利用CRISPR/Cas9技术创制高番茄红素番茄新材料

doi:10.16420/j.issn.0513-353x.2023-0782

园艺学报 ›› 2024, Vol. 51 ›› Issue (2): 253-265.doi: 10.16420/j.issn.0513-353x.2023-0782

• 遗传育种·种质资源·分子生物学 • 上一篇下一篇

利用CRISPR/Cas9技术创制高番茄红素番茄新材料

杨亮¹^,²^,³^,⁴, 刘欢¹^,³^,⁴, 马燕勤¹^,²^,³^,⁴, 李菊¹^,²^,³^,⁴, 王海娥¹^,⁴, 周玉洁¹^,³^,⁴, 龙海成¹^,³^,⁴, 苗明军¹^,²^,³^,⁴, 李志¹^,²^,³^,⁴^,^*(), 常伟²^,³^,⁴^,⁵^,^*()

¹ 四川省农业科学院园艺研究所，成都 610066
² 四川省蔬菜工程技术研究中心，四川彭州 611934
³ 蔬菜种质与品种创新四川省重点实验室，成都 610066
⁴ 农业农村部西南地区园艺作物生物学与种质创制重点实验室，成都 610066
⁵ 四川省食用菌研究所，成都 610066

收稿日期:2023-12-07 修回日期:2024-01-06 出版日期:2024-02-25 发布日期:2024-02-26
通讯作者:
*E-mail：changwei972@126.com，
lz20031977@126.com
基金资助:
国家重点研发计划项目(2022YFD1100205); 四川省科技计划项目(2021YFYZ0022); 四川省科技计划项目(2023NSFSC0162); 现代农业产业技术体系建设专项四川省蔬菜创新团队项目[川农函(2019); 四川省农业科学院“1+9”揭榜挂帅科技攻关项目(1+9KJGG001)

Creating High Lycopene Fruit Using CRISPR/Cas9 Technology in Tomato

YANG Liang¹^,²^,³^,⁴, LIU Huan¹^,³^,⁴, MA Yanqin¹^,²^,³^,⁴, LI Ju¹^,²^,³^,⁴, WANG Hai'e¹^,⁴, ZHOU Yujie¹^,³^,⁴, LONG Haicheng¹^,³^,⁴, MIAO Mingjun¹^,²^,³^,⁴, LI Zhi¹^,²^,³^,⁴^,^*(), CHANG Wei²^,³^,⁴^,⁵^,^*()

¹ Horticulture Research Institute，Sichuan Academy of Agricultural Sciences，Chengdu 610066，China
² Sichuan Province Engineering Technology Research Center of Vegetables，Pengzhou，Sichuan 611934，China
³ Vegetable Germplasm Innovation and Variety Improvement Key Laboratory of Sichuan Province，Chengdu 610066，China
⁴ Key Laboratory of Horticultural Crops Biology and Germplasm Enhancement in Southwest Regions，Ministry of Agriculture and Rural Affairs，Chengdu 610066，China
⁵ Sichuan Institute of Edible Fungi，Chengdu 610066，China

Received:2023-12-07 Revised:2024-01-06 Published:2024-02-25 Online:2024-02-26

摘要/Abstract

摘要：

以大果型栽培番茄资源“T048”为材料，利用CRISPR/Cas9技术对滞绿基因SlSGR1进行定向编辑。对19株转基因阳性株系进行靶位点序列分析发现，共有10株发生了编辑事件，编辑效率为52.6%。通过测序分析发现，编辑类型涉及碱基的缺失、插入及替换，多数发生在PAM序列上游3 ~ 4 bp碱基处，同时也发现了两个编辑位点间共565 bp的序列倒位。表型分析发现，T1代纯合编辑株系叶片衰老减缓，果实表现为铁锈色，且番茄红素、叶绿素及β-胡萝卜素含量显著提高。对类胡萝卜素合成关键基因进行分析发现，纯合编辑株系果实中的SGR1表达量显著降低，而PSY1表达量显著提高。结果表明，通过CRISPR/Cas9技术对滞绿基因SlSGR1进行定向编辑可有效提升番茄果实中番茄红素、β-胡萝卜素等类胡萝卜素含量，进而为番茄营养品质育种提供新种质。

关键词: 番茄, 基因编辑, 番茄红素, 类胡萝卜素, SlSGR1

Abstract:

The large-fruited cultivated tomato resource“T048”was used as the material for targeted editing of the tomato stay-green gene SlSGR1 using CRISPR/Cas9 technology. Sequence analysis of the target sites of 19 positive transgenic plants revealed that a total of ten plants had editing events，with an editing efficiency of 52.6%. The types of editing events involved deletions，insertions and substitutions of bases，most of which occurred 3-4 bp upstream of the PAM sequence. A total of 565 bp of sequence inversion between the two editing sites was also detected. Phenotypic analysis revealed that the homozygous edited plants in the T1 generation showed slower senescence of the leaves and rusty red color fruits，with significantly higher content of lycopene，chlorophyll and β-carotenoid，compared with unedited fruits. Analysis of the key genes for carotenoid synthesis revealed that the expression of the SGR1 in the fruit of the homozygous edited plants was significantly lower than that of the unedited plants，whereas the expression of the PSY1 was significantly higher than that of the unedited one. This study demonstrated that targeted editing of the SlSGR1 gene by CRISPR/Cas9 technology can effectively enhance the content of carotenoids such as lycopene and β-carotene in tomato fruits，and thus provide new germplasm for tomato nutritional quality breeding.

Key words: tomato, geno editing, lycopene, carotenoid, SlSGR1

杨亮, 刘欢, 马燕勤, 李菊, 王海娥, 周玉洁, 龙海成, 苗明军, 李志, 常伟. 利用CRISPR/Cas9技术创制高番茄红素番茄新材料[J]. 园艺学报, 2024, 51(2): 253-265.

YANG Liang, LIU Huan, MA Yanqin, LI Ju, WANG Hai'e, ZHOU Yujie, LONG Haicheng, MIAO Mingjun, LI Zhi, CHANG Wei. Creating High Lycopene Fruit Using CRISPR/Cas9 Technology in Tomato[J]. Acta Horticulturae Sinica, 2024, 51(2): 253-265.

图/表 9

表1 本研究中所使用引物

Table 1 The primers used in this study

用途 Aim	引物名称 Primer name	引物序列（5′-3′） Primer sequence
载体构建Construction vector	SGR1-target1-F	ATATATGGTCTCGTTTGAGATGAAGTTGTTGCAGAGGTTTTAGAGCTAGAAATAG
载体构建Construction vector	SGR1-target2-R	ATTATTGGTCTCGAAACCCAGTGAGTGTTATGCCTTCCAAACTACACTGTTAGATTC
阳性克隆验证Identification of the positive clones	M13-F	GTAAAACGACGGCCAGT
阳性克隆验证Identification of the positive clones	SGR1-target2	CCAGTGAGTGTTATGCCTT
转基因株验证Identification of the transformation plants	Cas9-F	AGGTCACGGTTAAGCAGCTC
转基因株验证Identification of the transformation plants	Cas9-R	CCCTTCTGCGTGGTCTGATT
靶位点检测 Detection target	SGR1-Seqtarget-F	GTTTGATTTGCAGTTGCAAGGTT
靶位点检测 Detection target	SGR1-Seqtarget-R	AATGCTGCTTCCACAAACCC
Real-time PCR	SlACTIN-RT-F	GGGATGGAGAAGTTTGGTGGTGG
	SlACTIN-RT-R	CTTCGACCAAGGGATGGTGTAGC
	SGR1-RT-F	GGGTCCACTCAGAGATGCAA
	SGR1-RT-R	ACTCACTGGGGGAAAGCAAC
	PDS-RT-F	AAGGCGCTGTCTTATCAGGAAA
	PDS-RT-R	TAAACTACGCTTGCTTCCGACA
	ZDS-RT-F	ACCGTACAACTACGCTACAATGG
	ZDS-RT-R	CATCTGGCGTATAGAGGAGATTG
	PSY1-RT-F	TATTTGCTGGAAGGGTGACC
	PSY1-RT-R	TAGCTGAGCTCAATTCTGTC

图1 番茄SlSGR1靶位点位置（A）及pKSE401-SGR1载体（B）示意图

Fig. 1 Schematic representation of SlSGR1 with target sites（A）and the pKSE401-SGR1 vector（B）

图2 番茄SlSGR1基因编辑T0代阳性株系鉴定 M：DL2000 DNA marker；1 ~ 26：生根株系样本；WT：野生型；+：阳性对照。

Fig. 2 Identification of positive transgenic lines in SlSGR1 edited T0 generation plants M：DL2000 DNA marker；1-26：Rooted plants；WT：Wild type；+：Positive control.

图3 番茄SlSGR1基因Target1和Target2位点编辑效率及类型统计

Fig. 3 Statistics of editing frequency and pattern in Target1 and Target2 loci of SlSGR1 edited plants

图4 番茄SlSGR1基因编辑T0代10个阳性株系突变序列分析 Ho：纯合突变；He：杂合突变；Bi：双等位突变；Ch：嵌合突变；WT：野生型；d：碱基缺失；i：碱基插入；s：碱基替换。

Fig. 4 Mutation sequence analysis of ten SlSGR1 edited T0 generation plants Ho：Homozygous mutation；He：Heterozygous mutation；Bi：Biallelic mutation；Ch：Chimeric mutation；WT：Wild type；d：Base deletion；i：Base insertion；s：Base substitution.

图5 番茄SlSGR1基因编辑株系（sgr1-4、sgr1-12和sgr1-22）的T1代无外源T-DNA纯合材料筛选（A）及靶位点序列分析（B）

Fig. 5 Screening of T1 exogenous T-DNA-free homozygous lines in SlSGR1 edited plants （sgr1-4, sgr1-12 and sgr1-22）（A）and sequence analysis of their target sites（B）

图6 番茄SlSGR1基因编辑株系（sgr1-4-10、sgr1-12-1和sgr1-22-12）叶片和果实的表型

Fig. 6 Phenotypes of leaves and fruits in SlSGR1 edited plants（sgr1-4-10，sgr1-12-1 and sgr1-22-12）

表2 番茄SlSGR1基因编辑阳性株系的类胡萝卜素、叶绿素及营养品质

Table 2 Content of carotenoids，chlorophyll and nutritional quality traits in SlSGR1 edited plants

基因编辑株系 Genome editing plant		叶绿素/（μg • g^-1） Chlorophyll	β-胡萝卜素/（μg • g^-1） β-carotenoids	抗坏血酸/（μg · g^-1） Vitamin C	可溶性固形物/% Soluble solids
野生型Wild type	53.70 ± 9.54 b	4.28 ± 1.24 b	38.44 ± 3.98 b	251.64 ± 11.58 a	4.77 ± 0.40 a
sgr1-4-10	73.21 ± 9.56 a	29.60 ± 1.50 a	53.32 ± 2.74 a	246.43 ± 17.73 a	5.13 ± 0.85 a
sgr1-22-12	75.08 ± 9.11 a	37.20 ± 6.30 a	59.71 ± 1.13 a	254.73 ± 7.27 a	4.87 ± 0.15 a

图7 番茄SlSGR1基因编辑株系sgr1-4-10和sgr1-22-12果实类胡萝卜素合成途径关键基因的相对表达量 MG：绿熟期；Br：破色期；Br + 4：破色后4 d；Br + 10：破色后10 d。不同小写字母表示同一时期材料间差异显著（P < 0.05）。

Fig. 7 Relative expression of key genes of carotenoid synthesis pathway in four fruit development stages of different genome editing tomato plants MG：Mature green stage；Br：Break stage；Br + 4：Four days after break stage；Br + 10：Ten days after break stage.Columns with different lowercase letters indicate significant differences among lines in the same stage（P < 0.05）.

参考文献 39

[1]	Barry C S, Pandey P. 2009. A survey of cultivated heirloom tomato varieties identifies four new mutant alleles at the green-flesh locus. Molecular Breeding, 24 (3):269-276.
[2]	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.
[3]	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
[4]	D’Ambrosio C, Stigliani A L, Giorio G. 2018. CRISPR/Cas 9 editing of carotenoid genes in tomato. Transgenic Research, 27 (4):367-378.
[5]	Dahan-Meir T, Filler-Hayut S, Melamed-Bessudo C, Bocobza S, Czosnek H, Aharoni A, Levy A A. 2018. Efficient in planta gene targeting in tomato using geminiviral replicons and the CRISPR/Cas9 system. Plant Journal, 95 (1):5-16.
[6]	Galpaz N, Ronen G, Khalfa Z, Zamir D, Hirschberg J. 2006. A chromoplast-specific carotenoid biosynthesis pathway is revealed by cloning of the tomato white-flower locus. Plant Cell, 18 (8):1947-1960. doi: 10.1105/tpc.105.039966 pmid: 16816137
[7]	Gao C X. 2021. Genome engineering for crop improvement and future agriculture. Cell, 184 (6):1621-1635. doi: 10.1016/j.cell.2021.01.005 pmid: 33581057
[8]	Hörtensteiner S. 2009. Stay-green regulates chlorophyll and chlorophyll-binding protein degradation during senescence. Trends in Plant Science, 14 (3):155-162. doi: 10.1016/j.tplants.2009.01.002 pmid: 19237309
[9]	Jacobs T B, LaFayette P R, Schmitz R J, Parrott W A. 2015. Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology, 15:16. doi: 10.1186/s12896-015-0131-2 pmid: 25879861
[10]	Li Guobin, Cai Liangyu, Xiao Licheng, Wang Jiafa, Zhang Dedi, Zhang Junhong. 2023. Creating pink fruit and fruit without green shoulder using CRISPR/Cas 9 technology in tomato. Acta Horticulturae Sinica, 50 (5):985-999. (in Chinese) doi: 10.16420/j.issn.0513-353x.2022-0273
	李国斌, 蔡梁玉, 肖立成, 王家发, 张得迪, 张俊红. 2023. 利用CRISPR/Cas9技术创制番茄粉果和无绿肩材料. 园艺学报, 50 (5):985-999. doi: 10.16420/j.issn.0513-353x.2022-0273
[11]	Li J H, Zhang S J, Zhang R Z, Gao J, Qi Y P, Song G Q, Li W, Li Y L, Li G Y. 2020. Efficient multiplex genome editing by CRISPR/Cas 9 in common wheat. Plant Biotechnology Journal, 19 (3):427-429.
[12]	Liang G, Zhang H M, Lou D J, Yu D Q. 2016. Selection of highly efficient sgRNAs for CRISPR/Cas9-based plant genome editing. Scientific Reports, 6:21451. doi: 10.1038/srep21451 pmid: 26891616
[13]	Liu G W, Lin Q P, Jin S, Gao C X. 2022a. The CRISPR-Cas toolbox and gene editing technologies. Molecular Cell, 82 (2):333-347.
[14]	Liu H, Ding Y D, Zhou Y Q, Jin W Q, Xie K B, Chen L L. 2017. CRISPR-P 2.0:an improved CRISPR-Cas 9 tool for genome editing in plants. Molecular Plant, 10 (3):530-532.
[15]	Liu Y, Zhang C L, Wang X F, Li X M, You C X. 2022b. CRISPR/Cas 9 technology and its application in horticultural crops. Horticultural Plant Journal, 8 (4):395-407.
[16]	Liu Y H, Ou L J, Liu Z B, Lyu J H, Wang J, Song J S, Yang B Z, Chen W C, Yang S, Liu W, Zou X X. 2023. A novel single-base mutation in CaSGR1 confers the stay-green phenotype in pepper. Horticultural Plant Journal, 9 (2):293-305.
[17]	Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-∆∆CTmethod. Methods, 25 (4):402-408. doi: 10.1006/meth.2001.1262 pmid: 11846609
[18]	Lu Y, Wang J Y, Chen B, Mo S D, Lian L, Luo Y M, Ding D H, Ding Y H, Cao Q, Li Y C, Li Y, Liu G Z, Hou Q Q, Cheng T T, Wei J T, Zhang Y R, Chen G W, Song C, Hu Q, Sun S, Fan G Y, Wang Y T, Liu Z T, Song B A, Zhu J K, Li H R, Jiang L J. 2021. A donor-DNA-free CRISPR/Cas-based approach to gene knock-up in rice. Nature Plants, 7 (11):1445-1452. doi: 10.1038/s41477-021-01019-4 pmid: 34782773
[19]	Lu Y M, Zhu J K. 2017. Precise editing of a target base in the rice genome using a modified CRISPR/Cas9 system. Molecular Plant, 10 (3):523-525. doi: S1674-2052(16)30297-0 pmid: 27932049
[20]	Luo Z D, Zhang J H, Li J H, Yang C X, Wang T T, Ouyang B, Li H X, Giovannoni J, Ye Z B. 2013. A STAY-GREEN protein SlSGR1 regulates lycopene and beta-carotene accumulation by interacting directly with SlPSY 1 during ripening processes in tomato. New Phytologist, 198 (2):442-452.
[21]	Ono K, Kimura M, Matsuura H, Tanaka A, Ito H. 2019. Jasmonate production through chlorophyll a degradation by Stay-Green in Arabidopsis thaliana. Journal of Plant Physiology, 238:53-62.
[22]	Park S Y, Yu J W, Park J S, Li J, Yoo S C, Lee N Y, Lee S K, Jeong S W, Seo H S, Koh H J, Jeon J S, Park Y I, Paek N C. 2007. The senescence-induced staygreen protein regulates chlorophyll degradation. Plant Cell, 19 (5):1649-1664.
[23]	Ren G D, An K, Liao Y, Zhou X, Cao Y J, Zhao H F, Ge X C, Kuai B K. 2007. Identification of a novel chloroplast protein AtNYE 1 regulating chlorophyll degradation during leaf senescence in Arabidopsis. Plant Physiology, 144 (3):1429-1441.
[24]	Sakuraba Y, Schelbert S, Park S Y, Han S H, Lee B D, Andrès C B, Kessler F, Hörtensteiner S, Paek N C. 2012. Stay-green and chlorophyll catabolic enzymes interact at light-harvesting complex ii for chlorophyll detoxification during leaf senescence in Arabidopsis. Plant Cell, 24 (2):507-518.
[25]	Sato T, Shimoda Y, Matsuda K, Tanaka A, Ito H. 2018. Mg-dechelation of chlorophyll a by stay-green activates chlorophyll b degradation through expressing non-yellow coloring 1 in Arabidopsis thaliana. Journal of Plant Physiology, 222:94-102.
[26]	Shan Q W, Wang Y P, Li J, Zhang Y, Chen K L, Liang Z, Zhang K, Liu J X, Xi J J, Qiu J L, Gao C X. 2013. Targeted genome modification of crop plants using a CRISPR-Cas system. Nature Biotechnology, 31 (8):686-688. doi: 10.1038/nbt.2650 pmid: 23929338
[27]	Shimoda Y, Ito H, Tanaka A. 2016. Arabidopsis STAY-GREEN,mendel’s green cotyledon gene,encodes magnesium-dechelatase. Plant Cell, 28 (9):2147-2160.
[28]	Svitashev S, Schwartz C, Lenderts B, Young J K, Mark Cigan A. 2016. Genome editing in maize directed by CRISPR-Cas 9 ribonucleoprotein complexes. Nature Communication, 7:13274.
[29]	van Eck J, Keen P, Tjahjadi M. 2019. Agrobacterium tumefaciens-mediated transformation of tomato. Methods in Molecular Biology, 1864:225-234.
[30]	Wang G X, Zong M, Liu D, Wu Y G, Tian S M, Han S, Guo N, Duan M M, Miao L M, 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.
[31]	Wang S J, Wang G, Li H L, Li F, Wang J B. 2023. Agrobacterium tumefaciens-mediated transformation of embryogenic callus and CRISPR/Cas9-mediated genome editing in‘Feizixiao’litchi. Horticultural Plant Journal, 9 (5):947-957.
[32]	Xie Z K, Wu S D, Chen J Y, Zhu X Y, Zhou X, Hörtensteiner S, Ren G D, Kuai B K. 2019. The C-terminal cysteine-rich motif of NYE1/SGR1 is indispensable for its function in chlorophyll degradation in Arabidopsis. Plant Molecular Biology, 101 (3):257-268.
[33]	Xing H L, Dong L, Wang Z P, Zhang H Y, Han C Y, Liu B, Wang X C, Chen Q J. 2014. A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biology,14327.
[34]	Yang Mengxia, Liu Xiaolin, Cao 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 a CRISPR/Cas 9 system for multiplex gene editing in tomato. 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
[35]	Yang T X, Deng L, Zhao W, Zhang R X, Jiang H L, Ye Z B, Li C B, Li C Y. 2019. Rapid breeding of pink-fruited tomato hybrids using the CRISPR/Cas9 system. Journal of Genetics and Genomics, 46 (10):505-508. doi: S1673-8527(19)30167-5 pmid: 31734133
[36]	Zhang Aiping, Liu Jiangna, Yan Jianjun, Zhang Xiying, Bai Yunfeng. 2022. Progress and prospect of genome editing in tomato. Acta Horticulturae Sinica, 49 (1):221-232. (in Chinese) doi: 10.16420/j.issn.0513-353x.2020-0976
	张爱萍, 刘江娜, 闫建俊, 张西英, 白云凤. 2022. 番茄基因编辑研究进展和前景. 园艺学报, 49 (1):221-232. doi: 10.16420/j.issn.0513-353x.2020-0976
[37]	Zhang C S, Liu C L, Weng J F, Cheng B J, Liu F, Li X H, Xie C X. 2017. Creation of targeted inversion mutations in plants using an RNA-guided endonuclease. Crop Journal, 5 (1):83-88. doi: 10.1016/j.cj.2016.08.001
[38]	Zhang F, Wen Y, Guo X. 2014. CRISPR/Cas 9 for genome editing:progress,implications and challenges. Human Molecular Genetics, 23 (R1):R40-R46.
[39]	Zheng Aihong, Zhang Fen, Jiang Min, Yuan Qiao, Jiang Leiyu, Chen Qing, Tang Haoru, Sun Bo. 2019. Targeted editing of BoaZDS by CRISPR/Ca 9 technology in Chinese kale. Acta Horticulturae Sinica, 46 (1):57-64. (in Chinese)
	郑爱红, 张芬, 江敏, 袁巧, 江雷雨, 陈清, 汤浩茹, 孙勃. 2019. 利用CRISPR/Ca9技术靶向编辑芥蓝BoaZDS. 园艺学报, 46 (1):57-64. doi: 10.16420/j.issn.0513-353x.2018-0426

利用CRISPR/Cas9技术创制高番茄红素番茄新材料

Creating High Lycopene Fruit Using CRISPR/Cas9 Technology in Tomato

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 39

相关文章 15

编辑推荐

Metrics

本文评价

访问总数:　当日访问总数:　当前在线人数:

[1]	赵佳莹, 曾周婷, 岑欣颖, 施姣淇, 李效贤, 沈晓霞, 俞振明, . 铁皮石斛CCO基因家族鉴定及其在花发育中的表达分析[J]. 园艺学报, 2024, 51(9): 2075-2088.
[2]	韩荧, 段颖, 牛一杰, 李衍素, 贺超兴, 孙敏涛, 王君, 李强, 陈双臣, 闫妍. 腐殖酸生物降解地膜提高番茄品质的转录代谢机制研究[J]. 园艺学报, 2024, 51(8): 1758-1772.
[3]	段敏杰, 李怡斐, 王春萍, 杨小苗, 黄任中, 黄启中, 张世才. 辣椒果实类胡萝卜素调控因子转录组和靶向代谢组分析[J]. 园艺学报, 2024, 51(8): 1773-1791.
[4]	龚小雅, 李贤, 周新刚, 吴凤芝. 分蘖洋葱伴生番茄诱导的根际微生物对根结线虫病的影响[J]. 园艺学报, 2024, 51(8): 1913-1926.
[5]	王晨雨, 刘孟军, 王立新, 刘志国, . CRISPR/Cas9技术研究进展及其在园艺植物中的应用进展[J]. 园艺学报, 2024, 51(7): 1439-1454.
[6]	孟思达, 韩磊磊, 相恒佐, 朱美玉, 冯珍, 叶云珠, 孙美华, 李艳冰, 赵利萍, 谭昌华, 齐明芳, 李天来. 番茄心室数的调控机制研究进展[J]. 园艺学报, 2024, 51(7): 1649-1664.
[7]	江睿, 周胜军, 朱育强, 王欣, 谭继宏, 王华森, 张鹏, . 瓜类作物CRISPR/Cas9基因编辑技术应用研究进展[J]. 园艺学报, 2024, 51(7): 1683-1694.
[8]	马星云, 范冰丽, 唐光彩, 贾芝琪, 李营, 薛东齐, 张世文. DXR调控番茄叶绿体发育、花色与果实着色机制初探[J]. 园艺学报, 2024, 51(6): 1241-1255.
[9]	张文静, 徐大勇, 吴倩琳, 杨佛, 信丙越, 曾昕, 李峰. 拮抗番茄灰霉病的贝莱斯芽孢杆菌XDY66基因组分析[J]. 园艺学报, 2024, 51(6): 1413-1425.
[10]	王永珍, 张剑国, 刘彩虹, 李思蓓, 吕甜甜. 番茄新品种‘圆红212’[J]. 园艺学报, 2024, 51(6): 1435-1436.
[11]	刘泽营, 孙帅, 刘志强, 崔霞, 李仁. 番茄尖果脐突变体的生理特性及其候选基因分析[J]. 园艺学报, 2024, 51(5): 982-992.
[12]	李品, 甘宁, 陈家伟, 项思翔, 沈静漪, 欧阳波, 卢永恩. 番茄自然群体磷利用效率分析及耐低磷种质筛选[J]. 园艺学报, 2024, 51(5): 993-1004.
[13]	王文娇, 邢军杰, 申成丞, 李斌. 黄瓜蜡质基因CsCER1调控因子CsCOL5的筛选及其功能分析[J]. 园艺学报, 2024, 51(5): 1005-1016.
[14]	杨婷, 席德慧, 夏明, 李佳楠. α-苦瓜素基因提高番茄对烟草花叶病毒抗性的机理研究[J]. 园艺学报, 2024, 51(5): 1126-1136.
[15]	胡志峰, 邵景成, 张莉. 番茄新品种‘陇番15号’[J]. 园艺学报, 2024, 51(4): 917-918.

利用CRISPR/Cas9技术创制高番茄红素番茄新材料

Creating High Lycopene Fruit Using CRISPR/Cas9 Technology in Tomato

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 39

相关文章 15

编辑推荐

Metrics

本文评价

访问总数: 当日访问总数: 当前在线人数:

访问总数:　当日访问总数:　当前在线人数: