Effect of Different Rootstocks on Fruit Quality and Expression of Genes Related to Gluconeogenesis in 'Shine Muscat' Grapes (Vitis vinfera)
SHEN Le-Yi1, WANG Li-Ru2, XU Yue1,3, CHEN Tian-Chi1, XU Tao1, GUO Yan-Fei1, FANG Cong-Ling2, FAN Lin-Jie2, WU Yue-Yan1,*
1 College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315000, China; 2 Cixi City Forestry Technology Extension Center, Ningbo 315000, China; 3 Institute of Virus and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
Abstract：Grapes (Vitis vinfera) are one of the four major fruits in the world, and their excellent cultivation depends to a large extent on grafting, which not only improves the resistance of the fruit trees, but also influences the fruit quality. The objective of this study was to investigate the effects of different rootstocks on the fruit quality and the expression of gluconeogenesis related genes in 'Shine Muscat' grapes, and to screen out rootstock combinations with excellent overall quality in Jiangsu and Zhejiang regions. In this study, samples of 'Shine Muscat' grapes grafted on 12 rootstocks were used to determine various quality indicators and to analyze the expression of phosphoenolpyruvate carboxykinase (PEPCK), pyruvate phosphate dikinase (PPDK), fructose-1,6-bisphosphatase (FBP) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes by real-time fluorescence quantitative PCR. The results showed that grafting of 12 rootstocks affected the quality index of 'Shine Muscat' grapes to varying degrees, especially the content of amino acids and sugar-acid fractions. Among the 12 rootstock combinations, '8B' rootstock grafted on 'Shine Muscat' gave the largest fruit size and hardness, '1103P' and '5BB' rootstocks grafted on 'Shine Muscat' gave the largest fruit solid-acid ratio, and 'Beda' rootstock grafted on 'Shine Muscat' gave the highest content of various amino acids in the fruit; '101-14' and '5BB' rootstocks had the highest glucose and fructose contents in the fruit after grafting 'Shine Muscat', while '3309C' rootstock grafted 'Shine Muscat' had the lower glucose and fructose contents in the fruit among the 12 rootstock combinations, but the content of acids such as tartaric acid, malic acid and citric acid was higher. Rootstock grafting had an effect on the expression of sugar anabolism-related genes, but it had the greatest effect on the expression of PEPCK gene, and the expression levels of PEPCK, PPDK, FBP and GAPDH were the highest in fruits of 'Shine Muscat' grafted on '101-14' rootstock. The expression of PEPCK genes in different rootstock combinations was significantly and positively correlated with glucose and fructose content, and it was hypothesized that different rootstocks affect the regulation of PEPCK gene in grape flesh, thus affecting conversion of organic acids to sugars and eventually the sugar accumulation. The top 3 rootstock combinations ranked in order by principal component analysis were 8B/SM, Beda/SM, and K3/SM. This study provides a theoretical basis for quality rootstock selection.
[1] 崔鹏飞, 魏灵珠, 程建徽, 等. 2021. 不同砧木对'天工翠玉'葡萄生长和果实品质的影响[J]. 浙江农业学报, 33(1): 52-61. (Cui P F, Wei L Z, Cheng J H, et al.2021. Effects of different rootstocks on the growth and fruit quality of 'Tiangong Cuiyu' grapes[J]. Zhejiang Journal of Agriculture, 33(1): 52-61.) [2] 付涛, 吴月燕, 王立如, 等. 2014. '鄞红'葡萄及其突变体果实氨基酸和香气分析[J]. 核农学报, 28(11): 2038-2050. (Fu T, Wu Y Y, Wang L R, et al.2014. Amino acid and aroma analysis of 'Yinhong' grape and its mutant fruits[J]. Journal of Nuclear Agriculture, 28(11): 2038-2050.) [3] 李明山, 魏灵珠, 沈碧薇, 等. 2021. 砧木对'天工玉柱'葡萄生长及果实品质的影响[J]. 中外葡萄与葡萄酒, 238(4): 13-19. (Li M S, Wei L Z, Shen B W, et al.2021. Effect of rootstocks on the growth and fruit quality of 'Tiangong Yuzhu' grapes[J]. Chinese and Foreign Vines and Wines, 238(4): 13-19.) [4] 李善菊, 曲晨艳, 师守国. 2022. 不同砧木对'阳光玫瑰'白兰地品质的影响[J]. 中国酿造, 41(8): 89-96. (Li S J, Qu C Y, Shi S G.2022. Effect of different rootstocks on the quality of Sunshine Rose Brandy[J]. China Brewing, 41(8): 89-96.) [5] 林茜, 韩晓华, 高营营, 等. 2020. 7种砧木对南宁'阳光玫瑰'葡萄生长及果实品质的影响[J]. 中国南方果树, 49(1): 100-104. (Lin X, Han X H, Gao Y Y, et al.2020. Effects of seven rootstocks on the growth and fruit quality of Nanning 'Shine Muscat' grapes[J]. Fruit Trees of Southern China, 49(1): 100-104.) [6] 刘翔宇, 李娟, 黄敏, 等. 2015. 柑橘砧木对'砂糖橘'果实糖积累的影响[J]. 中国农业科学, 48(11): 2217-2228. (Liu X Y, Li J, Huang M, et al.2015. Effect of citrus rootstocks on sugar accumulation in 'sugar orange fruits'[J]. Chinese Agricultural Science, 48(11): 2217-2228.) [7] 潘治利, 罗元奇, 艾志录, 等. 2016. 不同小麦品种醇溶蛋白的组成与速冻水饺面皮质构特性的关系[J]. 农业工程学报, 32(4): 242-248. (Pan Z L, Luo Y Q, Ai Z L, et al.2016. Relationship between the composition of alcohol-soluble proteins of different wheat varieties and the cortical properties of frozen dumplings[J]. Journal of Agricultural Engineering, 32(4): 242-248.) [8] 沈碧薇, 魏灵珠, 崔鹏飞, 等. 2022. 不同砧木对'瑞都红玉'葡萄生长结果与果实品质的影响[J]. 果树学报, 37(3): 350-361. (Shen B W, Wei L Z, Cui P F, et al.2022. Effects of different rootstocks on the growth and fruit quality of 'Ruidu Red Jade' grapes[J]. Journal of Fruit Trees, 37(3): 350-361.) [9] 沈乐意, 王锦阳, 陈天池, 等. 2023. 施用不同营养液对葡萄果实糖异生途径相关基因表达的影响[J]. 核农学报, 37(7): 1299-1306. (Shen L Y, Wang J Y, Chen T C, et al.2023. Effects of various nutrient solutions on the expression of gluconeogenesis-related genes in grape fruit[J]. Journal of Nuclear Agricultural Sciences, 37(7): 1299~1306.) [10] 王莎, 程大伟, 顾红, 等. 2019. 植物生长调节剂对'阳光玫瑰'葡萄果实无核及品质的影响[J]. 果树学报, 36(12): 1675-1682. (Wang S, Cheng D W, Gu H, et al.2019. Effect of plant growth regulators on nucleation and quality of 'Shine Muscat' grape fruit[J]. Journal of Fruit Trees, 36(12): 1675-1682.) [11] 王松, 罗恵格, 陈彦蓓, 等. 2020. 不同砧木对'阳光玫瑰'葡萄生长及果实品质的影响初报[J]. 中国南方果树, 49(5): 103-106. (Wang S, Luo H G, Chen Y B, et al.2020. Preliminary report on the effect of different rootstocks on the growth and fruit quality of 'Shine Muscat' grapes[J]. Southern China Fruit Tree, 49(5): 103-106.) [12] 王晓林, 吉姣姣, 高建平. 2018. 党参CpGAPDH基因的克隆及表达分析[J]. 中国中药杂志, 43(4): 712-720. (Wang X L, Ji J J, Gao J P.2018. Cloning and expression analysis of CpGAPDH gene from Radix Codonopsis pilosulae[J]. Chinese Journal of Traditional Chinese Medicine, 43(4): 712-720.) [13] 魏灵珠, 沈碧薇, 程建徽, 等. 2020. 砧木对'新雅'葡萄生长及果实品质的影响[J]. 果树学报, 37(9): 1346-1357. (Wei L Z, Shen B W, Cheng J H, et al.2020. Effect of rootstocks on the growth and fruit quality of 'Xinyia' grapes[J]. Journal of Fruit Trees, 37(9): 1346-1357.) [14] 邢朝斌, 吴鹏, 陈龙, 等. 2012. 刺五加GAPDH基因的克隆及序列分析[J]. 中草药, 43(1): 155-158. (Xing Z B, Wu P, Chen L, et al.2012. Cloning and sequence analysis of the GAPDH gene of Acanthopanax ginseng[J]. Chinese herbal medicine, 43(1): 155-158.) [15] 颜孙安, 姚清华, 林香信, 等. 2021. 成熟度对'红地球'葡萄氨基酸营养价值的影响[J]. 果树学报, 38(1): 64-72. (Yan S A, Yao Q H, Lin X X, et al.2021. Effect of maturity on the nutritional value of amino acids in 'red globe' grapes[J]. Journal of Fruit Tree Science, 38(1): 64-72.) [16] 岳彩凤, 康国章, 刘超, 等. 2008. 小麦GAPDH基因克隆及序列分析[J]. 中国农学通报, 24(4): 94-98. (Yue C F, Kang G Z, Liu C, et al.2008. Cloning and sequence analysis of wheat GAPDH gene[J]. Chinese Agronomy Bulletin, 24(4): 94-98.) [17] 张霞, 苏豫梅, 邓莉, 等. 2016. 拟南芥胞质甘油醛-3-磷酸脱氢酶基因的表达模式分析[J]. 生物技术, 26(5): 469-472. (Zhang X, Su Y M, Deng L, et al.2016. Expression pattern analysis of cytoplasmic glyceraldehyde-3-phosphate dehydrogenase gene in Arabidopsis thaliana[J]. Biotechnology, 26(5): 469-472.) [18] Attia F, Garcia F G M, Besnard E, et al.2007. Effect of rootstock on organic acids in leaves and berries and on must and wine acidity of two red wine grape cultivars malbec and negrette grown hydroponically[J]. Acta Horticulturae, 754: 473-482. [19] Chou M Y, Li K T.2014. Rootstock and seasonal variations affect anthocyanin accumulation and quality traits of 'Kyoho' grape berries in subtropical double cropping system[J]. Vitis, 53(4): 193-199. [20] Coelho E M, Da Silva Padilha C V, Miskinis G A, et al.2018. Simultaneous analysis of sugars and organic acids in wine and grape juices by HPLC: Method validation and characterization of products from northeast Brazil[J]. Journal of Food Composition and Analysis, 66: 160-167. [21] Eastmond P J, Astley H M, Parsley K, et al.2015. Arabidopsis uses two gluconeogenic gateways for organic acids to fuel seedling establishment[J]. Nature Communications, 6: 6659. [22] Famiani F, Moscatello S, Ferradini N, et al.2014. Occurrence of a number of enzymes involved in either gluconeogenesis or other processes in the pericarp of three cultivars of grape (Vitis vinifera L.) during development[J]. Plant Physiology and Biochemistry, 84: 261-270. [23] Famiani F, Paoletti A, Battistelli A, et al.2016. Phosphoenolpyruvate carboxykinase, pyruvate orthophosphate dikinase and isocitrate lyase in both tomato fruits and leaves, and in the flesh of peach and some other fruits[J]. Journal of Plant Physiology, 202: 34-44. [24] Famiani F, Paoletti A, Proietti P, et al.2018. The occurrence of phosphoenolpyruvate carboxykinase (PEPCK) in the pericarp of different grapevine genotypes and in grape leaves and developing seeds[J]. The Journal of Horticultural Science and Biotechnology, 93(5): 456-465. [25] Guo L X, Shi C Y, Liu X, et al.2016. Citrate accumulation-related gene expression and/or enzyme activity analysis combined with metabolomics provide a novel insight for an orange mutant[J]. Scientific Reports, 6: 29343. [26] Huang Y X, Yin Y G, Sanuki A, et al.2015. Phosphoenolpyruvate carboxykinase (PEPCK) deficiency affects the germination, growth and fruit sugar content in tomato (Solanum lycopersicum L.)[J]. Plant Physiology and Biochemistry, 96: 417-425. [27] Jin Z X, Sun T Y, Sun H, et al.2016. Modifications of 'Summer Black' grape berry quality as affected by the different rootstocks[J]. Scientia Horticulturae, 210: 130-137. [28] Jin Z, Sun H, Sun T, et al.2016. Modifications of 'gold finger' grape berry quality as affected by the different rootstocks[J]. Journal of Agricultural and Food Chemistry, 64(21): 4189-4197. [29] Jogaiah S, Oulkar D P, Banerjee K, et al.2012. Biochemically induced variations during some phenological stages in 'Thompson Seedless' grapevines grafted on different rootstocks[J]. South African Journal of Enology and Viticulture, 34(1): 36-45. [30] Le P H, Verscheure L, Le T T, et al.2020. Implementation of HPLC analysis for γ-aminobutyric acid (GABA) in fermented food matrices[J]. Food Analytical Methods, 13(5): 1190-1201. [31] Lee J, Steenwerth K L.2011. Rootstock and vineyard floor management influence on 'Cabernet Sauvignon' grape yeast assimilable nitrogen (YAN)[J]. Food Chemistry, 127(3): 926-933. [32] Lee S K, Jeon J S, Bornke F, et al.2008. Loss of cytosolic fructose-1,6-bisphosphatase limits photosynthetic sucrose synthesis and causes severe growth retardations in rice (Oryza sativa)[J]. Plant Cell and Environment, 31(12): 1851-1863. [33] Li M, Guo Z, Jia N, et al.2019. Evaluation of eight rootstocks on the growth and berry quality of 'Marselan' grapevines[J]. Scientia Horticulturae, 248: 58-61. [34] Li M, Li P, Ma F, et al.2018. Sugar metabolism and accumulation in the fruit of transgenic apple trees with decreased sorbitol synthesis[J]. Horticulture Research, 5: 60. [35] Lo'ay A A, El-khateeb A Y.2017. Evaluation the effect of rootstocks on postharvest berries quality of 'Flame Seedless' grapes[J]. Scientia Horticulturae, 220: 299-302. [36] Nie Z, Wan C, Chen C, et al.2019. Comprehensive evaluation of the postharvest antioxidant capacity of 'Majiayou' pomelo harvested at different maturities based on PCA[J]. Antioxidants, 8(5): 136. [37] Rojas-Gonzalez J A, Soto-Suarez M, García-Diaz A, et al.2015. Disruption of both chloroplastic and cytosolic FBPase genes results in a dwarf phenotype and important starch and metabolite changes in Arabidopsis thaliana[J]. Journal of Experimental Botany, 66(9): 2673-2689. [38] Schmittgen T D, Livak K J.2008. Analyzing real-time PCR data by the comparative CT method[J]. Nature Protocols, 3(6): 1101-1108. [39] Stefania S, Malagoli M, Stefano D A.2022. Foliar application of silicon in Vitis vinifera: Targeted metabolomics analysis as a tool to investigate the chemical variations in berries of four grapevine cultivars[J]. Plants, 11(21): 2998. [40] Thorbjurnsen T, Asp T, Jurgensen K, et al.2002. Starch biosynthesis from triose-phosphate in transgenic potato tubers expressing plastidic fructose-1,6-bisphosphatase[J]. Planta, 214(4): 616-624. [41] Verma S K, Singh S K, Krishna H.2010. The effect of certain rootstocks on the grape cultivar 'Pusa Urvashi' (Vitis vinifera L.)[J]. International Journal of Fruit Science, 10(1): 16-28. [42] Walker R P, Battistelli A, Moscatello S, et al.2011. Phosphoenolpyruvate carboxykinase in cherry (Prunus avium L.) fruit during development[J]. Journal of Experimental Botany, 62(15): 5357-5365. [43] Walker R P, Battistelli A, Moscatello S, et al.2015. Phosphoenolpyruvate carboxykinase and gluconeogenesis in grape pericarp[J]. Plant Physiology and Biochemistry, 97: 62-69. [44] Walker R P, Chen Z H, Famiani F.2021. Gluconeogenesis in plants: A key interface between organic acid/amino acid/lipid and sugar metabolism[J]. Molecules, 26(17): 5129. [45] Wei Q J, Ma Q L, Zhou G F, et al.2021. Identification of genes associated with soluble sugar and organic acid accumulation in 'Huapi' kumquat (Fortunella crassifolia Swingle) via transcriptome analysis[J]. Journal of the Science of Food and Agriculture, 101(10): 4321-4331.
