Comparative gene expression analysis of selected MYB and ZAT transcription factors in the roots of Vitis riparia and Vitis rupestris derived grapevine rootstocks under drought stress

Öz

Drought stress is one of the most important abiotic stress factors limiting yield and quality in viticulture. In particular, drought tolerance in grapevine rootstocks largely varies depending on genetic background, with genotypes derived from Vitis riparia generally exhibiting sensitivity to drought, whereas those derived from Vitis rupestris display a more tolerant profile. In this study, a total of seven grapevine rootstocks were used as plant material, including V. berlandieri × V. rupestris (110R, 1103P, 99R, and 140Ru) and V. berlandieri × V. riparia hybrids (SO4, 5BB, and 161-49C), which differ in drought stress tolerance. The aim of the study was to comparatively evaluate the Myb (Myb1 and Myb14) and ZAT10 transcription factor genes in response to drought stress at the gene expression level by Real-Time PCR. Following a drought stress treatment applied by withholding irrigation for 16 days, measurements of leaf water potential (Ψ, MPa) revealed that the greatest decreases were observed in the SO4 and 140Ru rootstocks, respectively. In the statistical analyses performed for the three gene regions, all three genes showed significant increases in expression in the 161-49C rootstock, with the highest expression level observed for the Myb1 gene in 161-49C rootstock, exhibiting a 25-fold increase. However, based on the gene expression results, no clear distinction was observed between drought-sensitive Vitis riparia and drought-tolerant Vitis rupestris origins.

Anahtar Kelimeler

• Grape rootstock
• Drought stress
• Real-Time PCR
• Gene expression

Vitis riparia ve Vitis rupestris kökenli asma anaçları köklerinde kuraklık stresine yanıtta bazı MYB ve ZAT transkripsiyon faktörlerinin karşılaştırmalı gen ifade analizi

Öz

Kuraklık stresi, bağcılıkta verim ve kaliteyi sınırlayan en önemli abiyotik stres faktörlerinden birisidir. Özellikle asma anaçlarında kuraklık toleransı, büyük ölçüde farklı genetik kökene bağlı olarak değişmekte olup, Vitis riparia kökenli genotipler genellikle kuraklığa hassas, Vitis rupestris kökenli genotipler ise daha tolerant bir profil sergilemektedir. Bu çalışmada, kuraklık stresi toleransı farklılık gösteren V. berlandieri × V. rupestris (110R, 1103P, 99R, 140Ru) ile V. berlandieri × V. riparia (SO4, 5BB ve 161-49C) melezi toplam 7 adet asma anacı kök dokusu bitkisel materyal olarak kullanılmıştır. Çalışmada, bu anaçların kuraklık stresine yanıtta Myb (Myb1 ve Myb14) ve ZAT10 transkripsiyon faktörü genlerinin, Real-Time PCR yöntemi kullanılarak gen ifadesi düzeyinde karşılaştırmalı olarak değerlendirilmesi amaçlanmıştır. 16 gün sulamama şeklinde uygulanan kuraklık stresi sonrasında yapılan yaprak su potansiyeli (Ψ, MPa) ölçümlerinde, su potansiyelindeki en büyük azalmalar sırası ile SO4 ve 140Ru anaçlarında gözlemlenmiştir. 3 gen bölgesine yönelik gerçekleştirilen istatistiksel analizlerde ise, 3 genin tümünde 161-49C anacı belirgin seviyelerde gen ifade artışı gösterirken, en yüksek gen ifade artışı da, 25 kat ile 161-49C anacında Myb1 geninde gözlemlenmiştir. Gen ifadesi sonuçları bakımından, kuraklık stresine hassas Vitis riparia ve kuraklık stresine tolerant Vitis rupestris kökenlerine göre net bir ayrım gözlemlenmemiştir.

Anahtar Kelimeler

• Üzüm anaç
• Kuraklık stresi
• Real-Time PCR
• Gen ifadesi

Etik Beyan

Bu makalede insan veya hayvan deneklerle herhangi bir çalışma bulunmaması nedeniyle etik onaya gerek duyulmamaktadır.

Teşekkür

Çalışmaya özverili desteklerinden dolayı, Ankara Üniversitesi Biyoteknoloji Enstitüsü, Bitki Biyoteknolojisi öğretim üyesi sayın Prof. Dr. Ali Ergül’e teşekkürlerimi sunarım.

Kaynakça

1. Alexander, R. D., Wendelboe-Nelson, C., & Morris, P. C. (2019). The barley transcription factor HvMYB1 is a positive regulator of drought tolerance. Plant Physiology and Biochemistry, 142, 246-253. https://doi.org/10.1016/j.plaphy.2019.07.014
2. Bianchi, D., Caramanico, L., Grossi, D., Brancadoro, L., & De Lorenzis, G. (2020). How do novel M-rootstock (Vitis spp.) genotypes cope with drought? Plants, 9(10), 1385. https://doi.org/10.3390/plants9101385
3. Bianchi, D., Ricciardi, V., Pozzoli, C., Grossi, D., Caramanico, L., Pindo, M., Stefani, E., Cestaro, A., Brancadoro, L., & De Lorenzis, G. (2023). Physiological and transcriptomic evaluation of drought effect on own rooted and grafted grapevine rootstock (1103P and 101-14MGt). Plants, 12(5), 1080. https://doi.org/10.3390/plants12051080
4. Bustin, S. A., Benes, V., Garson, J. A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M. W., Shipley, G. L., Vandesompele, J., & Wittwer, C. T. (2009). The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments. Clinical Chemistry, 55(4), 611-622. https://doi.org/10.1373/clinchem.2008.112797
5. Chaves, M. M., Zarrouk, O., Francisco, R., Costa, J. M., Santos, T., Regalado, A. P., Rodrigues, M. L., & Lopes, C. M. (2010). Grapevine under deficit irrigation: hints from physiological and molecular data. Annals of Botany, 105(5), 661-676. https://doi.org/10.1093/aob/mcq030
6. Chen, Y., Fei, Y., Howell, K., Chen, D., Clingeleffer, P., & Zhang, P. (2024). Rootstocks for grapevines now and into the future: selection of rootstocks based on drought tolerance, soil nutrient availability, and soil pH. Australian Journal of Grape and Wine Research, 2024(1), Article 6704238. https://doi.org/10.1155/2024/6704238
7. Ciftci-Yilmaz, S., & Mittler, R. (2008). The zinc finger network of plants. Cellular and Molecular Life Sciences, 65(7–8), 1150–1160. https://doi.org/10.1007/s00018-007-7473-4
8. Cramer, G. R., Ergül, A., Grimplet, J., Tillett, R. L., Tattersall, E. A. R., Bohlman, M. C., Vincent, D., Sonderegger, J., Evans, J., Osborne, C., Quilici, D., Schlauch, K. A., Schooley, D. A., Cushman, J. C., & Cushman, M. A. (2007). Water and salinity stress in grapevines: early and late changes in transcript and metabolite profiles. Functional & Integrative Genomics, 7(2), 111-134. https://doi.org/10.1007/s10142-006-0039-y

9. Çakır Aydemir, B., Yüksel Özmen, C., Kibar, U., Mutaf, F., Büyük, P. B., Bakır, M., & Ergül, A. (2020). Salt stress induces endoplasmic reticulum stress-responsive genes in a grapevine rootstock. PLoS ONE, 15(7), e0236424. https://doi.org/10.1371/journal.pone.0236424
10. Dubos, C., Stracke, R., Grotewold, E., Weisshaar, B., Martin, C., & Lepiniec, L. (2010). MYB transcription factors in Arabidopsis. Trends in Plant Science, 15(10), 573-581. https://doi.org/10.1016/j.tplants.2010.06.005
11. El Banna, M. F., AL-Huqail, A. A., Farouk, S., Belal, B. E. A., El-Kenawy, M. A., & Abd El Khalek, A. F. (2022). Morpho-Physiological and Anatomical Alterations of Salt-Affected Thompson Seedless Grapevine (Vitis vinifera L.) to Brassinolide Spraying. Horticulturae, 8(7), Article 568. https://doi.org/10.3390/horticulturae8070568
12. Gambetta, G. A., Herrera, J. C., Dayer, S., Feng, Q., Hochberg, U., & Castellarin, S. D. (2020). The physiology of drought stress in grapevine: towards an integrative definition of drought tolerance. Journal of Experimental Botany, 71(16), 4658-4676. https://doi.org/10.1093/jxb/eraa245
13. Han, J., Dai, J., Chen, Z., Li, W., Li, X., Zhang, L., Wang, W., Zhang, Z., Fan, P., Wang, Y., Wang, C., Han, D., Zhang, S., & Li, X. (2024). Overexpression of a ‘Beta’ MYB factor gene, VhMYB15, increases salinity and drought tolerance in Arabidopsis thaliana. International Journal of Molecular Sciences, 25(3), 1534. https://doi.org/10.3390/ijms25031534
14. Han, J., Xia, J., Wang, C., Wang, Z., Zhao, Z., Wang, Z., & Zhang, Y. (2025). Stage specific transcriptomic responses of wheat to heat, drought, and combined stress: AP2/ERF regulatory networks and functional characterization of TaEREBP1 L. BMC Plant Biology, 25(1), 1594.https://doi.org/10.1186/s12870-025-07615-2
15. Hoagland, D. R., & Arnon, D. I. (1950). The water culture method for growing plants without soil. University of California, Berkeley, CA, USA, Circular 347, 1-32.
16. Hosseini, E., Razi, H., Tahmasebi, A., Alemzadeh, A., & Ebrahimie, E. (2025). Comparative transcriptomic and biochemical analyses reveal candidate regulatory genes associated with early drought response signatures in contrasting rapeseed (Brassica napus) cultivars. Scientific Reports, 16, 966 . https://doi.org/10.1038/s41598-025-30408-4
17. Hrmova, M., & Hussain, S. S. (2021). Plant transcription factors involved in drought and associated stresses. International Journal of Molecular Sciences, 22(11), 5662. https://doi.org/10.3390/ijms22115662
18. Hussain, Q., Asim, M., Zhang, R., Khan, R., Farooq, S., & Wu, J. (2021). Transcription factors interact with ABA through gene expression and signaling pathways to mitigate drought and salinity stress. Biomolecules, 11(8), 1159. https://doi.org/10.3390/biom11081159
19. Irani, H., Valizadeh-Kaji, B., & Naeini, M. R. (2021). Biostimulant-induced drought tolerance in grapevine is associated with physiological and biochemical changes. Chemical and Biological Technologies in Agriculture, 8(1), 5. https://doi.org/10.1186/s40538-021-00210-7
20. Jiao, S., Huang, Y., Lu, S., Wang, H., Li, Y., Mao, J., & Chen, B. (2025). Transcriptomic changes and VvMYBPA1 function analysis reveal the molecular mechanism of drought tolerance in grapevine. Journal of Integrative Agriculture, 25 (4), 1501-1518. https://doi.org/10.1016/j.jia.2025.12.028
21. Kaya, C. (2025). Molecular crosstalk of transcription factors and hormones in drought tolerance genes in plants. Turkish Journal of Botany, 49(3), 145-164. https://doi.org/10.55730/1300-008X.2852
22. Krishankumar, S., Hunter, J. J., Alyafei, M., Hamed, F., Subramaniam, S., Ramlal, A., Yousuf, A., Al Jabri, H., Al Mahruqi, H., Al Harthy, S., Al Kharusi, L., Al Habsi, K., Al Yahyai, R., Al Mahmoudi, H., Al Rashdi, A., Al Harrasi, S., Al Mahri, A., Al Mahrouqi, A., Al Hinai, M., & Amiri, K. M. (2025). Physiological, biochemical and elemental responses of grafted grapevines under drought stress: insights into tolerance mechanisms. BMC Plant Biology, 25(1), 385. https://doi.org/10.1186/s12870-025-06374-4
23. Labarga, D., Mairata, A., Puelles, M., Santana, S., García-Escudero, E., & Pou, A. (2021, October). Rootstocks modulate the physiology of the scion responses to water deficit in grapevines. In XI International Symposium on Grapevine Physiology and Biotechnology (Vol. 1390, pp. 49-56).
24. Li, X. L., Meng, D., Li, M. J., Zhou, J., Yang, Y. Z., Zhou, B. B., Yang, M. M., Li, M., Ma, F. W., Xu, K. N., & Zhang, J. K. (2023). Transcription factors MhDREB2A/MhZAT10 play a role in drought and cold stress response crosstalk in apple. Plant Physiology, 192(3), 2203-2220. https://doi.org/10.1093/plphys/kiad147
25. Liu, Y., Khan, A. R., & Gan, Y. (2022). C2H2 zinc finger proteins response to abiotic stress in plants. International Journal of Molecular Sciences, 23(5), 2730. https://doi.org/10.3390/ijms23052730
26. Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2⁻ΔΔCT method. Methods, 25(4), 402-408. https://doi.org/10.1006/meth.2001.1262
27. Lukšić, K., Mucalo, A., Smolko, A., Brkljačić, L., Marinov, L., Hančević, K., Kovačević Ganić, K., Žulj Mihaljević, M., Maletić, E., Pejić, I., Poljuha, D., Radić, T., Osrečak, M., Žufić, V., Zdunić, G., & Kozina, B. (2023). Biochemical response and gene expression to water deficit of Croatian grapevine cultivars (Vitis vinifera L.) and a specimen of Vitis sylvestris. Plants, 12(19), 3420. https://doi.org/10.3390/plants12193420
28. Mullins, M. G., Bouquet, A., & Williams, L. E. (1992). Biology of the Grapevine. Cambridge University Press.
29. Nguyen, X. C., Kim, S. H., Lee, K., Kim, K. E., Liu, X. M., Han, H. J., Hoang, M. H., Lee, S. W., Hong, J. C., & Moon, Y. H. (2012). Identification of a C2H2-type zinc finger transcription factor (ZAT10) from Arabidopsis as a substrate of MAP kinase. Plant Cell Reports, 31(5), 737-745. https://doi.org/10.1007/s00299-011-1196-5
30. Ollat, N., Bordenave, L., Tandonnet, J. P., Boursiquot, J. M., & Marguerit, E. (2014, October). Grapevine rootstocks: origins and perspectives. In I International Symposium on Grapevine Roots (Vol. 1136, pp. 11–22).
31. Özmen, C. Y., Baydu, F. Y., Hasanzadeh, M., Öktem, M., Babaoğlu, G., Kibar, U., & Ergül, A. (2023). Assessment of drought stress responsive genes expression profiles and proline accumulation in a diverse set of grapevine rootstocks. Turkish Journal of Agriculture and Forestry, 47, 319-334. https://doi.org/10.55730/1300-011X.3089
32. Özmen, C. Y., Yılmaz Baydu, F., & Ergül, A. (2025). Comparative analysis of Cabernet Sauvignon (Vitis vinifera L.) and Kober 5BB (V. berlandieri × V. riparia) root transcriptomes reveals multiple processes associated with drought tolerance in grapevines. Horticulturae, 11(9), 1092. https://doi.org/10.3390/horticulturae11091092
33. Pérez-Álvarez, E. P., Intrigliolo, D. S., Martínez-Moreno, A., García-Sánchez, F., Parra, M., Alfosea-Simon, M., & Buesa, I. (2023). Ecophysiological responses of grapevine rootstocks to water deficit. Vitis- Journal of Grapevine Research, 62(2), https://doi.org/10.5073/VITIS.2023.62.59-74
34. Prinsi, B., Simeoni, F., Galbiati, M., Meggio, F., Tonelli, C., Scienza, A., & Espen, L. (2021). Grapevine rootstocks differently affect physiological and molecular responses of the scion under water deficit condition. Agronomy, 11(2), 289. https://doi.org/10.3390/agronomy11020289
35. Ren, C., Li, Z., Song, P., Wang, Y., Liu, W., Zhang, L., Wang, W., Zhang, Z., Fan, P., Wang, C., Han, D., Zhang, S., & Li, X. (2023). Overexpression of a grape MYB transcription factor gene VhMYB2 increases salinity and drought tolerance in Arabidopsis thaliana. International Journal of Molecular Sciences, 24(13), 10743. https://doi.org/10.3390/ijms241310743
36. Rodriguez-Dominguez, C. M., Carins-Murphy, M. R., Sebastian-Azcona, J., & Brodribb, T. J. (2025). Stem water potential measurements obtained using standard methodologies diverge under extreme drought in tomato and grapevine. Plant Physiology, 199(4), kiaf591. https://doi.org/10.1093/plphys/kiaf591
37. Romero, P., Acosta, J. M. N., Pérez-Pérez, J. G., Dodd, I. C., Antolinos, V., Marín, J. L., García-Tejero, I. F., Botía, P., Navarro, J. M., & Ordaz, P. B. (2025). The rootstock imparts different drought tolerance strategies in long-term field-grown deficit irrigated grapevines. Scientia Horticulturae, 346, 114167. https://doi.org/10.1016/j.scienta.2025.114167
38. Jogaiah, S., Ramteke, S. D., Sharma, J., & Upadhyay, A. K. (2014). Moisture and salinity stress induced changes in biochemical constituents and water relations of different grape rootstock cultivars. International Journal of Agronomy, 2014, Article 789087. https://doi.org/10.1155/2014/789087
39. Serra, I., Strever, A., Myburgh, P. A., & Deloire, A. (2013). Review: the interaction between rootstocks and cultivars (Vitis vinifera L.) to enhance drought tolerance in grapevine. Australian Journal of Grape and Wine Research, 20(1), 1-14. https://doi.org/10.1111/ajgw.12005
40. Shinozaki, K., & Yamaguchi-Shinozaki, K. (2007). Gene networks involved in drought stress response and tolerance. Journal of Experimental Botany, 58(2), 221-227. https://doi.org/10.1093/jxb/erl164
41. Tang, Y., & Xia, P. (2025). WRKY transcription factors: key regulators in plant drought tolerance. Plant Science, 359, 112647. https://doi.org/10.1016/j.plantsci.2025.112647
42. Tattersall, E. A., Ergul, A., AlKayal, F., DeLuc, L., Cushman, J. C., & Cramer, G. R. (2005). Comparison of methods for isolating high-quality RNA from leaves of grapevine. American Journal of Enology and Viticulture, 56, 400–406.
43. Toumi, I., M’Sehli, W., Bourgou, S., Jallouli, N., Bensalem-Fnayou, A., Ghorbel, A., & Mliki, A. (2007). Response of ungrafted and grafted grapevine cultivars and rootstocks (Vitis sp.) to water stress. Oeno One, 41(2), 85-94. https://doi.org/10.20870/oeno-one.2007.41.2.1127
44. Wang, Z., Yin, S., Wei, Y., Chen, B., & Li, W. (2025). Transcriptome and WGCNA analysis revealed the molecular mechanism of drought resistance in new sugarcane varieties. Frontiers in Plant Science, 16, 1687280. https://doi.org/10.3389/fpls.2025.1687280
45. Wenter, A., Zanotelli, D., Tagliavini, M., & Andreotti, C. (2019, June). Thresholds of soil and plant water availability that affect leaf transpiration, stomatal conductance and photosynthesis in grapevines. In IX International Symposium on Irrigation of Horticultural Crops (Vol. 1335, pp. 605-612).
46. Wyrzykowska, A., Bielewicz, D., Plewka, P., Sołtys-Kalina, D., Wasilewicz-Flis, I., Marczewski, W., Szczerba, H., Ciesielski, A., Szweykowska-Kulińska, Z., & Jarmołowski, A. (2022). The MYB33, MYB65, and MYB101 transcription factors affect Arabidopsis and potato responses to drought by regulating the ABA signaling pathway. Physiologia Plantarum, 174(5), e13775. https://doi.org/10.1111/ppl.13775
47. Yıldırım, K., Yağcı, A., Sucu, S., & Tunç, S. (2018). Responses of grapevine rootstocks to drought through altered root system architecture and root transcriptomic regulations. Plant Physiology and Biochemistry, 127, 256-268. https://doi.org/10.1016/j.plaphy.2018.04.021
48. Yilmaz Baydu, F. (2026). Comparative expression analysis of Myb1, Myb14, and Myb15 genes in leaf tissues of grapevine rootstocks under long-term drought stress. International Journal of Agriculture, Environment and Food Sciences, 10(2), 514-521. https://doi.org/10.31015/jaefs.2026.2.24
49. Zhu, N., Duan, B., Zheng, H., Mu, R., Zhao, Y., Ke, L., & Sun, Y. (2023). An R2R3 MYB gene GhMYB3 functions in drought stress by negatively regulating stomatal movement and ROS accumulation. Plant Physiology and Biochemistry, 197, 107648. https://doi.org/10.1016/j.plaphy.2023.107648