vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJGB-22-62vavilov-3471Research ArticleГЕНЕТИЧЕСКИЕ РЕСУРСЫ И БИОКОЛЛЕКЦИИPLANT GENETICSЗависимость содержания крахмала и редуцирующих сахаров от уровня экспрессии генов β-амилаз StBAM1 и StBAM9 и ингибитора амилаз StAI при длительном низкотемпературном хранении клубней картофеляDependence of the content of starch and reducing sugars on the level of expression of the genes of β-amylases StBAM1 and StBAM9 and the amylase inhibitor StAI during long-term low-temperature storage of potato tubershttps://orcid.org/0000-0002-3124-525XКулаковаА. В.KulakovaA. V.

Москва

Moscow

https://orcid.org/0000-0002-2943-5118ЕфремовГ. И.EfremovG. I.

Москва

Moscow

gleb_efremov@mail.ruhttps://orcid.org/0000-0003-4692-3727ЩенниковаА. В.ShchennikovaA. V.

Москва

Moscow

https://orcid.org/0000-0002-6091-0765КочиеваЕ. З.KochievaE. Z.

Москва

Moscow

Институт биоинженерии, Федеральный исследовательский центр «Фундаментальные основы биотехнологии» Российской академии наукРоссияInstitute of Bioengineering, Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of SciencesRussian Federation202209102022266507514Copyright © Кулакова А.В., Ефремов Г.И., Щенникова А.В., Кочиева Е.З., 20222022Кулакова А.В., Ефремов Г.И., Щенникова А.В., Кочиева Е.З.Kulakova A.V., Efremov G.I., Shchennikova A.V., Kochieva E.Z.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/3471

Картофель (Solanum tuberosum L.) – первая по важности незерновая крахмалоносная культура с уровнем потенциальной урожайности 38–48 т/га и содержанием крахмала 13.2–18.7 %. Клубни картофеля хранятся при низкой температуре (2–4 °С), что обеспечивает состояние физиологического покоя. Недостат- ком такого хранения являются распад крахмала и, как следствие, накопление редуцирующих сахаров (холо- довое осахаривание), в том числе за счет роста активности β-амилаз, гидролизующих крахмал до мальтозы. В настоящем исследовании проведен сравнительный анализ динамики экспрессии генов β-амилаз (StBAM1, StBAM9) и ингибитора амилаз (StAI), а также содержания крахмала и редуцирующих сахаров в процессе дли- тельного низкотемпературного хранения (сентябрь, февраль, апрель) клубней пяти сортов картофеля (На- дежда, Барин, Красавчик, Утро и Северное сияние). Гены β-амилаз – StBAM9 и один из двух гомологов StBAM1 (с наибольшей степенью гомологии с AtBAM1) – выбраны на основе данных филогенетического анализа. Оценка экспрессии этих генов, а также гена ингибитора амилаз показала тенденцию к снижению уровня транскрипции для всех анализируемых сортов. Обнаружено, что содержание крахмала в процессе хранения клубней также существенно падает. В то же время количество редуцирующих сахаров увеличивается в период сентябрь–апрель, тогда как в период февраль–апрель их содержание не меняется (Красавчик), снижа- ется (Барин, Северное сияние) или продолжает расти (Утро, Надежда). Можно предположить, что активность генов StBAM1 и StBAM9 коррелирует с количеством крахмала (положительно) и моносахаридов (отрица- тельно). А уровень экспрессии StAI, в свою очередь, находится в прямой зависимости от уровня экспрессии StBAM1. При этом зависимость между степенью предрасположенности сорта к холодовому осахариванию и профилем экспрессии генов StBAM1, StBAM9 и StAI отсутствует.

Solanum tuberosum L. is the most important non-grain starch crop with a potential yield of 38–48 t/ha and a starch content of 13.2–18.7 %. Potato tubers are stored at a low temperature (2–4 °C) in a state of physiological dormancy. A disadvantage of this type of storage is the degradation of starch and the accumulation of reducing sugars (cold-induced sweetening), including due to an increase in the activity of β-amylases that hydrolyze starch to maltose. In this study, a comparative analysis of the β-amylase (StBAM1, StBAM9) and amylase inhibitor (StAI ) gene expression, as well as starch and reducing sugar content in tubers during long-term low-temperature storage (September, February, April) was performed using potato cultivars Nadezhda, Barin, Krasavchik, Severnoe siyanie and Utro. The β-amylase genes, StBAM9 and one of the two StBAM1 homologs (with the highest degree of homology with AtBAM1), were selected based on phylogenetic analysis data. Evaluation of the expression of these genes and the amylase inhibitor gene showed a tendency to decrease in transcription for all analyzed cultivars. The starch content also significantly decreased during tuber storage. The amount of reducing sugars increased in the September–April period, while in February–April, their content did not change (Krasavchik), decreased (Barin, Severnoe siyanie) or continued to grow (Utro, Nadezhda). It can be assumed that the gene activity of StBAM1 and StBAM9 correlates with the amount of starch (positively) and monosaccharides (negatively). The level of StAI expression, in turn, may be directly dependent on the level of StBAM1 expression. At the same time, there is no relationship between the degree of cultivar predisposition to cold-induced sweetening and the expression profile of the StBAM1, StBAM9, and StAI genes.

Solanum tuberosumсорта картофеляхранение клубнейкатаболизм крахмалаэкспрессия генаβ-амилазаSolanum tuberosumpotato cultivarstuber storagestarch catabolismgene expressionβ-amylaseThe study was supported by the Federal Targeted Program for Agricultural Development of the Russian Federation for 2017–2025 (subprogram “Development of Selection and Seed Production of Potatoes in the Russian Federation”)ReferencesBello-Perez L.A., Flores-Silva P.C., Agama-Acevedo E., Tovar J. Starch digestibility: past, present, and future. J. Sci. Food Agric. 2020;100(14):5009-5016. DOI 10.1002/jsfa.8955.Bello-Perez L.A., Flores-Silva P.C., Agama-Acevedo E., Tovar J. Starch digestibility: past, present, and future. J. Sci. Food Agric. 2020;100(14):5009-5016. DOI 10.1002/jsfa.8955.Benkeblia N., Alexopoulos A.A., Passam H.C. Physiology and biochemistry regulation of dormancy and sprouting in potato tuber (Solanum tuberosum L.). Fruit Vegetable Cereal Sci. Biotechnol. 2008;2(1):54-68.Benkeblia N., Alexopoulos A.A., Passam H.C. Physiology and biochemistry regulation of dormancy and sprouting in potato tuber (Solanum tuberosum L.). Fruit Vegetable Cereal Sci. Biotechnol. 2008;2(1):54-68.Dyachenko E.A., Kulakova A.V., Meleshin A.A., Shchennikova A.V., Kochieva E.Z. Amylase inhibitor SbAI in potato species: structure, variability and expression pattern. Russ. J. Genet. 2021;57(1):36-46. DOI 10.1134/S102279542101004X.Dyachenko E.A., Kulakova A.V., Meleshin A.A., Shchennikova A.V., Kochieva E.Z. Amylase inhibitor SbAI in potato species: structure, variability and expression pattern. Russ. J. Genet. 2021;57(1):36-46. DOI 10.1134/S102279542101004X.Fettke J., Eckermann N., Kötting O., Ritte G., Steup M. Novel starchrelated enzymes and carbohydrates. Cell. Mol. Biol. 2007;52: 883-904.Fettke J., Eckermann N., Kötting O., Ritte G., Steup M. Novel starchrelated enzymes and carbohydrates. Cell. Mol. Biol. 2007;52: 883-904.Fischer M., Schreiber L., Colby T., Kuckenberg M., Tacke E., Hofferbert H.R., Schmidt J., Gebhardt C. Novel candidate genes influencing natural variation in potato tuber cold sweetening identified by comparative proteomics and association mapping. BMC Plant Biol. 2013;13:113. DOI 10.1186/1471-2229-13-113.Fischer M., Schreiber L., Colby T., Kuckenberg M., Tacke E., Hofferbert H.R., Schmidt J., Gebhardt C. Novel candidate genes influencing natural variation in potato tuber cold sweetening identified by comparative proteomics and association mapping. BMC Plant Biol. 2013;13:113. DOI 10.1186/1471-2229-13-113.Guo L., Tao H., Cui B., Janaswamy S. The effects of sequential enzyme modifications on structural and physicochemical properties of sweet potato starch granules. Food Chem. 2019;277:504-514. DOI 10.1016/j.foodchem.2018.11.014.Guo L., Tao H., Cui B., Janaswamy S. The effects of sequential enzyme modifications on structural and physicochemical properties of sweet potato starch granules. Food Chem. 2019;277:504-514. DOI 10.1016/j.foodchem.2018.11.014.Hopkins R.H., Jelinek B., Harrison L.E. The action of β-amylase on potato amylose. Biochem. J. 1948;43(1):32-38. DOI 10.1042/bj0430032.Hopkins R.H., Jelinek B., Harrison L.E. The action of β-amylase on potato amylose. Biochem. J. 1948;43(1):32-38. DOI 10.1042/bj0430032.Hou J., Liu T., Reid S., Zhang H., Peng X., Sun K., Du J., Sonnewald U., Song B. Silencing of α-amylase StAmy23 in potato tuber leads to delayed sprouting. Plant Physiol. Biochem. 2019;139:411-418. DOI 10.1016/j.plaphy.2019.03.044.Hou J., Liu T., Reid S., Zhang H., Peng X., Sun K., Du J., Sonnewald U., Song B. Silencing of α-amylase StAmy23 in potato tuber leads to delayed sprouting. Plant Physiol. Biochem. 2019;139:411-418. DOI 10.1016/j.plaphy.2019.03.044.Hou J., Zhang H., Liu J., Reid S., Liu T., Xu S., Tian Z., Sonnewald U., Song B., Xie C. Amylases StAmy23, StBAM1 and StBAM9 regulate cold-induced sweetening of potato tubers in distinct ways. J. Exp. Bot. 2017;68:2317-2331. DOI 10.1093/jxb/erx076.Hou J., Zhang H., Liu J., Reid S., Liu T., Xu S., Tian Z., Sonnewald U., Song B., Xie C. Amylases StAmy23, StBAM1 and StBAM9 regulate cold-induced sweetening of potato tubers in distinct ways. J. Exp. Bot. 2017;68:2317-2331. DOI 10.1093/jxb/erx076.Lin Q., Xie Y., Guan W., Duan Y., Wang Z., Sun C. Combined transcriptomic and proteomic analysis of cold stress induced sugar accumulation and heat shock proteins expression during postharvest potato tuber storage. Food Chem. 2019;297:124991. DOI 10.1016/j.foodchem.2019.124991.Lin Q., Xie Y., Guan W., Duan Y., Wang Z., Sun C. Combined transcriptomic and proteomic analysis of cold stress induced sugar accumulation and heat shock proteins expression during postharvest potato tuber storage. Food Chem. 2019;297:124991. DOI 10.1016/j.foodchem.2019.124991.Lopez-Pardo R., de Galarreta J.I.R., Ritter E. Selection of housekeeping genes for qRT-PCR analysis in potato tubers under cold stress. 31(1):39-45. DOI 10.1007/s11032-012-9766-z. Mol. Breed. 2013;Lopez-Pardo R., de Galarreta J.I.R., Ritter E. Selection of housekeeping genes for qRT-PCR analysis in potato tubers under cold stress. 31(1):39-45. DOI 10.1007/s11032-012-9766-z. Mol. Breed. 2013;Monroe J.D., Storm A.R. Review: The Arabidopsis β-amylase (BAM) gene family: Diversity of form and function. Plant Sci. 2018;276: 163-170. DOI 10.1016/j.plantsci.2018.08.016.Monroe J.D., Storm A.R. Review: The Arabidopsis β-amylase (BAM) gene family: Diversity of form and function. Plant Sci. 2018;276: 163-170. DOI 10.1016/j.plantsci.2018.08.016.Nielsen T.H., Deiting U., Stitt M. A [beta]-amylase in potato tubers is induced by storage at low temperature. Plant Physiol. 1997;113(2): 503-510. DOI 10.1104/pp.113.2.503.Nielsen T.H., Deiting U., Stitt M. A [beta]-amylase in potato tubers is induced by storage at low temperature. Plant Physiol. 1997;113(2): 503-510. DOI 10.1104/pp.113.2.503.Riyaphan J., Jhong C.H., Lin S.R., Chang C.H., Tsai M.J., Lee D.N., Sung P.J., Leong M.K., Weng C.F. Hypoglycemic efficacy of docking selected natural compounds against α-glucosidase and α-amylase. Molecules. 2018;23(9):2260. DOI 10.3390/molecules23092260.Riyaphan J., Jhong C.H., Lin S.R., Chang C.H., Tsai M.J., Lee D.N., Sung P.J., Leong M.K., Weng C.F. Hypoglycemic efficacy of docking selected natural compounds against α-glucosidase and α-amylase. Molecules. 2018;23(9):2260. DOI 10.3390/molecules23092260.Shoaib N., Liu L., Ali A., Mughal N., Yu G., Huang Y. Molecular functions and pathways of plastidial starch phosphorylase (PHO1) in starch metabolism: current and future perspectives. Int. J. Mol. Sci. 2021;22(19):10450. DOI 10.3390/ijms221910450.Shoaib N., Liu L., Ali A., Mughal N., Yu G., Huang Y. Molecular functions and pathways of plastidial starch phosphorylase (PHO1) in starch metabolism: current and future perspectives. Int. J. Mol. Sci. 2021;22(19):10450. DOI 10.3390/ijms221910450.Slugina M.A., Meleshin A.A., Kochieva E.Z., Shchennikova A.V. The opposite effect of low temperature on the Pho1a starch phosphorylase gene expression in Solanum tuberosum L. tubers and Petota species leaves. Am. J. Potato Res. 2020;97:78-87. DOI 10.1007/s12230-019-09758-z.Slugina M.A., Meleshin A.A., Kochieva E.Z., Shchennikova A.V. The opposite effect of low temperature on the Pho1a starch phosphorylase gene expression in Solanum tuberosum L. tubers and Petota species leaves. Am. J. Potato Res. 2020;97:78-87. DOI 10.1007/s12230-019-09758-z.Solomos T., Mattoo A.K. Starch-sugar metabolism in potato (Solanum tuberosum L.) tubers in response to temperature variations. In: Razdan M.K., Mattoo A.K. (Eds.). Gene Improvement of Solanaceous Crops. New York: Sci. Publ. Inc., 2005;1:209-234.Solomos T., Mattoo A.K. Starch-sugar metabolism in potato (Solanum tuberosum L.) tubers in response to temperature variations. In: Razdan M.K., Mattoo A.K. (Eds.). Gene Improvement of Solanaceous Crops. New York: Sci. Publ. Inc., 2005;1:209-234.Sonnewald S., Sonnewald U. Regulation of potato tuber sprouting. Planta. 2014;239(1):27-38. DOI 10.1007/s00425-013-1968-z.Sonnewald S., Sonnewald U. Regulation of potato tuber sprouting. Planta. 2014;239(1):27-38. DOI 10.1007/s00425-013-1968-z.Tai H.H., Lagüe M., Thomson S., Aurousseau F., Neilson J., Murphy A., Bizimungu B., Davidson C., Deveaux V., Bègue Y., Wang H.Y., Xiong X., Jacobs J.M.E. Tuber transcriptome profiling of eight potato cultivars with different cold-induced sweetening responses to cold storage. Plant Physiol. Biochem. 2020;146:163-176. DOI 10.1016/j.plaphy.2019.11.001.Tai H.H., Lagüe M., Thomson S., Aurousseau F., Neilson J., Murphy A., Bizimungu B., Davidson C., Deveaux V., Bègue Y., Wang H.Y., Xiong X., Jacobs J.M.E. Tuber transcriptome profiling of eight potato cultivars with different cold-induced sweetening responses to cold storage. Plant Physiol. Biochem. 2020;146:163-176. DOI 10.1016/j.plaphy.2019.11.001.Tang X., Zhang N., Si H., Calderón-Urrea A. Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress. Plant Methods. 2017;13(85):85. DOI 10.1186/s13007-017-0238-7.Tang X., Zhang N., Si H., Calderón-Urrea A. Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress. Plant Methods. 2017;13(85):85. DOI 10.1186/s13007-017-0238-7.Taylor M.A., Ross H.A., McRae D., Stewart D., Roberts I., Duncan G., Wright F., Millam S., Davies H.V. A potato alpha-glucosidase gene encodes a glycoprotein-processing alpha-glucosidase II-like activity. Demonstration of enzyme activity and effects of down-regulation in transgenic plants. Plant J. 2000;24(3):305-316. DOI 10.1046/j.1365-313x.2000.00873.x.Taylor M.A., Ross H.A., McRae D., Stewart D., Roberts I., Duncan G., Wright F., Millam S., Davies H.V. A potato alpha-glucosidase gene encodes a glycoprotein-processing alpha-glucosidase II-like activity. Demonstration of enzyme activity and effects of down-regulation in transgenic plants. Plant J. 2000;24(3):305-316. DOI 10.1046/j.1365-313x.2000.00873.x.Thalmann M., Coiro M., Meier T., Wicker T., Zeeman S.C., Santelia D. The evolution of functional complexity within the β-amylase gene family in land plants. BMC Evol. Biol. 2019;19(1):66. DOI 10.1186/s12862-019-1395-2.Thalmann M., Coiro M., Meier T., Wicker T., Zeeman S.C., Santelia D. The evolution of functional complexity within the β-amylase gene family in land plants. BMC Evol. Biol. 2019;19(1):66. DOI 10.1186/s12862-019-1395-2.Vajravijayan S., Pletnev S., Mani N., Pletneva N., Nandhagopal N., Gunasekaran K. Structural insights on starch hydrolysis by plant β-amylase and its evolutionary relationship with bacterial enzymes. Int. J. Biol. Macromol. 2018;113:329-337. DOI 10.1016/j.ijbiomac.2018.02.138.Vajravijayan S., Pletnev S., Mani N., Pletneva N., Nandhagopal N., Gunasekaran K. Structural insights on starch hydrolysis by plant β-amylase and its evolutionary relationship with bacterial enzymes. Int. J. Biol. Macromol. 2018;113:329-337. DOI 10.1016/j.ijbiomac.2018.02.138.Van Harsselaar J.K., Lorenz J., Senning M., Sonnewald U., Sonnewald S. Genome-wide analysis of starch metabolism genes in potato (Solanum tuberosum L.). BMC Genomics. 2017;18(1):37. DOI 10.1186/s12864-016-3381-z.Van Harsselaar J.K., Lorenz J., Senning M., Sonnewald U., Sonnewald S. Genome-wide analysis of starch metabolism genes in potato (Solanum tuberosum L.). BMC Genomics. 2017;18(1):37. DOI 10.1186/s12864-016-3381-z.Vinje M.A., Walling J.G., Henson C.A., Duke S.H. Comparative gene expression analysis of the β-amylase and hordein gene families in the developing barley grain. Gene. 2019;693:127-136. DOI 10.1016/j.gene.2018.12.041.Vinje M.A., Walling J.G., Henson C.A., Duke S.H. Comparative gene expression analysis of the β-amylase and hordein gene families in the developing barley grain. Gene. 2019;693:127-136. DOI 10.1016/j.gene.2018.12.041.Zeeman S.C., Delatte T., Messerli G., Umhang M., Stettler M., Mettler T., Streb S., Reinhold H., Kötting O. Starch breakdown: recent discoveries suggest distinct pathways and novel mechanisms. Funct. Plant Biol. 2007;34(6):465-473. DOI 10.1071/FP06313.Zeeman S.C., Delatte T., Messerli G., Umhang M., Stettler M., Mettler T., Streb S., Reinhold H., Kötting O. Starch breakdown: recent discoveries suggest distinct pathways and novel mechanisms. Funct. Plant Biol. 2007;34(6):465-473. DOI 10.1071/FP06313.Zhang H., Hou J., Liu J., Xie C., Song B. Amylase analysis in potato starch degradation during cold storage and sprouting. Potato Res. 2014a;57:47-58. DOI 10.1007/s11540-014-9252-6.Zhang H., Hou J., Liu J., Xie C., Song B. Amylase analysis in potato starch degradation during cold storage and sprouting. Potato Res. 2014a;57:47-58. DOI 10.1007/s11540-014-9252-6.Zhang H., Liu J., Hou J., Yao Y., Lin Y., Ou Y., Song B., Xie C. The potato amylase inhibitor gene SbAI regulates cold-induced sweetening in potato tubers by modulating amylase activity. Plant Biotechnol. J. 2014b;12(7):984-993. DOI 10.1111/pbi.12221.Zhang H., Liu J., Hou J., Yao Y., Lin Y., Ou Y., Song B., Xie C. The potato amylase inhibitor gene SbAI regulates cold-induced sweetening in potato tubers by modulating amylase activity. Plant Biotechnol. J. 2014b;12(7):984-993. DOI 10.1111/pbi.12221.Zhang H., Yao Y., Chen S., Hou J., Yu Y., Liu T., Du J., Song B., Xie C. SbRFP1 regulates cold-induced sweetening of potato tubers by inactivation of StBAM1. Plant Physiol. Biochem. 2019;136:215-221. DOI 10.1016/j.plaphy.2019.01.019.Zhang H., Yao Y., Chen S., Hou J., Yu Y., Liu T., Du J., Song B., Xie C. SbRFP1 regulates cold-induced sweetening of potato tubers by inactivation of StBAM1. Plant Physiol. Biochem. 2019;136:215-221. DOI 10.1016/j.plaphy.2019.01.019.Zhao L., Yang T., Xing C., Dong H., Qi K., Gao J., Tao S., Wu J., Wu J., Zhang S., Huang X. The β-amylase PbrBAM3 from pear (Pyrus betulaefolia) regulates soluble sugar accumulation and ROS homeostasis in response to cold stress. Plant Sci. 2019;287:110184. DOI 10.1016/j.plantsci.2019.110184.Zhao L., Yang T., Xing C., Dong H., Qi K., Gao J., Tao S., Wu J., Wu J., Zhang S., Huang X. The β-amylase PbrBAM3 from pear (Pyrus betulaefolia) regulates soluble sugar accumulation and ROS homeostasis in response to cold stress. Plant Sci. 2019;287:110184. DOI 10.1016/j.plantsci.2019.110184.

The authors declare that there are no conflicts of interest present.