بررسی تغییرات ناشی از منبع دانه‌‌‌گرده بر اندازه میوه و الگوی بیانی miR396a و ژن هدف آن در مراحل نموی میوه Prunus arabica

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه باغبانی، دانشکده کشاورزی، دانشگاه شهرکرد، ایران، شهرکرد

2 گروه اصلاح نباتات و بیوتکنولوژی، دانشکده کشاورزی، دانشگاه شهرکرد

چکیده

هدف: بادام (Prunus dulcis MILL) از میوه‌های بسیار ارزشمندی است که به صورت تجاری در مناطق معتدل جهان کشت می‌شود. امروزه میوه‌های بزرگتر بازار فروش و سود بیشتری دارند. با توجه به تأثیر منابع گرده بر ویژگی‌های میوه بادام از جمله اندازه هسته، در این تحقیق پدیده زنیا در سطح مورفولوژیک و مولکولی جهت شناسایی و تایید ژن‌های مرتبط با اندازه میوه، مورد مطالعه قرار گرفته است.
مواد و روش‌ها: برای این منظور بادام اربیکا با گونه Prunus orientalis و رقم سفید با دانه ریز و دو رقم مامایی و پویا با دانه درشت گرده‌افشانی شد و نمونه‌برداری از مراحل مختلف رشد میوه برای مطالعات مورفولوژیک و مولکولی انجام شد. ابعاد میوه با کولیس دیجیتال و وزن با ترازو دیجیتال اندازه‌گیری شد.
نتایج: نتایج خصوصیات مورفولوژیک بذر نشان داد که بین تلاقی‌های مختلف تفاوت معنی‌داری وجود دارد، به طوری که گرده‌افشانی با رقم پویا باعث افزایش ویژگی‌های مورفولوژیکی مانند طول، عرض، ضخامت و وزن هسته والد مادری نسبت به P. Orientalis شد. همچنین برای بررسی نقش زنیا بر اندازه میوه در سطح مولکولی، میزان بیان ژن‌های Pdu-miR396a-BEN1 به عنوان ژن‌های دخیل در فرآیند رشد و نمو گیاهان، در مراحل مختلف رشد مغز بادام حاصل از تلاقی‌های مختلف بررسی و نقش احتمالی آن‌ها در کنترل اندازه هسته توسط RT-qPCR تایید شد. به طوری که Pdu-miR396a کاهش بیان معنی‌داری را در میوه بزرگ حاصل از تلاقی اربیکا × پویا در مقایسه با دیگر تلاقی‌ها داشت. همچنین روند بیان این ژن کاهشی بوده و با پیشرفت مراحل نموی و بزرگ شدن اندازه میوه، بیان آن کاهش پیدا کرده است. علاوه بر این همبستگی منفی بین بیان Pdu-miR396a و پروتئین هدف آن (BRI1-5 ENHANCED 1 (BEN1) ، مشاهده شد. به طوری که PduBEN1 افزایش بیان معنی‌داری را در میوه بزرگ حاصل از تلاقی اربیکا × پویا در مقایسه با دیگر تلاقی‌ها داشت. بنابراین، Pdu-miR396a-BEN1 به طور بالقوه در تنظیم اندازه دانه بادام توسط والدین مادری و پدری نقش دارد.
نتیجه گیری: با توجه به نتایج بدست آمده می‌توان نتیجه گرفت که Pdu-miR396a-BEN1 به طور بالقوه در تنظیم اندازه دانه بادام نقش اساسی دارد و توسط والدین مادری و پدری بر اندازه میوه تاثیر می‌گذارد بنابراین بررسی بیان این ژن در کنار بررسی های مورفولوژیکی می‌تواند نشان دهنده و تایید کننده پدیده زنیا در بادام باشد.

کلیدواژه‌ها

عنوان مقاله [English]

Investigation of Pollen Source-Induced Variation in Fruit Size and Expression Pattern of Mir396a and its Target Gene During Fruit Developmental Stage in Prunus Arabica

نویسندگان [English]

  • Marjan Jafari 1
  • Behrouz Shiran 2
1 Department of Horticulture, Faculty of Agriculture, Shahrekored University, P.O. Box 115, Shahrekord, Iran
2 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Shahrekord University
چکیده [English]

Objective
Almond (Prunus dulcis Mill.) represents a high-value fruit crop that is widely cultivated in temperate agro-climatic regions. Market preference and economic returns are often strongly linked to fruit size, with larger almonds generally commanding greater commercial interest. Given that the pollen parent can influence fruit characteristics, including kernel dimensions, this study applied the concept of xenia to evaluate its impact at both morphological and molecular levels, with the aim of identifying and substantiating genes involved in the regulation of fruit size.
Materials and Methods
To address this objective, Arabica almond trees were pollinated using four distinct almond genotypes: Mamaee and Pooya, characterized by large kernel size, and the Sefid cultivar along with Prunus orientalis, both producing small kernels. Fruits were sampled at multiple developmental stages and subjected to morphological and molecular assessments. Measurements of fruit length and width were obtained using a digital caliper, while fruit weight was recorded with a digital balance.
Results
Morphological analysis revealed significant differences among the crosses. Pollination with Pooya led to an increase in maternal morphological traits such as kernel length, width, thickness, and weight compared to P. orientalis. At the molecular level, the expression of Pdu-miR396a and BEN1—genes involved in plant growth and development—was examined in kernels at different developmental stages using RT-qPCR. The results showed that Pdu-miR396a expression was significantly reduced in large fruits obtained from the P. arabica × Pooya cross compared to other combinations. Moreover, its expression decreased progressively during fruit development, correlating with kernel enlargement. A negative correlation was observed between Pdu-miR396a and its target protein BRI1-5 ENHANCED 1 (BEN1), where PduBEN1 exhibited significantly higher expression in large fruits from the P. arabica × Pooya cross compared to others. These findings suggest that the Pdu-miR396a–BEN1 module potentially regulates almond kernel size through interactions between maternal and paternal parents.
Conclusion
The findings demonstrate that the Pdu-miR396a–BEN1 regulatory module is a key factor in the control of almond kernel size, with contributions from both maternal and paternal genotypes to fruit development. Accordingly, the combined use of gene expression profiling and morphological assessment offers a robust approach for supporting and verifying the influence of xenia in almond production.

کلیدواژه‌ها [English]

  • Xenia
  • gene
  • Almond kernel
  • miRNA
  • RT-qPCR

مراجع

Alizadeh-Saltah, S. (2007). Compatibility of pollination and pollen tube growth of Shahrud 12 and 21 almond (Prunus amygdalus Batsch) cultivars (Master’s thesis, Faculty of Agriculture, Tarbiat Modares University).
Amiri Roudbar, M., Mohammadabadi, M. R., Ayatollahi Mehrgardi, A., Abdollahi-Arpanahi, R., Momen, M., Morota, G., Brito Lopes, F., Gianola, D., & Rosa, G. J. M. (2020). Integration of single nucleotide variants and whole-genome DNA methylation profiles for classification of rheumatoid arthritis cases from controls. Heredity, 124(5), 658–674. https://doi.org/10.1038/s41437-020-0301-4
Barazandeh, A., Mohammadabadi, M. R., Ghaderi-Zefrehei, M., & Nezamabadi-Pour, H. (2016). Genome-wide analysis of CpG islands in some livestock genomes and their relationship with genomic features. Czech Journal of Animal Science, 61(11), 487–495. https://doi.org/10.17221/78/2015-CJAS
Bordbar, F., Mohammadabadi, M., Jensen, J., Xu, L., Li, J., & Zhang, L. (2022). Identification of candidate genes regulating carcass depth and hind leg circumference in Simmental beef cattle using Illumina Bovine Beadchip and next-generation sequencing analyses. Animals (Basel), 12(9), 1103. https://doi.org/10.3390/ani12091103
Browicz, K., & Zohary, D. (1996). The genus Amygdalus L.(Rosaceae): species relationships, distribution and evolution under domestication. Genetic Resources and Crop Evolution, 43(3), 229-247. https://doi.org/10.1007/ Bf00123275
Cao, D., Wang, J., Ju, Z., Liu, Q., Li, S., Tian, H., Fu, D., Zhu, H., Luo, Y., & Zhu, B. (2016). Regulations on growth and development in tomato cotyledon, flower and fruit via destruction of miR396 with short tandem target mimic. Plant Science, 247, 1–12. https://doi.org/10.1016/j.plantsci.2016.02.012
Correa, J. P. O., Silva, E. M., & Nogueira, F. T. S. (2018). Molecular control by non-coding RNAs during fruit development: From gynoecium patterning to fruit ripening. Frontiers in Plant Science, 9, 1760. https://doi.org/10.3389/fpls.2018.01760
Denney, J.O., 1992. Xenia includes metaxenia. HortScience, 27(7), pp.722-728.
Dicenta, F., Ortega, E., Cánovas, J. A., & Egea, J. (2002). Self‐pollination vs. cross‐pollination in almond: pollen tube growth, fruit set and fruit characteristics. Plant Breeding, 121(2), 163-167.  https://doi.org/10.1046/j.1439-0523.2002.00689.x
Ehlenfeldt, M. K. (2003). Investigations of metaxenia in northern highbush blueberry (Vaccinium corymbosum L.) cultivars. Journal of the American Pomological Society, 57(1), 26.
Hewezi, T., & Baum, T. J. (2012). Complex feedback regulations govern the expression of miRNA396 and its GRF target genes. Plant signaling & behavior, 7(7), 749-751. https://doi.org/10.4161/psb.20420
Hwang, H., Ryu, H., & Cho, H. (2021). Brassinosteroid signaling pathways interplaying with diverse signaling cues for crop enhancement. Agronomy, 11(3), 556.  https://doi.org/10.3390/agronomy11030556
Jafari, M., Shiran, B., Rabiei, G., Ravash, R., Sayed Tabatabaei, B. E., & Martinez-Gomez, P. (2021). Identification and verification of seed development related miRNAs in kernel almond by small RNA sequencing and qPCR. Plos one, 16(12), e0260492. https://doi.org/10.1371/journal.pone.0260492
Jafari, M., Shiran, B., Rabiei, G., Ravash, R., Sayed Tabatabaei, B. E., Martínez-García, P. J., & Martínez-Gómez, P. (2022). Identification of genes associated with kernel size in almond [Prunus dulcis (Mill.) DA Webb] using RNA-Seq. Plant Growth Regulation, 97(2), 357-373. https://doi.org/10.1007/s10725-022-00801-4\
Jiang, W. B., & Lin, W. H. (2013). Brassinosteroid functions in Arabidopsis seed development. Plant signaling & behavior, 8(10), e25928. https://doi.org/10.4161/psb.25928
Kester, D. E., Gradziel, T. M., & Grasselly, C. (1991). Almonds (Prunus). Genetic Resources of Temperate Fruit and Nut Crops 290, 701-760.Khadivi, A. (2011). Fruit growing (general and private) (2nd ed.). Agricultural Education and Extension Publications. [In Persian].
Khabiri, A., Toroghi, R., Mohammadabadi, M., & Tabatabaeizadeh, S. E. (2023). Introduction of a Newcastle disease virus challenge strain (sub-genotype VII.1.1) isolated in Iran. Veterinary Research Forum, 14(4), e221. https://doi.org/10.30466/vrf.2022.548152.3373
Li, N., & Li, Y. (2015). Maternal control of seed size in plants. Journal of Experimental Botany, 66(4), 1087-1097. https://doi.org/10.1093/jxb/eru549
Liebsch, D., & Palatnik, J. F. (2020). MicroRNA miR396, GRF transcription factors and GIF co-regulators: a conserved plant growth regulatory module with potential for breeding and biotechnology. Current opinion in plant biology, 53, 31-42.
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.
Militaru, M., Butac, M., Sumedrea, D., & Chiţu, E. (2015). Effect of metaxenia on the fruit quality of scab resistant apple varieties. Agriculture and Agricultural Science Procedia, 6, 151-156.
Mohammadabadi, M., Babenko, I. O., Borshch, O., Kalashnyk, O., Ievstafiieva, Y., & Buchkovska, V. (2024). Measuring the relative expression pattern of the UCP2 gene in different tissues of the Raini Cashmere goat. Agricultural Biotechnology Journal, 16(3), 317-332. https://doi.org/10.22103/jab.2024.24337.1627
Muzaffar, S., & Bisht, K. (2017). Recent developments in understanding the mechanism and functions of microRNAs. Journal of Applied Biology & Biotechnology Vol, 5(02), 001-007. DOI: 10.7324/JABB.2017.50201
Olfati, J. A., Sheykhtaher, Z., Qamgosar, R., Khasmakhi-Sabet, A., Peyvast, G. H., Samizadeh, H., & Rabiee, B. (2010). Xenia and metaxenia on cucumber fruit and seed characteristics. International Journal of Vegetable Science, 16(3), 243-252. https://doi.org/10.1080/19315260903584167
Ortega, E., Egea, J., Cánovas, J., & Dicenta, F. (2002). Pollen tube dynamics following half-and fully-compatible pollinations in self-compatible almond cultivars. Sexual Plant Reproduction, 15(1), 47-51. https://doi.org/10.1007/s00497-002-0137-5
Piotto, F. A., Batagin-Piotto, K. D., Almeida, M. D., & Oliveira, G. C. X. (2013). Interspecific xenia and metaxenia in seeds and fruits of tomato. Scientia Agricola, 70, 102-107.
Rubio-Piña, J. A., & Zapata-Pérez, O. (2011). Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants. Electronic journal of Biotechnology, 14(5), 11-11. DOI: 10.2225/vol14-issue5-fulltext-10    
Sabir, A. (2015). Xenia and metaxenia in grapes: differences in berry and seed characteristics of maternal grape cv.‘Narince’(Vitis vinifera L.) as influenced by different pollen sources. Plant Biology, 17(2), 567-573.  https://doi.org/10.1111/plb.12266
Shahsavari, M., Mohammadabadi, M., Khezri, A., Borshch, O., Babenko, O., Kalashnyk, O., ... & Kondratiuk, V. (2022). Effect of fennel (Foeniculum vulgare) seed powder consumption on insulin-like growth factor 1 gene expression in the liver tissue of growing lambs. Gene Expression, 21(2), 21-26.https://doi.org/10.14218/GE.2022.00017
Shmida, A., & Cohen, O. (1989). Rare plants in Israel. Amygdalus arabica. Teva Va ‘aretz, 31, 13-14.
Shemer, A., Biton, I., Many, Y., Vaknin, Y., Lavee, S., Avidan, B., & Ben-Ari, G. (2014). The olive cultivar ‘Picual’is an optimal pollen donor for ‘Barnea’. Scientia Horticulturae, 172, 278-284.
Shokri, S., Khezri, A., Mohammadabadi, M., & Kheyrodin, H. (2023). The expression of MYH7 gene in femur, humeral muscle, and back muscle tissues of fattening lambs of the Kermani breed. Agricultural Biotechnology Journal, 15(2), 217-236. https://doi.org/10.22103/jab.2023.21524.1486
Varkonyi-Gasic, E., Wu, R., Wood, M., Walton, E. F., & Hellens, R. P. (2007). Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant methods, 3(1), 12. doi:10.1186/1746-4811-3-12
Vazifeh-Shenas, M. R. (2015). Evaluation of pollination methods and xenia and metaxenia effects in pomegranate (Doctoral dissertation, Faculty of Agriculture, Ferdowsi University of Mashhad). [In Persian].
Yang, G., Zhou, R., Tang, T., & Shi, S. (2008). Simple and efficient isolation of high-quality total RNA from Hibiscus tiliaceus, a mangrove associate and its relatives. Preparative Biochemistry & Biotechnology, 38(3), 257-264. https://doi.org/10.1080/10826060802164991
Yew, C. W., & Kumar, S. V. (2010, July). MicroRNA regulates gene expression during fruit development in pineapple. In VII International Pineapple Symposium 902 (pp. 177-184).
Zhang, H., Yin, L., Wang, H., Wang, G., Ma, X., Li, M., ... & Yi, H. (2017). Genome-wide identification of Hami melon miRNAs with putative roles during fruit development. PLoS One, 12(7), e0180600. https://doi.org/10.1371/journal.pone.0180600.
مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 2
خرداد 1405
صفحه 1-20

فایل ها

هم رسانی

ارجاع به این مقاله

آمار