A study of PTF1 interaction with phosphorus stress inducing genes and its influence on root architecture of transgenic Arabidopsis

Aqsa Akhtar, Muhammad Irfan, Asma Maqbool, Kauser Abdulla Malik

Abstract


Background: Phosphorus; an essential macronutrient needed by the plant for its robust growth is inaccessible to the plant as required. Hence, a need arises to develop smart crops capable of utilizing maximum phosphorus from soil. PTF1(Inorganic Phosphorus Starvation Induced Transcription Factor 1) is overexpressed during phosphorus stress and regulates number of genes to combat this abiotic stress.

Methods: The current study is the first ever reported case of transforming Arabidopsis thaliana with plant expression binary vector pSB219 harboring PTF1 via floral dip method and analyzing phosphorus stress induced genes interaction, through yeast-one-hybrid. Yeast-one-hybrid analysis was performed on four selected genes namely LPR1 (Low Phosphate Response), PDR2 (Phosphate Deficiency Response 2), PHT1;2 (Phosphate Transporter) and RNS1 (Ribonuclease). The positive transformed lines were expression analyzed for PTF1 by real time PCR and further studied for their root morphology.

Results: The results clearly showed direct interaction of LPR1 with PTF1 while other genes, although being overexpressed, were indirectly regulated. Transformation efficiency of 1% was achieved and a maximum 2.5-fold increase in PTF1 expression was observed. Root morphological studies exhibited significantly enhanced root hair and lateral surface area when grown in phosphorus deficient MS medium.

Conclusion: The results of the current study may pave path for improved comprehension of gene interactions and root architecture modifications under phosphorus limiting conditions.

Keywords: Phosphorus; Arabidopsis; Yeast one hybrid; Roots 


Full Text:

PDF

References


Baker A, Ceasar SA, Palmer AJ, Paterson JB, Qi W, et al. Replace, reuse, recycle: improving the sustainable use of phosphorus by plants. Journal of experimental botany, (2015); 66(12): 3523-3540.

Tam THY, Catarino B, Dolan L. Conserved regulatory mechanism controls the development of cells with rooting functions in land plants. Proceedings of the National Academy of Sciences, (2015); 112(29): E3959-E3968.

Taliman NA, Dong Q, Echigo K, Raboy V, Saneoka H. Effect of phosphorus fertilization on the growth, photosynthesis, nitrogen fixation, mineral accumulation, seed yield, and seed quality of a soybean low-phytate line. Plants, (2019); 8(5): 119.

Sun B, Gao Y, Wu X, Ma H, Zheng C, et al. The relative contributions of pH, organic anions, and phosphatase to rhizosphere soil phosphorus mobilization and crop phosphorus uptake in maize/alfalfa polyculture. Plant and Soil, (2020); 447(1): 117-133.

Antoniadis V, Koliniati R, Efstratiou E, Golia E, Petropoulos S. Effect of soils with varying degree of weathering and pH values on phosphorus sorption. Catena, (2016); 139214-219.

Abid N, Khatoon A, Maqbool A, Irfan M, Bashir A, et al. Transgenic expression of phytase in wheat endosperm increases bioavailability of iron and zinc in grains. Transgenic research, (2017); 26(1): 109-122.

Lu M, Ying S, Zhang D-F, Shi Y-S, Song Y-C, et al. A maize stress-responsive NAC transcription factor, ZmSNAC1, confers enhanced tolerance to dehydration in transgenic Arabidopsis. Plant Cell Reports, (2012); 31(9): 1701-1711.

Yan H, Jia H, Chen X, Hao L, An H, et al. The Cotton WRKY Transcription Factor GhWRKY17 Functions in Drought and Salt Stress in Transgenic Nicotiana benthamiana Through ABA Signaling and the Modulation of Reactive Oxygen Species Production. Plant and Cell Physiology, (2014); 55(12): 2060-2076.

Li Z, Liu C, Zhang Y, Wang B, Ran Q, et al. The bHLH family member ZmPTF1 regulates drought tolerance in maize by promoting root development and abscisic acid synthesis. Journal of experimental botany, (2019); 70(19): 5471-5486.

Li X, Wu B, Kong Y, Zhang C. GmPTF1, a novel transcription factor gene, is involved in conferring soybean tolerance to phosphate starvation. Genetics and Molecular Research, (2014); 13(1): 926-937.

Yi K, Wu Z, Zhou J, Du L, Guo L, et al. OsPTF1, a novel transcription factor involved in tolerance to phosphate starvation in rice. Plant physiology, (2005); 138(4): 2087-2096.

Janes G, von Wangenheim D, Cowling S, Kerr I, Band L, et al. Cellular patterning of Arabidopsis roots under low phosphate conditions. Frontiers in plant science, (2018); 9735.

Lopato S, Bazanova N, Morran S, Milligan AS, Shirley N, et al. Isolation of plant transcription factors using a modified yeast one-hybrid system. Plant Methods, (2006); 2(1): 3.

Breton G, Kay SA, Pruneda-Paz JL (2016) Identification of Arabidopsis transcriptional regulators by yeast one-hybrid screens using a transcription factor ORFeome. Environmental Responses in Plants: Springer. pp. 107-118.

Reece-Hoyes JS, Walhout AM. Yeast one-hybrid assays: a historical and technical perspective. Methods, (2012); 57(4): 441-447.

Castrillo G, Turck F, Leveugle M, Lecharny A, Carbonero P, et al. Speeding cis-trans regulation discovery by phylogenomic analyses coupled with screenings of an arrayed library of Arabidopsis transcription factors. PloS one, (2011); 6(6): e21524.

Xu JM, Wang ZQ, Wang JY, Li PF, Jin JF, et al. Low phosphate represses histone deacetylase complex1 to regulate root system architecture remodeling in Arabidopsis. New Phytologist, (2020); 225(4): 1732-1745.

Abel S. Phosphate sensing in root development. Current opinion in plant biology, (2011); 14(3): 303-309.

Liu X, Zhao X, Zhang L, Lu W, Li X, et al. TaPht1; 4, a high-affinity phosphate transporter gene in wheat (Triticum aestivum), plays an important role in plant phosphate acquisition under phosphorus deprivation. Functional Plant Biology, (2013); 40(4): 329-341.

Jost R, Pharmawati M, Lapis-Gaza HR, Rossig C, Berkowitz O, et al. Differentiating phosphate-dependent and phosphate-independent systemic phosphate-starvation response networks in Arabidopsis thaliana through the application of phosphite. Journal of experimental botany, (2015); 66(9): 2501-2514.

Dong J, Piñeros MA, Li X, Yang H, Liu Y, et al. An Arabidopsis ABC transporter mediates phosphate deficiency-induced remodeling of root architecture by modulating iron homeostasis in roots. Molecular plant, (2017); 10(2): 244-259.

Ma Z, Bielenberg D, Brown KM, Lynch JP. Regulation of root hair density by phosphorus availability in Arabidopsis thaliana. Plant, Cell & Environment, (2001); 24(4): 459-467.

Clough SJ, Bent AF. Floral dip: a simplified method for Agrobacterium -mediated transformation of Arabidopsis thaliana. The Plant Journal, (1998); 16(6): 735-743.

Li C, Tao Y, Yang Y, Xiang Y, Li G. In Vitro Analysis of DNA–Protein Interactions in Gene Transcription Using DNAzyme-Based Electrochemical Assay. Analytical chemistry, (2017); 89(9): 5003-5007.

Niranjani G, Murugan R. Generalized theory on the mechanism of site-specific DNA–protein interactions. Journal of Statistical Mechanics: Theory and Experiment, (2016); 2016(5): 053501.

Gordân R, Shen N, Dror I, Zhou T, Horton J, et al. Genomic regions flanking E-box binding sites influence DNA binding specificity of bHLH transcription factors through DNA shape. Cell reports, (2013); 3(4): 1093-1104.

Sun X, Wang Y, Sui N. Transcriptional regulation of bHLH during plant response to stress. Biochemical and biophysical research communications, (2018); 503(2): 397-401.

Kang K, Chung JH, Kim J. Evolutionary Conserved Motif Finder (ECMFinder) for genome-wide identification of clustered YY1-and CTCF-binding sites. Nucleic acids research, (2009); 37(6): 2003-2013.

Wang T, Kettenbach AN, Gerber SA, Bailey-Kellogg C. MMFPh: a maximal motif finder for phosphoproteomics datasets. Bioinformatics, (2012); 28(12): 1562-1570.

Schmidt MC, Rocha AM, Padmanabhan K, Chen Z, Scott K, et al. Efficient α, β-motif finder for identification of phenotype-related functional modules. BMC Bioinformatics, (2011); 12(1): 440.

Kasas S, Dietler G. DNA-protein interactions explored by atomic force microscopy; 2018. Elsevier. pp. 231-239.

Frykholm K, Nyberg LK, Westerlund F. Exploring DNA–protein interactions on the single DNA molecule level using nanofluidic tools. Integrative Biology, (2017); 9(8): 650-661.

Sewell JA, Bass JIF (2018) Options and considerations when using a yeast one-hybrid system. Two-Hybrid Systems: Springer. pp. 119-130.

Fukushima A, Iwasa M, Nakabayashi R, Kobayashi M, Nishizawa T, et al. Effects of combined low glutathione with mild oxidative and low phosphorus stress on the metabolism of Arabidopsis thaliana. Frontiers in plant science, (2017); 81464.

Cai Q, Ji C, Yan Z, Jiang X, Fang J. Anatomical responses of leaf and stem of Arabidopsis thaliana to nitrogen and phosphorus addition. Journal of Plant Research, (2017); 130(6): 1035-1045.

Wang F, Ding D, Li J, He L, Xu X, et al. Characterisation of genes involved in galactolipids and sulfolipids metabolism in maize and Arabidopsis and their differential responses to phosphate deficiency. Functional Plant Biology, (2020); 47(4): 279-292.

Fabiańska I, Gerlach N, Almario J, Bucher M. Plant-mediated effects of soil phosphorus on the root-associated fungal microbiota in Arabidopsis thaliana. New Phytologist, (2019); 221(4): 2123-2137.

Gutiérrez-Alanís D, Ojeda-Rivera JO, Yong-Villalobos L, Cárdenas-Torres L, Herrera-Estrella L. Adaptation to phosphate scarcity: tips from Arabidopsis roots. Trends in plant science, (2018); 23(8): 721-730.

Kisko M, Shukla V, Kaur M, Bouain N, Chaiwong N, et al. Phosphorus transport in Arabidopsis and wheat: Emerging strategies to improve P pool in seeds. Agriculture, (2018); 8(2): 27.

Wu L, Kobayashi Y, Wasaki J, Koyama H. Organic acid excretion from roots: a plant mechanism for enhancing phosphorus acquisition, enhancing aluminum tolerance, and recruiting beneficial rhizobacteria. Soil Science and Plant Nutrition, (2018); 64(6): 697-704.

Kong Y, Wang B, Du H, Li W, Li X, et al. GmEXLB1, a soybean expansin-like B gene, alters root architecture to improve phosphorus acquisition in Arabidopsis. Frontiers in Plant Science, (2019);

Janes G (2018) Factors regulating cortex cell file proliferation under low phosphorus stress in Arabidopsis thaliana roots: University of Nottingham.




DOI: http://dx.doi.org/10.62940/als.v9i1.1211

Refbacks

  • There are currently no refbacks.