Molecular screening of squash and patisson squash collection samples using markers of the Pm-0 gene, which controls resistance to powdery mildew

Abstract

Powdery mildew (PM) is one of the most harmful diseases of cucurbits. Modern commercial varieties of squash Cucurbita pepo L. var. giraumonas Duch and patisson C. pepo var. melopepo L. received powdery mildew resistance genes from wild species. The Pm-0 resistance gene belongs to the linkage group 10; two CAPS markers were developed for its mapping [15]. The main intragenic marker NBS_S9_1495924/HaeIII is localized in the NBS-LRR region, the additional marker S9_1539675/MspI shows complete co-segregation with resistance to PM [15]. In the present study, these markers were used for molecular screening of an experimental set of squash and patisson samples (differ in resistance to powdery mildew) from the VIR gene bank collection and perspective breeding lines of the Krymsk Experiment Breeding Station – branch of VIR. In total, 80 samples were investigated. Samples, carrying fragments of both CAPS markers (17) and fragments of only one marker (31) were found. Presence of two markers of the Pm-0 gene (NBS_S9_1495924/HaeIII и S9_1539675/MspI) has the strong correlation with resistance to PM (r = 0.837).

For three powdery mildew-resistant samples, which has demonstrated presence of both markers of the Pm-0 gene, additional analysis of individual plants was done. Resistant plants with Pm-0 gene markers were self-pollinated to create resistant lines. Thus, molecular screening allowed to preserve the valuable trait of resistance during maintaining of the squash collection samples.

Keywords

Cucurbita pepo L. / powdery mildew / resistance / Pm-0 gene / CAPS markers

References

[1] Piskunova TM, Muteva ZF. The VIR collection — a source of initial breeding material for the perspective directions of breeding of a vegetable marrow and pumpkin. Vegetable crops of Russia. 2016;3:18–23. (In Russ.) DOI: 10.18619/2072-9146-2016-3-18-23

[2] Пискунова Т.М., Мутьева З.Ф. Коллекция ВИР — источник исходного материала для перспективных направлений селекции кабачка и тыквы. Овощи России. 2016. № 3. С. 18–23. DOI: 10.18619/2072-9146-2016-3-18-23

[3] Aliu S, Rusinovci I, Fetahu S, et al. Nutritive and Mineral composition in a collection of Cucurbita pepo L. grown in Kosova. Food and Nutrition Sciences. 2012;3(5):634–638. DOI: 10.4236/fns.2012.35087

[4] Aliu S., Rusinovci I., Fetahu S., et al. Nutritive and Mineral composition in a collection of Cucurbita pepo L. grown in Kosova // Food and Nutrition Sciences. 2012. Vol. 3, No. 5. P. 634–638. DOI: 10.4236/fns.2012.35087

[5] Palamarmuk II. Effectiveness of patisson (Сucurbita pepo var. melopepo L.) for different terms of crops in the conditions of forest step. Bulletin of Uman National University of Horticulture. 2019;1:25–28. DOI: 10.31395/2310-0478-2019-1-25-28

[6] Palamarmuk II. Effectiveness of patisson (Сucurbita pepo var. melopepo L.) for different terms of crops in the conditions of forest step // Bulletin of Uman National University of Horticulture. 2019. No. 1. P. 25–28. DOI: 10.31395/2310-0478-2019-1-25-28

[7] Varivoda OP, Maslennikova ES. Assessment and selection of source material for creating melon hybrids with integrated resistance to anthracnose and powdery mildew. Vegetable Crops of Russia. 2019;5:20–24. (In Russ.) DOI: 10.18619/2072-9146-2019-5-20-24

[8] Варивода О.П., Масленникова Е.С. Оценка и подбор исходного материала для создания гибридов дыни с комплексной устойчивостью к антракнозу и мучнистой росе // Овощи России. 2019. № 5. С. 20–24. DOI: 10.18619/2072-9146-2019-5-20-24

[9] Lebeda A, Krístkova E, Sedlakova B, et al. New concept for determination and denomination of pathotypes and races of cucurbit powdery mildew. In: Proceedings of Cucurbitaceae 2008, IX EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae. Pitrat M. ed. 2008. P. 125–134.

[10] Lebeda A., Krístkova E., Sedlakova B., et al. New concept for determination and denomination of pathotypes and races of cucurbit powdery mildew // Proceedings of Cucurbitaceae 2008, IX EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae. Pitrat M., ed. 2008. P. 125–134.

[11] Lebeda A, Sedlakova B, Krístkova E, Vysoudil M. Long-lasting changes in the species spectrum of cucurbit powdery mildew in the Czech Republic — influence of climate changes or random effect. Plant Protection Science. 2009;45(10):41–47. DOI: 10.17221/2807-PPS

[12] Lebeda A., Sedlakova B., Krístkova E., Vysoudil M. Long-lasting changes in the species spectrum of cucurbit powdery mildew in the Czech Republic — influence of climate changes or random effect? // Plant Protect Sci. 2009. Vol. 45, No. 10. P. 41–47. DOI: 10.17221/2807-PPS

[13] Treivas LYu, Bolezni i vrediteli ovoshchnykh kultur: atlas-opredelitel. Moscow: Fiton XXI; 2019. 192 p. (In Russ).

[14] Трейвас Л.Ю. Болезни и вредители овощных культур: атлас-определитель. Москва: Фитон XXI, 2019. 192 c.

[15] Ermolaeva LV, Khmelinskaya TV, Piskunova TM, et al. Ustoichivost ovoshchnykh kul’tur k boleznyam. In: Realizatsiya metodologicheskikh i metodicheskikh idei professora B.A. Dospekhova v sovershenstvovanii adaptivno-landshaftnykh sistem zemledeliya. Moscow; Suzdal, 2017. P. 91–95. (In Russ.)

[16] Ермолаева Л.В., Хмелинская Т.В., Пискунова Т.М., и др. Устойчивость овощных культур к болезням // Реализация методологических и методических идей профессора Б.А. Доспехова в совершенствовании адаптивно-ландшафтных систем земледелия. Коллективная монография: в 2 т. Москва; Суздаль. 2017. C. 91–95.

[17] Abbaskulieva SG. Muchnistaya rosa tykvennykh (Cucurbitaceae) rastenii i mery borby s nei // Uspekhi sovremennoi nauki i obrazovaniya. 2017;2:(3):85–88. (In Russ.)

[18] Аббаскулиева С.Г. Мучнистая роса тыквенных (Сucurbitaceae) растений и меры борьбы с ней // Успехи современной науки и образования 2017. Т. 2, № 3. С. 85–88.

[19] Medvedev AV, Kuzmin SV, Bukharov AF. Squash selection for powdery mildew resistance in the south of Russia. Agrarian Science. 2019;(3):91–95. (In Russ.) DOI: 10.32634/0869-8155-2019-326-3-91-95

[20] Медведев А.В., Кузьмин С.В., Бухаров А.Ф. Cелекция кабачка на устойчивость к мучнистой росе на юге России // Аграрная наука. 2019. № 3. С. 91–95. DOI: 10.32634/0869-85-2019-326-3-91-95

[21] Medvedev AV. Selektsiya ogurtsov na ustoichivost k muchnistoi rose [dissertation abstract]. Leningrad; 1974. 20 p. (In Russ.)

[22] Медведев А.В. Селекция огурцов на устойчивость к мучнистой росе: автореф. дис. … канд. с.-х. наук. Ленинград, 1974. 20 c.

[23] Peresypkin VF. Sel’skokhozyaistvennaya fitopatologiya. Moscow; 1989. 480 p. (In Russ.)

[24] Пересыпкин В.Ф. Сельскохозяйственная фитопатология. Москва, 1989. 480 c.

[25] Expert Analytical Center for Agribusiness. [Internet]. Available from: https://ab-centre.ru/news/kabachki-ploschadi-i-sbory-po-regionam-rf-v-2007-2021-gg. [cited 2022 August 29] (In Russ.)

[26] Экспертно-аналитический центр агробизнеса [Электронный ресурс]. Режим доступа: https://ab-centre.ru/news/kabachki-ploschadi-i-sbory-po-regionam-rf-v-2007-2021-gg. Дата обращения 29.08.2022.

[27] Medvedev AV, Medvedevа NI, Kuzmin SV. Against cucumber mosaic virus No. 1. Potato and Vegetables. 2016;9:39–40. (In Russ.)

[28] Медведев А.В., Медведева Н.И., Кузьмин С.В. Против вируса огуречной мозаики № 1 // Картофель и овощи. 2016. № 9, С. 39–40.

[29] Vielba-Fernandez A, Polonio A., Ruiz-Jimenez L, et al. Fungicide resistance in powdery mildew fungi. Microorganisms. 2020;8(9):1431. DOI: 10.3390/microorganisms8091431

[30] Vielba-Fernández A., Polonio Á., Ruiz-Jiménez L., et al. Fungicide resistance in powdery mildew fungi // Microorganisms. 2020 Vol. 8, No. 9. P. 1431. DOI: 10.3390/microorganisms8091431

[31] Holdsworth W, LaPlant K, Bell D, et al. Cultivar-based introgression mapping reveals wild species-derived Pm-0, the major powdery mildew resistance locus in squash. PLoS ONE. 2016;11(12): e0167715. DOI: 10.1371/journal.pone.0167715

[32] Holdsworth W., LaPlant K., Bell D., et al. Cultivar-based introgression mapping reveals wild species-derived Pm-0, the major powdery mildew resistance locus in squash // PLoS ONE. 2016. Vol. 11, No. 12. P. e0167715. DOI: 10.1371/journal.pone.0167715

[33] Guo WL, Chen BH, Guo YY, et al. Improved powdery mildew resistance of transgenic Nicotiana benthamiana overexpressing the Cucurbita moschata CmSGT1 Gene. Frontiers in Plant Science. 2019;10:955. DOI: 10.3389/fpls.2019.00955

[34] Guo W.L., Chen B.H., Guo Y.Y., et al. Improved powdery mildew resistance of transgenic Nicotiana benthamiana overexpressing the Cucurbita moschata CmSGT1 Gene // Frontiers in Plant Science. 2019. No. 10. P. 955. DOI: 10.3389/fpls.2019.00955

[35] Sowell F., Corley W. Resistance of Cucurbita plant introductions to powdery mildew. HortScience. 1973;8(6):492–493. DOI: 10.21273/HORTSCI.8.6.492

[36] Sowell F., Corley W. Resistance of Cucurbita plant introductions to powdery mildew // HortScience. 1973. Vol. 8, No. 6. P. 492–493. DOI: 10.21273/HORTSCI.8.6.492

[37] Lebeda A, Kristova E. Genotypic variation in field resistance of Cucurbita pepo cultivars to powdery mildew (Erysiphe cichoracearum). Genetic Resources and Crop Evolution. 1996;43(1):79–84. DOI: 10.1007/BF00126944

[38] Lebeda A., Křistova E. Genotypic variation in field resistance of Cucurbita pepo cultivars to powdery mildew (Erysiphe cichoracearum) // Genetic Resources and Crop Evolution. 1996. No. 43, No. 1. P. 79–84. DOI: 10.1007/BF00126944

[39] Cohen R, Hanan A, Paris H. Single-gene resistance to powdery mildew in zucchini squash (Cucurbita pepo). Euphytica. 2003;130(3):433–441. DOI: 10.1023/A:1023082612420

[40] Cohen R., Hanan A., Paris H. Single-gene resistance to powdery mildew in zucchini squash (Cucurbita pepo) // Euphytica. 2003. No. 130, No. 3. P. 433–441. DOI: 10.1023/A:1023082612420

[41] Contin ME, Munger HM. Inheritance of powdery mildew resistance in interspecific crosses with Cucurbita martinezii. HortScience. 1977;12(4):397.

[42] Contin M.E., Munger H.M. Inheritance of powdery mildew resistance in interspecific crosses with Cucurbita martinezii // HortScience. 1977. No. 12, No. 4. P. 397.

[43] Jahn M, Munger H, McCreight J. Breeding cucurbit crops for powdery mildew resistance. In: The powdery mildews, a comprehensive treatise. Ed. by R.R. Belanger, W.R. Bushnell, A.J. Dik, T.L.W. Carver. Amer Phytopathol Soc., St. Paul, Minn. 2002. P. 239–248.

[44] Jahn M., Munger H., McCreight J. Breeding cucurbit crops for powdery mildew resistance // The powdery mildews, a comprehensive treatise. Ed. by R.R. Belanger, W.R. Bushnell, A.J. Dik, T.L.W. Carver. Amer. Phytopathol. Soc., St. Paul, Minn, 2002. P. 239–248.

[45] Rhodes AM. Inheritance of powdery mildew resistance in the genus Cucurbita. The Plant Disease Reporter. 1964;(48):54–55.

[46] Rhodes A.M. Inheritance of powdery mildew resistance in the genus Cucurbita // The Plant Disease Reporter. 1964. No. 48. P. 54–55.

[47] Paris HS, Cohen S. Oligogenic inheritance for resistance to zucchini yellow mosaic virus in Cucurbita pepo. Annals of Applied Biology. 2000;(136):209–214.

[48] Paris H.S., Cohen S. Oligogenic inheritance for resistance to zucchini yellow mosaic virus in Cucurbita pepo // Annals of Applied Biology. 2000. No. 136. P. 209–214.

[49] Sazonova LV, Vlasova EG, Krivchenko VI, et al. Izuchenie i podderzhanie kollektsii ovoshchnykh rastenii: (metodicheskie ukazaniya). Leningrad: VIR; 1981. 192 p. (In Russ.)

[50] Сазонова Л.В., Власова Э.Г., Кривченко В.И., и др. Изучение и поддержание коллекции овощных растений: (методические указания). Ленинград: ВИР, 1981. 192 c.

[51] Kuzmin SV, Medvedev AV, Bukharov AF. Production of hybrid seeds of the vegetable marrow at free pollination. Vegetable Crops of Russia. 2018;(1):32–36. (In Russ.) DOI: 10.18619/2072-9146-2018-1-32-36

[52] Кузьмин С.В., Медведев А.В., Бухаров А.Ф. Получение гибридных семян кабачка при свободном опылении // Овощи России. 2018. № 1. С. 32–36. DOI: 10.18619/2072-9146-2018-1-32-36

[53] Cornell Vegetables. Disease-resistant cucurbit varieties [Internet]. Available from: https://www.vegetables.cornell.edu/pest-management/disease-factsheets/disease-resistant-vegetable-varieties/disease-resistant-cucurbit-varieties [cited 2022 Oct 15].

[54] Cornell Vegetables. Disease-resistant cucurbit varieties [Электронный ресурс]. Режим доступа: https://www.vegetables.cornell.edu/pest-management/disease-factsheets/disease-resistant-vegetable-varieties/disease-resistant-cucurbit-varieties. Дата обращения: 15.10.2022.

[55] Piskunova TM. Studying the global collection of pumpkin, marrow, pattypan and crookneck squashes and its maintenance in viable conditions: guidelines. Saint Petersburg: VIR; 2020. 48 p. (In Russ.) DOI: 10.30901/978-5-907145-21-4

[56] Пискунова Т.М. Изучение и поддержание в живом виде мировой коллекции тыквы, кабачка, патиссона, крукнека: (методические указания). Санкт-Петербург: ВИР, 2020. 48 с. DOI: 10.30901/978-5-907145-21-4

[57] Antonova OYu, Klimenko NS, Rybakov DA, et al. SSR analysis of modern Russian potato varieties using DNA samples of nomenclatural standards. Plant Biotechnology and Breeding. 2020;3(4):77–96. (In Russ.) DOI: 10.30901/2658-6266-2020-4-o2

[58] Антонова О.Ю., Клименко Н.С., Рыбаков Д.А., и др. SSR-анализ современных российских сортов картофеля с использованием ДНК номенклатурных стандартов // Биотехнология и селекция растений. 2020. Т. 3, № 4. С. 77–96. DOI: 10.30901/2658-6266-2020-4-o2

[59] Inglis PW, Pappas MdCR, Resende LV, Grattapaglia D. Fast and inexpensive protocols for consistent extraction of high quality DNA and RNA from challenging plant and fungal samples for high-throughput SNP genotyping and sequencing applications. PLoS ONE. 2018;13(10): e0206085. DOI: 10.1371/journal.pone.0206085

[60] Inglis P.W., Pappas Md.C.R., Resende L.V., Grattapaglia D. Fast and inexpensive protocols for consistent extraction of high quality DNA and RNA from challenging plant and fungal samples for high-throughput SNP genotyping and sequencing applications // PLoS One. 2018 Vol. 13, No. 10. P. e0206085. DOI: 10.1371/journal.pone.0206085

[61] Ivanter EV, Korosov AV. Vvedenie v kolichestvennuyu biologiyu. Petrozavodsk: PetrGU; 2011. 302 p. (In Russ.)

[62] Ивантер Э.В., Коросов А.В. Введение в количественную биологию. Петрозаводск: ПетрГУ, 2011. 302 c.

[63] Paris HS, Padley LD. Gene List for Cucurbita species. Cucurbit Genet Coop Rep. 2014;(37):1–14.

[64] Paris H.S., Padley L.D. Gene List for Cucurbita species // Cucurbit Genet Coop. Rep. 2014. No. 37. P. 1–14.

[65] Ogden AB. Utilization of exotic germplasm in a Cucurbita breeding program [Dissertation]. University of New Hampshire: Durham; 2021.

[66] Ogden A.B. Utilization of exotic germplasm in a Cucurbita breeding program [Dissertation]. University of New Hampshire: Durham, 2021.

[67] Margaritopoulou T, Kizis D, Kotopoulis D, et al. Corrigendum to: Enriched HeK4me3 marks at Pm-0 resistance-related genes prime courgette against Podosphaera xanthii. Plant Physiol. 2022;188(1):576–592. DOI: 10.1093/plphys/kiab502

[68] Margaritopoulou T., Kizis D., Kotopoulis D., et al. Corrigendum to: Enriched HeK4me3 marks at Pm-0 resistance-related genes prime courgette against Podosphaera xanthii // Plant Physiol. 2022 Vol. 188. No. 1. P. 576–592. DOI: 10.1093/plphys/kiab502

[69] Totsky IV, Rozanova IV, Safonova AD, et al. Geno-typing of potato samples from the GenAgro ICG SB RAS collection using DNA markers of genes conferring resistance to phytopathogens. Vavilov Journal of Genetics and Breeding 2021;25(6):677–686. DOI: 10.18699/VJ21.077

[70] Totsky I.V., Rozanova I.V., Safonova A.D., et al. Geno-typing of potato samples from the GenAgro ICG SB RAS collection using DNA markers of genes conferring resistance to phytopathogens // Vavilov Journal of Genetics and Breeding. 2021. Vol. 25, No. 6. P. 677–686. DOI: 10.18699/VJ21.077

[71] Klimenko NS, Antonova OY, Kostina LI, et al. Marker-associated selection of russian potato varieties with using markers of resistance genes to the golden potato cyst nematode (pathotype RO1). Proceedings on Applied Botany, Genetics and Breeding. 2017;178(4):66–75. (In Russ.) DOI: 10.30901/2227-8834-2017-4-66-75

[72] Клименко Н.С., Антонова О.Ю., Костина Л.И., и др. Маркер-опосредованная селекция отечественных сортов картофеля с маркерами генов устойчивости к золотистой картофельной нематоде (патотип RO1) // Труды по прикладной ботанике, генетике и селекции. 2017. Т. 178, № 4. С. 66–75. DOI: 10.30901/2227-8834-2017-4-66-75

[73] Schultz L, Cogan NOI, McLean K, et al. Evaluation and implementation of a potential diagnostic molecular marker for H1-conferred potato cyst nematode resistance in potato (Solanum tuberosum L.). Plant Breeding. 2012;131(2):315–321. DOI: 10.1111/j.1439-0523.2012.01949.x

[74] Schultz L., Cogan N.O.I., McLean K., et al. Evaluation and implementation of a potential diagnostic molecular marker for H1-conferred potato cyst nematode resistance in potato (Solanum tuberosum L.) // Plant Breeding. 2012. Vol. 131, No. 2. P. 315–321. DOI: 10.1111/j.1439-0523.2012.01949.x

[75] Biryukova VA, Shmyglya IV, Meleshin AA, et al. Study of Genetic Collections of the All-Russian Research Institute of potato farming by means of molecular markers. Dostizheniya Nauki i Tekhniki APK. 2016;30(10):22–26. (In Russ.)

[76] Бирюкова В.А., Шмыгля И.В., Мелёшин А.А., и др. Изучение генетических коллекций ВНИИ картофельного хозяйства с помощью молекулярных маркеров // Достижения науки и техники АПК. 2016. Т. 30, № 10. С. 22–26.

[77] Njuguna W. Development and use of molecular tools in Fragaria [Dissertation]. Corvallis, Oregon, USA: Oregon State University; 2010.

[78] Njuguna W. Development and use of molecular tools in Fragaria [Dissertation]. Corvallis, Oregon, USA: Oregon State University, 2010.

[79] Lyzhin AS, Lukyanchuk IV, Zhbanova EV, et al. Polymorphism of the Rca2 anthracnose resistance gene in strawberry cultivars (Fragaria × ananassa). Proceedings on Applied Botany, Genetics and Dreeding. 2019;180(1):73–77. (In Russ.) DOI: 10.30901/2227-8834-2019-1-73-77

[80] Лыжин А.С., Лукъянчук И.В., Жбанова Е.В. Полиморфизм сортов земляники (Fragaria × ananassa) по гену устойчивости к антракнозу Rca2 // Труды по прикладной ботанике, генетике и селекции. 2019. Т. 180, № 1. С. 73–77. DOI: 10.30901/2227-8834-2019-1-73-77

[81] Lukyanchuk IV, Lyzhin AS, Kozlova II. Analysis of the genetic collection of strawberries (Fragaria L.) for the Rca2 and Rpf1 genes with molecular markers. Vavilov Journal of Genetics and Breeding. 2018;22(7):795–799. (In Russ.) DOI: 10.18699/VJ18.423

[82] Лукьянчук И.В., Лыжин А.С., Козлова И.И. Анализ генетической коллекции клубники (Fragaria L.) для генов Rca2 и Rpf1 с молекулярными маркерами // Вавиловский журнал генетики и селекции. 2018. Т. 22, № 7. С. 795–799. DOI: 10.18699/VJ18.423

[83] Sturzeanu M, Coman M, Ciuca M, et al. Molecular characterization of allelic status of the Rpf1 and Rca2 genes in six cultivars of strawberries. Acta Hortic. 2016;1139:107–112. DOI: 10.17660/ActaHortic.2016.1139.19

[84] Sturzeanu M., Coman M., Ciuca M., et al. Molecular characterization of allelic status of the Rpf1 and Rca2 genes in six cultivars of strawberries // Acta Hortic. 2016. Vol. 1139 P. 107–112. DOI: 10.17660/ActaHortic.2016.1139.19

[85] Khrabrov IE, Antonova OYu, Shapovalov MI, Semenova LG. Molecular screening of the VIR strawberry varieties collection for the presence of a marker for the anthracnose black rot resistance gene Rca2. Plant Biotechnology and Breeding. 2021;4(4):15–24. (In Russ.) DOI: 10.30901/2658-6266-2021-4-o3

[86] Храбров И.Э., Антонова О.Ю., Шаповалов М.И., Семенова Л.Г. Молекулярный скрининг сортовой коллекции земляники ВИР на наличие маркера гена устойчивости к антракнозной черной гнили Rca2 // Биотехнология и селекция растений. 2021. Т. 4, № 4. С. 15–24. DOI: 10.30901/2658-6266-2021-4-o3

[87] Milczarek D, Flis B, Przetakiewicz A. Suitability of molecular markers for selection of potatoes resistant to Globodera spp. American Journal of Potato Research 2011;88:245–255. DOI: 10.1007/s12230-011-9189-0

[88] Milczarek D., Flis B., Przetakiewicz A. Suitability of molecular markers for selection of potatoes resistant to Globodera spp // American Journal of Potato Research. 2011 Vol. 88. P. 245–255. DOI: 10.1007/s12230-011-9189-0

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Правительство Российской ФедерацииGovernment of the Russian Federation(0481-2022-0008)

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