甘薯液泡转化酶基因(IbVINs)克隆及结构和功能分析

doi:10.3969/j.issn.1674-7968.2026.03.015

Abstract
Figure/Table
References
Related Citation (13)

Download: PDF (25260 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Sweet potato (Ipomoea batatas) is an important food, industrial, and feed crop. The contents and composition of starch and soluble sugars are key determinants of sweet potato quality. Vacuolar invertase (VIN) genes are closely associated with starch-sugar metabolism; however, their functions and regulatory mechanisms in the starch-sugar metabolism of sweet potato storage roots remain to be systematically elucidated. This study conducted a systematic investigation of the vacuolar invertase gene family in sweet potato to elucidate its role in starch-sugar metabolism and carbon partitioning of storage roots. Three IbVIN members were cloned from 7 sweet potato genotypes, their sequences were analyzed bioinformatically, phylogenetic tree was built, and protein secondary/tertiary structures and conserved domains were predicted. Expression patterns were profiled across cultivars, developmental stages, and tissues using qPCR, starch and soluble sugars were quantified by high-performance liquid chromatography (HPLC), and correlation analyses were performed. This study also cloned the 1.5~2.5 kb upstream promoter regions of IbVINs, predicted cis-acting elements, and carried out subcellular localization assays. In Arabidopsis thaliana, Agrobacterium-mediated transgenic lines overexpressing IbVIN1 were generated, and starch and soluble sugar contents were measured in leaves and seeds. The results showed that IbVIN1, IbVIN2, and IbVIN3 all contained the conserved GH32 domain and the catalytic motifs NDPNG, RDP, and WEC, with structural differences among the proteins. The 3 genes were highly expressed in storage roots but weakly expressed in stems, peaking during early-mid root swelling (65~80 d after transplanting), indicating tissue- and stage-specific patterns. At 95 d after transplanting, IbVIN1 and IbVIN2 correlated negatively with starch content and positively with hexoses content, while IbVIN3 correlated positively with hexoses content. At 125 d, all 3 genes showed significant negative correlations with hexoses content, suggesting dynamic regulation of carbon allocation. All 3 promoter types harbored light-, hormone-, and stress-responsive elements, with typeⅡuniquely containing the low-temperature responsive element (LTR) and typeⅢ containing an auxin-responsive element (AuxRE). Subcellular localization confirmed that all 3 proteins resided in the vacuole. Overexpression of IbVIN1 in A. thaliana significantly decreased leaf starch and increased leaf soluble sugars, while promoting the accumulation of both starch and soluble sugars in seeds. Collectively, these findings reveal that the IbVIN family exhibits spatiotemporal expression and stage-dependent roles in carbon partitioning during storage-root development, clarifies its regulatory function in starch-sugar metabolism, and provides a theoretical basis and candidate targets for sweet potato quality improvement and molecular breeding.

Key words： Sweet potato Vacuolar invertase (VIN) Starch-sugar metabolism

Received: 05 September 2025

ZTFLH:	S531
	Q943.2

Corresponding Authors: ^*1065679162@qq.com

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	HAN Hao-Hao
	DUAN Li-Jun
	WEI Yan-Mei
	HAN Dong-Chen
	ZHAO Ren-Na
	DING Yan
	YAN Feng-Ying

Cite this article:

HAN Hao-Hao,DUAN Li-Jun,WEI Yan-Mei, et al. Cloning, Structural and Functional Analysis of Vacuolar Invertase Genes (IbVINs) in Sweet Potato (Ipomoea batatas)[J]. 农业生物技术学报, 2026, 34(3): 627-645.

URL:

https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2026.03.015 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2026/V34/I3/627

[1] 蔡柳, 曾璐, 张婷婷, 等. 2011. 薯类原料生物转化燃料乙醇研究进展[J]. 中国酿造, 30(3): 1-5.
(Cai L, Zeng L, Zhang T T, et al.2011. Research progress on bio-conversion of tuber raw materials to fuel ethanol[J]. China Brewing, 30(3): 1-5.)
[2] 李强, 赵海, 靳艳玲, 等. 2022. 中国甘薯产业助力国家粮食安全的分析与展望[J]. 江苏农业学报, 38(6): 1484-1491.
(Li Q, Zhao H, Jin Y L, et al.2022. Analysis and prospect of the Chinese sweet-potato industry in safeguarding national food security[J]. Jiangsu Agricultural Sciences, 38(6): 1484-1491.)
[3] 刘庆昌. 2004. 甘薯在我国粮食和能源安全中的重要作用[J]. 科技导报, 22(9): 21-22.
(Liu Q C.2004. The important role of sweet potato in China's food and energy security[J]. Science & Technology Review, 22(9): 21-22.)
[4] 刘钊君. 2019. 木薯液泡转化酶基因MeVINV1的抗逆功能鉴定[D]. 硕士学位论文, 海南大学, 导师: 胡新文, pp. 73.
(Liu Z J. 2019. Functional analysis of stress resistance of cassava vacuolar invertase MeVINV1[D]. Thesis for M.S., Hainan University, Supervisor: Hu X W, pp. 73.)
[5] 陆璐, 陶雅军, 罗学娅, 等. 2019. 糖苷水解酶32家族结构与功能的研究进展[J]. 中国酿造, 38: 14-19.
(Lu L, Tao Y J, Luo X Y, et al.2019. Research progress on the structure and function of glycoside hydrolase family 32[J]. China Brewing, 38: 14-19.)
[6] 马代夫, 李强, 曹清河, 等. 2012. 中国甘薯产业及产业技术的发展与展望[J]. 江苏农业学报, 28(5): 969-973.
(Ma D F, Li Q, Cao Q H, et al.2012. Development and prospects of China's sweet-potato industry and related technologies[J]. Jiangsu Journal of Agricultural Sciences, 28(5): 969-973.)
[7] 石晓雯, 贺立恒, 焦晋华, 等. 2018. 甘薯二倍体近缘野生种三裂叶薯MYB转录因子全基因组分析及逆境胁迫响应[J]. 核农学报, 32(7): 1338-1348.
(Shi X W, He L H, Jiao J H, et al.2018. Genome-wide analysis of MYB transcription factors in the diploid wild relative Ipomoea trifida of sweet potato and their response to abiotic stress[J]. Journal of Nuclear Agricultural Sciences, 32(7): 1338-1348.)
[8] 王文质, 以凡, 杜述荣, 等. 1989. 甘薯淀粉含量换算公式及换算表[J]. 作物学报, 1: 94-96.
(Wang W Z, Yi F, Du S R, et al.1989. Conversion formula and table for starch content in sweet potato[J]. Acta Agronomica Sinica, 1: 94-96.)
[9] 王欣, 李强, 曹清河, 等. 2021. 中国甘薯产业和种业发展现状与未来展望[J]. 中国农业科学, 54(3): 483-492.
(Wang X, Li Q, Cao Q H, et al.2021. Current status and future prospects of China's sweet-potato industry and seed sector[J]. Scientia Agricultura Sinica, 54(3): 483-492.)
[10] 闫庆祥, 黄东益, 李开绵, 等. 2010. 利用改良CTAB法提取木薯基因组DNA[J]. 中国农学通报, 26(4): 30-32.
(Yan Q X, Huang D Y, Li K M, et al.2010. Genomic DNA extraction in cassava by modified CTAB method[J]. Chinese Agricultural Science Bulletin, 26(4): 30-32.)
[11] 张绍铃, 贾璐婷, 王利斌, 等. 2019. 园艺作物果实液泡糖、酸转运与转化研究进展[J]. 南京农业大学学报, 42(4): 583-593.
(Zhang S L, Jia L T, Wang L B, et al.2019. Recent advance on vacuolar sugar and acid transportation and conversion in horticultural fruit[J]. Journal of Nanjing Agricultural University, 42(4): 583-593.)
[12] Alberto F, Bignon C, Sulzenbacher G, et al.2004. The three-dimensional structure of invertase (β-fructosidase) from Thermotoga maritima reveals a bimodular arrangement and an evolutionary relationship between retaining and inverting glycosidases[J]. Journal of Biological Chemistry, 279(18): 18903-18910.
[13] Barratt D H P, Derbyshire P, Findlay K, et al.2009. Normal growth of Arabidopsis requires cytosolic invertase but not sucrose synthase[J]. Proceedings of the National Academy of Sciences of the USA, 106(31): 13124-13129.
[14] Bhaskar P B, Wu L, Busse J S, et al.2010. Suppression of the vacuolar invertase gene prevents cold-induced sweetening in potato[J]. Plant Physiology, 154(2): 939-948.
[15] Birchler J A, Yang H.2022. The multiple fates of gene duplications: Deletion, hypofunctionalization, subfunctionalization, neofunctionalization, dosage-balance constraints, and neutral variation[J]. The Plant Cell, 34(7): 2466-2474.
[16] Chen L, Liu X H, Huang X J, et al.2019. Functional characterization of a drought-responsive invertase inhibitor from maize (Zea mays L.)[J]. International Journal of Molecular Sciences, 20(17): 4081.
[17] Clough S J, Bent A F.1998. Floral dip: Aa simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J]. The Plant Journal, 16(6): 735-743.
[18] Davies C, Robinson S P.1996. Sugar accumulation in grape berries: Cloning of two putative vacuolar invertase cDNAs and their expression in grapevine tissues[J]. Plant Physiology, 111(1): 275-283.
[19] Huang Y H, Picha D H, Kilili A W, et al.1999. Changes in invertase activities and reducing sugar content in sweetpotato stored at different temperatures[J]. Journal of Agricultural and Food Chemistry, 47(12): 4927-4931.
[20] Katayama K, Kobayashi A, Sakai T, et al.2017. Recent progress in sweetpotato breeding and cultivars for diverse applications in Japan[J]. Breeding Science, 67(1): 3-14.
[21] Klann E M, Hall B, Bennett A B.1996. Antisense acid invertase (TIV1) gene alters soluble sugar composition and size in transgenic tomato fruit[J]. Plant Physiology, 112(4): 1321-1330.
[22] Kunz H H, Häusler R E, Fettke J, et al.2010. The role of plastidial glucose-6-phosphate/phosphate translocators in vegetative tissues of Arabidopsis thaliana mutants impaired in starch biosynthesis[J]. Plant Biology, 12(1): 115-128.
[23] La Bonte D R, Picha D H, Johnson H A.2000. Carbohydrate-related changes in sweetpotato storage roots during development[J]. Journal of the American Society for Horticultural Science, 125(2): 200-204.
[24] Lammens W, Le Roy K, Schroeven L, et al.2009. Structural insights into glycoside hydrolase family 32 and 68 enzymes: Functional implications[J]. Journal of Experimental Botany, 60(3): 727-740.
[25] Lee B R, Cho J H, Wi S G, et al.2021. The sucrose-to-hexose ratio is a significant determinant for fruit maturity and is modulated by invertase and sucrose re-synthesis during fruit development and ripening in Asian pear (Pyrus pyrifolia) cultivars[J]. Horticultural Science and Technology, 39(2): 141-151.
[26] Lee D W, Lee S K, Rahman M M, et al.2019. The role of rice vacuolar invertase2 in seed size control[J]. Molecules and Cells, 42(10): 711-720.
[27] Li Z H, Lu X, Gao Y, et al.2011. Polyploidization and epigenetics[J]. Chinese Science Bulletin, 56(3): 245-252.
[28] Liu Q.2017. Improvement for agronomically important traits by gene engineering in sweetpotato[J]. Breeding Science, 67(1): 15-26.
[29] Lombardo V A, Osorio S, Borsani J, et al.2011. Metabolic profiling during peach fruit development and ripening reveals the metabolic networks that underpin each developmental stage[J]. Plant Physiology, 157(4): 1696-1710.
[30] Park S C, Kim Y H, Ji C Y, et al.2012. Stable internal reference genes for the normalization of real-time PCR in different sweet-potato cultivars subjected to abiotic stress conditions[J]. PLOS ONE, 7(12): e51502.
[31] Peixoto B, Baena-González E.2022. Management of plant central metabolism by SnRK1 protein kinases[J]. Journal of Experimental Botany, 73(20): 7068-7082.
[32] Picha D H.1985. HPLC determination of sugars in raw and baked sweet potatoes[J]. Journal of Food Science, 50(4): 1189-1190.
[33] Ruan Y L.2014. Sucrose metabolism: Gateway to diverse carbon use and sugar signaling[J]. Annual Review of Plant Biology, 65: 33-67.
[34] Schmittgen T D, Livak K J.2008. Analyzing real-time PCR data by the comparative C(T) method[J]. Nature Protocols, 3(6): 1101-1108.
[35] Sturm A.1996. Molecular characterization and functional analysis of sucrose-cleaving enzymes in carrot (Daucus carota L.)[J]. Journal of Experimental Botany, 47(Special Issue): 1187-1192.
[36] Sturm A.1999. Invertases: Primary structures, functions, and roles in plant development and sucrose partitioning[J]. Plant Physiology, 121(1): 1-8.
[37] Takahata Y, Noda T, Sato T.1996. Relationship between acid invertase activity and hexose content in sweet potato storage roots[J]. Journal of Agricultural and Food Chemistry, 44(8): 2063-2066.
[38] Tauzin AS, Sulzenbacher G, Lafond M, et al.2014. Functional characterization of a vacuolar invertase from Solanum lycopersicum: Post-translational regulation by N-glycosylation and a proteinaceous inhibitor[J]. Biochimie, 101: 39-49.
[39] Tiessen A, Prescha K, Branscheid A, et al.2003. Evidence that SNF1-related kinase and hexokinase are involved in separate sugar-signalling pathways modulating post-translational redox activation of ADP-glucose pyrophosphorylase in potato tubers[J]. The Plant Journal, 35(4): 490-500.
[40] Tymowska-Lalanne Z, Kreis M.1998. The plant invertases: Physiology, biochemistry and molecular biology[J]. Advances in Botanical Research, 28: 71-117.
[41] Van den Ende W, Lammens W, Van Laere A, et al.2009. Donor and acceptor substrate selectivity among plant glycoside hydrolase family 32 enzymes[J]. The FEBS Journal, 276(20): 5788-5798.
[42] Wan H J, Wu L M, Yang Y J, et al.2018. Evolution of sucrose metabolism: The dichotomy of invertases and beyond[J]. Trends in Plant Science, 23(2): 163-177.
[43] Wang L T, Wang A Y, Hsieh C W, et al.2005. Vacuolar invertases in sweet potato: Molecular cloning, characterization and analysis of gene expression[J]. Journal of Agricultural and Food Chemistry, 53(10): 3672-3678.
[44] Yang J, Moeinzadeh M H, Kuhl H, et al.2017. Haplotype-resolved sweet-potato genome traces back its hexaploidization history[J]. Nature Plants, 3: 696-703.
[45] Yang Z, Zhu P, Kang H, et al.2022. Comparative analysis of salt responsive microRNAs in two sweetpotato (Ipomoea batatas (L.) Lam.) cultivars with different salt stress resistance[J]. Frontiers in Plant Science, 13: 879819.
[46] Yelle S, Hewitt J D, Robinson N L, et al.1991. Sink metabolism in tomato fruit: III. Analysis of carbohydrate assimilation in a wild species[J]. Plant Physiology, 95(4): 1026-1035.