Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (4): 696-707.doi: 10.3864/j.issn.0578-1752.2021.04.003

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Using CRISPR/Cas9-mediated Targeted Mutagenesis of ZmFKF1 Delayed Flowering Time in Maize

YANG Min1(),XU HuaWei2,WANG CuiLing2,YANG Hu1,WEI YueRong1()   

  1. 1Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences/ Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization (MOA)/Guangdong Province Key Laboratory of Tropical and Subtropical Fruit Tree Research, Guangzhou 510640
    2College of Agriculture, Henan University of Science and Technology, Luoyang 471000, Henan
  • Received:2020-07-25 Accepted:2020-09-23 Online:2021-02-16 Published:2021-02-16
  • Contact: YueRong WEI E-mail:minyang_0123@126.com;weid18@163.com

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Abstract:

【Objective】FKF1 is a key gene that plays an important role in many plant flowering pathways. In order to study the function of FKF1 in maize, ZmFKF1 editing mutants were obtained by using CRISPR/cas9 technology. In this study, we used these mutants as materials to clarify the role of ZmFKF1 in maize flowering pathway through phenotypic analysis and the expression analysis of key flowering genes. The result provide a theoretical basis for molecular breeding and genetic improvement of maize.【Method】 ZmFKF1 gene was cloned from B104, and its structure was determined by sequence alignment. The targeted sequences of ZmFKF1 were designed according to the principle of CRISPR/Cas9 technology, then these targeted sequences were compared and analyzed in maize reference genome, and the non-specific target site was excluded. Finally, ZmFKF1-T1 on exon 1 of ZmFKF1 was selected to construct CRISPR/cas9 gene editing expression vector. At the same time, B104 immature embryos were selected as explants to transform by Agrobacterium-mediated genetic transformation, the resistant calli were obtained through resistance screening, and then buds and roots were induced. The T0 generation ZmFKF1 gene editing positive plants were obtained and verified by using the specific primers of cas9. The target site amplification and sequencing analysis was used to determine whether the T1 transgenic lines had mutation at the expected target site of ZmFKF1 and the type of mutation, and screened the homozygous lines of ZmFKF1 site mutation. After obtaining the above materials, the flowering phenotypes of these materials were statistically analyzed with wild type as control. At the same time, qRT-PCR was used to detect the expression of key genes that related to flowering pathway in the above materials and further verify the phenotype. 【Result】The target sequence was designed on exon1 of ZmFKF1 to construct gene editing expression vector. The transgenic lines were obtained by genetic transformation realized site directed mutagenesis of ZmFKF1. A total of 18 T0 generation ZmFKF1 gene-edited lines were obtained, and 6 of them were generated mutation on the exon1 with two different mutations types, including single base insertion and multiple bases deletion. The phenotype analysis shown that the flowering time of three T2 generation ZmFKF1 homozygous lines was delayed compared with wild type B104. Furthermore, the expression levels of three related flowering pathway genes (ZmGI, conz1 and ZmZCN8) in the mutants were significantly lower than those in the wild type B104 (P＜0.05), which was consistent with the late flowering phenotype of the mutants. 【Conclusion】CRISPR/Cas9 technology can be used to edit ZmFKF1 to obtain gene editing mutants, and the flowering time of these mutants is significantly delayed compared to wild type.

Key words: CRISPR/Cas9, gene editing, maize, ZmFKF1, flowering time

Table 1

Cloning and identification primers used in this study"

引物名称 Primer name 引物序列 Primer sequence (5′-3′) 用途 Usage
ZmFKF1-F CACCATGTTTGACGACGCGGGGGC ZmFKF1克隆
Cloning of ZmFKF1 gene
ZmFKF1-R GACTTTGCCATGCGAATCTACAATGC
FKF1-U6b-F GTTGAGGTGGACGCGGAGCCGGGA 载体构建及鉴定
Vector construction and verification
FKF1-U6b-R AAACTCCCGGCTCCGCGTCCACCT
U-F CTCCGTTTTACCTGTGGAATCG
gR-R CGGAGGAAAATTCCATCCAC
Pps-R TTCAGAggtctcTaccgACTAGTCACGCGTATGGAATCGGCAGCAAA
Pgs-L AGCGTGggtctcGctcgACGCGTATCCATCCACTCCAAGC
SP-L1 GCGGTGTCATCTATGTTACTAG
SP-R TGCAATAACTTCGTATAGGCT
Cas9-F CTGACGCTAACCTCGACAAG 鉴定Cas9
PCR detection for Cas9
Cas9-R CCGATCTAGTAACATAGATGACACC
ZmCri-F CCCTACCTGGCAGCAGAATC 靶点检测及测序引物
Target detection and sequencing primer
ZmCri-R AGCAGCCCAAATCTGTCAGTC
ZmUbi-F GTCTCCGTGGTGGTCAGTAAGT qRT-PCR检测引物
Primers used for qRT-PCR assay
ZmUbi-R GACACGAACAGCAGATACTTTGAC
ZmGI-F ACTTTCCCAACCAACGC
ZmGI-R GGGCAGCTACAACAATTTCA
conz1-F ATCGTCAATGGAGGTGGGTAT
conz1-R GACCCGACGAGAACAGGC
ZmZCN8-F ACTTGCACTGGATGGTGACAGATA
ZmZCN8-R CGGCCTCTCATAAAATATTAGCTCTT

Fig. 1

Target site of the gRNA in the ZmFKF1 gene, phylogenetic tree analysis of FKF1 amino acid sequences in nine species and the diagram of T-DNA region of pYLCRISPR/Cas9-ZmFKF1-T1 plasmid A: Position of the target in ZmFKF1 gene locus; B: Phylogenetic tree analysis of FKF1 amino acid sequences in 9 species; C: The diagram of T-DNA region of pYLCRISPR/Cas9-ZmFKF1-T1 plasmid"

Fig. 2

Transformation process of immature embryo of B104 A: Immature embryo of B104; B: Primary callus; C: Resistant callus; D: Resistant regeneration buds; E: Resistant regeneration plantlets"

Fig. 3

Identification and sequencing analysis of ZmFKF1 editing lines A: Identification of positive ZmFKF1 gene editing lines; B: Sequencing results of ZmFKF1 editing homozygote"

Table 2

Mutation analysis on the exon1 of ZmFKF1 gene"

株系 Line 序列 Sequence (5′-3′) 突变类型 Mutation type 是否纯合 Homozygous or not
B104 AGGTGGACGCGGAGCCGGGATGG --- ---
C1-6 AGGTGGACGCGGAGCTATGG +1（T）,-4（CGGG） 否 No
C2-12 AGGTGGACGCGGAGCCGAGGATGG +1（A） 是 Yes
C3-3 AGGTGGACGCGGAGCCG -9（GGATGGCCG） 否 No
C5-7 AGGTGGACGCGGAGCCGGGATGG -20（AGGTGGACGCGGAGCCGGGA） 否 No
C6-2 AGGTGGACGCGGAGCCATGG -3（GGG） 是 Yes
C7-1 AGGTGGACGCGGAGCCG -6（GGGATG） 是 Yes

Fig. 4

Prediction of ZmFKF1 protein domain and analysis of ZmFKF1 coding amino acid sequence in mutants A: Prediction results of ZmFKF1 protein domain; B: Amino acid sequence prediction analysis and comparison results of ZmFKF1 in ZmFKF1gene editing mutants"

Fig. 5

Phenotype and flowering related genes expression analysis of ZmFKF1 editing lines A: The phenotype of C7-1 compared to ‘B104’ at 67 days after sowing; B: The phenotype of C7-1 compared to ‘B104’ at 70 days after sowing; C: Expression analysis of flowering related genes in different genotype lines. ZmUbi primers were used for PCR as an internal control, the result of relative genes expression was calculated by 2-ΔΔCT method"

Table 3

Statistical results of flowering time in different genotype lines"

基因型
Genotype		 抽雄时间
Male phase		 散粉时间
Pollen phase		 吐丝时间
Female phase
B104 62.70±1.37 67.87±1.04 70.53±1.25
C2-12 63.43±1.04* 69.13±1.36* 72.57±1.65*
C6-2 64.07±0.98* 69.37±1.56* 71.90±1.22*
C7-1 63.96±1.30* 69.04±1.81* 72.33±1.12*
[1] 库丽霞, 孙朝辉, 王翠玲, 张君, 张伟强, 陈彦惠. 玉米光周期敏感相关性状发育动态QTL定位. 作物学报, 2010,36(4):602-611.
KU L X, SUN Z H, WANG C L, ZHANG J, ZHANG W Q, CHEN Y H. QTL analysis of the photoperiod sensitivity-related traits at different developmental stages in maize (Zea mays L.). Acta Agronomica Sinica, 2010,36(4):602-611. (in Chinese)
[2] OIL M, KHARSHIING E V. Photoreceptor mediated plant growth responses: Implications for photoreceptor engineering toward improved performance in crops. Frontiers in Plant Science, 2017,8:1181-1194.
[3] NELSON D C, LASSWELL J, ROGG L E, COHEN M A, BARTEL B. FKF1, a clock-controlled gene that regulates the transition to flowering in Arabidopsis. Cell, 2000,101(3):331-340.
[4] SAWA M, NUSINOW D A, KAY S A, IMAIZUMI T. FKF1 and GIGANTEA complex formation is required for day-length measurement in Arabidopsis. Science, 2007,318(5848):261-265.
[5] IMAIZUMI T. FKF1 F-Box protein mediates cyclic degradation of a repressor of CONSTANS in Arabidopsis. Science, 2005,309(5732):293-297.
[6] GORALOGIA G S, LIU T, ZHAO L, PANIPINTO P M, IMAIZUMI T. CYCLING DOF FACTOR 1 represses transcription through the TOPLESS co-repressor to control photoperiodic flowering in Arabidopsis. The Plant Journal, 2017,92(2):244-262.
[7] KOBAYASHI Y, WEIGEL D. Move on up, it’s time for change- mobile signals controlling photoperiod-dependent flowering. Genes & Development, 2007,21(19):2371-2384.
[8] HWANG D Y, PARK S, LEE S, IMAIZUMI T, SONG Y H. GIGANTEA regulates the timing stabilization of CONSTANS by altering the interaction between FKF1 and ZEITLUPE. Molecular Cells, 2019,42(10):693-701.
[9] CORBESIER L, VINCENT C, JANG S, FORNARA, F, FAN Q Z, SEARLE I, GIAKOUNTIS A, FARRONA S, GISSOT L, TURNBULL C, COUPLAND G. FT protein movement contributes to long-distance signaling in floral induction of Arabidopsis. Science, 2007,316:1030-1033.
[10] D’ALOIA M, BONHOMME D, BOUCHÉ F, TAMSEDDAKK, ORMENESE S, TORTI S, COUPLAND G, PERILLEUX C. Cytokinin promotes flowering of Arabidopsis via transcriptional activation of the FT paralogue TSF. The Plant Journal, 2011,65(6):972-979.
[11] SONG Y H, SMITH R W, TO B J, MILLAR A J, IMAIZUMI T. FKF1 conveys timing information for CONSTANS stabilization in photoperiodic flowering. Science, 2012,336(6084):1045-1049.
[12] HAN S H, YOO S C, LEE B D, AN G. PAEK N C. Rice FLAVIN-BINDING, KELCH REPEAT, F-BOX1 (OsFKF1) promotes flowering independent of photoperiod. Plant Cell Environ, 2015,38(12):2527-2540.
[13] MOUTIQ R, RIBAUT J M, EDMEADES G, KRAKOWSKY M D, LEE M. Quantitative trait loci for photoperiod response in maize. Maize Genetics Conference Abstracts, 2002(43):147.
[14] WANG C L, CHENG F F, SUN Z H, TANG J H, WU L C, KU L X, CHEN Y H. Genetic analysis of photoperiod sensitivity by molecular markers in a tropical by temperate maize recombinant in bred population. Theoretical and Applied Genetics, 2008,117:1129-1139.
[15] WANG X T, WU L J, ZHANG S F, WU L C, KU L X, WEI X M, XIE L L, CHEN Y H. Robust expression and association of ZmCCA1 with circadian rhythms in maize. Plant Cell Report, 2011,30:1261-1272.
[16] CHARDON F, VIRLON B, MOREAU L, FALQUE M, JOETS J, DECOUSSET L, MURIGNEUX A, CHARCOSSET A. Genetic architecture of flowering time in maize as inferred from QTL meta-analysis and synteny conservation with the rice genome. Genetics, 2004,162:2169-2185.
[17] COLES N D, MCMULLEN M D, BALINT-KURTI P J, PRATT R C , HOLLAND J B . Genetic control of photoperiod sensitivity in maize revealed by joint multiple population analysis. Genetics, 2010,184(3):799-812.
[18] DUCROCQ S, GIAUFFRET C, MADUR D, COMBES V, DUMAS F, JOUANNE S, COUBRICHE D, JAMIN P, MOREAU L, CHARCOSSET A. Fine mapping and haplotype structure analysis of a major flowering time quantitative trait locus on maize chromosome 10. Genetics, 2009,183(4):1555-1563.
[19] MU G Q, LIANG Y, ZHANG Z M, WU Y Q, LIU S J, PENG H, ZHANG S Z, PAN G T. Mapping quantitative trait loci associated with photoperiod sensitivity in maize (Zea mays L.). Agricultural Sciences in China, 2009,8(1):24-30.
[20] HUNG H Y, SHANNON L M, TIAN F, BRADBURY P J, HOLLAND J B. ZmCCT and the genetic basis of day-length adaptation underlying the post domestication spread of maize. Proceedings of the National Academy of Sciences of the United States of America, 2012,109(28):1913-1921.
[21] YANG Q, LI Z, LI W. CACTA-like transposable element in ZmCCT attenuated photoperiod sensitivity and accelerated the post domestication spread of maize. Proceedings of the National Academy of Sciences of the United States of America, 2013,110:16969-16974.
[22] 赵淑靓. 玉米伪应答调节基因ZmPRR73的克隆与表达分析[D]. 洛阳: 河南科技大学, 2018.
ZHAO S J. Gene cloning and expression analysis of pseudo-response regulator 73(ZmPRR73) in maize[D]. Luoyang: Henan University of Science and Technology, 2018. (in Chinese)
[23] 刘玲. 玉米光周期途径中蓝光响应节律基因ZmFKF1的分子进化与关联分析[D]. 雅安: 四川农业大学, 2014.
LIU L. Molecular evolution and association analysis of ZmFKF1, a clock-controlled gene in maize[D]. Yaan: Sichuan Agricultural University, 2014. (in Chinese)
[24] LI J F, NORVILLE J E, AACH J, MCCORMACK M, ZHANG D D, BUSH J, CHURCH G M, SHEEN J. Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nature Biotechnology, 2013,31(8):688-691.
[25] MAO Y F, ZHANG H, XU N F, ZHANG B T, GOU F, ZHU J K. Application of the CRISPR-Cas system for efficient genome engineering in plants. Molecular Plant, 2013,6(6):2008-2011.
[26] WANG Y P, CHENG X, SHAN Q W, ZHANG Y, LIU J X, GAOC X, QIU J L. Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew. Nature Biotechnology, 2014,32(9):947-951.
[27] LI C, UNVER T, ZHANG B H. A high-efficiency CRISPR/Cas9 system for targeted mutagenesis in cotton (Gossypium hirsutum L.). Scientific Reports, 2017,7:43902.
[28] ITO Y, NISHIZAWA-YOKOI A, ENDO M, MIKAMI M, TOKI S. CRISPR/Cas9-mediated mutagenesis of the RIN locus that regulates tomato fruit ripening. Biochemical & Biophysical Research Communications, 2015,467(1):76-82.
[29] ZHOU X H, JACOBS T B, XUE L J, HARDING S A, TSAI C J. Exploiting SNPs for biallelic CRISPR mutations in the outcrossing woody perennial Populus reveals 4-coumarate: CoA ligase specificity and redundancy. New Phytologist, 2015,208(2):298-301.
[30] MALNOY M, VIOLA R, JUNG M H, KOO O J, KIM S, KIM J S, VELASCO R, KANCHISWAMY C N. DNA-free genetically edited grapevine and apple protoplast using CRISPR/Cas9 ribonucleoproteins. Frontiers in Plant Science, 2016,7:1904.
[31] TIAN S W, JIANG L J, GAO Q, ZHANG J, ZONG M, ZHANG H Y, REN Y, GUO S G, GONG G Y, LIU F, XU Y. Efficient CRISPR/Cas9-based gene knockout in watermelon. Plant Cell Reports, 2017,36(3):399-406.
[32] 杨禄山, 郭晔, 胡洋, 文颖强. 利用CRISPR/Cas9 系统敲除葡萄中VviEDR2提高对白粉病的抗性. 园艺学报, 2020,47(4):623-634.
YANG L S, GUO Y, HU Y, WEN Y Q. CRISPR/Cas9-mediated mutagenesis of VviEDR2 results in enhanced resistance to powdery mildew in grapevine (Vitis vinifera). Acta Horticulturae Sinica, 2020,47(4):623-634. (in Chinese)
[33] 郑爱红, 张芬, 江敏, 袁巧, 江雷雨, 陈清, 汤浩茹, 孙勃. 利用CRISPR/Cas9技术靶向编辑芥蓝BoaZDS. 园艺学报, 2019,46(1):57-64.
ZHENG A H, ZHANG F, JIANG M, YUAN Q, JIANG L Y, CHEN Q, TANG H R, SUN B. Targeted editing of BoaZDS by CRISPR/Ca9 technology in Chinese kale. Acta Horticulturae Sinica, 2019,46(1):57-64. (in Chinese)
[34] 邹修平, 范迪, 彭爱红, 何永睿, 许兰珍, 雷天刚, 姚利晓, 李强, 罗克明, 陈善春. CRISPR/Cas9介导柑橘CsLOB1基因启动子的多位点编辑. 园艺学报, 2019,46(2):337-344.
ZOU X P, FAN D, PENG A H, HE Y R, XU L Z, LEI T G, YAO L X, LI Q, LUO K M, CHEN S C. CRISPR/Cas9-mediated editing of multiple sites in the citrus CsLOB1 promoter. Acta Horticulturae Sinica, 2019,46(2):337-344. (in Chinese)
[35] FENG C, YUAN J, WANG R, LIU Y, BIRCHLER J A, HAN F P. Efficient targeted genome modification in maize using CRISPR/Cas9 system. Journal of Genetics & Genomics, 2016,43(1):37-43.
[36] 曾栋昌, 马兴亮, 谢先荣, 祝钦泷, 刘耀光. 植物crispr/cas9多基因编辑载体构建和突变分析的操作方法. 中国科学: 生命科学, 2018,48(7):783-794.
ZENG D C, MA X L, XIE X R, ZHU Q L, LIU Y G. A protocol for CRISPR/Cas9-based multi-gene editing and sequence decoding of mutant sites in plants. Scientia Sinica Vitae, 2018,48(7):783-794. (in Chinese)
[37] 王新涛. 玉米光周期敏感基因ZmGI的克隆和功能分析[D]. 郑州: 河南农业大学, 2009.
WANG X T. Cloning and characterization of photoperiod sensitive gene ZmGI in maize. Zhengzhou: Henan Agricultural University, 2009. (in Chinese)
[38] MILLER T A, MUSLIN E H, DORWEILER J E. A maize CONSTANS-like gene, conz1, exhibits distinct diurnal expression patterns in varied photoperiods. Planta, 2008,227(6):1377-1388.
[39] MENG X, MUSZYNSKI M G, DANILEVSKAYA O N. The FT-like ZCN8 gene functions as a floral activator and is involved in photoperiod sensitivity in maize. The Plant Cell, 2011,23(3):942-960.
[40] LEBLANC C, ZHANG F, MENDEZ J, LOZANO Y, CHATPAR K, IRISH V F, JACOB Y. Increased efficiency of targeted mutagenesis by CRISPR/Cas9 in plants using heat stress. The Plant Journal, 2018,93(2):377-386.
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