猕猴桃维生素C的合成规律及关键基因鉴定研究

doi:10.11913/PSJ.2095-0837.2022.50657

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摘要

本研究以3个不同杂交猕猴桃 (Actinidia)群体的果实为材料，测定果实不同发育时期的维生素C(Vc)含量；采用实时荧光定量法PCR法分析猕猴桃Vc合成关键基因的表达情况；对发掘的关键基因GGP开展蛋白结构、理化性质和亲水性和疏水性分析，并在杂交群体中验证GGP基因对于后代果实Vc合成的调控作用。结果显示，当高Vc的猕猴桃物种作为亲本时，杂交子代Vc含量在早期(花后60 d)达到最大值，再随着果实发育而逐渐降低；当中低Vc含量的猕猴桃物种作为亲本，其后代果实Vc含量一直较低且发育规律与母本相似。Vc合成相关基因中，仅有GGP基因在高Vc杂交群体中的表达量均显著高于低Vc杂交群体。在GGP的3个同源基因中，GGP1和GGP3的蛋白质二级、三级结构、理化性质以及亲、疏水性高度相似；但GGP2与前者存在较大的差异。在3个杂交群体后代果实中采用关联分析验证GGP1和GGP3的功能发现，GGP3基因对杂交后代果实的Vc合成可能具有最为重要的作用。研究结果表明，不同猕猴桃杂交群体Vc含量的合成规律存在差异，GGP3基因可能对不同杂交群体后代果实的Vc合成具有重要影响。

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关键词 ：猕猴桃, 维生素C, 合成规律, 基因鉴定, GGP基因

Abstract：

We investigated the developmental patterns of vitamin C (Vc) in hybrid Actinidia species to explore key genes regulating Vc synthesis and provide references and data for the creation of Actinidia germplasms with high Vc content. The fruits of three different hybrid kiwifruit populations were used as test material to investigate Vc synthesis patterns by measuring Vc content in fruit at different developmental stages. Key genes for Vc synthesis in Actinidia were determined using quantitative real-time polymerase chain reaction (qRT-PCR). The protein structure, physicochemical properties, and hydrophilicity of the GGP gene, and the regulation of Vc synthesis in fruits from inbred populations were analyzed. Results showed that Vc inheritance from different cross combinations differed:when Actinidia species with high Vc were used as parents, Vc content in hybrid offspring reached a maximum in the early stage (60 d after flowering) and then decreased gradually with fruit development. In contrast, Vc content in the fruit of offspring from parents with low Vc content (A. rufa cv. 'Guihai') was low and the development pattern was similar to that of the female (A. rufa). The relative expression level of the GGP gene among other Vc synthesis-related genes was significantly higher in the high Vc inbred population than in the low Vc inbred population. Thus, as a key gene for Vc synthesis, GGP was selected for further analysis, with three homologous genes identified (GGP1, GGP3, and GGP2). The α-helix site region in the secondary structure of GGP1 and GGP3 was relatively conserved, and the tertiary spatial structure, physical and chemical properties, and hydrophilicity were also highly similar. In contrast, GGP2 was quite different. Association analysis was used to verify the functions of GGP1 and GGP3 in the offspring fruit of the three hybrid populations, which suggested that GGP3 may play the most important role in Vc synthesis. Our results indicated that the genetic trends of Vc content in the different Actinidia hybrid populations differed. Furthermore, Vc synthesis in the hybrid fruit from middle and high Vc populations reached a maximum in the early and middle stages of fruit development. GGP3 may have an important effect on Vc synthesis in the fruit of offspring from different hybrid populations.

Key words： Actinidia Vitamin C Synthesis rule Gene identification GGP

收稿日期: 2022-02-22

ZTFLH:

Q943.2

基金资助:

国家自然科学基金(C150102)；湖北省猕猴桃自然科技资源库项目(ZKJXPJ202100000)。

通讯作者: 李大卫,E-mail:lidawei@wbgcas.cn;钟彩虹,E-mail:zhongch@wbgcas.cn E-mail: lidawei@wbgcas.cn;zhongch@wbgcas.cn

作者简介: 刘晓莹(1993-),女,博士研究生,研究方向为植物分子生物学(E-mail:liuxiaoying17@mails.ucas.ac.cn)。

引用本文:

刘晓莹, 解潇冬, 成畅, 汪文杰, 吕海燕, 李大卫, 钟彩虹. 猕猴桃维生素C的合成规律及关键基因鉴定研究[J]. 植物科学学报, 2022, 40(5): 657-668. Liu Xiao-Ying, Xie Xiao-Dong, Cheng Chang, Wang Wen-Jie, Lü Hai-Yan, Li Da-Wei, Zhong Cai-Hong. Synthesis patterns of vitamin C in Actinidia and identification of key genes. Plant Science Journal, 2022, 40(5): 657-668.

链接本文:

http://www.plantscience.cn/CN/10.11913/PSJ.2095-0837.2022.50657 或 http://www.plantscience.cn/CN/Y2022/V40/I5/657

[1] 车秀琴.维生素C与人体健康[J].科学之友, 2013(1):112-113. Che XQ. Vitamin C and human health[J]. Friend of Science Amateurs, 2013(1):112-113.
[2] 李明媚.维生素C,维持人体健康的多面手[J].江苏卫生保健, 2020(8):50.
[3] Wheeler GL, Jones MA, Smirnoff N. The biosynthetic pathway of vitamin C in higher plants[J]. Nature, 1998, 393(6683):365-369.
[4] Linster CL, Adler LN, Webb K, Christensen KC, Brenner C, Clarke S. A second GDP-L-galactose phosphorylase in Arabidopsis enroute to vitamin C:covalent intermediate and substrate requirements for the conserved reaction[J]. J Biol Chem, 2008, 283(27):18483-18492.
[5] Wheeler G, Ishikawa T, Pornsaksit V, Smirnoff N. Evolution of alternative biosynthetic pathways for vitamin C following plastid acquisition in photosynthetic eukaryotes[J]. eLife, 2015, 4:e06369.
[6] Li MJ, Ma FW, Liu J, Li J. Shading the whole vines during young fruit development decreases ascorbate accumulation in kiwi[J]. Physiol Plant, 2010, 140(3):225-237.
[7] Laing WA, Bulley S, Wright M, Cooney J, Jensen D, et al. A highly specific L-galactose-1-phosphate phosphatase on the path to ascorbate biosynthesis[J]. Proc Natl Acad Sci USA, 2004, 101(48):16976-16981.
[8] Bulley SM, Laing W. The Kiwifruit Genome:Ascorbic acid-related genes[M]. Switzerland:Springer Nature, 2016:163-177.
[9] 陈璐,王小玲,张小丽,高柱.猕猴桃AsA生物合成途径及调控机制研究进展[J].江西科学, 2021, 39(2):224-232. Chen L, Wang XL, Zhang XL, Gao Z. Advance in biosynthesis pathway and regulation mechanism of AsA in kiwifruit[J]. Jiangxi Science, 2021, 39(2):224-232.
[10] Li SH, Wang J, Yu YW, Wang FR, Dong JG, Huang RF. D27E mutation of[STXFX]VTC1[STXFZ] impairs the interaction with CSN5B and enhances ascorbic acid biosynthesis and seedling growth in Arabidopsis[J]. Plant Mol Biol, 2016, 92(4/5):473-482.
[11] Lim B, Smirnoff N, Cobbett CS, Golz JF. Ascorbate-deficient[STXFX]vtc2[STXFZ] mutants in Arabidopsis do not exhibit decreased growth[J]. Front Plant Sci, 2016, 7:1025.
[12] Laing W, Norling C, Brewster D, Wright M, Bulley S. Ascorbate concentration in Arabidopsis thaliana and expression of ascorbate related genes using RNAseq in response to light and the diurnal cycle[J]. BioRxiv, 2017.https://doi.org/10.1101/138008.[BFY]
[13] Li Y, Chu ZN, Luo JY, Zhou YH, Cai YJ, et al. The C2H2 zinc-finger protein SlZF3 regulates AsA synthesis and salt tolerance by interacting with CSN5B[J]. Plant Biotechnol J, 2018, 16(6):1201-1213.
[14] Imai T, Ban Y, Yamamoto T, Moriguchi T. Ectopic overexpression of peach GDP-D-mannose pyrophosphorylase and GDP-D-mannose-3', 5'-epimerase in transgenic tobacco[J]. Plant Cell Tiss Org, 2012, 111(1):1-13.
[15] Wang LY, Meng X, Yang DY, Ma NN, Wang GD, Meng QW. Overexpression of tomato GDP-L-galactose phosphorylase gene in tobacco improves tolerance to chilling stress[J]. Plant Cell Rep, 2014, 33(9):1441-1451.
[16] Hu TX, Ye J, Tao PW, Li HX, Zhang JH, et al. The tomato HD-ZipⅠ[STXFZ] transcription factor SlHZ24 modulates ascorbate accumulation through positive regulation of the D-mannose/L-galactose pathway[J]. Plant J, 2016, 85(1):16-29.
[17] Chen WF, Hu TX, Ye J, Wang B, Liu GZ, et al. A CCAAT-binding factor, SlNFYA10, negatively regulates ascorbate accumulation by modulating the D-mannose/L-galactose pathway in tomato[J]. Hortic Res, 2020, 7(1):200.
[18] 李超汉,李青竹,史庆华,白龙强,郭晓青,等.超表达番茄GDP-D-甘露糖焦磷酸化酶基因马铃薯对温度胁迫的响应[J].中国农业科学, 2011, 44(23):4952-4961. Li CH, Li QZ, Shi QH, Bai LQ, Guo XQ, et al. Effect of tomato GMPase overexpression on tolerance of potato plants to temperature stress[J]. Scientia Agricultura Sinica, 2011, 44(23):4952-4961.
[19] Bulley SM, Rarram M, Hoser D, Otto W, Nickel N, et al. Gene expression studies in kiwifruit and gene over-expression in Arabidopsis indicates that GDP-L-galactose guanyltransferase is a major control point of vitamin C biosynthesis[J]. J Exp Bot, 2009, 60(3):765-778.
[20] Bulley SM, Wright M, Rommens C, Yan H, Rassam M, et al. Enhancing ascorbate in fruits and tubers through over-expression of the L-galactose pathway gene GDP-L-galactose phosphorylase[J]. Plant Biotechnol J, 2012, 10(4):390-397.
[21] Mellidou I, Chagne D, Laing WA, Keulemans J, Davey MW. Allelic variation in paralogs of GDP-L-galactose phosphorylase is a major determinant of vitamin C concentrations in apple fruit[J]. Plant Physiol, 2012, 160(3):1613-1629.
[22] Li J, Li MJ. Liang D, Cui M, Mm FW. Expression patterns and promoter characteristics of the gene encoding Act-inidia deliciosa L-galactose-1-phosphate phosphatase involved in the response to light and abiotic stresses[J]. Mol Biol Rep, 2013, 40(2):1473-1485.
[23] 郭春苗,李明军,马锋旺,梁东,张敏.苹果L-半乳糖-1-磷酸磷酸酶cDNA克隆及其表达[J].中国农业科学, 2011, 44(4):762-770. Guo CM, Li MJ, Ma FW, Liang D, Zhang M. cDNA cloning and expression analysis of L-galactose-1-phosphate phosphatase in apple[J]. Scientia Agricultura Sinica, 2011, 44(4):762-770.
[24] 黄宏文,龚俊杰,王圣梅,何子灿,张忠慧,李建强.猕猴桃属(Actinidia)植物的遗传多样性[J].生物多样性, 2000(1):1-12. Huang HW, Gong JJ, Wang SM, He ZC, Zhang HZ, Li JQ. Genetic diversity in the genus Actinidia[J]. Biodiversity Science, 2000(1):1-12.
[25] 黄宏文,钟彩虹,姜正旺,李新伟,姚小洪,等.猕猴桃属分类、资源、驯化、栽培[M].北京:科学出版社, 2013:1-10.
[26] Liu XY, Xie XD, Zhong CH, Li DW. Comparative transcriptome analysis revealed the key genes regulating ascorbic acid synthesis in Actinidia[J]. Int J Mol Sci, 2021, 22:12894.
[27] 潘春华.维C之王:猕猴桃[J].绿化与生活, 2012(3):52.
[28] 李国秀,刘小宁,李劼.高效液相色谱法测定猕猴桃中的VC含量[J].保鲜与加工, 2016, 16(5):89-93. Li GX, Liu XN, Li J. Determination of VC content in kiwifruit by high performance liquid chromatography[J]. Storage and Process, 2016, 16(5):89-93.
[29] Liu XY, Wu RM, Bulley SM, Zhong CH, Li DW. Kiwifruit MYBS1-like and GBF3 transcription factors influence L-ascorbic acid biosynthesis by activating transcription of[STXFX]GDP-L-galactose phosphorylase 3[STXFZ] [J]. New Phytol, 2022, 243(5):1782-1800.
[30] Wang ZP, Wang SB, Li DW, Zhang Q, Li L, et al. Optimized paired-sgRNA/Cas9 cloning and expression cassette triggers high-efficiency multiplex genome editing in kiwifruit[J]. Plant Biotechnol J, 2018, 16(8):1424-1433.
[31] Jumper J, Evans R, Pritzel A, Green T, Figurnov M, et al. Highly accurate protein structure prediction with AlphaFold[J]. Nature, 2021, 596(7873):583-589.
[32] 张蕾,王彦昌,黄宏文.猕猴桃(Actinidia Lindl.)叶片与果实维生素C含量的相关性研究[J].武汉植物学研究, 2010, 28(6):750-755. Zhang L, Wang YC, Huang HW. Investigation of the correlations between the leaf and fruit vitamin C content in Actinidia Lindl.[J]. Plant Science Journal, 2010, 28(6):750-755.
[33] 钟彩虹,张鹏,姜正旺,王圣梅,韩飞,等.中华猕猴桃和毛花猕猴桃果实碳水化合物及维生素C的动态变化研究[J].植物科学学报, 2011, 29(3):370-376. Zhong CH, Zhang P, Jiang ZW, Wang SM, Han F, et al. Dynamic changes of carbohydrate and vitamin C in fruits of Actinidia chinensis and A. eriantha during growing season[J]. Plant Science Journal, 2011, 29(3):370-376.
[34] 魏丽红.软枣猕猴桃果实发育与营养成分变化规律研究[J].湖北农业科学, 2017, 56(11):2070-2073. Wei LH. Studies on fruit development and nutritional ingredients change regularity of Actinidia arguta[J]. Hubei Agricultural Sciences, 2017, 56(11):2070-2073.
[35] Li MJ, Ma FW, Liang D, Li J, Wang YL. Ascorbate biosynthesis during early fruit development is the main reason for its accumulation in kiwi[J]. PLoS One, 2010, 5(12):e14281.
[36] Li J, Li MJ, Liang D, Ma FW, Lei YS. Comparison of expression pattern, genomic structure, and promoter analysis of the gene encoding GDP-l-galactose phosphorylase from two Actinidia species[J]. Sci Hortic, 2014, 169:206-213.