CLC number:
On-line Access: 2022-04-11
Received: 2022-02-25
Revision Accepted: 2022-03-11
Crosschecked: 2022-04-19
Cited: 0
Clicked: 1454
Citations: Bibtex RefMan EndNote GB/T7714
https://orcid.org/0000-0001-9734-3090
Wenbo PAN, Zhentao CHENG, Zhiguo HAN, Hong YANG, Wanggen ZHANG, Huawei ZHANG. Efficient genetic transformation and CRISPR/Cas9-mediated genome editing of watermelon assisted by genes encoding developmental regulators[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B2200119 @article{title="Efficient genetic transformation and CRISPR/Cas9-mediated genome editing of watermelon assisted by genes encoding developmental regulators", %0 Journal Article TY - JOUR
生长调节基因介导的西瓜高效遗传转化与CRISPR/Cas9基因编辑体系创新点:通过比较多个生长调节基因和农杆菌菌株,选取生长调节基因AtGRF5和农杆菌菌株GV3101,在西瓜中建立了高效且不依赖于基因型的转化体系,实现了西瓜突变体的快速创制。 方法:通过测试生长调节因子基因对西瓜转化率的影响,发现多个生长调节基因可以提高西瓜的遗传转化效率。其中AtGRF5可将西瓜的转化效率0.88%左右提高至24.73%。此外,筛选了不同的农杆菌菌株,发现使用GV3101表达AtGRF5可获得最高的转化效率。进一步使用AtGRF5辅助转化的方式对西瓜进行基因编辑,成功在T0代对PDS基因进行敲除并获得纯合突变体。 结论:使用农杆菌GV3101过表达AtGRF5可将西瓜的转化效率从0.88%左右提高至24.73%,且在两个基因型的西瓜中作用类似。该方法也可结合基因编辑技术用于西瓜突变体的快速获取。 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
Reference[1]DebernardiJM, TricoliDM, ErcoliMF, et al., 2020. A GRF-GIF chimeric protein improves the regeneration efficiency of transgenic plants. Nat Biotechnol, 38(11):1274-1279. [2]FengQ, XiaoL, HeYZ, et al., 2021. Highly efficient, genotype-independent transformation and gene editing in watermelon (Citrullus lanatus) using a chimeric CLGRF4-GIF1 gene. J Integr Plant Biol, 63(12):2038-2042. [3]GuoSG, ZhaoSJ, SunHH, et al., 2019. Resequencing of 414 cultivated and wild watermelon accessions identifies selection for fruit quality traits. Nat Genet, 51(11):1616-1623. [4]KongJX, Martin-OrtigosaS, FinerJ, et al., 2020. Overexpression of the transcription factor GROWTH-REGULATING FACTOR5 improves transformation of dicot and monocot species. Front Plant Sci, 11:572319. [5]LoweK, WuE, WangN, et al., 2016. Morphogenic regulators Baby boom and Wuschel improve monocot transformation. Plant Cell, 28(9):1998-2015. [6]Méndez-Hernández1 HA, Ledezma-Rodríguez M, Avilez-Montalvo RN,et al., 2019. Signaling overview of plant somatic embryogenesis. Front Plant Sci, 10:77. [7]NanasatoY, TabeiY, 2020. A method of transformation and current progress in transgenic research on cucumbers and Cucurbita species. Plant Biotechnol (Tokyo), 37(2):141-146. [8]ParkSM, LeeJS, JegalS, et al., 2005. Transgenic watermelon rootstock resistant to CGMMV (cucumber green mottle mosaic virus) infection. Plant Cell Rep, 24(6):350-356. [9]TanYY, DuH, WuX, et al., 2020. Gene editing: an instrument for practical application of gene biology to plant breeding. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(6):460-473. http://doi.org/10.1631/jzus.B1900633 [10]TianSW, JiangLJ, GaoQ, et al., 2017. Efficient CRISPR/Cas9-based gene knockout in watermelon. Plant Cell Rep, 36(3):399-406. [11]WangJF, WangYP, ZhangJ, et al., 2021. The NAC transcription factor CLNAC68 positively regulates sugar content and seed development in watermelon by repressing CLINV and CLGH3.6. Hortic Res, 8:214. [12]WangYP, WangJF, GuoSG, et al., 2021. CRISPR/Cas9-mediated mutagenesis of CLBG1 decreased seed size and promoted seed germination in watermelon. Hortic Res, 8:70. [13]XingHL, DongL, WangZP, et al., 2014. A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol, 14:327. [14]ZhangXY, XuGC, ChengCH, et al., 2021. Establishment of an Agrobacterium-mediated genetic transformation and CRISPR/Cas9-mediated targeted mutagenesis in hemp (Cannabis sativa L.). Plant Biotechnol J, 19(10):1979-1987. Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou
310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE |
Open peer comments: Debate/Discuss/Question/Opinion
<1>