甘蓝型油菜带节子叶再生体系的建立及遗传转化应用.PDF

中国农学通报 2017,33(34):40-45 Chinese Agricultural Science Bulletin 甘蓝型油菜带节子叶再生体系的建立及遗传转化应用 廖志强，王小武，孙 亮，魏志英 （江西省宜春市农业科学研究所，江西宜春336000） 摘 要：优良再生体系的建立是农杆菌介导植物遗传转化的基础。本研究建立以甘蓝型油菜含部分子 叶节的子叶为外植体的再生体系，该再生体系为：切取5 日苗龄甘蓝型油菜含部分子叶节的带柄子叶为 外植体，置于添加3 mg/L 6-BA 的MS 基本培养基中培养5 天，外植体再生出丛生不定芽，不定芽再生率 为100% 。基于新建立的带节子叶再生体系，通过农杆菌介导，成功地将用pFGC5941 载体构建的 BnTFL 1基因干扰载体转入甘蓝型油菜中，从播种到得到生根抗性苗，整个转化周期只需要60~70 天，较 传统甘蓝型油菜以不带节子叶为外植体100~ 130天的转化周期大大缩短。 关键词：甘蓝型油菜；子叶节；子叶；再生体系；遗传转化 中图分类号：Q813.1+2 文献标志码：A 论文编号：cas EstablishmentofRegenerationSystemofCotyledonwithCotyledonaryNode fromBrassicanapusandTransformationApplication LiaoZhiqiang,WangXiaowu,SunLiang,WeiZhiying (AgriculturalScienceInstituteofYichun,YichunJiangxi336000) Abstract: Theestablishmentofagood regenerationsystemisthefoundationofagrobacterium-mediated genetictransformation.Withcotyledonsincludingpartcotyledonarynodefrom Brassica napus asexplants,a regeneration system was established. The content of this system was that cotyledons including part cotyledonarynodefromfivedaysageofrapeseedseedlingswerecutasexplantandthenwereculturedinthe MSmediasupplementedwith 3 mg/L 6-BA,andfivedayslater,shootregeneratedfromtheexplant.The frequency of shoot regeneration was 100%. Based on the new regeneration system of cotyledon with cotyledonarynode,a BnTFL1RNAivectorconstructedfrompFGC5941 vectorwastransformedinto Brassica napussuccessfullybyagrobacteriummediationintheresearch.Thewholetransformationperiodfromseeding togettingrootedresistantseedlingswas 60-70 days.Comparedwith 100-130 daysperiodoftraditional transformationbasedonregenerationcultureofcotyledonwithoutcotyledonarynode,thisnewtransformation methodobviouslyshor