JIANG Ruipeng,ZHU Xiaolin,WANG Baoqiang,et al.Resistance Analysis of Transgenic T1 Tomato to Yellow Leaf Curl Disease[J].Northern Horticulture,2024,(9):1-10.[doi:10.11937/bfyy.20233633]
转基因T1代番茄对黄化曲叶病抗性分析
- Title:
- Resistance Analysis of Transgenic T1 Tomato to Yellow Leaf Curl Disease
- 文章编号:
- 1001-0009(2024)09-0001-10
- Keywords:
- transgenic tomato; T1; TYLCV; disease resistance
- 分类号:
- S 641.2
- 文献标志码:
- A
- 摘要:
- 以转黄化曲叶病（TYLCV）编码外壳蛋白（capsid protein，CP）的V1、编码复制激发蛋白（replication-associated protein，Rep）C1基因的T1代J6番茄为试材，采用盆栽法，研究转基因T1代番茄接种TYLCV后不同时期的生长特性、抗病相关酶活性和抗病相关基因表达量的影响，以期明确转V1、C1基因的T1代J6番茄植株对TYLCV的抗性。结果表明：通过RNAi技术获得的转V1、C1基因的T1代转基因J6番茄植株成功转化且供试番茄材料转化率为87.5%；T1代转基因植株抗病性研究发现：转基因番茄可以有效缓解病害对其生长的抑制，光合特性得到一定程度的恢复；其抗病相关酶PAL、PPO活性下降；MDA和抗氧化相关酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和过氧化物酶（POD）活性显著降低；抗病相关基因的表达量Ty-1、Ty-2、Ty-3显著上调。说明T1代转基因番茄对TYLCV具有一定的抗性，且J6+V1的抗病性强于J6+C1。
- Abstract:
- In order to clarify the resistance of T1 generation J6 tomato plants transformed with V1 and C1 genes to TYLCV，the V1 gene encoding capsid protein (CP) of TYLCV and the J6 gene encoding replication-associated protein (Rep) C1 were used as test materials to study the effects of TYLCV on the growth characteristics，disease resistance-related enzyme activities and disease resistance-related gene expression at different stages after inoculation with TYLCV by pot experiment.The results showed that the T1 transgenic J6 tomato plants with V1 and C1 genes obtained by RNAi technology were successfully transformed and the transformation rate of the tested tomato materials was 87.5%.The study of disease resistance of T1 generation transgenic plants found that transgenic tomatoes could limitedly alleviate the inhibition of disease on their growth，and the photosynthetic characteristics were restored to a certain extent.The activities of PAL and PPO related to disease resistance decreased.The MDA content and activities of antioxidant enzymes superoxide dismutase (SOD)，catalase (CAT)，ascorbate peroxidase (APX) and peroxidase (POD) were significantly decreased.The expression levels of resistance-related genes Ty-1，Ty-2 and Ty-3 were significantly up-regulated.The results showed that the T1 generation of transgenic tomatoes had certain resistance to TYLCV，and the resistance of ‘J6+V1’ was stronger than that of ‘J6+C1’.
参考文献/References:
［1］王光正,吕剑,毛娟,等.油菜素内酯对低温胁迫下番茄幼苗生理指标的影响［J］.甘肃农业大学学报,2021,56(4):69-75.［2］金宁,肖雪梅,郁继华,等.不同品种番茄果实矿质元素含量评价［J］.甘肃农业大学学报,2020,55(4):76-84.［3］李云洲,岳宁波,陈相儒,等.RNA干扰V1和C1提高番茄植株对番茄黄化曲叶病毒(TYLCV)的抗性［J］.核农学报,2021,35(9):2027-2034.［4］李明骏,王吕鑫,郎鑫源,等.双生病毒复制增强蛋白C3/AC3的研究进展［J］.植物保护学报,2023,50(3):593-601.［5］魏艳.烟粉虱化学感受蛋白在番茄黄化曲叶病毒传播中的作用［D］.荆州:长江大学,2023.［6］CUI J,JIANG N,ZHOU X,et al.Tomato MYB49 enhances resistance to Phytophthora infestans and tolerance to water deficit and salt stress［J］.Planta,2018,248(6):1487-1503.［7］REZK A,ABDALLAH N,ABDEL A,et al.Transgene-mediated RNA silencing of TYLCV genes affecting the accumulation of viral DNA in plants［J］.Arab J Biotech,2006,9(1):143-158.［8］王贤,刘放,魏小红,等.不同种质番茄材料抗番茄黄化曲叶病毒病特性研究［J］.浙江农业学报,2021,33(11):2085-2097.［9］张琛,陈思博,胡本进,等.化感水稻内生真菌发酵产物的抑菌活性评价及其对水稻保护性酶活性的影响［J］.核农学报,2019,33(12):2347-2353.［10］邹芳斌,司龙亭,李新,等.黄瓜枯萎病抗性与防御系统几种酶活性关系的研究［J］.华北农学报,2008,23(3):181-184.［11］JI J Y,YANG J,ZHANG B W,et al.Sodium pheophorbide a controls cherry tomato gray mold (Botrytis cinerea) by destroying fungal cell structure and enhancing disease resistance-related enzyme activities in fruit［J］.Pesticide Biochemistry and Physiology,2020,166:104581.［12］邹虎成,李玉洪,覃柳兰,等.抗番茄黄化曲叶病毒病基因ty-5的功能标记开发与利用［J］.中国蔬菜,2022(5):23-28.［13］金宝霞.番茄黄化曲叶病毒DNA复制蛋白基因的沉默研究［D］.兰州:甘肃农业大学,2022.［14］王伟杰.番茄黄化曲叶病毒V1和C2基因沉默的遗传转化及抗病性研究［D］.兰州:甘肃农业大学,2022.［15］王贤.外源NO对番茄黄化曲叶病毒病抗性的诱导及转录组分析［D］.兰州:甘肃农业大学,2022.［16］CHAHID K,LAGLAOUI A,ZANTAR S,et al.Antioxidant-enzyme reaction to the oxidative stress due to alpha-cypermethrin,chlorpyriphos,and pirimicarb in tomato (Lycopersicon esculentum Mill.)［J］.Environmental Science and Pollution Research,2015,22(22):18115-18126.［17］YANG Y,LIU Q,WANG G X,et al.Germination,osmotic adjustment,and antioxidant enzyme activities of gibberellin-pretreated Picea asperata seeds under water stress［J］.New Forests,2010,39(2):231-243.［18］GONG B,WEN D,VANDENLANGENBERG K,et al.Comparative effects of NaCl and NaHCO3 stress on photosynthetic parameters,nutrient metabolism,and the antioxidant system in tomato leaves［J］.Scientia Horticulturae,2013,157:1-12.［19］HEATH R L,PACKER L.Photoperoxidation in isolated chloroplasts.I.Kinetics and stoichiometry of fatty acid peroxidation［J］.Archives of Biochemistry and Biophysics,1968,125(1):189-198.［20］GAYOSO C,POMAR F,NOVO-UZAL E,et al.The Ve-mediated resistance response of the tomato to Verticillium dahliae involves H2O2,peroxidase and lignins and drives PAL gene expression［J］.BMC Plant Biology,2010（10）:232.［21］SPAGNA G,BARBAGALLO R N,CHISARI M,et al.Characterization of a tomato polyphenol oxidase and its role in browning and lycopene content［J］.Journal of Agricultural and Food Chemistry,2005,53(6):2032-2038.［22］高晓萍,张婧,牛天航,等.甜菜碱对高温胁迫下茄子幼苗生理特性的影响［J］.浙江农业学报,2023,35(9):2097-2108.［23］孙伟博,宫新栋,周燕,等.转玉米PEPC和PPDK基因杨树苗期的光合生理特性［J］.林业科学,2020,56(7):33-43.［24］陈兰兰，王丽，吴亚娟，等．植物响应干旱胁迫的分子和微生态机制［J/OL］．分子植物育种，(2023-04-07)［2024-09-20］．https：//kns．cnki．net/kcms/detail/46.1068．S．20230406.1634.006．html． ［25］邱思鑫,阮宏春,宋美仙,等.内生解淀粉芽孢杆菌TB2菌株活性物质诱导辣椒果抗疫病的生化机理［J］.热带作物学报,2010,31(10):1813-1820.［26］骆霞,滕元旭,张雪蒙,等.外源氯化血红素对NaCl胁迫下加工番茄幼苗生理特性的影响［J］.北方园艺,2023(18):1-8.［27］权建华,陈修斌,鄂利锋,等.番茄抗病育种研究概况及展望［J］.蔬菜,2020(1):43-47.［28］田兆丰,刘伟成,厚凌宇,等.不同番茄品种对番茄黄化曲叶病毒的抗病性分析［J］.中国农业科学,2016,49(14):2850-2856.［29］刘放.不同番茄材料黄化曲叶病毒病抗性研究［D］.兰州:甘肃农业大学,2020.［30］王琳珊,宋层孝,宋建军,等.番茄黄化曲叶病毒抗病基因Ty-2的SCAR标记筛选［J］.北方园艺,2015(15):94-99.
相似文献/References:
[1]肖宇,张剑侠,周起,等.中国野生华东葡萄芪合成酶基因启动子转化番茄的研究[J].北方园艺,2013,37(01):89.
XIAO Yu,ZHANG Jian-xia,ZHOU Qi,et al.Research on the STS Gene Promoter of Chinese Wild Vitis?pseudoreticulata Transformed Tomato[J].Northern Horticulture,2013,37(9):89.
备注/Memo
第一作者简介：姜睿鹏（1995-），男，硕士研究生，研究方向为生物技术与工程等。E-mail：405620525@qq.com.责任作者：魏小红(1964-)，女，博士，教授，博士生导师，现主要从事植物逆境生理与分子生理等研究工作。E-mail:weixh@gsau.edu.cn.基金项目：国家自然科学基金资助项目（32060401）；甘肃省自然科学基金资助项目（21JR7RA808）。收稿日期：2023-10-20