| 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T1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T2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。 |
| 关键词: 黄籽甘蓝型油菜；油菜育种 BnaTT2基因 基因编辑技术 |
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| 基金项目: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） |
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| Creation of yellow-seed Brassica napus with BnaTT2 gene deletion |
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AN Ran,ZHANG Yanfeng,CHEN Nana,et al
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| Abstract: |
| 【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 T1 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 T2 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 T2 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. |
| Key words: yellow-seed Brassica napus L. rape breeding BnaTT2 genome editing technology |
