تنظیم بیان ژن سوکروز-فسفات فسفاتاز در گندم نان و گونه های اجدادی آن تحت تنش شوری

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد گروه بیوتکنولوژی دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، کرمان، ایران.

2 استادیار گروه بیوتکنولوژی دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، کرمان، ایران

3 دانشیار گروه بیوتکنولوژی دانشکده کشاورزی، دانشگاه شهید باهنر کرمان، کرمان، ایران

چکیده

چکیده
هدف
سوکروز یا ساکارز محصول اصلی فتوسنتز در گیاهان می‏باشد که نقش‏های مهمی در رشد، نمو، ذخیره انرژی، انتقال سیگنال و سازگاری با تنش‌های محیطی دارد. ژن سوکروز فسفات فسفاتاز (SPP)، آخرین مرحله در مسیر بیوسنتز سوکروز که یک واکنش غیر برگشت پذیر است را کاتالیز می‌کند. در این پژوهش ژن‌های اورتولوگ SPP در گندم نان و گونه‌های اجدادی آن شناسایی و انتخاب شد و تغییرات میزان بیان آنها در پاسخ به تنش شوری مورد ارزیابی قرار گرفت.
مواد و روش‌ها
بذور گندم نان (Triticum aestivum CV. Rooshan) ، گندم دوروم (Triticum turgidum CV. Hana) و آجیلوپس (Aegilops tauschii) از موسسه تحقیقات اصلاح و تهیه نهال و بذر کرج تهیه شد. استخراج RNA از بافت‌های ریشه و برگ گیاهان کنترل و تحت تیمار (NaCl 200 mM) انجام شد و تغییرات بیان با روش qRT-PCR مورد ارزیابی قرار گرفت. به منظور مطالعه رخدادهای تکاملی و شناسایی روابط فیلوژنی بین اورتولوگ‌ها و پارالوگ‌های ژن SPP، درخت ژنی رسم شد. با استفاده از ابزار آنلاین psRNATarget امکان تنظیم پس از رونویسی با واسطه miRNA ها نیز مورد ارزیابی قرار گرفت.
نتایج
نتایج آنالیز فیلوژنی نشان داد که بر روی کروموزوم‌های 1 و 5 گندم نان، گندم دوروم و آجیلوپس به ترتیب 6، 4 و 2 ژن پارالوگ SPP تکامل یافته است. هم‌چنین مشخص شد که 16 miRNA متفاوت می‌توانند برخی از رونوشت ژن‌های SPP را در گندم نان و آجیلوپس، به عنوان ژن هدف شناسایی نمایند. نتایج تغییرات بیان ژن نیز نشان داد که در بافت ریشه و برگ گندم نان در پاسخ به تنش شوری بیان ژن SPP افزایش یافت اما در گندم دوروم کاهش بیان ژن SPP مشاهده شد. واکنش گیاه آجیلوپس به تنش شوری متفاوت بود به طوری که در بافت برگ کاهش بیان ولی در بافت ریشه افزایش بیان ژن SPP مشاهده شد.
نتیجه‌گیری
تنظیم پس از رونویسی با واسطه miRNA ها می‌تواند نقش مهمی در تنظیم متابولیسم پیچیده سوکروز و بطور ویژه تنظیم بیان ژن SPP داشته باشد. نتایج بیان ژن نشان داد که گیاه گندم نان تحت شرایط تنش شوری تولید سوکروز را افزایش داد در حالی که در گندم دوروم که حساسیت بیشتری به تنش شوری دارد تحت شرایط تنش شوری تولید سوکروز مختل و یا کاهش یافت که این کاهش تولید سوکروز در کنار سایر اثرات مخرب تنش شوری در کاهش عملکرد نقش خواهد شد.

کلیدواژه‌ها


عنوان مقاله [English]

Expression regulation of sucrose-phosphate phosphatase gene in bread wheat and its progenitors under salt stress

نویسندگان [English]

  • Elham Obeidnejad 1
  • Mehdi Mansouri 2
  • Hamid Reza Kavousi 3
1 MSc. Student,, Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
2 Assistant Professor, Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
3 Associate Professor, Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran
چکیده [English]

Abstract
Objective
Sucrose, the final product of photosynthesis, plays important roles in growth, development, storage, signal transduction, and acclimation to environmental stresses in higher plants. Sucrose-phosphate phosphatase (SPP) catalyzes the final irreversible step in the sucrose biosynthesis pathway. In this study, the SPP orthologous genes in wheat and progenitors were selected, and the expression regulation of the orthologous SPP genes were analyzed in response to salinity stress.
Materials and methods
Seeds of bread wheat (Triticum aestivum CV. Rooshan), durum wheat (Triticum turgidum CV. Hana), and Aegilops tauschii were provided by the Seed and Plant Improvement Institute, Karaj, Iran. In order to quantify the expression of SPP genes, qRT-PCR was performed on total RNA extracted from leaf and root tissues of control and salt-treated (NaCl 200 mM) samples. A gene tree was constructed to calculate the evolutionary and phylogenetic relationships of the SPP protein family in bread wheat and its progenitors. The possibility of post-transcriptional regulation of the SPP genes was also evaluated using the online tool psRNATarget.
Results
The results of the phylogenetic analysis showed that 6, 4, and 2 SPP paralogous genes have evolved on chromosomes 1 and 5 of bread wheat, durum wheat, and Ae. tauschii, respectively. Additionally, We predicted 16 different microRNAs can target some of the SPP gene transcripts in bread wheat and Ae. tauschii. Gene expression results also indicated that SPP gene expression increased in the root and leaf tissues of bread wheat in response to salinity stress, while it decreased in durum wheat. The response of Ae. tauschii to salinity stress was different, with decreased expression observed in leaf tissue but increased expression in root tissue.
Conclusion
Post-transcriptional regulation through miRNAs can play an important role in the regulation of the complex sucrose metabolism, especially in the regulation of SPP gene expression. Gene expression results showed that under salinity stress conditions, bread wheat increases sucrose production, while in durum wheat, which is more sensitive to salinity stress, sucrose production is disrupted or decreased. This reduction in sucrose production can lead to a decrease in yield.

کلیدواژه‌ها [English]

  • Sucrose
  • Gene expression
  • Salinity stress
Annunziata MG, Ciarmiello LF, Woodrow P et al. (2017) Durum Wheat Roots Adapt to Salinity Remodeling the Cellular Content of Nitrogen Metabolites and Sucrose. Front Plant Sci 7, 2035.
Bonnet E, Wuyts J, Rouzé P et al. (2004) Detection of 91 potential conserved plant microRNAs in Arabidopsis thaliana and Oryza sativa identifies important target genes. Proc Natl Acad Sci 101, 11511-11516.
Chen X, Xia J, Xia Z et al. (2015) Potential functions of microRNAs in starch metabolism and development revealed by miRNA transcriptome profiling of cassava cultivars and their wild progenitor. BMC Plant Biol 15, 33.
Consortium IWGS, Mayer KF, Rogers J et al. (2014) A chromosome-based draft sequence of the hexaploid bread wheat (Triticum aestivum) genome. Science 345, 1251788.
Dubcovsky J, Dvorak J (2007) Genome plasticity a key factor in the success of polyploid wheat under domestication. Science 316, 1862-1866.
Duncan R (2000) Plant tolerance to acid soil constraints: genetic resources, breeding methodology, and plant improvement. In: plant-Environment Interactions. CRC Press. pp. 19-56.
Eckardt NA (2014) Genome dominance and interaction at the gene expression level in allohexaploid wheat. Plant Cell 26, 1834.
Fileccia V, Bertolini E, Ruisi P et al. (2017) Identification and characterization of durum wheat microRNAs in leaf and root tissues. Funct Integr Genomics 17, 583-598.
Guo R, Yang Z, Li F et al. (2015) Comparative metabolic responses and adaptive strategies of wheat (Triticum aestivum) to salt and alkali stress. BMC Plant Biol 15, 170.
Halford N, Curtis T, Muttucumaru N et al. (2011) Sugars in crop plants. Ann Appl Biol 158, 1-25.
Han R, Jian C, Lv J et al. (2014) Identification and characterization of microRNAs in the flag leaf and developing seed of wheat (Triticum aestivum L.). BMC Genom 15, 1-14.
Hou G, Du C, Gao H et al. (2020) Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels. BMC Plant Biol 20, 87.
Jiang S-Y, Chi Y-H, Wang J-Z et al. (2015) Sucrose metabolism gene families and their biological functions. Sci Rep 5, 17583.
Jibiki M, Kuno Y, Shinoyama H et al. (1993) Isolation and properties of large cell strains from a methanol-utilizing yeast, Candida sp. N-16 by colchicine treatment. J Gen Appl Microbiol 39, 439-442.
Jing F, Miao Y, Zhang P et al. (2022) Characterization of TaSPP-5A gene associated with sucrose content in wheat (Triticum aestivum L.). BMC Plant Biol 22, 58.
Leloir LF, Cardini C (1955) The biosynthesis of sucrose phosphate. J Biol Chem 214, 157-165.
Li Y-F, Wei K, Wang M et al. (2019) Identification and temporal expression analysis of conserved and novel microRNAs in the leaves of winter wheat grown in the field. Front Genet 10, 779.
Li Y, Wang L-F, Bhutto SH et al. (2021) Blocking miR530 Improves Rice Resistance, Yield, and Maturity. Front Plant Sci 12, 729560.
Lunn JE (2003) Sucrose-phosphatase gene families in plants. Gene 303, 187-196.
Mansouri M, Naghavi MR, Alizadeh H et al. (2019) Transcriptomic analysis of Aegilops tauschii during long-term salinity stress. Funct Integr Genomics 19, 13-28.
Meng F, Liu H, Wang K et al. (2013) Development-associated microRNAs in grains of wheat (Triticum aestivum L.). BMC Plant Biol 13, 140.
Munns R, James RA (2003) Screening methods for salinity tolerance: a case study with tetraploid wheat. Plant Soil 253, 201-218.
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59, 651-681.
Murat F, Zhang R, Guizard S et al. (2013) Shared subgenome dominance following polyploidization explains grass genome evolutionary plasticity from a seven protochromosome ancestor with 16K protogenes. Genome Biol Evol 6, 12-33.
Orcutt DM, Nilsen ET (2000) Physiology of Plants Under Stress: Soil and Biotic Factors. Wiley.
Peng T, Sun H, Du Y et al. (2013) Characterization and expression patterns of microRNAs involved in rice grain filling. PloS one 8, e54148.
Pessarakli M, Szabolcs I (1999) Soil salinity and sodicity as particular plant/crop stress factors. Handbook of plant and crop stress 1, 1-16.
Petersen G, Seberg O, Yde M et al. (2006) Phylogenetic relationships of Triticum and Aegilops and evidence for the origin of the A, B, and D genomes of common wheat (Triticum aestivum). Mol Phylogenet Evol 39, 70-82.
Sharifi-Sirchi G, Beheshti B, Hosseinipour A et al. (2011) Priming against Asiatic citrus canker and monitoring of PR genes expression during resistance induction. Afr J Biotechnol 10, 3818-3823.
Storey KB, Tanino KK (2012) Temperature adaptation in a changing climate: nature at risk. CABI.
Vilella AJ, Severin J, Ureta-Vidal A et al. (2009) EnsemblCompara GeneTrees: Complete, duplication-aware phylogenetic trees in vertebrates. Genome Res 19, 327-335.
Xie F, Frazier TP, Zhang B (2011) Identification, characterization and expression analysis of MicroRNAs and their targets in the potato (Solanum tuberosum). Gene 473, 8-22.
Xu T, Zhang L, Yang Z et al. (2021) Identification and functional characterization of plant MiRNA under salt stress shed light on salinity resistance improvement through MiRNA manipulation in crops. Front Plant Sci 12, 665439.
Xue L-J, Zhang J-J, Xue H-W (2008) Characterization and expression profiles of miRNAs in rice seeds. Nucleic Acids Res 37, 916-930.
Yang C, Zhao L, Zhang H et al. (2014) Evolution of physiological responses to salt stress in hexaploid wheat. Proc Natl Acad Sci 111, 11882-11887.
Zhang Y-C, Yu Y, Wang C-Y et al. (2013) Overexpression of microRNA OsmiR397 improves rice yield by increasing grain size and promoting panicle branching. Nat Biotechnol 31, 848-852.