Analysis of Carotenoid Contents and Key Regulatory Genes Expression in Wheat (Triticum aestivum) Grains from Different Varieties
LI Ji-Hu1,2, SONG Guo-Qi1,2, GENG Jia-Tao1,3, LI Wei1,2, ZHANG Shu-Juan1,2, ZHANG Rong-Zhi1,2, GAO Jie1,2, LI Yu-Lian1,2, LI Gen-Ying1,2,*
1 Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, China; 2 National Engineering Research Center for Wheat and Maize, Jinan 250100, China; 3 College of Life Sciences, Shandong Normal University, Jinan 250358, China
Abstract：Deep analysis of carotenoid contents and key gene expression patterns in wheat (Triticum aestivum) grains is of great significance for breeding wheat varieties with high carotenoid content. In this study, carotenoid components in grains of 3 yellow-flour varieties (JM1, JM1803, and JM8040) and 3 white-flour varieties (JN17, JM22, and JM44) were comparatively analyzed using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS). Results showed a total of 7 xanthophylls, 10 xanthophyll esters and 3 carotenes were identified across the 6 varieties. 8 components, including lutein and zeaxanthin, were widely detected in all varieties. JM1, JM1803, and JM8040 contained more components, including 4 unique components such as violaxanthin, lutein dimyristate, β-carotene, and phytoene. Quantitative analysis revealed that total carotenoid content in JM1, JM1803, and JM8040 reached 9.30, 8.97, and 5.67 μg/g, respectively, significantly higher than other varieties (P<0.05). These 3 varieties also exhibited significantly higher total xanthophylls, total xanthophyll esters, total carotenoids, and most of carotenoid components. Further analysis of the expression patterns of key carotenoid biosynthesis genes (PSY1, PDS, ZDS, LCYE, and LCYB) in developing grains showed that although the expression trends of the 5 genes were consistent across varieties, expression levels of PSY1, PDS, and ZDS in JM1, JM1803, and JM8040 were significantly higher at 14 and 21 d after flowering (DAF)(P<0.05). Additionally, expression leve of LCYE in these varieties was significantly higher at 7 DAF, and expression level of LCYB was significantly elevated at 7, 14, and 28 DAF (P<0.05). Moreover, these gene expression levels exhibited an extremely significant positive correlation with contents of total carotenoid, xanthophylls, xanthophyll esters, and carotenes (P<0.01). The high expression of these key genes may contribute to the elevated carotenoid content in these 3 yellow-flour varieties. This study provides a theoretical foundation for understanding carotenoid biosynthesis pathways and breeding wheat varieties with enhanced carotenoid content.
[1] 丛玲. 2008. 小麦类胡萝卜素生物合成关键酶基因的克隆与遗传转化研究[D]. 博士学位论文, 华中科技大学, 导师: 何光源, 杨广笑, pp. 61-65. (Cong L.2008. Cloning and genetic transformation of genes encodingkey enzymes in carotenoids biosynthsis pathway in wheat (Triticum aestivum L.) [D]. Thesis for Ph.D., Huazhong University of Science and Technology, Supervisor: He G Y, Yang G X, pp. 61-65.) [2] 刘源, 赵冉, 卢振芳, 等. 2025. 植物类胡萝卜素生物代谢途径及其功能研究进展[J]. 生物技术通报, 41(5): 23-31. (Liu Y, Zhao R, Lu Z F, et al.2025. Research progress in the biological metabolic pathway and functions of plant carotenoids[J]. Biotechnology Bulletin, 41(5): 23-31.) [3] 孙鑫, 崔正勇, 李新华, 等. 2023. 不同小麦品种类胡萝卜素含量比较分析[J]. 农学学报, 13(6): 1-4. (Sun X, Cui Z Y, Li X H, et al.2023. Carotenoids' contents in different wheat varieties: Comparative analysis[J]. Journal of Agriculture, 13(6): 1-4.) [4] 王志忠, 燕丽, 郑文寅, 等. 2017. 不同生态区域小麦品种籽粒类胡萝卜素含量及品质性状研究[J]. 南京农业大学学报, 40(1): 20-26. (Wang Z Z, Yan L, Zheng W Y, et al.2017. Research on carotenoid contents and quality traits of wheat varieties in different ecological regions[J], Journal of Nanjing Agricultural University, 40(1): 20-26.) [5] 吴媛媛, 周健, 包晓婷, 等. 2015. 基因型和环境对小麦类胡萝卜素含量及其品质性状的影响[J]. 麦类作物学报, 35(9): 1257-1261. (Wu Y Y, Zhou J, Bao X T, et al.2015. Effect of genotypes and environments to carotenoid contents and some quality traits of wheat varieties[J]. Journal of Triticeae Crops, 35(9): 1257-1261.) [6] 徐昌杰, 张上隆. 2000. 植物类胡萝卜素的生物合成及其调控[J]. 植物生理学通讯, 36(1): 64-70. (Xu C J, Zhang S L.2000. Biosynthesis and regulation of plant carotenoids[J]. Plant Physiology Communications, 36(1): 64-70.) [7] 张上隆, 陈昆松. 2007. 果实品质形成与调控的分子生理[M]. 中国农业出版社, 北京. pp. 107-148. (Zhang S L, Chen K S.2007. Molecular Physiology of Fruit Quality Development and Regulation[M]. China Agriculture Press, Beijing. pp. 107-148.) [8] 张钰玉, 张晓科, 蒋雷, 等. 2013. 陕西小麦黄色素含量基因TaZds-A1和TaZds-D1的组成与分布[J]. 麦类作物学报, 33(6): 1144-1148. (Zhang Y Y, Zhang X K, Jiang L, et al.2013. Detection of genes TaZds-A1 and TaZds-D1 for yellow pigment content and their distribution in Shaanxi wheat cultivars and lines[J]. Journal of Triticeae Crops, 33(6): 1144-1148.) [9] 庄文锋, 黎飞飞, 赵恒, 等. 2024. 叶黄素合成关键基因及小麦生物强化研究展望[J]. 分子植物育种, 22(13): 4377-4383. (Zhuang W F, Li F F, Zhao H, et al.2024. Key genes for lutein synthesis and prospects for research on wheat bio-fortification[J]. Molecular Plant Breeding, 22(13): 4377-4383.) [10] Adom K K, Sorrells M E, Liu R H.2003. Phytochemical profiles and antioxidant activity of wheat varieties[J]. Journal of Agricultural and Food Chemistry, 51(26): 7825-7834. [11] Arnoux P, Morosinotto T, Saga G, et al.2009. A structural basis for the pH-dependent xanthophyll cycle in Arabidopsis thaliana[J]. Plant Cell, 21(7): 2036-2044. [12] Bartleyg E, Scolnik P A.1995. Plant carotenoids: Pigments for photoprotection, visual attraction and human health[J]. Plant Cell, 7(7): 1027-1038. [13] Britton G, Liaaen-Jensen S, Pfander H.2004. Carotenoids Handbook[M]. Springer Verlag, Basel. pp. 13-30. [14] Cazzonelli C I, Pogson B J.2010. Source to sink: Regulation of carotenoid biosynthesis in plants[J]. Trends in Plant Science, 15(5): 266-274. [15] Cong L, Wang C, Li Z, et al.2010. cDNA cloning and expression analysis of wheat (Triticum aestivum L.) phytoene and ζ-carotene desaturase genes[J]. Molecular Biology Reports, 37(7): 3351-3361. [16] Dong C, Qu G, Guo J, et al.2022. Rational design of geranylgeranyl diphosphate synthase enhances carotenoid production and improves photosynthetic efficiency in Nicotiana tabacum[J]. Science Bulletin, 67(3): 315-327. [17] Farre G, Bai C, Twyman R M, et al.2011. Nutritious crops producing multiple carotenoids-a metabolic balancing act[J]. Trends in Plant Science, 16(10): 532-540. [18] Faustino J F, Ribeiro-Silva A, Dalto R F, et al.2016. Vitamin A and the eye: An old tale for modern times[J]. Arquivos Brasileiros de Oftalmologia, 79(1): 56-61. [19] Flowerika, Neha T, Siddharth T.2021. Correlation of carotenoid accumulation and expression pattern of carotenoid biosynthetic pathway genes in Indian wheat varieties[J]. Journal of Cereal Science, 102: 103303. [20] Fratianni A, Irano M, Panfili G, et al.2005. Estimation of color of durum wheat. Comparison of WSB, HPLC, and reflectance colorimeter measurements[J]. Journal of Agricultural and Food Chemistry, 53: 2373-2378. [21] Guzman I, Hamby S, Romero J, et al.2010. Variability of carotenoid biosynthesis in orange colored Capsicum spp[J]. Plant Science, 179(12): 49-59. [22] He Z H, Yang J, Zhang Y, et al.2004. Pan bread and dry white Chinese noodle quality in Chinese winter wheats[J]. Euphytica, 139: 257-267. [23] He X Y, Zhang Y L, He Z H, et al.2008. Characterization of phytoene synthase 1 gene (Psy1) located on common wheat chromosome 7A and development of a functional marker[J]. Theoretical and Applied Genetics, 116: 213-221. [24] Howitt C A, Cavanagh C R, Bowerman A F, et al.2009. Alternative splicing, activation of cryptic exons and amino acid substitutions in carotenoid biosynthetic genes are associated with lutein accumulation in wheat endosperm[J]. Functional Integrative Genomics, 9(3): 363-376. [25] Hussain A, Larsson H, Kuktaite, R, et al.2015. Carotenoid content in organically produced wheat: Relevance for human nutritional health on consumption[J]. International Journal of Environmental Research and Public Health, 12(11): 14068-14083. [26] Inbaraj B S, Lu H, Hing C F, et al.2008. Determination of carotenoids and their esters in fruits of Lycium barbarum Linnaeus by HPLC-DAD-APCI-MS[J]. Journal of Pharmaceutical and Biomedical Analysis, 47(4-5): 812-818. [27] Landrum J T, Bone R A.2004. Dietary lutein & zeaxanthin: Reducing the risk for macular degeneration[J]. Agro Food Industry Hi-Tech, 15(6): 22-25. [28] Lewis E R, Nogueira M, Enfissi E M A, et al.2021. The esterification of xanthophylls in Solanum lycopersicum (tomato) chromoplasts: The role of a non-specific acyltransferase[J]. Phytochemistry, 191: 112912. [29] Liang L, Li Y, Mao X, et al.2024. Metabolomics applications for plant-based foods origin tracing, cultivars identification and processing: Feasibility and future aspects[J]. Food Chemistry, 449: 139227. [30] Miskelly D M.1984. Flour components affecting paste and noodle colour[J]. Journal of the Science of Food and Agriculture, 35(4): 463-471. [31] Paolacci A R, Tanzarella O A, Porceddu E, et al.2009. Identification and validation of reference genes for quantitative RT-PCR normalization in wheat[J]. BMC Molecular Biology, 10: 11. [32] Pozniak C J, Knox R E, Clarke F R, et al.2007. Identification of QTL and association of a phytoene synthase gene with endosperm colour in durum wheat[J]. Theoretical and Applied Genetics, 114(3): 525-537. [33] Pu X, Li Z, Tian Y, et al.2020. The honeysuckle genome provides insight into the molecular mechanism of carotenoid metabolism underlying dynamic flower coloration[J]. New Phytologist, 227(3): 930-943. [34] Rivera S M, Christou P, Canela-Garayoa R.2014. Identification of carotenoids using mass spectrometry[J]. Mass Spectrometry Reviews, 33(5): 353-372. [35] Rodrigues D B, MariuttiL R B, Mercadante A Z.2016. Two-step cleanup procedure for the identification of carotenoid esters by liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry[J]. Journal of Chromatography A, 1457: 116-124. [36] Rodriguez-Concepcion M, Avalos J, Bonet ML, et al.2018. A global perspective on carotenoids: Metabolism, biotechnology, and benefits for nutrition and health[J]. Progress in Lipid Research, 70: 62-93. [37] Trono D.2019. Carotenoids in cereal food crops: Composition and retention throughout grain storage and food processing[J]. Plants, 8(12): 551. [38] Wang J W, He X Y, He Z H, et al.2009. Cloning and phylogenetic analysis of phytoene synthase 1 (Psy1) genes in common wheat and related species[J]. Hereditas, 146(5): 208-256. [39] Winters A, Heywood S, Farrar K, et al.2009. Identification of an extensive gene cluster among a family of PPOs in Trifolium pratense L. (red clover) using a large insert BAC library[J]. BMC Plant Biology, 9: 94. [40] Xia Y, Chen W, Xiang W, et al.2021. Integrated metabolic profiling and transcriptome analysis of pigment accumulation in Lonicera japonica flower petals during colour-transition[J]. BMC Plant Biology, 21(1): 98. [41] Yeum K J, Russell R M.2002. Carotenoid bioavailability and bioconversion[J]. Annual Review of Nutrition, 22: 483-504. [42] Zhang C Y, Dong C H, He X Y, et al.2011. Allelic variants at the TaZds-D1 Locus on wheat chromosome 2DL and their association with yellow pigment content[J]. Crop Science, 51(4): 1580-1590.
