BnaTT2基因缺失黄籽甘蓝型油菜材料的创制  

Creation of yellow-seed Brassica napus with BnaTT2 gene deletion

作　　者：安然 张彦锋[1] 陈娜娜[1] 韦世豪[1] 朱彦涛[1] 穆建新[1] AN Ran;ZHANG Yanfeng;CHEN Nana;WEI Shihao;ZHU Yantao;MU Jianxin(Hybrid Rapeseed Research Center of Shaanxi Province,Yangling,Shaanxi 712100,China)

机构地区：[1]陕西省杂交油菜研究中心,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2022年第7期35-40,47,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：中国科学院“西部之光”人才培养引进计划(XAB2018AW16);陕西省创新能力支撑计划项目(2020TD-051);陕西省油菜分子设计育种科技资源共享平台项目(2021PT-036)。

摘　　要：【目的】创制可以稳定遗传的黄籽甘蓝型油菜种质,为黄籽油菜材料的创制与利用奠定理论与实践基础。【方法】以甘蓝型油菜BnaTT2基因为研究对象,以甘蓝型油菜K407为受体材料,构建相应单靶点的CRISPR/Cas9基因编辑载体,利用农杆菌介导法将载体转化入油菜植株,测定获得的黄籽油菜基因编辑材料TT2-Mutant的籽粒色度和千粒质量,并用红外分析仪测定其脂肪酸组分相对含量和含油量。【结果】对甘蓝型油菜T_(1)代转基因株系靶位点的序列检测和突变类型分析结果表明,BnaTT2基因靶位点被成功编辑,编辑类型为单碱基的插入或缺失。获得的T_(2)代BnaTT2基因缺失油菜材料籽粒千粒质量(4.74 g)相比对照材料K407(3.62 g)增大;种子颜色变黄,黄色度等级为4.8;籽粒中的含油量极显著升高6.10%,亚油酸和亚麻酸相对含量分别极显著提升3.13%和4.99%,芥酸相对含量则极显著降低1.53%。【结论】BnaTT2基因敲除后可导致相应功能缺失,从而改变油菜籽的种皮颜色,并对其脂肪酸组成产生影响。【Objective】This study created yellow-seed Brassica napus germplasm that can be inherited stably to provide foundation for the creation and utilization of yellow-seed rape materials.【Method】The Brassica napus BnaTT2 gene was selected for constructing corresponding single-target CRISPR/Cas9 gene editing vector using B.napus K407 as the acceptor.The vector was then transformed into the rape plant using the Agrobacterium-mediated method.The rape gene editing material TT2-Mutant was used to measure its seed color and thousand-grain quality,and the content of fatty acids was measured with an infrared analyzer.【Result】The target site sequence detection and mutation type analysis of the T_(1) generation transgenic line showed that the target site of the BnaTT2 gene was successfully edited,and the editing type was single-base insertion or deletion.The obtained T_(2) generation BnaTT2 gene-deficient rape material had larger thousand-grain weight(4.74 g)than the control(3.62 g).The seed turned yellow with a yellowness grade of 4.8.The biological analysis of the homozygous T_(2) generation of yellow rapeseed showed that the oil content was very significantly increased(6.1%),contents of linoleic acid and linolenic acid were significantly increased by 3.13%and 4.99%,while the content of erucic acid was very significantly reduced by 1.53%.【Conclusion】The functional mutation caused by the knockout of BnaTT2 gene could change the color of rapeseed seed coat and affect fatty acid composition.

关 键 词：黄籽甘蓝型油菜 油菜育种 BnaTT2基因 基因编辑技术 

分 类 号：S565.403.2[农业科学—作物学]