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| Creating Potato (Solanum tuberosum) Mutants with Enhanced Resistance to Cold-induced Sweetening by Editing the Vacuolar Invertase Gene (StVInv) |
| ZHU Xu1,*, LI Nan1,*, MU Shu-Jing1, ZHANG Ling1, GUAN Ke-Xing1, LI Chuang2, LI Yi-Dan3, WANG Zhong-Wei2,**, HE Hong-Xia1,** |
1 Jilin Provincial Key Laboratory of Agricultural Biotechnology/Institute of Agricultural Biotechnology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, China; 2 Economic plants Research Instiute, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, China; 3 Staple Food Engineering Research Center, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, China |
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Abstract Low-temperature saccharification refers to the process in which starch in the tubers of potato (Solanum tuberosum) is rapidly converted into reducing sugars when the potato is stored at low temperatures. It is a physiological response of potato to low-temperature stress, but severely impairs the quality of fried potato products. In order to reduce the accumulation of reducing sugars in potato tubers caused by low-temperature storage and improve the quality offried-potato products, this study used CRISPR/Cas9 technology to knock down the expression of the vacuolar invertase gene (StVInv), and created StVInv gene mutants. The results of qPCR and physiological and biochemical indicators testing indicated that after 21 d of storage at 4 ℃, the expression level of the StVInv gene in the mutant tubers was significantly lower than that in the wild type (P<0.0001). The vacuolar invertase activity was reduced by 33.68% to 60.4% compared with the wild type, and the reducing sugar content was decreased by 55.30% to 56.75% compared with the wild type. The color of the fried-potato chips was significantly improved compared with the wild type. This study provides technical support for the use of biotechnology to improve the deep-fried processing traits of potatoes.
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Received: 26 June 2025
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Corresponding Authors:
**13756127666@163.com; hongxia_365@163.com
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[1] 金成. 2015. 油炸马铃薯制品高丙烯酰胺暴露的成因及危害物综合防控[D]. 博士学位论文, 浙江大学, 导师:张英, pp. 4-11. (Jin C.2015. Mechanism of high level of acrylamide and simultaneous control of Maillard reaction-derived contaminants in fried potato foods[D]. Thesis for Ph. D., Zhejiang University, Supervisor:Zhang Y, pp. 4-11.) [2] 李传龙, 李楠, 朱旭, 等. 2021. 马铃薯地方主栽品种“春薯4号”遗传转化体系的建立[J]. 吉林农业大学学报, 43(03):289-96. (Li C L, Li N, Zhu X, et al.2021. Establishment of genetic transformation system of a main local potato cultivar Chunshu 4[J]. Journal of Jilin Agricultural University, 43(03):289-96.) [3] 李远欣. 2008. 马铃薯新品种“春薯4号”丰产栽培技术[J].福建农业,(11):8. (Li Y Y. 2008. High-yield cultivation techniques for the new potato variety “Spring Potato No. 4”[J]. FuJian Agriculture, (11):8.) [4] 刘娟, 梁延超, 余斌, 等. 2018. 马铃薯薯条色泽和质地特性及薯条加工型品系筛选[J]. 中国农业科学, 50(22):4247-4265. (Liu J. Liang Y C, Yu B, et al.2018. Screening for french fries processing potato lines according to colour qualities and texture properties[J]. Scientia Agricultura Sinica, 50(22):4247-4265.) [5] 刘勋. 2010. 马铃薯转化酶及其抑制子基因家族分析及与低温糖化关系研究[D]. 博士学位论文, 华中农业大学, 导师:柳俊, pp. 36-39. (Liu X.2010. Analysis of potato acid invertase and invertase inhibitor gene families and their relationship with cold-induecd sweetening of tubers[D]. Thesis for Ph.D., Huazhong Agricultural University, Supervisor:Liu J, pp. 36-39.) [6] 罗文彬, 许国春, 李华伟, 等. 2023.马铃薯营养品质变异特征及优异品系发掘[J]. 植物遗传资源学报, 24(02):505-513. (Luo W B, Xu G C, Li H w, et al.2023. Features and variations of potato nutritional components and identification of elite high-quality lines[J]. Journal of Plant Genetic Resources, 24(02):505-513.) [7] 欧庸彬, 姚春光, 柳寒, 等. 2008. 马铃薯高代系炸片色泽分析与加工品质评价[J]. 中国马铃薯, (05):274-277. (Ou Y B, Yao C G, Liu H, et al. Evaluation for processing quality of potato progenies[J]. Chinese Potato Journal, (05):274-277.) [8] 欧庸彬. 2013. 马铃薯液泡酸性转化酶基因的表达调控机制研究[D]. 博士学位论文, 华中农业大学, 导师:柳俊, pp. 6-18. (Ou Y B.2013. Expression regulatory mechanisms of the potato vacuolar acid invertase gene[D]. Thesis for Ph.D., Huazhong Agricultural University, Supervisor:Liu J, pp. 6-18.) [9] 孙世磊. 2022. 玉米光合相关转录因子的鉴定、突变体创制及功能分析[D]. 博士学位论文, 山东农业大学, 导师:李平华, pp.52-56. (Sun S L.2022. Identification, mutant creation, and functional analysis of photosynthesis-related transcription factors in maize[D]. Thesis for Ph.D., Shandong Agricultural University, Supervisor:Li P H, pp. 52-56.) [10] 王丹. 2016. 专用型马铃薯优良新品系选育研究[D]. 博士学位论文, 内蒙古农业大学, 导师:于卓, pp. 11. (Wang D. 2016. Research on breeding excellent new strains of special potato[D]. Thesis for Ph.D, Inner Mongolia Agricultural University, Supervisor:Yu Z, pp. 11.) [11] 韦孟, 许慧珍, 张宁, 等. 2024. 中国马铃薯加工业现状调查分析及发展对策[J]. 中国马铃薯, 38(02):176-185. (Wei M, Xu H Z, Zhang N, et al.2024. Investigation on current situation of potato processing industry and relevant countermeasure in China[J]. Chinese Potato Journal, 38(02):176-185.) [12] 吴超, 方小婷, 石鑫鑫, 等. 2024. 马铃薯块茎低温糖化研究进展及展望[J]. 中国马铃薯, 38(02):168-75. (Wu C, Fang X T, Shi X X, et al.2024. Research progress and prospects on cold-induced sweetening in potato tubers[J]. Chinese Potato Journal, 38(02):168-175.) [13] 吴艳倩, 徐建飞, 马丽娜, 等. 2012. 马铃薯抗低温糖化育种研究进展[J]. 中国马铃薯, 26(02):108-115. (Wu Y Q, Xu J F, Ma L N, et al.2012. Advance in potato breeding for resistance to cold-induced sweetening[J]. Chinese Potato Journal, 26(02):108-115.) [14] 徐建飞, 金黎平. 2017. 马铃薯遗传育种研究:现状与展望[J].中国农业科学, 50(06):990-1015. (Xu J F, Jin L P.2017. Advances and perspectives in research of potato genetics and breeding[J]. Scientia Agricultura Sinica, 50(06):990-1015.) [15] 徐威, 朱春宝, 朱宝泉, 等. 2003. 利用电转化和三亲杂交方法高效转化根癌农杆菌[J]. 沈阳药科大学学报, (06):451-454. (Xu W, Zhu C B, Zhu B Q, et al. 2003. Efficient transformation of agrobacterium tumefaciens using electroporation and triparental mating[J]. Journal of Shenyang Pharmaceutical University, (06):451-454.) [16] 杨调调, 何志勇, 秦昉, 等. 2017. 美拉德反应对产品风味品质的影响及其衍生危害物研究进展[J]. 食品安全质量检测学报, 8(03):854-861. (Yang D D, He Z Y, Qin F, et al.Research progress of the effects of Maillard reaction on flavor and quality of products as well as derivatized harmful substances[J]. Journal of Food Safety and Quality, 8(03):854-861.) [17] 苑彬, 金慧, 骈琳, 等. 2024.美拉德反应对食品品质与安全的影响及其产物检测研究进展[J]. 轻工学报, 39(02):60-68. (Yuan B, Jin H, Pian L, et al.2024.A review of the impact of Maillard reaction on food quality and safety and the detection of its products[J]. Journal of Light Industry, 39(02):60-68.) [18] 张若芳, 金黎平, 谢开云, 等. 2015. GB/T 31784-2015, 马铃薯商品薯分级与检验规程[S]. 北京:中华人民共和国国家质量监督检验检疫总局. (Zhang R F, Jin L P, Xie K Y, et al. 2015. GB/T 31784-2015, Code of practice for grading and inspecting of commercial potatoes[S]. Beijing:General Administration of Quality Supervision, Inspection and Quarantine Supervision, Inspection and Quarantine of the Peoples Republic of China.) [19] Gikundi E N, Buzera A, Orina I, et al.2024. Impact of the temperature reconditioning of cold-stored potatoes on the color of potato chips and french fries[J]. Foods, 13(5):652. [20] Liu H, Ding Y D, Zhou Y Q, et al.2017. CRISPR-P 2.0:An improved CRISPR-CAS9 tool for genome editing in plants[J]. Molecular Plant, 10(3):530-532. [21] 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. [22] Maillard L C.1912. Action des acides amines surles sucres:Formation des melanoidines par voie methodique[J]. Compte‐rendu de l'Académie des Sciences, 154:66-68. [23] Marquez G, Anon M C.1986. Influence of reducing sugars and amino acids in the color development of fried potatoes[J]. Journal of Food Science, 51(1):157-160. [24] Nicot N, Hausman J F, Hoffmann L, et al.2005. Housekeeping gene selection for real-time RT-PCR normalization in potato during biotic and abiotic stress[J]. Function Plant Biology, 56(421):2907-2914. [25] Pritchard M K, Adam L R.1994. Relationships between fry color and sugar concentration in stored russet burbank and shepody potatoes[J]. American Potato Journal, 71:59-68. [26] Shumbe L, Soares E, Muhovski Y, et al.2024. Mutation of the vinv 50 UTR regulatory region reduces acrylamide levels in processed potato to reach EU food-safety standards[J]. Plant Biotechnology Journal, 22(10):2738-2740. [27] Sturm A.1999. Invertases. Primary structures, functions, and roles in plant development and sucrose partitioning[J]. Plant Physiology, 121(1):1-7. [28] Wiberley-Bradford A E, Busse J S, Jiang J, et al.2014. Sugar metabolism, chip color, invertase activity, and gene expression during long-term cold storage of potato (Solanum tuberosum) tubers from wild-type and vacuolar invertase silencing lines of Katahdin[J]. BMC Research Notes, 7(1):1-19. [29] Zhu X B, Chen A, Butler N M, et al.2024. Molecular dissection of an intronic enhancer governing cold-induced expression of the vacuolar invertase gene in potato[J]. Plant Cell, 36(5):1985-1999. |
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