DXR调控番茄叶绿体发育、花色与果实着色机制初探

doi:10.16420/j.issn.0513-353x.2023-0774

摘要/Abstract

摘要：

在植物的生长发育过程中，类胡萝卜素对叶、花、果的着色有着重要影响，是果实发育过程中的主要色素物质。1-脱氧-D-木酮糖-5-磷酸还原异构酶基因（DXR）参与了类胡萝卜素的合成，构建番茄DXR基因编辑株系和超量表达株系，发现基因编辑株系叶片呈现黄色甚至白色，叶片的叶绿素含量和最大PSⅡ的光能转换效率显著降低，细胞叶绿体发育受到影响，类囊体数量减少；花瓣边缘白化，红熟期果实颜色变为橙色甚至黄色，果实番茄红素、类黄酮等含量有不同程度降低。而超量表达株系与野生型对照相比无明显差异，但可溶性固形物含量明显增多。通过转录组测序发现，SlDXR与类黄酮生物合成、类胡萝卜素生物合成等途径相关。推测SlDXR可能通过调控萜类化合物的合成来影响番茄果实着色。

关键词: 番茄, DXR, 基因编辑, 叶绿素, 番茄红素, 果实颜色

Abstract:

In the growth and development of plants，carotenoids play a crucial role in leaf，flower，and fruit pigmentation，serving as the primary pigment compounds during fruit maturation. The 1-deoxyd-D-xylose-5-phosphate reduction isomerase gene（DXR）is involved in the synthesis of carotenoids. The SlDXR gene editing and overexpression lines of tomato were constructed. It was found that gene editing lines exhibited yellow or even white leaves，the chlorophyll content and maximum photoenergy conversion efficiency of PSⅡ decreased significantly，the chloroplast development was affected，and the number of thylakoids was reduced. Meanwhile，the petal edge of gene editing lines turned white and the fruit color changed to orange or even yellow in the red ripening stage. The content of lycopene，flavonoid and other pigments in these fruits decreased to different degrees. There was no significant difference between the overexpression linesand the wild type（the control），however，the soluble solids content was significantly increased. Transcriptome sequencing showed that SlDXR was related to flavonoid biosynthesis，carotenoid biosynthesis and other pathways. These findings suggested that SlDXR could affect the coloring of tomato by regulating the synthesis of terpenoids.

Key words: tomato, DXR, gene editing, chlorophyll, lycopene, fruit color

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.

MA Xingyun, FAN Bingli, TANG Guangcai, JIA Zhiqi, LI Ying, XUE Dongqi, ZHANG Shiwen. Preliminary Study on the Mechanism of DXR Regulating Chloroplast Development Flower Color and Fruit Coloring in Tomato[J]. Acta Horticulturae Sinica, 2024, 51(6): 1241-1255.

图/表 11

表1 qRT-PCR所用引物序列

Table 1 Sequence of primers used for qRT-PCR

基因序列 Primer name	正向引物序列（5′-3′） Forward primer sequence	反向引物序列（5′-3′） Reverse primer sequence
Actin	GTCCTCTTCCAGCCATCCAT	ACCACTGAGCACAATGTTACCG
Solyc03g114340（DXR）	AAGGCAGAGTTGGGATGGTC	TTGTCCGGATTCTCAGCGAC
Solyc04g012120（14-3-3）	GAGCTAGGAGAGCATCGTGG	GCAACTTGAGGATGCCGTTAC
Solyc03g034180（14-3-3）	TGATGATGCCATCGCAGAGC	TCATCCGTGGTATCAGAAGTCC
Solyc01g079620（MYB12）	GGGCATCAAGAGAGGCAGAT	AACGACCTCCAAGAGCCTTC
Solyc11g013110（FLS）	AAGATTTGGCCTCCTCCTGC	TCCAAACCAAGCCCAAGTGA
Solyc04g040190（LCYB）	GTGCTCGATGCAACTGGCTT	GGTGCTCTTCCACTTCAGCC
Solyc12g008980（LCYE）	TGGTGTATGGGAGGACGAGT	AACTCTTCCATAGGCACGGC
Solyc05g010320（CHI）	GAATCCTCTGGGCTCCAACA	ACACACCGATCGCAGTAAAC

表2 表达载体所用引物序列

Table 2 Sequence of primers used for expression vectors

引物名称 Primer name	正向引物序列（5′-3′） Forward primer sequence	反向引物序列（5′-3′） Reverse primer sequence
CP098-SlDXR	AATCTAACAGTGTAGTTTGTGAGCCAGGAAGGCAGAGTTGTTTTAGAGCTAGAAATAGC	CTATTTCTAGCTCTAAAACTGCTCCCCAGGTATAATCTCCAAACTACACTGTTAGATTC
pHellgate8-SlDXR	CATTTGGAGAGGACACGCTCGAGATGGCCCTCAATTTGCTTTCTC	TCTCATTAAAGCAGGACTCTAGATACAAGAGCTGGACTCAAACCAGAT
pCAMBIA1302-SlDXR-GFP	ACGGGGGACTCTTGACCATGGATGGCCCTCAATTTGCTTTCTC	AAGTTCTTCTCCTTTACTAGTTACAAGAGCTGGACTCAAACCAGAT
pMV2-SlDXR	TGCATCCAACGCGTTGGGAGCTCGCCTGCATCACACCCGTAT	GCCTTCGCCATTCTAGACTCGAGGCTATGTGTCTCGTTGTGGC

图1 SlDXR 在番茄各个组织中的表达量各组织与根中相对表达量相比。

Fig. 1 Relative expression of SlDXR in various tissues of tomato Comparison of relative expression levels between tissues and roots. t-test，* P < 0.05；** P < 0.01

图2 ‘Micro-Tom’番茄野生型（WT）和转基因（pMV2-SlDXR）株系的GUS表达分析

Fig. 2 GUS expression analysis of‘Micro-Tom’tomato wild type（WT）and transgenic（pMV2-SlDXR）lines

图3 番茄SlDXR在烟草中的亚细胞定位叠加1为GFP荧光与叶绿体荧光的叠加图；叠加2为明场、GFP荧光和叶绿体荧光的叠加图。

Fig. 3 Subcellular localization of tomato SlDXR in tobacco Superposition 1 represents the superposition of GFP fluorescence and chloroplast fluorescence；Superimposed 2 generations of brighfield，GFP fluorescence and chloroplast fluorescence.

图4 番茄野生型（WT）、T0代SlDXR编辑株系（CR）和超量表达株系（OE）株系的鉴定 A：基因编辑载体CP098阳性检测；B：超量表达载体pHellsgate8阳性检测；C：CRISPR/Cas9基因编辑SlDXR的靶点和突变位点；D：CR株系的SlDXR基因序列。

Fig. 4 Identification and gene expression detection of tomato wild type（WT），T0 generation SlDXR edited lines（CR）and overexpressed lines（OE） A：Positive detection of gene editing vector CP098；B：Positive detection of overexpression lines vector pHellsgate8；C：Target and mutation sites of CRISPR/ Cas9 gene editing SlDXR；D：SlDXR gene sequence of CR lines.

图5 SlDXR相对表达量

Fig. 5 The relative expression of SlDXR ** P < 0.01.

图6 番茄野生型（WT）、SlDXR编辑株系（CR）和超量表达株系（OE）的表型与光合相关指标测定 A：T0代阳性植株；B：T2代叶片叶绿素荧光成像；C：叶片最大PSⅡ的光能转换效率；D：叶片叶绿素含量。** P < 0.01。

Fig. 6 Phenotypic and photosynthetic related indexes determination of tomato wild type（WT），SlDXR edited（CR）and overexpressed（OE）lines A：T0 generation positive plants；B：T2 generation leaf chlorophyll fluorescence imaging；C：The light energy conversion efficiency of the maximum PSⅡof the leaves；D：Chlorophyll content in leaves. ** P < 0.01.

图7 番茄野生型（WT）和SlDXR基因编辑株系（CR）叶片中叶绿体结构观察

Fig. 7 Chloroplast structure in the leaves of tomato wild type（WT）and SlDXR gene editing lines（CR）

图8 番茄野生型（WT）、SlDXR编辑株系（CR）和超量表达株系（OE）果实表型及其相关生理指标

Fig. 8 Fruit phenotype and related physiological indexes of tomato wild type（WT），SlDXR edited（CR）and overexpression lines（OE） * P < 0.05；** P < 0.01.

图9 SlDXR相关基因在番茄野生型（WT）、SlDXR编辑株系（CR）和超量表达株系（OE）叶片（A、B）和红熟果实（C ~ G）中的表达量

Fig. 9 Relative expression levels of SlDXR-related genes in tomato wild type（WT），SlDXR edited（CR）and overexpressed（OE）lines in leaves（A，B）and red ripe fruits（C-G） ** P < 0.01；*** P < 0.001.

参考文献 37

[1]	Ballester A R, Molthoff J, de Vos R, Hekkert B T, Orzaez D, Fernández-Moreno J P, Tripodi P, Grandillo S, Martin C, Heldens J, Ykema M, Granell A, Bovy A. 2010. Biochemical and molecular analysis of pink tomatoes:deregulated expression of the gene encoding transcription factor SlMYB 12 leads to pink tomato fruit color. Plant Physiol, 152 (1):71-84. doi: 10.1104/pp.109.147322 pmid: 19906891
[2]	Carretero-Paulet L, Cairó A, Botella-Pavía P, Besumbes O, Campos N, Boronat A, Rodríguez-Concepción M. 2006. Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase. Plant Mol Biol, 62 (4-5):683-695. pmid: 16941216
[3]	Chen Tongqiang, Zhang Tianzhu, Wang Xiaozhuo. 2022. Research progress of the regulation of light on lycopene biosynthesis in tomato fruit. Acta Horticulturae Sinica, 49 (4):907-923. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0063
	陈同强, 张天柱, 王晓卓. 2022. 光照对番茄果实中番茄红素生物合成的调控研究进展. 园艺学报, 49 (4):907-923. doi: 10.16420/j.issn.0513-353x.2021-0063
[4]	Dai Xiang, Liu Weiwei, Lin Guanghao. 2022. Analysis of pigment content in blueberry fruit. Zhejiang Agricultural Sciences, 63 (12):2917-2920. (in Chinese)
	戴翔, 刘伟伟, 林光号. 2022. 蓝莓果实色素含量的分析. 浙江农业科学, 63 (12):2917-2920. doi: 10.16178/j.issn.0528-9017.20220153
[5]	Ding Fei. 2022. Epigenetic mechanism analysis of SlFolB regulation of chloroplast development in tomato[Ph. D. Dissertation]. Zhengzhou: Henan Agricultural University. (in Chinese)
	丁飞. 2022. SlFolB调控番茄叶绿体发育的表观遗传机理解析[博士论文]. 郑州: 河南农业大学.
[6]	Eisenreich W, Schwarz M, Cartayrade A, Arigoni D, Zenk M H, Bacher A. 1998. The deoxyxylulose phosphate pathway of terpenoid biosynthesis in plants and microorganisms. Chem Biol, 5:R221-R233.
[7]	Gao Hong-yan, Mei Xiao-hong, Su Ye-yang, Zhu Ben-zhong, Luo Yun-bo. 2005. Studies on lycopene and its biosynthetic pathway. Food Science and Technology,(8):42-45. (in Chinese)
	高红岩, 梅晓宏, 苏夜阳, 朱本忠, 罗云波. 2005. 番茄红素及其生物合成途径的研究. 食品科技,(8):42-45.
[8]	Gomez Roldan M V, Outchkourov N, van Houwelingen A, Lammers M,Romero de la Fuente I,Ziklo N,Aharoni A,Hall R D,Beekwilder J. 2014. An O-methyltransferase modifies accumulation of methylated anthocyanins in seedlings of tomato. Plant Journal, 80 (4):695-708.
[9]	Guo Ya-fei, Wang Jun-ya, Guo Fei, Ni De-jiang. 2018. Cloning and expression analysis of 1-deoxyd-xylose-5-phosphate synthetase gene CsDXS1 in tea plant. Bulletin of Biotechnology, 34 (1):144-152. (in Chinese)
	郭亚飞, 王君雅, 郭飞, 倪德江. 2018. 茶树1-脱氧-D-木酮糖-5-磷酸合成酶基因CsDXS1的克隆与表达分析. 生物技术通报, 34 (1):144-152. doi: 10.13560/j.cnki.biotech.bull.1985.2017-0652
[10]	Haba P D, Mata L D, Molina E, Aguera E. 2014. High temperature promotes early senescence in primary leaves of sunflower(Helianthus annuus L.)plants. Can J Plant Sci, 94:659-668.
[11]	Hasunuma T, Takeno S, Hayashi S, Sendai M, Bamba T, Yoshimura S, Tomizawa K, Fukusaki E, Miyake C. 2008. Overexpression of 1-Deoxy-D-xylulose-5-phosphate reductoisomerase gene in chloroplast contributes to increment of isoprenoid production. J Biosci Bioeng, 105 (5):518-526. doi: 10.1263/jbb.105.518 pmid: 18558344
[12]	Jiang J, Huang H, Gao Q, Li Y, Xiang H, Zeng W, Xu L, Liu X, Li J, Mi Q, Deng L, Yang W, Zhang J, Yang G, Li X. 2023. Effects of editing DFR genes on flowers,leaves,and roots of tobacco. BMC Plant Biol, 23 (1):349.
[13]	Lange B M, Rujan T, Martin W, Croteau R. 2000. Isoprenoid biosynthesis:the evolution of two ancient and distinct pathways across genomes. Proc Natl Acad Sci USA, 97:13172-13177. doi: 10.1073/pnas.240454797 pmid: 11078528
[38]	Zhang Li, Chen Feng-feng, Wang Hong-xia, Liao Le-qin, Zhang Peng, Zhou Quan-lu, Kang Le. 2023. Progress of metabolic regulation of carotenoids in sweet potato. Chinese Journal of Agricultural Biotechnology, 31 (8):1719-1729. (in Chinese)
	张丽, 陈丰酆, 王红霞, 廖乐琴, 张鹏, 周全卢, 康乐. 2023. 甘薯类胡萝卜素的代谢调控研究进展. 农业生物技术学报, 31 (8):1719-1729.
[39]	Zhang Qiaoli, Chen Di, Song Yanping, Zhu Hongliang, Luo Yunbo, Qu Guiqin. 2023. Review on transcriptional regulation of chlorophyll metabolism network in tomato fruits. Acta Horticulturae Sinica, 50 (9):2031-2047. (in Chinese) doi: 10.16420/j.issn.0513-353x.2022-0110
	张巧丽, 陈笛, 宋艳萍, 朱鸿亮, 罗云波, 曲桂芹. 2023. 番茄果实叶绿素代谢转录调控网络研究进展. 园艺学报, 50 (9):2031-2047. doi: 10.16420/j.issn.0513-353x.2022-0110
[40]	Zhang Shiwen. 2018. Study on heat resistance gene LrgB and fruit firmness regulation gene in tomato[Ph. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	张世文. 2018. 番茄抗热基因LrgB及果实硬度调控基因的研究[博士论文]. 武汉: 华中农业大学.
[41]	Zhang Wei,Liang Cheng-wei. 2014. Advances in the synthesis of isoprene in plants. Shandong Chemical Industry, 43 (5):57-58. (in Chinese)
	张伟, 梁成伟. 2014. 植物类异戊二烯合成途径的研究进展. 山东化工, 43 (5):57-58.
[42]	Zhang Wencai. 2023. Study on expression of terpenoids and flavonoids and their synthesis-related genes in different elevations of Tibetan Balsa flower[M. D. Dissertation]. Nyingzhi: Xizang University. (in Chinese)
	张文才. 2023. 不同海拔藏波罗花中萜类和黄酮类化合物及其合成相关基因表达的研究[硕士论文]. 林芝: 西藏大学.
[43]	Zhang Xinye, Wang Yuxin, Sun Yanxiang, Zhu Shu, Wang Congyan, Li Wenjing. 2021. Determination of flavonoid content in carrot and identification of DcCHS gene family. Journal of Northwest Agricultural Sciences, 30 (4):572-581. (in Chinese)
	张新业, 王雨欣, 孙艳香, 朱姝, 王聪艳, 李文静. 2021. 胡萝卜类黄酮含量的测定及DcCHS基因家族的鉴定分析. 西北农业学报, 30 (4):572-581.
[44]	Zhou Ying, Li Bingying, Li Xuebao. 2012. Regulation of 14-3-3 proteins on plant development. Acta Botanica Sinica, 47 (1):55-64. (in Chinese)
	周颖, 李冰樱, 李学宝. 2012. 14-3-3蛋白对植物发育的调控作用. 植物学报, 47 (1):55-64. doi: 10.3724/SP.J.1259.2012.00055
[14]	Laule O, Fürholz A, Chang H S, Zhu T, Wang X, Heifetz P B, Gruissem W, Lange M. 2003. Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana. Proc Natl Acad Sci USA, 100 (11):6866-6871.
[15]	Lichtenthaler H K. 2000. Non-mevalonate isoprenoid biosynthesis:enzymes,genes and inhibitors. Biochem Soc Trans, 28 (6):785-789.
[16]	Li Y, Cui W, Qi X, Qiao C, Lin M, Zhong Y, Hu C, Fang J. 2019. Chalcone synthase encoding AeCHS is involved in normal petal coloration in Actinidia eriantha. Genes (Basel), 10 (12):949.
[17]	Lu Chenfei, Gao Yuexia, Huang He, Dai Silan. 2022. Carotenoid metabolism and regulation in plants. Acta Horticulturae Sinica, 49 (12):2559-2578. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0531
	陆晨飞, 高月霞, 黄河, 戴思兰. 2022. 植物类胡萝卜素代谢及调控研究进展. 园艺学报, 49 (12):2559-2578. doi: 10.16420/j.issn.0513-353x.2021-0531
[18]	Luo Jin-feng, Ren Mei-yan, Chen Jing-xin, Ding Xiao-wen. 2011. Research progress on the physiological function of lycopene and methods for maintaining its stability. Food Science, 32 (19):279-283. (in Chinese) doi: 10.7506/spkx1002-6630-201119062
	罗金凤, 任美燕, 陈敬鑫, 丁晓雯. 2011. 番茄红素的生理功能及保持其稳定性方法的研究进展. 食品科学, 32 (19):279-283. doi: 10.7506/spkx1002-6630-201119062
[19]	Luo Zhidan. 2015. Functional identification of tomato fruit color and texture regulation genes SlSGR1 and nsLTP[Ph. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	罗志丹. 2015. 番茄果实颜色与质地调控基因SlSGR1和nsLTP的功能鉴定[博士论文]. 武汉: 华中农业大学.
[20]	Ouyang Bo. 2003. Transformation of several disease-course related protein genes in tomato[Ph. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	欧阳波. 2003. 几种病程相关蛋白基因转化番茄的研究[博士论文]. 武汉: 华中农业大学.
[21]	Owens D K, Alerding A B, Crosby K C, Bandara A B, Westwood J H, Winkel B S. 2008. Functional analysis of a predicted flavonol synthase gene family in Arabidopsis. Plant Physiol, 147 (3):1046-1061.
[22]	Pogson B J, Rissler H M. 2000. Genetic manipulation of carotenoid biosynthesis and photoprotection. Philos Trans R Soc Lond B Biol Sci, 355 (1402):1395-1403.
[23]	Qiu Jianfang. 2023. Drought and cold tolerance of tomato transcription factor PHYTOCLOCK1( SlPCL1)gene[M. D. Dissertation]. Chongqing: Southwest University. (in Chinese)
	邱见方. 2023. 番茄转录因子PHYTOCLOCK1( SlPCL1) 基因的耐旱和耐寒功能研究[硕士论文]. 重庆: 西南大学.
[24]	Rodríguez-Concepción M, Ahumada I, Diez-Juez E, Sauret-Güeto S, Lois L M, Gallego F, Carretero-Paulet L, Campos N, Boronat A. 2001. 1-Deoxy-D-xylulose 5-phosphate reductoisomerase and plastid isoprenoid biosynthesis during tomato fruit ripening. Plant Journal, 27 (3):213-222. doi: 10.1046/j.1365-313x.2001.01089.x pmid: 11532167
[25]	Rohmer M. 1999. The discovery of a mevalonate-independent pathway for isoprenoid biosynthesis in bacteria,algae and higher plants. Natural Product Reports, 16 (5):565-574. doi: 10.1039/a709175c pmid: 10584331
[26]	Ruan Meiying. 2014. Research on Main pigment accumulation,fruit color heredity and germplasm innovation of purple tomato fruit[M. D. Dissertation]. Jinhua: Zhejiang Normal University. (in Chinese)
	阮美颖. 2014. 紫色番茄果实主要色素积累、果色遗传规律及种质创新研究[硕士论文]. 金华: 浙江师范大学.
[27]	Schijlen E G, de Vos C H, Martens S, Jonker H H, Rosin F M, Molthoff J W, Tikunov Y M, Angenent G C, van Tunen A J, Bovy A G. 2007. RNA interference silencing of chalcone synthase,the first step in the flavonoid biosynthesis pathway,leads to parthenocarpic tomato fruits. Plant Physiol, 144 (3):1520-1530. doi: 10.1104/pp.107.100305 pmid: 17478633
[28]	Song W, Wei F, Gao S, Dong C, Hao J, Jin L, Li F, Wei P, Guo J, Wang R. 2022. Functional characterization and comparison of lycopene epsilon-cyclase genes in Nicotiana tabacum. BMC Plant Biol, 22 (1):252.
[29]	Wang Jin, Wang Xinyu, Shen Yuanbo, Zhang Qinghua, Lou Qianqi, Zhang Shijie, Zhao Pan, Liang Yan. 2022. Regulation of chloroplast development in tomato fruit and its application. Acta Horticulturae Sinica, 49 (12):2669-2682. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0913
	王晋, 王新宇, 沈渊博, 张清花, 娄茜棋, 张世杰, 赵攀, 梁燕. 2022. 番茄果实叶绿体发育调控及其应用的研究进展. 园艺学报, 49 (12):2669-2682. doi: 10.16420/j.issn.0513-353x.2021-0913
[30]	Wang X W, Shao C X, Liu L B, Wang Y, An Y Q, Li H, Ding Y W, Jing Y P, Li X J, Xiao J W. 2023. PtrDJ1C,an atypical member of the DJ-1 superfamily, is essential for early chloroplast development and lignin deposition in poplar. Horticultural Plant Journal, 9 (5):1039-1054.
[31]	Xing S, Miao J, Li S, Qin G, Tang S, Li H, Gu H, Qu L J. 2010. Disruption of the 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR)gene results in albino,dwarf and defects in trichome initiation and stomata closure in Arabidopsis. Cell Research, 20 (6):688-700.
[32]	Xu Meng. 2018. Discovery and functional analysis of important regulatory factors for early development of tomato fruit[M. D. Dissertation]. Beijing: Chinese Academy of Agricultural Sciences. (in Chinese)
	徐萌. 2018. 番茄果实早期发育重要调控因子的发掘与功能分析[硕士论文]. 北京: 中国农业科学院.
[33]	Yang J, Adhikari M N, Liu H, Xu H, He G, Zhan R, Wei J, Chen W. 2012. Characterization and functional analysis of the genes encoding 1-deoxy-D-xylulose-5-phosphate reductoisomerase and 1-deoxy-D-xylulose-5-phosphate synthase,the two enzymes in the MEP pathway,from Amomum villosum Lour. Molecular Biology Reports, 39 (8):8287-8296.
[34]	Yang J, Worley E, Udvardi M. 2014. A NAP-AAO 3 regulatory module promotes chlorophyll degradation via ABA biosynthesis in Arabidopsis leaves. Plant Cell, 26 (12):4862-4874.
[35]	Yang Xiao-fei, Guo Fang-qing. 2014. Research progress on leaf senescence mechanism under high temperature stress. Chinese Journal of Plant Physiology, 50 (9):1285-1292. (in Chinese)
	杨小飞, 郭房庆. 2014. 高温逆境下植物叶片衰老机理研究进展. 植物生理学报, 50 (9):1285-1292.
[36]	Yin Y C, Zhang X D, Gao Z Q, Hu T, Liu Y. 2019. The research progress of chalcone isomerase(CHI)in plants. Mol Biotechnol, 61 (1):32-52.
[37]	Zhang D, Yang Y, Chen G, Tian X, Wang Z, Fan S, Xin Z. 2021. Nondestructive evaluation of soluble solids content in tomato with different stage by using Vis/NIR technology and multivariate algorithms. Spectrochim Acta A Mol Biomol Spectrosc, 248:119139.