Abstract：The solute carrier family 25 (SLC25) members participate in regulation of anthocyanin biosynthesis. The SLC25 family member SLC25A4 is adenine nucleotide translocase 1 (ANT1), which plays important roles in growth and development, and response to stresses of plant. To enrich the research of SLC25A4 in wheat (Triticum aestivum) and explore biological functions of SLC25A4 in the regulation of anthocyanin biosynthesis, the expression of SLC25A4 was analyzed based on transcriptome sequence results at different developmental stages of 'Guizi 1' wheat grains and was further verified by qPCR. The results demonstrated that the expression of TaSLC25A4-7A, TaSLC25A4-7B and TaSLC25A4-7D1 genes was significantly higher at 25 days post anthesis (DPA) and 35 DPA than that at 10 DPA (P<0.05), particularly qPCR analysis showed that the dynamic change of TaSLC25A4-7A gene expression was consistent with that of anthocyanin accumulation. Furthermore, TaSLC25A4-7A gene overexpression transgenic tobacco (Nicotiana tabacum) lines were obtained, qPCR results showed that the expression level of anthocyanin synthesis related genes (NtDFR, Nt4CL, NtCHI and NtF3H) was significantly higher in transgenic lines than that in wild type (P<0.05), which demonstrated that TaSLC25A4-7A involved in the biological process of regulating anthocyanin synthesis. The results of carbonyl cyanide m-chlorophenylhydrazone (CCCP) treatment to transgenic lines and wild type demonstrated that the TaSLC25A4-7A overexpression transgenic lines were more sensitive to CCCP. The above results demonstrated that TaSLC25A4 gene (especially TaSLC25A4-7A) is involved in anthocyanin synthesis, which provides reference for further exploring the biological function of the TaSLC25A4 gene in anthocyanin accumulation and stresses response of wheat.
[1] 安畅, 李鲁华, 杨文荟, 等 . 2021. 小麦生长素内输基因 GzLAX3-1B 的克隆及生物信息学分析[J]. 分子植物育种 ( 网络首发).(An C, Li L H, Yang W H, et al. 2021. Cloning and bioinformatics analysis of auxin input gene GzLAX3-1B of 'Guizimai No. 1'[J]. Molecular Plant Breeding, https://kns.cnki.net/kcms/detail/46.1068.S.20210625.1116.006.html.)
[2] 巨伟, 王红日, 薛春芝, 等 . 2021. 面包面条优质小麦转录组学分析[J]. 山东农业科学, 53(5): 8-18.
(Ju W, Wang H R, Xue C Z, et al.2021. Transcriptomic analysis of wheat cultivars in bread- and noodle-making quality[J]. Shandong Agricultural Sciences, 53(5): 8-18.)
[3] 李小兰 .2018. 逆境信号增强贵紫麦 1 号花青素合成的分子机制研究[D]. 博士学位论文, 贵州大学, 导师: 张明生, pp. 64-95.
(Li X L.2018. The molecular mechanism on stress signals enhancing anthocyanin biosynthesis in Triticum aestivum L. cv. Guizi 1[D]. Thesis for Ph. D., Guizhou University, Supervisor: Zhang M S, pp. 64-95.)
[4] 张广旭, 王康君, 谭一罗, 等 . 2021. 小麦穗部产量性状研究进展与展望[J]. 农业与技术, 41(08): 809-813.
(Zhang G X, Wang K J, Tan Y L, et al.2021. Research progress and prospect of spike yield characters in wheat[J]. Agri‐ culture and Technology, 41(08): 809-813.)
[5] 张艳俊, 杨毅清, 袁心悦, 等 . 2020. 基于转录组测序对小麦抗叶锈性相关基因的筛选与分析[J]. 河北农业大学学报, 43(1): 11-16.
(Zhang Y J, Yang Y Q, Yuan X Y, et al.2020. Screening and analysis of wheat resistance-related genes to leaf rust based on transcriptome sequencing[J]. Journal of HeBei Agricultural University, 43(1): 11-16.)
[6] Aquila H, Misra D, Eulitz M, et al.1982. Complete amino ac‐ id sequence of the ADP/ATP carrier from beef heart mi‐ tochondria[J]. Hoppe-Seyler's Zeitschrift fur Physiolo‐ gische Chemie, 363(3): 345-349.
[7] Cavalcanti R N, Santos D T, Meireles M A A.2011. Non-ther‐ mal stabilization mechanisms of anthocyanins in model and food systems-an overview[J]. Food Research Inter‐ national, 44(2): 499-509.
[8] Cress B F, Leitz Q D, Kim D C, et al.2017. CRISPRi-mediated metabolic engineering of E. coli for O-methylated an‐ thocyanin production[J]. Microbial Cell Factories, 16(1): 10.
[9] Dufay J N, Fernández-Murray J P, McMaster C R.2017. SLC25 family member genetic interactions identify a role for HEM25 in yeast electron transport chain stabili‐ ty[J]. G3-Genes Genomes Genetics, 7(6): 1861-1873.
[10] Eisenhut M, Planchais S, Cabassa C, et al.2013. Arabidopsis ABOUT DE SOUFFLE is a putative mitochondrial trans‐ porter involved in photorespiratory metabolism and is required for meristem growth at ambient CO2 levels[J]. The Plant Journal, 73(5): 836-839.
[11] Feng N, Song GY, Guan JT, et al.2017. Transcriptome profil‐ ing of wheat inflorescence development from spikelet initiation to floral patterning identified stage-specific regulatory genes[J]. Plant Physiology, 174(3): 1779-1794. Gao A, Hu M, Gong Y, et al. 2020. Pm21 CC domain activity modulated by intramolecular interactions is implicated in cell death and disease resistance[J]. Molecular Plant Pathology, 21(7): 975-984.
[12] Haferkamp I, Hackstein J H, Voncken F G, et al.2002. Functional integration of integration of mitochondrial and hy‐ drogenosomal ADP/ATP carriers in the Escherichia coli membrane reveals different biochemical characteristics for plants, mammals and anaerobic chytrids[J]. Europe‐ an Journal of Biochemistry, 269(13): 3172-3181.
[13] Hoffmann C, Plocharski B, Haferkamp I, et al.2013. From en‐ doplasmic reticulum to mitochondria: Absence of the Arabidopsis ATP antiporter endoplasmic reticulum ade‐ nylate transporter1 perturbs photorespiration[J]. The Plant Cell, 25(7): 2647-2660.
[14] Kharenko O A, Boyd J, Nelson K M, et al.2011. Identifica‐ tion and characterization of interactions between abscis‐ ic acid and mitochondrial adenine nucleotide transloca‐ tors[J]. Biochemical Journal, 437(1): 117-123.
[15] Leroch M, Neuhaus H E, Kirchberger S, et al.2008. Identification of a novel adenine nucleotide transporter in the endoplasmic reticulum of Arabidopsis[J]. The Plant Cell, 20(2): 438-451.
[16] Li X L, LÜ X, Wang X H, et al.2020. Biotic and abiotic stress-responsive genes are stimulated to resist drought stress in purple wheat[J]. Journal of Integrative Agricul‐ ture, 19(1): 33-50.
[17] Li X L, Qian X K, Lǚ X, et al.2018. Upregulated structural and regulatory genes involved in anthocyanin biosynthe‐ sis for coloration of purple grains during the middle and late grain-filling stages[J]. Plant Physiology and Bio‐ chemistry, 130: 235-247.
[18] Liu D, Li S M, Chen W J, et al.2016 Transcriptome analysis of purple pericarps in common wheat (Triticum aestivum L.)[J]. PLOS ONE, 11(5): e0155428.
[19] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J]. Methods, 25(4): 402-408.
[20] Lohmann A, Schöttler M A, Bréhélin C, et al.2006. Deficien‐ cy in phylloquinone (Vitamin K1) methylation affects prenyl quinone distribution, photosystem I abundance, and anthocyanin accumulation in the Arabidopsis AtmenG mutant[J]. Journal of Biological Chemistry, 281(52):40461-40472.
[21] Millar A H, Heazlewood J L.2003. Genomic and proteomic analysis od mitochondrial carrier proteins in Arabidopsis[J]. Plant Physiology, 131(2): 443-453.
[22] Monné M, Daddabbo L, Gagneul D, et al.2018. Uncoupling proteins 1 and 2 (UCP1 and UCP2) from Arabidopsis thaliana are mitochondrial transporters of aspartate, glu‐ tamate and dicarboxylates[J]. Journal of Biological Chemistry, 293(11): 4213-4227.
[23] Nishi Y, Fujimoto S, Sasaki M, et al.2011. Role of mitochon‐ drial phosphate carrier in metabolism-secretion coupling in rat insulinoma cell line INS-1[J]. Biochemical Jour‐ nal, 435(2): 421-430.
[24] Palmieri F.2004. The mitochondrial transporter family (SLC25): Physiological and pathological implications[J]. Pflugers Archiv-European Journal of Physiology, 447(5): 689-709.
[25] Palmieri F.2013. The mitochondrial transporter family SLC25: Identification, properties and physiopathology[J]. Molecular Aspects of Medicine, 34(2-3): 465-484.
[26] Palmieri F.2014. Mitochondrial transporters of the SLC25 family and associated diseases: A review[J]. Journal of Inherited Metabolic Disease, 37(4): 565-575.
[27] Pasqualone A, Bianco A M, Paradiso V M, et al.2015. Produc‐ tion and characterization of functional biscuits obtained from purple wheat[J]. Food Chemistry, 180: 64-70.
[28] Ray D, Mueller N, West P, et al.2013. Yield trends are insuffi‐ cient to double global crop production by 2050[J]. PLOS ONE, 8: e66428.
[29] Rieder B, Neuhaus H E.2011. Identification of an Arabidopsis plasma membrane-located ATP transporter important for anther development[J]. The Plant Cell, 23(5): 1932-1944.
[30] Shin D H, Choi M G, Kang C S, et al.2016. A wheat R2R3- MYB protein PURPLE PLANT1 (TaPL1) functions as a positive regulator of anthocyanin biosynthesis[J]. Bio‐ chemical and Biophysical Research Communications,469(3): 686-691.
[31] Solfanelli C, Poggi A, Loreti E, et al.2006. Sucrose-specific induction of the anthocyanin biosynthetic pathway in Arabidopsis[J]. Plant Physiology, 140(2): 637-646.
[32] Su P, Yan J, Li W, et al.2020. A member of wheat class Ⅲperoxidase gene family, TaPRX-2A, enhanced the tolerance of salt stress[J]. BMC Plant Biology, 20(1): 392. Wang X M, Chen S Y, Shi X, et al. 2019a. Hybrid sequencing reveals insight into heat sensing and signaling of bread wheat[J]. The Plant Journal, 98(6): 1015-1032.
[33] Wang F, Dong Y X, Tang X Z, et al.2018. Comparative tran‐ scriptome analysis revealing the effect of light on antho‐ cyanin biosynthesis in purple grains of wheat[J]. Journal of Agricultural Food Chemistry, 66(13): 3465-3476.
[34] Wang J, Qin Q, Pan J J, et al.2019b. Transcriptome analysis in roots and leaves of wheat seedlings in response to low-phosphorus stress[J]. Scientific Reports, 9: 19802.
[35] Zhang X, Zheng X, Ke S, et al.2016. ER-localized adenine nucleotide transporter ER-ANT1: An integrator of ener‐ gy and stress signaling in rice[J]. Plant Molecular Biolo‐ gy, 92(6): 701-715.
[36] Zhang Y, Fu Y, Wang Q, et al.2020. Transcriptome analysis reveals rapid defence responses in wheat induced by phytotoxic aphid Schizaphis graminum feeding[J]. BMC Genomics, 21(1): 339.
[37] Zhao S, Xi X, Zong Y, et al.2019. Overexpression of Th- MYC4E enhances anthocyanin biosynthesis in common wheat[J]. International Journal of Molecular Sciences, 21(1): 137.
[38] Zheng X W, Shi M M, Wang J, et al.2020. Isoform sequenc‐ ing provides insight into freezing response of common wheat (Triticum aestivum L.)[J]. Frontiers in Genetics,11: 462.
[39] Zilić S, Serpen A, Akıllıoğlu G, et al.2012. Phenolic com‐ pounds, carotenoids, anthocyanins, and antioxidant ca‐ pacity of colored maize (Zea mays L.) kernels[J]. Jour‐ nal of Agricultural and Food Chemistry, 60(5): 1224-1231.
