[1]张光智,史寒琪,吴保唐,等.基于CRISPR/Cas9技术提高番茄果实中番茄红素含量[J].江苏农业科学,2026,54(1):27-34.
　Zhang Guangzhi,et al.Improving lycopene content in tomato fruits based on CRISPR/Cas9 technology[J].Jiangsu Agricultural Sciences,2026,54(1):27-34.
点击复制

基于CRISPR/Cas9技术提高番茄果实中番茄红素含量()

《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第54卷
期数:
2026年第1期
页码:
27-34
栏目:
生物技术
出版日期:
2026-01-05

文章信息/Info

Title:
Improving lycopene content in tomato fruits based on CRISPR/Cas9 technology
作者:
张光智; 史寒琪; 吴保唐; 刘涛; 祝建波
石河子大学生命科学学院农业生物技术重点实验，新疆石河子 832003
Author(s):
Zhang Guangzhi; et al
关键词:
番茄; CRISPR/Cas9; SlLCY-E; SlLCY-B; 番茄红素
Keywords:
-
分类号:
S641.201
DOI:
-
文献标志码:
A
摘要:
旨在采用CRISPR/Cas9基因编辑技术对番茄中番茄红素代谢相关的关键基因进行定向编辑，从而提高番茄果实中番茄红素的含量。以石番43亲本为试验材料，基于CRISPR/Cas9系统，构建SlLCY-E、SlLCY-B基因的双元表达载体，用农杆菌介导的遗传转化法获得15株番茄再生苗。经PCR检测发现，有9株番茄植株呈阳性，阳性转化率为60%；对阳性植株的靶序列进行测序发现，有7株番茄发生了基因编辑，编辑效率约为77.8%，其中有4株番茄在2个靶位点均发生了基因突变。此外，SlLCY-E、SlLCY-B基因之间的编辑效率也存在差异，分别为55.6%、667%。基因编辑类型包括碱基缺失、插入。对成熟期番茄进行检测发现，在突变株果实中，番茄红素的含量均高于野生型果实，且不影响其他农艺性状，其中，12号植株果实中的番茄红素含量最高，为野生型的3.2倍。qRT-PCR结果表明，突变株中SlLCY-E、SlLCY-B基因的相对表达量多较野生型显著降低。综上，利用CRISPR/Cas9基因编辑技术抑制了番茄红素环化酶基因的表达，成功提高了果实中番茄红素的含量，同时证明SlLCY-E、SlLCY-B基因表达量与番茄红素积累量呈负相关。
Abstract:
-

参考文献/References:

［1］Wang S N，Zhu F. Tamarillo(Solanum betaceum)：chemical composition，biological properties，and product innovation［J］. Trends in Food Science & Technology，2020，95：45-58.
［2］Ellinger S，Ellinger J，Stehle P. Tomatoes，tomato products and lycopene in the prevention and treatment of prostate cancer：do we have the evidence from intervention studies？［J］. Current Opinion in Clinical Nutrition and Metabolic Care，2006，9(6)：722-727.
［3］Xie Z H，Yang F. The effects of lycopene supplementation on serum insulin-like growth factor 1 (IGF-1) levels and cardiovascular disease：a dose-response meta-analysis of clinical trials［J］. Complementary Therapies in Medicine，2021，56：102632.
［4］Ilahy R，Hdider C，Lenucci M S，et al. Phytochemical composition and antioxidant activity of high-lycopene tomato (Solanum lycopersicum L.) cultivars grown in Southern Italy［J］. Scientia Horticulturae，2011，127(3)：255-261.
［5］Liu H R，Cao X M，Azam M，et al. Metabolism of carotenoids and β-ionone are mediated by carotenogenic genes and PpCCD4 under ultraviolet B irradiation and during fruit ripening［J］. Frontiers in Plant Science，2022，13：814677.
［6］Yuan Y，Ren S Y，Liu X F，et al. SlWRKY35 positively regulates carotenoid biosynthesis by activating the MEP pathway in tomato fruit［J］. The New Phytologist，2022，234(1)：164-178.
［7］Sukegawa S，Saika H，Toki S. Plant genome editing：ever more precise and wide reaching［J］. The Plant Journal，2021，106(5)：1208-1218.
［8］Xue C Y，Greene E C. DNA repair pathway choices in CRISPR-Cas9-mediated genome editing［J］. Trends in Genetics，2021，37(7)：639-656.
［9］Zhou J M，Luan X C，Liu Y X，et al. Strategies and methods for improving the efficiency of CRISPR/Cas9 gene editing in plant molecular breeding［J］. Plants，2023，12(7)：1478.
［10］Li M J，Li H Y，Zhu Q D，et al. Knockout of the sugar transporter OsSTP15 enhances grain yield by improving tiller number due to increased sugar content in the shoot base of rice (Oryza sativa L.)［J］. The New Phytologist，2024，241(3)：1250-1265.
［11］Wan D Y，Guo Y，Cheng Y，et al. CRISPR/Cas9-mediated mutagenesis of VvMLO3 results in enhanced resistance to powdery mildew in grapevine (Vitis vinifera)［J］. Horticulture Research，2020，7：116.
［12］Yang T X，Deng L，Zhao W，et al. Rapid breeding of pink-fruited tomato hybrids using the CRISPR/Cas9 system［J］. Journal of Genetics and Genomics，2019，46(10)：505-508.
［13］滕秀兰，蒲陆梅，王兴民. 分光光度法测定番茄红素［J］. 理化检验(化学分册)，2013，49(11)：1385-1386，1388.
［14］Livak K J，Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method［J］. Methods，2001，25(4)：402-408.
［15］Lakshmi Jayaraj K，Thulasidharan N，Antony A，et al. Targeted editing of tomato carotenoid isomerase reveals the role of 5′UTR region in gene expression regulation［J］. Plant Cell Reports，2021，40(4)：621-635.
［16］Ma X L，Zhang Q Y，Zhu Q L，et al. A robust CRISPR/Cas9 system for convenient，high-efficiency multiplex genome editing in monocot and dicot plants［J］. Molecular Plant，2015，8(8)：1274-1284.
［17］Liu D Y，Qiu T，Ding X H，et al. Rapid construction of multiple sgRNA vectors and knockout of the Arabidopsis IAA2 gene using the CRISPR/Cas9 genomic editing technology［J］. Yi Chuan，2016，38(8)：756-764.
［18］周池. 番茄中番茄红素环化酶基因的编辑及其功能研究［D］. 长沙.：湖南农业大学，2021：55-56.
［19］El-Mounadi K，Morales-Floriano M L，Garcia-Ruiz H.Principles，applications，and biosafety of plant genome editing using CRISPR-Cas9［J］. Frontiers in Plant Science，2020，11：56.
［20］朱宗财，王志军，高能，等. CRISPR/Cas9基因编辑技术在植物抗病性改良中的应用综述［J］. 江苏农业科学，2024，52(3)：1-11.
［21］Li R，Li R，Li X D，et al. Multiplexed CRISPR/Cas9-mediated metabolic engineering of γ-aminobutyric acid levels in Solanum lycopersicum［J］. Plant Biotechnology Journal，2018，16(2)：415-427.
［22］Li J，Scarano A，Gonzalez N M，et al. Biofortified tomatoes provide a new route to vitamin D sufficiency［J］. Nature Plants，2022，8(6)：611-616.
［23］Li X D，Wang Y N，Chen S，et al. Lycopene is enriched in tomato fruit by CRISPR/Cas9-mediated multiplex genome editing［J］. Frontiers in Plant Science，2018，9：559.

相似文献/References:

[1]何从亮,毛久庚,甘小虎,等.玻璃温室番茄长季节基质袋栽培技术[J].江苏农业科学,2013,41(04):158.
[2]李永灿,余文贵,陈怀谷,等.番茄灰霉病菌产毒条件优化[J].江苏农业科学,2013,41(05):94.
　Li Yongcan,et al.Optimization of toxigenic conditions of tomato Botrytis cinerea[J].Jiangsu Agricultural Sciences,2013,41(1):94.
[3]赵秋月,甘潇,张广臣.Na2CO3胁迫对番茄幼苗生长的影响[J].江苏农业科学,2013,41(05):128.
　Zhao Qiuyue,et al.Effect of Na2CO3 stress on growth of tomato seedlings[J].Jiangsu Agricultural Sciences,2013,41(1):128.
[4]耿德刚,徐俊伟,戈振超,等.温室大棚番茄滴灌试验研究及效益分析[J].江苏农业科学,2013,41(05):132.
　Geng Degang,et al.Drip irrigation experimental and benefit analysis on greenhouse tomato[J].Jiangsu Agricultural Sciences,2013,41(1):132.
[5]杜中平,聂书明.不同配方基质对番茄生长特性、光合特性及产量的影响[J].江苏农业科学,2013,41(05):138.
　Du Zhongping,et al.Effects of different substrates on growth，photosynthetic characteristics and yield of tomato[J].Jiangsu Agricultural Sciences,2013,41(1):138.
[6]赵河,毛秀杰,叶景学.抗叶霉病不同基因型番茄的光合特性[J].江苏农业科学,2014,42(11):185.
　Zhao He,et al(8).Photosynthetic characteristics of different genotypes of tomato with resistance to leaf mold[J].Jiangsu Agricultural Sciences,2014,42(1):185.
[7]陈素娟,孙娜娜.不同基质配比对番茄秧苗生长的影响[J].江苏农业科学,2013,41(06):128.
　Chen Sujuan,et al.Effect of different substrate compositions on growth of tomato seedling[J].Jiangsu Agricultural Sciences,2013,41(1):128.
[8]孙禛禛,吴秋霞,温新宇,等.转反义LetAPX基因番茄抗氧化酶活性在苗期、花期、果期的变化[J].江苏农业科学,2015,43(12):188.
　Sun Zhenzhen,et al.Study on antioxidant enzymes activity during seedling，flowering and fruiting of tomato with antisense LetAPX gene[J].Jiangsu Agricultural Sciences,2015,43(1):188.
[9]李晓慧,张恩让,何玉安,等.亚高温及外源物质调节下番茄的生理响应[J].江苏农业科学,2013,41(07):135.
　Li Xiaohui,et al.Physiological response of tomato under the regulation of sub-high temperature and exogenous substances[J].Jiangsu Agricultural Sciences,2013,41(1):135.
[10]李建宏,张楠,张泽,等.番茄红素提取与测定方法的优化[J].江苏农业科学,2013,41(08):259.
　Li Jianhong,et al.Optimization of lycopene extraction and determination method[J].Jiangsu Agricultural Sciences,2013,41(1):259.

备注/Memo

备注/Memo:
收稿日期：2024-10-17
基金项目：新疆生产建设兵团财政科技计划-重点领域科技攻关计划(编号：2023AB006-02)。
作者简介：张光智(2001—)，男，河南新乡人，硕士研究生，主要从事植物基因工程研究。E-mail：zhangguangzhi2021@126.com。
通信作者：祝建波，博士，研究员，主要从事植物抗逆分子研究。E-mail：zjbshz@126.com。
更新日期/Last Update: 2026-01-05