叶脉针孔注射法分别侵染葡萄幼嫩、成熟叶片,观察表型并测定原花青素含量,实时荧光定量PCR测定被侵染叶片中VvANR及其相关基因表达。结果表明,真空侵染的幼嫩、成熟叶片分别于第6、第7天出现漂白,侵染15 d后,幼嫩叶片几乎全部漂白,成熟叶片大面积漂白。主叶脉针孔注射侵染的幼嫩、成熟叶片分别于第17、第19天出现漂白;侵染25 d后,幼嫩和成熟叶片发生全叶漂白,且漂白均沿主叶脉逐渐扩展至整个叶面;对照均未发生漂白现象。pTRV2-ANR侵染后叶片中原花青素含量极显著降低,VvANR基因表达量极显著降低,且原花青素生物合成相关基因VvLAR1、VvLAR2、VvMYBPA1、VvMYBPA2、VvUFGT表达量显著降低,除VvUFGT表达量呈显著差异外,其余均呈极显著差异;而VvANS、VvDFR表达量轻微上调,且与对照差异不显著。本研究建立了葡萄叶片VIGS体系,为基因功能快速验证提供了新方法;同时,揭示了葡萄叶片中原花青素含量、VvANR表达及与相关基因的关系,本研究为完善葡萄原花青素积累机制奠定了一定的理论基础。 | 杨波 刘海霞 牛铁泉 张鹏飞 梁长梅 赵旗峰 温鹏飞 | 2021 | 核农学报2021，35，4： | 8 |
| 3 | A detached petal disc assay and virus-induced gene silencing facilitate the study of Botrytis cinerea resistance in rose flowers显示文摘Fresh-cut roses(Rosa hybrida)are one of the most important ornamental crops worldwide,with annual trade in the billions of dollars.Gray mold disease caused by the pathogen Botrytis cinerea is the most serious fungal threat to cut roses,causing extensive postharvest losses.In this study,we optimized a detached petal disc assay(DPDA)for artificial B.cinerea inoculation and quantification of disease symptoms in rose petals.Furthermore,as the identification of rose genes involved in B.cinerea resistance could provide useful genetic and genomic resources,we devised a virusinduced gene silencing(VIGS)procedure for the functional analysis of B.cinerea resistance genes in rose petals.We used RhPR10.1 as a reporter of silencing efficiency and found that the rose cultivar‘Samantha’showed the greatest decrease in RhPR10.1 expression among the cultivars tested.To determine whether jasmonic acid and ethylene are required for B.cinerea resistance in rose petals,we used VIGS to silence the expression of RhLOX5 and RhEIN3(encoding a jasmonic acid biosynthesis pathway protein and an ethylene regulatory protein,respectively)and found that petal susceptibility to B.cinerea was affected.Finally,a VIGS screen of B.cinerea-induced rose transcription factors demonstrated the potential benefits of this method for the high-throughput identification of gene function in B.cinerea resistance.Collectively,our data show that the combination of the DPDA and VIGS is a reliable and highthroughput method for studying B.cinerea resistance in rose. | Xiaoqian Cao Huijun Yan Xintong Liu Dandan Li Mengjie Sui Jie Wu Hongqiang Yu Zhao Zhang | 2019 | Horticulture Research2019，6，1： | 5 |
| 4 | Overview and detectability of the genetic modifications in ornamental plants显示文摘The market of ornamental plants is extremely competitive,and for many species genetic engineering can be used to introduce original traits of high commercial interest.However,very few genetically modified(GM)ornamental varieties have reached the market so far.Indeed,the authorization process required for such plants has a strong impact on the profitability of the development of such products.Considering the numerous scientific studies using genetic modification on ornamental species of interest,a lot of transformed material has been produced,could be of commercial interest and could therefore be unintentionally released on the market.The unintentional use of GM petunia in breeding programs has indeed recently been observed.This review lists scientific publications using GM ornamental plants and tries to identify whether these plants could be detected by molecular biology tools commonly used by control laboratories. | Anne-Laure Boutigny Nicolas Dohin David Pornin Mathieu Rolland | 2020 | Horticulture Research2020，7，1： | 3 |
| 5 | VIGS技术在观赏植物中的应用显示文摘后基因组时代,基因功能研究备受关注,病毒诱导基因沉默(virus-induced gene silencing,VIGS)技术便是一种高效的基因功能鉴定方法。VIGS虽然在模式植物以及经济作物上已经有了大量的研究基础,但在观赏植物领域研究较少。观赏植物具有一定的经济、生态和社会价值,在对其基因功能进行研究时,利用高效的技术方法有利于更好地提高其观赏性、耐胁迫等品质,实现这些价值。所以,为降低观赏植物栽培成本,提高质量,将VIGS技术用于观赏植物研究,可有效解决传统方法效率低,存在技术瓶颈等问题并实现其价值的统一。本综述对VIGS这项简便、快速的技术的分子机理、体系的优化与发展以及其在观赏植物生长发育、遗传改良、抗胁迫等方面基因功能研究上的应用进行总结,希望该技术在不断优化的同时能够在观赏植物基因功能研究中得到更广泛的运用,为观赏植物基因功能鉴定、品质改良和定向育种提供新思路。 | 吴燕 刘咪 陈明 姜宇 孙静 | 2020 | 分子植物育种2020，18，3： | 2 |
| 6 | VIGS技术初步鉴定RgMLO6和RlMLO7基因对白粉病的负调控功能显示文摘为了探讨蔷薇科植物MLO基因在抗白粉病中的作用,研究应用病毒诱导的基因沉默技术(virus induced gene silencing,VIGS)抑制了大花香水月季RgMLO6基因和长尖叶蔷薇RlMLO7基因的表达,随后接种白粉菌对这2个基因进行抗性鉴定.研究发现在VIGS载体转化植株叶片20 d后,RgMLO6和RlMLO7基因的相对表达量显著下降了80%~90%,沉默效果明显.分别对2个基因沉默后的嫩叶进行白粉病抗性鉴定,大花香水月季和长尖叶蔷薇的抗性水平较对照组均提高.显微镜观察白粉菌接种2个基因沉默后植株叶片中菌丝体的生长情况,整体表现出沉默植株叶表皮细胞上的白粉菌生长较对照组生长缓慢.结果表明RgMLO6与RlMLO7基因对蔷薇科植物的白粉病有负向调控作用. | 朱正锦 陈敏 田永贤 向贵生 张颢 邱显钦 | 2023 | 云南大学学报(自然科学版)2023，45，1： | 1 |
| 7 | 病毒诱导的基因沉默技术用于植物色素代谢机制的研究进展显示文摘色彩是评价园艺植物观赏性状的重要指标,而植物色素是影响植物色彩表型的关键因子。植物色素及其代谢产物在植物观赏器官颜色形成、植株生长发育调节及对逆境胁迫的响应等方面发挥着重要的作用,是植物研究领域长期关注的热点问题。病毒诱导基因沉默(virus-induced gene silencing,VIGS)是利用植物同源依赖性防御机制,特异性降低宿主内源性基因表达的一种重要基因组学工具,能够通过快速诱导植物基因沉默表型的产生,表征基因的功能,为缺乏遗传转化体系的植物的基因功能鉴定提供高效可行的替代方案。本文综述了VIGS技术在植物色素的生物合成、降解和调控机制上的应用现状,并探讨了VIGS技术在探究色素调控机制上的潜力和未来前景,以期进一步完善对不同植物色素的代谢过程和调控机制的理解,为改良植物色彩性状提供参考依据。 | 潘多 张嵩玥 刘芳伊 田清尹 杨秀莲 王良桂 岳远征 | 2023 | 生物工程学报2023，39，7： | 1 |
| 8 | Tobacco Rattle Virus-induced Phytoene Desaturase (PDS) Silencing in Centaurea cyanus显示文摘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. | Chengyan Deng Fan Zhang Jiaying Wang Yanfei Li He Huang Silan Dai | 2021 | Horticultural Plant Journal2021，7，2： | 1 |
| 9 | 基于三角梅的病毒诱导基因沉默体系的建立与优化显示文摘三角梅是紫茉莉科灌木,具有较高的观赏价值,是研究花色叶色调控的理想材料。建立快速高效的基因验证体系,是三角梅基因功能研究的关键。以三角梅‘金心双色’品种为材料,选用八氢番茄红素脱氢酶基因(PDS)为指示基因,探索不同接种部位和基因片段长度对烟草脆裂病毒(TRV)诱导三角梅叶片内源PDS mRNA沉默效果的影响,建立适用于三角梅的病毒诱导基因沉默体系(VIGS)。结果表明,摩擦注射嫩叶法相比顶端嫩茎沉默效果更明显;基因沉默片段长度在336 bp和457 bp均可诱导PDS基因沉默,后者效果更加明显。初步构建了三角梅VIGS体系,为后续基因功能研究提供了技术参考。 | 刘珍银 段郅臻 彭婷 王童欣 王健 | 2023 | 生物技术通报2023，39，7： | 0 |
| 10 | Peculiarity of transcriptional and H3K27me3 dynamics during peach bud dormancy显示文摘Bud dormancy facilitates the survival of meristems under harsh environmental conditions.To elucidate how molecular responses to chilling accumulation controlling dormancy in peach buds,chromatin immunoprecipitation sequencing to identify the H3K27me3 modifications and RNA sequencing of two peach cultivars with pronounced differences in chilling requirement were carried out,the results showed that genes associated with abscisic acid and gibberellic acid signal pathways play key roles in dormancy regulation.The results demonstrated that peach flower bud differentiation occurred continuously in both cultivars during chilling accumulation,which was correlated with the transcript abundance of key genes involved in phytohormone metabolism and flower bud development under adverse conditions.The more increased strength in high chillingrequirement cultivar along with the chilling accumulation at the genome-wide level.The function of the dormancy-associated MADS-box gene PpDAM6 was identified,which is involved in leaf bud break in peach and flower development in transgenic Nicotiana tabacum(NC89).In addition,PpDAM6 was positively regulated by PpCBF,and the genes of putative dormancy-related and associated with metabolic pathways were proposed.Taken together,these results constituted a theoretical basis for elucidating the regulation of peach bud dormancy transition. | Yalin Zhao Yong Li Ke Cao Weichao Fang Changwen Chen Xinwei Wang Jinlong Wu Wenwu Guo Lirong Wang | 2024 | Horticultural Plant Journal2024，10，1： | 0 |
| 11 | TRV介导的溪荪VIGS转化体系的构建与鉴定显示文摘病毒诱导基因沉默(Virus-induced gene silencing,VIGS)技术为缺乏稳定遗传转化体系的植物进行基因功能分析提供了可能,为解决缺少稳定遗传转化体系的植物进行基因功能验证的需求,以单子叶植物溪荪(Iris sanguinea)为材料,克隆IsPDS基因的特异性片段并构建其VIGS重组载体pTRV2-IsPDS,通过注射法侵染溪荪叶片,结果显示:pTRV1和pTRV2-IsPDS的菌液OD_(600)调至1.8~2.0后重悬,重悬液OD600调至0.8~1.0,通过注射器叶脉注射方式侵染溪荪叶片效果最佳。在室外温度为15~20℃的下午6~8时进行,用1 mL注射器针头扎伤溪荪叶片外表皮,沿着平行脉缓慢注射重悬液1mL至溪荪叶片维管束中,14 d左右会出现明显的白化表型。在出现表型变化的植株和空载组中均检测到TRV1和TRV2的病毒载体,白化植株的IsPDS表达量显著低于对照组。侵染液配制中通过提高携带病毒载体的农杆菌的浓度提高侵染效率,且接种后无需遮光。 | 刘桂伶 徐诺 史恭发 王玲 | 2024 | 植物研究2024，44，1： | 0 |
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