Research Progress of CRISPR/Cas9 Technology and Its Application in Horticultural Plants

doi:10.16420/j.issn.0513-353x.2023-0760

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (7): 1439-1454.doi: 10.16420/j.issn.0513-353x.2023-0760

• Reviews • Next Articles

Research Progress of CRISPR/Cas9 Technology and Its Application in Horticultural Plants

WANG Chenyu¹, LIU Mengjun¹^,², WANG Lixin¹^,^*(), LIU Zhiguo¹^,²^,^*()

¹ College of Horticulture，Hebei Agricultural University，Baoding，Hebei 071000，China
² Chinese Jujube Research Center，Hebei Agricultural University，Baoding，Hebei 071000，China

Received:2024-03-26 Revised:2024-05-22 Online:2024-07-25 Published:2024-07-19
Contact: WANG Lixin, LIU Zhiguo

Abstract

Abstract:

CRISPR/Cas9 is the immune defense system in bacteria and archaea. In 1987，a special repeat interval sequence was firstly discovered in Escherichia coli. Later，this repeat interval sequence was also found in more than 20 bacteria and archaea. In 2002，this special sequence was officially named as CRISPR. Subsequently，a series of studies using CRISPR/Cas9 technology for gene editing were carried out. CRISPR/Cas9 technology is the third generation gene editing technology，following ZFNs（zinc finger nucleases）technology and TALENs（transfer activator like effector nucleases）technology. This system has the advantages of simple operation design，high mutation efficiency，low cost，and has been successively applied in many horticultural plants such as citrus，grape，banana，strawberry，cucumber，and potato. This article reviews the principles and research progress of CRISPR/Cas9 technology，discusses the developmental history of various editors，including single base editor，double base editor，and guided editor，introduces the application of CRISPR/Cas9 technology in horticultural plants，and finally proposes the remaining problems and future prospects of CRISPR/Cas9 technology.

Key words: CRISPR/Cas9, gene editor, research progress, horticultural plants

WANG Chenyu, LIU Mengjun, WANG Lixin, LIU Zhiguo. Research Progress of CRISPR/Cas9 Technology and Its Application in Horticultural Plants[J]. Acta Horticulturae Sinica, 2024, 51(7): 1439-1454.

Figures/Tables 3

References 107

[1]	Anzalone A V, Randolph P B, Davis J R, Sousa A A, Koblan L W, Levy J M, Chen P J, Wilson C, Newby G A, Raguram A, Liu D R. 2019. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature, 576 (7785):149-157.
[2]	Ballester A R, Molthoff J, de Vos R, Hekkert B T L, Orzaez D, Fernaݩndez-Moreno J P, Tripodi P, Grandillo S, Martin C, Bovy A. 2010. Biochemical and molecular analysis of pink tomatoes:deregulated expression of the gene encoding transcription factor SlMYB12 leads to pink tomato fruit color. Plant Physiology, 152 (1):71-84. doi: 10.1104/pp.109.147322 pmid: 19906891
[3]	Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero A D, Horvath P. 2007. CRISPR provides acquired resistance against viruses in prokaryotes. Science, 315 (5819):1709-1712. doi: 10.1126/science.1138140 pmid: 17379808
[4]	Broggini G A L, Wöhner T, Fahrentrapp J, Kos T D, Flachowsky H, Peil A, Hanke M V, Richter K, Patocchi A, Gessler C. 2014. Engineering fire blight resistance into the apple cultivar‘Gala’using the FB_MR5CC-NBS-LRR resistance gene of Malus × robusta 5. Plant Biotechnology Journal, 12 (6):728-733.
[5]	Brooks C, Nekrasov V, Lippman Z B, van Eck J. 2014. Efficient gene editing in tomato in the first generation using the clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 system. Plant Physiology, 166 (3):1292-1297.
[6]	Brouns J J S, Jore M M, Lundgren M, Westra R E, Slijkhuis J H R, Snijders P L A, Dickman J M, Makarova S K, Koonin V E, Oost V D J. 2008. Small CRISPR RNAs guide antiviral defense in prokaryotes. Science, 321 (5891):960-964. doi: 10.1126/science.1159689 pmid: 18703739
[7]	Campa M, Piazz S, Righetti L, Oh C S, Conterno L, Borejsza-Wysocka E, Nagamangala K C, Beer S V, Aldwinckle H S, Malnoy M. 2019. HIPM is a susceptibility gene of Malus spp.:reduced expression reduces susceptibility to Erwinia amylovora. Molecular Plant-Microbe Interactions, 32 (2):167-175.
[8]	Chandrasekaran J, Brumin M, Wolf D, Leibman D, Klap C, Pearlsman M, Pearlsman M, Sherman A, Arazi T, Gal-On A. 2016. Development of broad virus resistance in non-transgenic cucumber using CRISPR/Cas9 technology. Molecular Plant Pathology, 17 (7):1140-1153. doi: 10.1111/mpp.12375 pmid: 26808139
[9]	Charrier A, Vergne E, Dousset N, Richer A, Petiteau A, Chevreau E. 2019. Efficient targeted mutagenesis in apple and first time edition of pear using the CRISPR-Cas9 system. Frontiers in Plant Science, 10:40. doi: 10.3389/fpls.2019.00040 pmid: 30787936
[10]	Chen Wenjun. 2019. Function analysis of FvCO4in the early development of strawberry fruit[M. D. Dissertation]. Shenyang: Shenyang Agricultural University. (in Chinese)
	陈文君. 2019. FvCO4在草莓果实早期发育中的功能鉴定[硕士论文]. 沈阳: 沈阳农业大学.
[11]	Choi J, Chen W, Suiter C C, Lee C, Chardon F M, Yang W, Leith A, Daza R M, Martin B, Shendure J. 2021. Precise genomic deletions using paired prime editing. Nature Biotechnology, 40 (2):218-226. doi: 10.1038/s41587-021-01025-z pmid: 34650269
[12]	Cong L, Ran F A, Cox D, Lin S, Barretto R, Habib N, Hsu P D, Wu X, Jiang W, Marraffini L A, Zhang F. 2013. Multiplex genome engineering using CRISPR/Cas systems. Science, 339 (6121):819-823. doi: 10.1126/science.1231143 pmid: 23287718
[13]	Feng Y, Kou X, Yuan S, Wu C, Zhao X, Xue Z, Li Q, Huang Z, Sun Y. 2023. CRISPR/Cas9-mediated SNAC9 mutants reveal the positive regulation of tomato ripening by SNAC9 and the mechanism of carotenoid metabolism regulation. Horticulture Research, 10 (4):uhad019.
[14]	Filler Hayut S, Melamed Bessudo C, Levy A A. 2017. Targeted recombination between homologous chromosomes for precise breeding in tomato. Nature Communications, 8 (1):15605.
[15]	Gao Q, Luo H, Li Y, Liu Z, Kang C. 2020. Genetic modulation of RAP alters fruit coloration in both wild and cultivated strawberry. Plant Biotechnol J, 18 (7):1550-1561. doi: 10.1111/pbi.13317 pmid: 31845477
[16]	Han J, Li X, Li W, Yang Q, Li Z, Cheng Z, Lv L, Zhang L, Han D. 2023. Isolation and preliminary functional analysis of FvICE1,involved in cold and drought tolerance in Fragaria vesca through overexpression and CRISPR/Cas9 technologies. Plant Physiology and Biochemistry, 196:270-280.
[17]	Hsu P D, Lander E S, Zhang F. 2014. Development and applications of CRISPR-Cas 9 for genome engineering. Cell, 157 (6):1262-1278.
[18]	Hu Y, Zhang J, Jia H, Sosso D, Li T, Frommer W B, Yang B, White F F, Wang N, Jones J. B. 2014. Lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease. Proceedings of the National Academy of Sciences, 111 (4):E521-E529.
[19]	Isaacson T, Ronen G, Zamir D, Hirschberg J. 2002. Cloning of tangerine from tomato reveals a carotenoid isomerase essential for the production of β-carotene and xanthophylls in plants. The Plant Cell, 14 (2):333-342.
[20]	Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A. 1987. Nucleotide sequence of the iap gene,responsible for alkaline phosphatase isozyme conversion in Escherichia coli,and identification of the gene product. Journal of Bacteriology, 169 (12):5429-5433. doi: 10.1128/jb.169.12.5429-5433.1987 pmid: 3316184
[21]	Jansen R, van Embden J D A, Gaastra W, Schouls L M. 2002. Identification of genes that are associated with DNA repeats in prokaryotes. Molecular Microbiology, 43 (6):1565-1575. doi: 10.1046/j.1365-2958.2002.02839.x pmid: 11952905
[22]	Jia Hui. 2019. Construction of CRISPR/Cas9 vector for grape resistance to powdery mildew VvBAK1 and VvLecRK1 and study on grape transformation[M. D. Dissertation]. Yangling: Northwest A & F University. (in Chinese)
	贾慧. 2019. 葡萄抗白粉病VvBAK1和VvLecRK1 CRISPR/Cas9载体构建及转化葡萄的研究[硕士论文]. 杨凌: 西北农林科技大学.
[23]	Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna J A, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science, 337 (6096):816-821. doi: 10.1126/science.1225829 pmid: 22745249
[24]	Jore M M, Lundgren M, van Duijn E, Bultema J B, Westra E R, Waghmare S P, Wiedenheft B, Pul U, Wurm R, Wagner R, Beijer M R, Barendregt A, Zhou K, Snijders A P L, Dickman M J, Doudna J A, Boekema E J, Heck A J R, Oost V D J, Brouns S J. 2011. Structural basis for CRISPR RNA-guided DNA recognition by Cascade. Nature Structural & Molecular Biology, 18 (5):529-536.
[25]	Gaudelli N M, Komor A C, Rees H A, Packer M S, Badran A H, Bryson D I, Liu D R. 2017. Programmable base editing of A • T to G • C in genomic DNA without DNA cleavage. Nature, 551 (7681):464-471.
[26]	González M N, Massa G A, Andersson M, Turesson H, Olsson N, Fält A S, Storani1 L, Oneto C, Hofvander P, Feingold S E. 2020. Reduced enzymatic browning in potato tubers by specific editing of a polyphenol oxidase gene via ribonucleoprotein complexes delivery of the CRISPR/Cas9 system. Frontiers in Plant Science, 10:1649.
[27]	Kaur N, Alok A, Kumar P, Kaur N, Awasthi P, Chaturvedi S, Pandey P, Pandey A, Pandey A K, Tiwari S. 2020. CRISPR/Cas 9 directed editing of lycopene epsilon-cyclase modulates metabolic flux for β-carotene biosynthesis in banana fruit. Metabolic Engineering, 59:76-86.
[28]	Kieu N P, Lenman M, Wang E S, Petersen B L, Andreasson E. 2021. Mutations introduced in susceptibility genes through CRISPR/Cas 9 genome editing confer increased late blight resistance in potatoes. Scientific Reports, 11 (1):4487.
[29]	Klap C, Yeshayahou E, Bolger A M, Arazi T, Gupta S K, Shabtai S, Usadel B, Salts Y, Barg R. 2017. Tomato facultative parthenocarpy results from SlAGAMOUS-LIKE 6 loss of function. Plant Biotechnology Journal, 15 (5):634-647.
[30]	Komor A C, Kim Y B, Packer M S, Zuris J A, Liu D R. 2016. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature, 533 (7603):420-424.
[31]	Komor A C, Badran H A, Liu D R. 2017. CRISPR-Based Technologies for the manipulation of eukaryotic genomes. Cell, 169 (3):559. doi: S0092-8674(17)30417-8 pmid: 28431253
[32]	Kost T D, Gessler C, Jänsch M, Flachowsky H, Patocchi A, Broggini G A. 2015. Development of the first cisgenic apple with increased resistance to fire blight. Public Library of Science ONE, 10 (12):e0143980.
[33]	Lang Z, Wang Y, Tang K, Tang D, Datsenka T, Cheng J, Zhang Y, Handa A K, Zhu J K. 2017. Critical roles of DNA demethylation in the activation of ripening-induced genes and inhibition of ripening-repressed genes in tomato fruit. Proceedings of the National Academy of Sciences, 114 (22):E4511-E4519.
[34]	LeBlanc C, Zhang F, Mendez J, Lozano Y, Chatpar K, Irish V F, Jacob Y. 2018. Increased efficiency of targeted mutagenesis by CRISPR/Cas 9 in plants using heat stress. The Plant Journal, 93 (2):377-386.
[35]	Lemmon Z H, Reem N T, Dalrymple J, Soyk S, Swartwood K E, Rodriguez-Leal D, Eck J V, Lippman Z B. 2018. Rapid improvement of domestication traits in an orphan crop by genome editing. Nature Plants, 4 (10):766-770. doi: 10.1038/s41477-018-0259-x pmid: 30287957
[36]	Li C, Zhang R, Meng X, Chen S, Zong Y, Lu C, Qiu J, Chen Y, Li J, Gao C. 2020. Targeted,random mutagenesis of plant genes with dual cytosine and adenine base editors. Nature Biotechnology, 38 (7):875-882.
[37]	Li J F, Norville J E, Aach J, McCormack M, Zhang D, Bush J, Church G M, Sheen J. 2013. Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nature Biotechnology, 31 (8):688-691.
[38]	Li Mengyuan. 2020. Study on the anti downy mildew function of grape disease progression related protein PR4b[M. D. Dissertation]. Yangling: Northwest A & F University. (in Chinese)
	李梦媛. 2020. 葡萄病程相关蛋白PR4b抗霜霉病功能研究[硕士论文]. 杨凌: 西北农林科技大学.
[39]	Li Qiuyue. 2019. Identification of citrus U-box gene family and study on CRISPR/Cas9 editing of CcPUB4 gene[M. D. Dissertation]. Chongqing: Southwest University. (in Chinese)
	李秋月. 2019. 柑橘U-box基因家族的鉴定和CRISPR/Cas9编辑CcPUB4基因的研究[硕士论文]. 重庆: 西南大学.
[40]	Li R, Liu C, Zhao R, Wang L, Chen L, Yu W, Zhang S, Sheng J, Shen L. 2019. CRISPR/Cas9-Mediated SlNPR1 mutagenesis reduces tomato plant drought tolerance. BioMed Central Plant Biology, 19 (1):1-13.
[41]	Li R, Zhang L, Wang L, Chen L, Zhao R, Sheng J, Shen L. 2018. CRISPR/Cas9-mediated SlCBF1 mutagenesis reduces tomato plant chilling tolerance. Journal of Agricultural and Food Chemistry, 66 (34):9042-9051.
[42]	Lin Qiupeng, Zhu Xiuli, Ma Linsha, Yao Pengcheng. 2024. Recent advances in prime editing system. Journal of South China Agricultural University, 45 (2):159-171. (in Chinese)
	林秋鹏, 朱秀丽, 马琳莎, 姚鹏程. 2024. 引导编辑系统研究进展. 华南农业大学学报, 45 (2):159-171.
[43]	Liu Jiangna, Zhang Xiying, Bai Yunfeng, Zhang Aiping. 2023. Using CRISPR/Cas 9 technology for targeted editing and processing of tomato SlACS2. Acta Agriculturae Boreali-occidentalis Sinica, 32 (6):910-918. (in Chinese)
	刘江娜, 张西英, 白云凤, 张爱萍. 2023. 利用CRISPR/Cas9技术定向编辑加工番茄SlACS2. 西北农业学报, 32 (6):910-918.
[44]	Liu Y, Zhang C, Wang X, Li X, You C. 2022. CRISPR/Cas 9 technology and its application in horticultural crops. Horticultural Plant Journal, 8 (4):395-407.
[45]	López-Casado G, Sánchez-Raya C, Ric-Varas P D, Paniagua C, Blanco-Portales R, Muñoz-Blanco J, Pose S, Matas A J, Mercado J A. 2023. CRISPR/Cas 9 editing of the polygalacturonase FaPG1 gene improves strawberry fruit firmness. Horticulture Research, 10 (3):uhad011.
[46]	Lucioli A, Tavazza R, Baima S, Fatyol K, Burgyan J, Tavazza M. 2022. CRISPR-Cas9 Targeting of the eIF4E1 gene extends the potato virus Y resistance spectrum of the Solanum tuberosum L. cv. Desirée. Frontiers in Microbiology, 13:873930.
[47]	Lu Y, Tian Y, Shen R, Yao Q, Zhong D, Zhang X, Zhu J. 2021. Precise genome modification in tomato using an improved prime editing system. Plant Biotechnology Journal, 19 (3):415. doi: 10.1111/pbi.13497 pmid: 33091225
[48]	Maioli A, Gianoglio S, Moglia A, Acquadro A, Valentino D, Milani A M, Prohens J, Orzaez D, Granell A, Lanteri S, Comino C. 2020. Simultaneous CRISPR/Cas9 editing of three PPO genes reduces fruit flesh browning in Solanum melongena L. Frontiers in Plant Science, 11:607161.
[49]	Mali P, Aach J, Stranges P B, Esvelt K M, Moosburner M, Kosuri S, Yang L, Church G M. 2013. CAS 9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nature Biotechnology, 31 (9):833-838.
[50]	Mojica F J, Díez-Villaseñor C S, García-Martínez J, Soria E. 2005. Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. Journal of Molecular Evolution, 60 (2):174-182. doi: 10.1007/s00239-004-0046-3 pmid: 15791728
[51]	Nakayasu M, Akiyama R, Lee H J, Osakabe K, Osakabe Y, Watanabe B, Sugimoto Y, Umemoto N, Saito K, Muranaka T, Mizutani M. 2018. Generation of α-solanine-free hairy roots of potato by CRISPR/Cas9 mediated genome editing of the St16DOX gene. Plant Physiology and Biochemistry, 131:70-77. doi: S0981-9428(18)30184-0 pmid: 29735370
[52]	Nekrasov V, Staskawicz B, Weigel D, Jones J D, Kamoun S. 2013. Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas 9 RNA-guided endonuclease. Nature Biotechnology, 31 (8):691-693.
[53]	Nekrasov V, Wang C, Win J, Lanz C, Weigel D, Kamoun S. 2017. Rapid generation of a transgene-free powdery mildew resistant tomato by genome deletion. Scientific Reports, 7 (1):482. doi: 10.1038/s41598-017-00578-x pmid: 28352080
[54]	Nguyen N H, Bui T P, Le N T, Nguyen C X, Le M T T, Dao N T, Phan Q,Le, T V, To H M T, Pham N B, Chu H H, Do P T. 2023. Disrupting Sc-uORFs of a transcription factor bZIP1 using CRISPR/Cas 9 enhances sugar and amino acid contents in tomato (Solanum lycopersicum). Planta, 257 (3):57. doi: 10.1007/s00425-023-04089-0 pmid: 36795295
[55]	Nishihara M, Higuchi A, Watanabe A, Tasaki K. 2018. Application of the CRISPR/Cas 9 system for modification of flower color in Torenia fournieri. BioMed Central Plant Biology, 18 (1):1-9.
[56]	Omori M, Yamane H, Osakabe K, Osakabe Y, Tao R. 2021. Targeted mutagenesis of CENTRORADIALIS using CRISPR/Cas 9 system through the improvement of genetic transformation efficiency of tetraploid highbush blueberry. The Journal of Horticultural Science and Biotechnology, 96 (2):153-161.
[57]	Qi Jingjing. 2017. Creating dwarf cucumber strains using CRISPR/Cas9 technology[M. D. Dissertation]. Nanjing: Nanjing Agricultural University. (in Chinese)
	戚晶晶. 2017. 利用CRISPR/Cas9技术创制矮生黄瓜株系[硕士论文]. 南京: 南京农业大学.
[58]	Qin Nannan. 2020. Cloning and functional validation of cucumber carpel number gene CsCLAVAATA3[Ph. D. Dissertation]. Jingzhong: Shanxi Agricultural University. (in Chinese)
	秦楠楠. 2020. 黄瓜心皮数基因CsCLAVATA3的克隆及功能验证[博士论文]. 晋中: 山西农业大学.
[59]	Ren C, Liu X, Zhang Z, Wang Y, Duan W, Li S, Liang Z. 2016. CRISPR/Cas9-mediated efficient targeted mutagenesis in Chardonnay(Vitis vinifera L.). Scientific Reports, 6 (1):32289.
[60]	Roldan M V G, Périlleux C, Morin H, Huerga-Fernandez S, Latrasse D, Benhamed M, Bendahmane A. 2017. Natural and induced loss of function mutations in SlMBP21 MADS-box gene led to jointless-2 phenotype in tomato. Scientific Reports, 7 (1):4402.
[61]	Shan Q, Wang Y, Li J, Zhang Y, Chen K, Liang Z, Zhang K, Liu J, Xi J J, Qiu J L, Gao C. 2013. Targeted genome modification of crop plants using a CRISPR-Cas system. Nat Biotechnol, 31(8):686-688. doi: 10.1038/nbt.2650 pmid: 23929338
[62]	Shang L, Tao J, Song J, Wang Y, Zhang X, Ge P, Li F, Dong H, Gai W, Grierson D, Ye Z, Zhang Y. 2024. CRISPR/Cas9-mediated mutations of FANTASTIC FOUR gene family for creating early flowering mutants in tomato. Plant Biotechnology Journal, 22 (3):774-784.
[63]	Shao X, Wu S, Dou T, Zhu H, Hu C, Huo H, He W, Deng G, Sheng O, Bi F, Gao H, Dong T, Li C, Yang Q, Yi G. 2020. Using CRISPR/Cas 9 genome editing system to create MaGA20ox2 gene-modified semi-dwarf banana. Plant Biotechnology Journal, 18 (1):17.
[64]	Shimatani Z, Kashojiya S, Takayama M, Terada R, Arazoe T, Ishii H, Teramura H, Yamamoto H, Komatsu H, Miura K, Ezura H, Nishida K, Ariizumi T, Kondo A. 2017. Targeted base editing in rice and tomato using a CRISPR-Cas 9 cytidine deaminase fusion. Nature Biotechnology, 35 (5):441-443. doi: 10.1038/nbt.3833 pmid: 28346401
[65]	Shnaider Y, Elad Y, Rav-David D, Pashkovsky E, Leibman D, Kravchik M, Shtarkman-Cohen M, Gal-On A, Spiegelman Z. 2023. Development of powdery mildew resistance in cucumber using CRISPR/Cas9-mediated mutagenesis of CsaMLO8. Phytopathology, 113 (5):786-790.
[66]	Su S, Xiao W, Guo W, Yao X, Xiao J, Ye Z, Wang N, Jiao K, Lei M, Peng Q, Hu X, Huang X, Luo D, Luo D. 2017. The CYCLOIDEA-RADIALIS module regulates petal shape and pigmentation,leading to bilateral corolla symmetry in Torenia fournieri(Linderniaceae). New Phytologist, 215 (4):1582-1593.
[67]	Tang X, Chen S, Yu H, Zheng X, Zhang F, Deng X, Xu Q. 2021. Development of a gRNA-tRNA array of CRISPR/Cas 9 in combination with grafting technique to improve gene-editing efficiency of sweet orange. Plant Cell Reports, 40 (12):2453-2456.
[68]	Tang X, Simon S, Ren Q, Jia X, Li M, Fan T, Yin D, Xiang S, Guo Y, Liu L, Zheng X, Qi Y, Zhang Y. 2020. Plant prime editors enable precise gene editing in rice cells. Molecular Plant, 13 (5):667-670. doi: S1674-2052(20)30072-1 pmid: 32222487
[69]	Tao W, Liu Q, Huang S, Wang X, Qu S,Guo, Ou D, Li G, Zhang Y, Xu X, Huang X. 2021. CABE-RY: A PAM-flexible dual-mutation base editor for reliable modeling of multi-nucleotide variants. Molecular Therapy-Nucleic Acids, 26:114-121. doi: 10.1016/j.omtn.2021.07.016 pmid: 34513298
[70]	Tasaki K, Higuchi A, Watanabe A, Sasaki N, Nishihara M. 2019. Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers. Scientific Reports, 9 (1):15831. doi: 10.1038/s41598-019-51808-3 pmid: 31676875
[71]	Tasaki K, Yoshida M, Nakajima M, Higuchi A, Watanabe A, Nishihara M. 2020. Molecular characterization of an anthocyanin-related glutathione S-transferase gene in Japanese gentian with the CRISPR/Cas9 system. BioMed Central Plant Biology, 20 (1):370.
[72]	Tian S, Jiang L, Cui X, Zhang J, Guo S, Li M, Zhang H, Re Yn, Gong G, Zong M, Liu F, Chen Q, Xu Y. 2018. Engineering herbicide-resistant watermelon variety through CRISPR/Cas9-mediated base-editing. Plant Cell Reports, 37 (9):1353-1356. doi: 10.1007/s00299-018-2299-0 pmid: 29797048
[73]	Tomlinson L, Yang Y, Emenecker R, Smoker M, Taylor J, Perkins S, Smith J, MacLean D, Olszewski N, Jones J D. 2019. Using CRISPR/Cas 9 genome editing in tomato to create a gibberellin-responsive dominant dwarf DELLA allele. Plant Biotechnology Journal, 17 (1):132-140. doi: 10.1111/pbi.12952 pmid: 29797460
[74]	Tripathi J N, Ntui O, Ron M, Muiruri S K, Britt A, Tripathi L. 2019. CRISPR/Cas 9 editing of endogenous banana streak virus in the B genome of Musa spp. overcomes a major challenge in banana breeding. Communications Biology, 2 (1):46. doi: 10.1038/s42003-019-0288-7 pmid: 31924917
[75]	Tripathi J N, Ntui V O, Shah T, Tripathi L. 2021. CRISPR/Cas9-mediated editing of DMR6 orthologue in banana(Musa spp.)confers enhanced resistance to bacterial disease. Plant Biotechnology Journal, 19 (7):1291. doi: 10.1111/pbi.13614 pmid: 33934462
[76]	Veillet F, Chauvin L, Kermarrec M P, Sevestre F, Merrer M, Terret Z, Szydlowski N, Devaux P, Gallois J, Chauvin J. 2019a. The Solanum tuberosum GBSSI gene: a target for assessing gene and base editing in tetraploid potato. Plant Cell Reports, 38 (9):1065-1080.
[77]	Veillet F, Perrot L, Chauvin L, Kermarrec M, Guyon-Debast A, Chauvin J, Nogué F, Mazier M. 2019b. Transgene-free genome editing in tomato and potato plants using agrobacterium-mediated delivery of a CRISPR/Cas 9 cytidine base editor. International Journal of Molecular Sciences, 20 (2):402.
[78]	Wang G X, Zong M, Liu D, Wu Y G, Tian S W, Han S, Guo N, Duan M M, Miao M L, Liu F. 2022. Efficient generation of targeted point mutations in the Brassica oleracea var. botrytis genome via a modified CRISPR/Cas9 system. Horticultural Plant Journal, 8 (4):527-530.
[79]	Wang Hao. 2019. The effect of knockout of MIR159 gene in Petunia on growth and development[M. D. Dissertation]. Chongqing: Southwest University. (in Chinese)
	王浩. 2019. 矮牵牛MIR159基因敲除对生长发育的影响[硕士论文]. 重庆: 西南大学.
[80]	Wang L, Chen L, Li R, Zhao R, Yang M, Sheng J, Shen L. 2017. Reduced drought tolerance by CRISPR/Cas9-mediated SlMAPK3 mutagenesis in tomato plants. Journal of Agricultural and Food Chemistry, 65 (39):8674-8682. doi: 10.1021/acs.jafc.7b02745 pmid: 28873302
[81]	Wang X, Tu M, Wang D, Liu J, Li Y, Li Z, Wang Y, Wang X. 2018. CRISPR/Cas9-mediated efficient targeted mutagenesis in grape in the first generation. Plant Biotechnology Journal, 16 (4):844-855. doi: 10.1111/pbi.12832 pmid: 28905515
[82]	Wang Y, Wang J, Guo S, Tian S, Zhang J, Ren Y, Li M, Gong G, Zhang H, Xu Y. 2021. CRISPR/Cas9-mediated mutagenesis of ClBG1 decreased seed size and promoted seed germination in watermelon. Horticulture Research, 8 (1):70.
[83]	Watanabe K, Kobayashi A, Endo M, Sage-Ono K, Toki S, Ono M. 2017. CRISPR/Cas9-mediated mutagenesis of the dihydroflavonol-4-reductase-B (DFR-B)locus in the Japanese morning glory Ipomoea(Pharbitis)nil. Scientific Reports, 7 (1):10028. doi: 10.1038/s41598-017-10715-1 pmid: 28855641
[84]	Watanabe K, Oda-Yamamizo C, Sage-Ono K, Ohmiya A, Ono M. 2018. Alteration of flower colour in Ipomoea nil through CRISPR/Cas9-mediated mutagenesis of carotenoid cleavage dioxygenase 4. Transgenic Research, 27 (1):25-38. doi: 10.1007/s11248-017-0051-0 pmid: 29247330
[85]	Wu J, Chen C, Xian G, Liu D, Lin L, Yin S, Sun Q, Fang Y, Zhang H, Wang Y. 2020. Engineering herbicide-resistant oilseed rape by CRISPR/Cas9-mediated cytosine base-editing. Plant Biotechnology Journal, 18 (9):1857. doi: 10.1111/pbi.13368 pmid: 32096325
[86]	Wu Ruotong. 2021. Molecular marker assisted selection and CsSH1 gene editing application for excellent traits of cucumber in Xishuangbanna[M. D. Dissertation]. Nanjing: Nanjing Agricultural University. (in Chinese)
	伍若彤. 2021. 西双版纳黄瓜优异性状的分子标记辅助选择及CsSH1基因编辑应用[硕士论文]. 南京: 南京农业大学.
[87]	Xu C, Park S J, van Eck J, Lippman Z B. 2016. Control of inflorescence architecture in tomato by BTB/POZ transcriptional regulators. Genes & Development, 30 (18):2048-2061.
[88]	Xu J, Kang B C, Naing A H, Bae S J, Kim J S, Kim H, Kim C K. 2020. CRISPR/Cas9-mediated editing of 1-aminocyclopropane-1-carboxylate oxidase 1 enhances Petunia flower longevity. Plant Biotechnology Journal, 18 (1):287-297.
[89]	Xu Yanhui. 2023. Functional and molecular mechanism analysis of Csi-miR159a-DUO1 module regulating citrus pollen development[Ph. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	徐彦辉. 2023. Csi-miR159a-DUO1模块调控柑橘花粉发育的功能及分子机制解析[博士论文]. 武汉: 华中农业大学.
[90]	Yan R, Wang Z, Ren Y, Li H, Liu N, Sun H. 2019. Establishment of efficient genetic transformation systems and application of CRISPR/Cas 9 genome editing technology in Lilium pumilum DC. Fisch. and Lilium longiflorum White Heaven. International Journal of Molecular Sciences, 20 (12):2920.
[91]	Yang Liang, Liu Huan, Li Ju, Li Zhi, Miao Mingjun. 2023. Rapid creation of tomato male sterile lines using CRISPR-Cas9 technology. Molecular Plant Breeding, 21 (11):3619-3627. (in Chinese)
	杨亮, 刘欢, 李菊, 李志, 苗明军. 2023. 利用CRISPR-Cas9技术快速创制番茄雄性不育系. 分子植物育种, 21 (11):3619-3627.
[92]	Yang Lushan, Guo Ye, Hu Yang, Wen Yingqiang. 2020. Knockout of VviEDR2 in grapes by CRISPR/Cas 9 system enhances resistance to powdery mildew. Acta Horticulturae Sinica, 47 (4):623-634. (in Chinese) doi: 10.16420/j.issn.0513-353x.2019-0660
	杨禄山, 郭晔, 胡洋, 文颖强. 2020. 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, 47 (4):623-634. doi: 10.16420/j.issn.0513-353x.2019-0660
[93]	Yang Mengxia, Liu Xiaolin, Cai Xue, Wei Kai, Ning Yu, Yang Pei, Li Shanshan, Chen Ziyue, Wang Xiaoxuan, Guo Yanmei, Du Yongchen, Li Junming, Liu Lei, Li Xin, Huang Zejun. 2023. Construction and application of tomato CRISPR/Cas 9 mediated multi gene editing technology system. Acta Horticulturae Sinica, 50 (6):1215-1229. (in Chinese) doi: 10.16420/j.issn.0513-353x.2022-0339
	杨孟霞, 刘晓林, 曹雪, 魏凯, 宁宇, 杨沛, 李珊珊, 陈紫月, 王孝宣, 国艳梅, 杜永臣, 李君明, 刘磊, 李鑫, 黄泽军. 2023. 番茄CRISPR/Cas9介导的多基因编辑技术体系构建与应用. 园艺学报, 50 (6):1215-1229. doi: 10.16420/j.issn.0513-353x.2022-0339
[94]	Yin Y, Qin K, Song X, Zhang Q, Zhou Y, Xia X, Yu J. 2018. BZR1 transcription factor regulates heat stress tolerance through FERONIA receptor-like Kinases-mediated reactive oxygen species signaling in tomato. Plant Cell Physiol, 59 (11):2239-2254.
[95]	Zhang B, Xu X, Huang R, Yang S, Li M, Guo Y. 2021. CRISPR/Cas9-mediated targeted mutation reveals a role for AN4 rather than DPL in regulating venation formation in the corolla tube of Petunia hybrida. Horticulture Research, 8 (1):116. doi: 10.1038/s41438-021-00555-6 pmid: 34059660
[96]	Zhang C, Liu Y, Wang B, Li H, Zhang J, Ma Y, Dai H, Wang Y, Zhang Z. 2023. CRISPR/Cas 9 targeted knockout FvPHO2 can increase phosphorus content and improve fruit quality of woodland strawberry. Scientia Horticulturae, 317:112078.
[97]	Zhang Cui. 2021. Optimization of chrysanthemum genetic transformation system and establishment of‘Shenma’mutant library using gene editing techniques[Ph. D. Dissertation]. Tai’an: Shandong Agricultural University. (in Chinese)
	张翠. 2021. 菊花遗传转化体系的优化和使用基因编辑技术建立‘神马’突变体库初探[博士论文]. 泰安: 山东农业大学.
[98]	Zhang F, LeBlanc C, Irish V F, Jacob Y. 2017. Rapid and efficient CRISPR/Cas 9 gene editing in Citrus using the YAO promoter. Plant Cell Reports, 36 (12):1883-1887. doi: 10.1007/s00299-017-2202-4 pmid: 28864834
[99]	Zhang M, Liu Q, Yang X, Xu J, Liu G, Yao X, Ren R, Xu J, Lou L. 2020a. CRISPR/Cas9-mediated mutagenesis of Clpsk1 in watermelon to confer resistance to Fusarium oxysporum f. sp. niveum. Plant Cell Reports, 39 (5):589-595.
[100]	Zhang Qianqian, Xu Yong, Tian Shouwei, Jiang Linjian, Cui Xiaxia. 2019. Creating new herbicide resistant watermelon germplasm using CRISPR/Cas 9 mediated single base editing technology. China Cucurbits and Vegetables, 32 (8):219-220. (in Chinese)
	张倩倩, 许勇, 田守蔚, 姜临建, 崔霞霞. 2019. 利用CRISPR/Cas9介导的单碱基编辑技术创制抗除草剂西瓜新种质. 中国瓜菜, 32 (8):219-220.
[101]	Zhang S, Wang L, Zhao Ri, Yu W, Li R, Li Y, Sheng J, Shen L. 2018. Knockout of SlMAPK3 reduced disease resistance to Botrytis cinerea in tomato plants. Journal of Agricultural and Food Chemistry, 66 (34):8949-8956.
[102]	Zhang X, Zhu B, Chen L, Xie L, Yu W, Wang Y, Li L, Yin S, Yang L, Hu H, Han H, Li Y, Wang L, Chen G, Ma X, Geng H, Huang W, Pang X, Yang Z, Wu Y, Stefan S, Kurita R, Yukio N, Yang L, Liu M, Li D. 2020b. Dual base editor catalyzes both cytosine and adenine base conversions in human cells. Nature Biotechnology, 38 (7):856-860.
[103]	Zhao D, Li J, Li S, Xin X, Hu M, Price M A, Rosser S, Bi C, Zhang X. 2021. Glycosylase base editors enable C-to-A and C-to-G base changes. Nature Biotechnology, 39 (1):35-40.
[104]	Zhao Y, Li M, Liu J, Xue X, Zhong J, Lin J, Ye B, Chen J, Qiao Y. 2023. Dual guide RNA-mediated concurrent C&G-to-T&A and A&T-to-G&C conversions using CRISPR base editors. Computational and Structural Biotechnology Journal, 21:856-868.
[105]	Zhou J, Li D, Wang G, Wang F, Kunjal M, Joldersma D, Liu Z. 2020. Application and future perspective of CRISPR/Cas 9 genome editing in fruit crops. Journal of integrative Plant Biology, 62 (3):269-286.
[106]	Zhu C, Zheng X, Huang Y, Ye J, Chen P, Zhang C, Zhao F, Xie Z, Zhang S, Wang N, Li H, Wang H, Tang X, Chai L, Xu Q, Deng X. 2019. Genome sequencing and CRISPR/Cas 9 gene editing of an early flowering mini-citrus (Fortunella hindsii). Plant Biotechnology Journal, 17 (11):2199-2210.
[107]	Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu J, Gao C. 2018. Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Nature Biotechnology, 36 (10):950-953.

应用 Application	种类 Species	目标基因 Target gene	目标性状 Target trait	参考文献 Reference
调节生长特性 Regulating growth characteristics	苹果 Malus × domestica	MdPDS	白化表型 Albino phenotype	Charrier et al.，2019
	香蕉 Musa nana Lour.	MaGA20ox2	半矮生 Semidwarf	Shao et al.，2020
	西瓜 Citrullus lanatus	ClBG1	改变种子大小和质量Changing seed size and quality	Wang et al.，2021
	蓝莓 Vaccinium spp.	CEN	促营养生长Promote nutritional growth	Omori et al.，2021
	番茄 Solanum lycopersicum	AGL6	单性结实 Parthenocarpy	Klap et al.，2017
		MBP21	果柄无分离层Fruit stalk without separation layer	Roldan et al.，2017
		SlTM6	雄性不育系 male sterile line	杨亮等，2023
	黄瓜 Cucumis sativus	CsVFB1	矮生 Dwarf	戚晶晶，2017
		CsCLAVATA3	改变心皮数Change the number of carpels	秦楠楠，2020
		Cs SH1	短下胚轴 Short hypocotyl	伍若彤，2021
	百合Lilium	LpPDS	白化表型 Albino phenotype	Yan et al.，2019
	菊花 Chrysanthemum	AN4	株形、叶片面积、冠径、株高和色素含量 Plant shape，leaf area，crown diameter，plant height，and pigment content	张翠，2021
改变开花时间及花色 Change flowering time and color	柑橘 Citrus reticulata Blanco	DUO1	改变花粉育性 Changing pollen fertility	徐彦辉，2023
	番茄 Solanum lycopersicum	SlFAF	调节开花 Adjusting flowering	Shang et al.，2024
	番茄 Solanum lycopersicum	TMF	花序简化 Simplified inflorescence	Xu et al.，2016
	日本牵牛花 Ipomoea nil	DFR-B	获得白色花 Obtain white flowers	Watanabe et al.，2017
		CCD4	白色花瓣变成淡黄色 White petals turn pale yellow	Watanabe et al.，2018
		Gt5GT，Gt3’GT，Gt5/3’AT	获得淡红色、紫罗兰色、暗粉红色和淡紫色花 Get light red，violet，dark pink，and light purple flowers	Tasaki et al.，2019
		GST1	获得白色和淡蓝色花Obtain white and light blue flowers	Tasaki et al.，2020
	矮牵牛 Petunia hybrid Vilm.	ACO1	延长花寿命Extend the lifespan of flowers	Xu et al.，2020
		AN4	花冠管脉络缺失Lacking veins in the corolla tube	Zhang et al.，2021
		miRl59b	延迟开花 Delayed flowering	王浩，2019
	蓝猪耳 Torenia fournieri	TfRAD1	改变花瓣形状和颜色Change petal shape and color	Su et al.，2017
	蓝猪耳 Torenia fournieri	F3H	花瓣改色 Petal color change	Nishihara et al.，2018
提高果实品质 Improve fruit quality	香蕉 Musa nana Lour.	LCYε	提高β胡萝卜素Increase β carotene	Kaur et al.，2020
	草莓 Fragaria × ananassa	FaPG1	提高硬度 Improve hardness	López-Casado et al.，2023
		FvCO4	果实发育密切相关Fruit development is closely related	陈文君，2019
		PHO2	提高果实中磷含量 Increase the phosphorus content in fruits	Zhang et al.，2023
		RAP	绿茎白果 Green stemmed white fruit	Gao et al.，2020
	毛酸浆 Physalis pubescens	SELF-PRUNING 5G	提高果实数量和质量 Improve the quantity and quality of fruits	Lemmon et al.，2018
	番茄 Solanum lycopersicum	CRITOSO	橙色果实 Orange fruit	Isaacson et al.，2002
		MYB12	粉红色果实 Pink fruit	Ballester et al.，2010
		PSY1	黄色果实 Yellow fruit	Filler et al.，2017
		RIN	果实成熟延迟 Delayed fruit ripening	Lang et al.，2017
		SlACS2	果实成熟延迟 Delayed fruit ripening	刘江娜等，2023
		SlbZIP1	提高糖和氨基酸含量 Increase sugar and amino acid content	Nguyen et al.，2023
		SNAC9	果实成熟延迟 Delayed fruit ripening	Feng et al.，2023
增强植株抗性 Enhance plant resistance	苹果 Malus × domestica	FB_MR5	火枯病症状下降Reduced symptoms of fire blight	Broggini et al.，2014
		FB_MR5	增强抗火枯病能力Enhance the ability to resist fire blight	Kost et al.，2015
		HIPM	火疫病减轻Alleviation of fire epidemic	Campa et al.，2019
	柑橘 Citrus reticulata Blanco	CcPUB4	增强耐盐性Enhance salt resistance	李秋月，2019
	柑橘 Citrus reticulata Blanco	CsLOB1	抗柑橘溃疡病Anti citrus ulcer disease	Hu et al.，2014
	葡萄 Vitis vinifera	VvBAK1，VvLecRK1	抗白粉病 Anti powdery mildew	贾慧，2019
		VvWRKY52	抗灰葡萄孢Resistance to botrytis cinerea	Wang et al.，2018
		VvPR4b	抗霜霉病Anti downy mildew disease	李梦媛，2020
		VviEDR2	抗白粉病 Anti powdery mildew	杨禄山等，2020
增强植株抗性 Enhance plant resistance	香蕉Musa nana Lour.	eBSV	抗条斑病毒 Anti stripe virus	Tripathi et al.，2019
	香蕉Musa nana Lour.	MusaDMR6	抗黄萎病 Verticillium wilt	Tripathi et al.，2021
	西瓜 Citrullus lanatus	ALS	抗除草剂 Herbicide resistant	张倩倩，2019
	西瓜 Citrullus lanatus	Clpsk1	抗尖孢镰刀菌Anti Fusarium oxysporum	Zhang et al.，2020a
	草莓 Fragaria × ananassa	FVICE1	抗严寒和干旱Resistance to severe cold and drought	Han et al.，2023
	番茄 Solanum lycopersicum	BZR1	参与调控耐热途径 Participate in regulating heat resistance pathways	Yin et al.，2018
		Mlo1	抗白粉病 Anti powdery mildew	Nekrasov et al.，2017
		MAPK3	抗灰霉病、耐旱Resistant to gray mold and drought	Zhang et al.，2018；Wang et al.，2017
		SlCBF1	防止植物受冻害Prevent plants from freezing damage	Li et al.，2018
		SINPR1	耐旱 Drought resistance	Li et al.，2019
	黄瓜 Cucumis sativus	eIF4E	抗黄瓜叶脉黄变病毒、西葫芦黄花叶病毒、番木瓜环斑花叶病毒Anti cucumber vein yellowing virus，zucchini yellow mosaic virus，papaya ring spot mosaic virus	Chandrasekaran et al.，2016
	黄瓜 Cucumis sativus	Mlo1	抗白粉病 Anti powdery mildew	Shnaider et al.，2023
	马铃薯 Solanum tuberosum	PVY	抗马铃薯Y病毒 Anti potato Y Virus	Lucioli et al.，2022
		SmelPPO	降低褐化 Reduce browning	González et al.，2020
		StCHLI	抗晚疫病 Anti late epidemic disease	Kieu et al.，2021
		St16DOX	减少糖苷生物碱Reduce glycoside alkaloids	Nakayasu et al.，2018
	茄子 Solanum melongena	SmelPPO	降低褐化 Reduce browning	Maioli et al.，2020

应用 Application	种类 Species	目标基因 Target gene	目标性状 Target trait	参考文献 Reference
调节生长特性 Regulating growth characteristics	苹果 Malus × domestica	MdPDS	白化表型 Albino phenotype	Charrier et al.，2019
	香蕉 Musa nana Lour.	MaGA20ox2	半矮生 Semidwarf	Shao et al.，2020
	西瓜 Citrullus lanatus	ClBG1	改变种子大小和质量Changing seed size and quality	Wang et al.，2021
	蓝莓 Vaccinium spp.	CEN	促营养生长Promote nutritional growth	Omori et al.，2021
	番茄 Solanum lycopersicum	AGL6	单性结实 Parthenocarpy	Klap et al.，2017
		MBP21	果柄无分离层Fruit stalk without separation layer	Roldan et al.，2017
		SlTM6	雄性不育系 male sterile line	杨亮等，2023
	黄瓜 Cucumis sativus	CsVFB1	矮生 Dwarf	戚晶晶，2017
		CsCLAVATA3	改变心皮数Change the number of carpels	秦楠楠，2020
		Cs SH1	短下胚轴 Short hypocotyl	伍若彤，2021
	百合Lilium	LpPDS	白化表型 Albino phenotype	Yan et al.，2019
	菊花 Chrysanthemum	AN4	株形、叶片面积、冠径、株高和色素含量 Plant shape，leaf area，crown diameter，plant height，and pigment content	张翠，2021
改变开花时间及花色 Change flowering time and color	柑橘 Citrus reticulata Blanco	DUO1	改变花粉育性 Changing pollen fertility	徐彦辉，2023
	番茄 Solanum lycopersicum	SlFAF	调节开花 Adjusting flowering	Shang et al.，2024
	番茄 Solanum lycopersicum	TMF	花序简化 Simplified inflorescence	Xu et al.，2016
	日本牵牛花 Ipomoea nil	DFR-B	获得白色花 Obtain white flowers	Watanabe et al.，2017
		CCD4	白色花瓣变成淡黄色 White petals turn pale yellow	Watanabe et al.，2018
		Gt5GT，Gt3’GT，Gt5/3’AT	获得淡红色、紫罗兰色、暗粉红色和淡紫色花 Get light red，violet，dark pink，and light purple flowers	Tasaki et al.，2019
		GST1	获得白色和淡蓝色花Obtain white and light blue flowers	Tasaki et al.，2020
	矮牵牛 Petunia hybrid Vilm.	ACO1	延长花寿命Extend the lifespan of flowers	Xu et al.，2020
		AN4	花冠管脉络缺失Lacking veins in the corolla tube	Zhang et al.，2021
		miRl59b	延迟开花 Delayed flowering	王浩，2019
	蓝猪耳 Torenia fournieri	TfRAD1	改变花瓣形状和颜色Change petal shape and color	Su et al.，2017
	蓝猪耳 Torenia fournieri	F3H	花瓣改色 Petal color change	Nishihara et al.，2018
提高果实品质 Improve fruit quality	香蕉 Musa nana Lour.	LCYε	提高β胡萝卜素Increase β carotene	Kaur et al.，2020
	草莓 Fragaria × ananassa	FaPG1	提高硬度 Improve hardness	López-Casado et al.，2023
		FvCO4	果实发育密切相关Fruit development is closely related	陈文君，2019
		PHO2	提高果实中磷含量 Increase the phosphorus content in fruits	Zhang et al.，2023
		RAP	绿茎白果 Green stemmed white fruit	Gao et al.，2020
	毛酸浆 Physalis pubescens	SELF-PRUNING 5G	提高果实数量和质量 Improve the quantity and quality of fruits	Lemmon et al.，2018
	番茄 Solanum lycopersicum	CRITOSO	橙色果实 Orange fruit	Isaacson et al.，2002
		MYB12	粉红色果实 Pink fruit	Ballester et al.，2010
		PSY1	黄色果实 Yellow fruit	Filler et al.，2017
		RIN	果实成熟延迟 Delayed fruit ripening	Lang et al.，2017
		SlACS2	果实成熟延迟 Delayed fruit ripening	刘江娜等，2023
		SlbZIP1	提高糖和氨基酸含量 Increase sugar and amino acid content	Nguyen et al.，2023
		SNAC9	果实成熟延迟 Delayed fruit ripening	Feng et al.，2023
增强植株抗性 Enhance plant resistance	苹果 Malus × domestica	FB_MR5	火枯病症状下降Reduced symptoms of fire blight	Broggini et al.，2014
		FB_MR5	增强抗火枯病能力Enhance the ability to resist fire blight	Kost et al.，2015
		HIPM	火疫病减轻Alleviation of fire epidemic	Campa et al.，2019
	柑橘 Citrus reticulata Blanco	CcPUB4	增强耐盐性Enhance salt resistance	李秋月，2019
	柑橘 Citrus reticulata Blanco	CsLOB1	抗柑橘溃疡病Anti citrus ulcer disease	Hu et al.，2014
	葡萄 Vitis vinifera	VvBAK1，VvLecRK1	抗白粉病 Anti powdery mildew	贾慧，2019
		VvWRKY52	抗灰葡萄孢Resistance to botrytis cinerea	Wang et al.，2018
		VvPR4b	抗霜霉病Anti downy mildew disease	李梦媛，2020
		VviEDR2	抗白粉病 Anti powdery mildew	杨禄山等，2020
增强植株抗性 Enhance plant resistance	香蕉Musa nana Lour.	eBSV	抗条斑病毒 Anti stripe virus	Tripathi et al.，2019
	香蕉Musa nana Lour.	MusaDMR6	抗黄萎病 Verticillium wilt	Tripathi et al.，2021
	西瓜 Citrullus lanatus	ALS	抗除草剂 Herbicide resistant	张倩倩，2019
	西瓜 Citrullus lanatus	Clpsk1	抗尖孢镰刀菌Anti Fusarium oxysporum	Zhang et al.，2020a
	草莓 Fragaria × ananassa	FVICE1	抗严寒和干旱Resistance to severe cold and drought	Han et al.，2023
	番茄 Solanum lycopersicum	BZR1	参与调控耐热途径 Participate in regulating heat resistance pathways	Yin et al.，2018
		Mlo1	抗白粉病 Anti powdery mildew	Nekrasov et al.，2017
		MAPK3	抗灰霉病、耐旱Resistant to gray mold and drought	Zhang et al.，2018；Wang et al.，2017
		SlCBF1	防止植物受冻害Prevent plants from freezing damage	Li et al.，2018
		SINPR1	耐旱 Drought resistance	Li et al.，2019
	黄瓜 Cucumis sativus	eIF4E	抗黄瓜叶脉黄变病毒、西葫芦黄花叶病毒、番木瓜环斑花叶病毒Anti cucumber vein yellowing virus，zucchini yellow mosaic virus，papaya ring spot mosaic virus	Chandrasekaran et al.，2016
	黄瓜 Cucumis sativus	Mlo1	抗白粉病 Anti powdery mildew	Shnaider et al.，2023
	马铃薯 Solanum tuberosum	PVY	抗马铃薯Y病毒 Anti potato Y Virus	Lucioli et al.，2022
		SmelPPO	降低褐化 Reduce browning	González et al.，2020
		StCHLI	抗晚疫病 Anti late epidemic disease	Kieu et al.，2021
		St16DOX	减少糖苷生物碱Reduce glycoside alkaloids	Nakayasu et al.，2018
	茄子 Solanum melongena	SmelPPO	降低褐化 Reduce browning	Maioli et al.，2020

种类 Species	碱基编辑器 Base editor	目标基因 Target gene	目标性状 Target trait	编辑效率/% Editing efficiency	参考文献 Reference
西瓜 Citrullus lanatus	BE3	ClALS	抗除草剂 Herbicide resistant	23.0	Tian et al.，2018
番茄 Solanum lycopersicum	Target-AID	DELLA，ETR1	株型 Plant type	50.5	Shimatani et al.，2017
	CBE	ALS	抗除草剂 Herbicide resistant	71.0	Veillet et al.，2019b
	pCXPE03	GAI，ALS2，PDS1	抗除草剂 Herbicide resistant	3.4 ~ 6.7	Lu et al.，2021
马铃薯 Solanum tuberosum	A3A-PBE	StGBSS-T6	淀粉含量 Starch content	6.5	Zong et al.，2018
马铃薯 Solanum tuberosum	CBE	StGBSS1	淀粉含量 Starch content	8.00 ~ 16.67	Veillet et al.，2019a
油菜 Brassica napus	CBE	BnALS1，BnALS3	抗除草剂 Herbicide resistant	1.8	Wu et al.，2020

种类 Species	碱基编辑器 Base editor	目标基因 Target gene	目标性状 Target trait	编辑效率/% Editing efficiency	参考文献 Reference
西瓜 Citrullus lanatus	BE3	ClALS	抗除草剂 Herbicide resistant	23.0	Tian et al.，2018
番茄 Solanum lycopersicum	Target-AID	DELLA，ETR1	株型 Plant type	50.5	Shimatani et al.，2017
	CBE	ALS	抗除草剂 Herbicide resistant	71.0	Veillet et al.，2019b
	pCXPE03	GAI，ALS2，PDS1	抗除草剂 Herbicide resistant	3.4 ~ 6.7	Lu et al.，2021
马铃薯 Solanum tuberosum	A3A-PBE	StGBSS-T6	淀粉含量 Starch content	6.5	Zong et al.，2018
马铃薯 Solanum tuberosum	CBE	StGBSS1	淀粉含量 Starch content	8.00 ~ 16.67	Veillet et al.，2019a
油菜 Brassica napus	CBE	BnALS1，BnALS3	抗除草剂 Herbicide resistant	1.8	Wu et al.，2020

Research Progress of CRISPR/Cas9 Technology and Its Application in Horticultural Plants

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