Abstract：The GATA transcription factor family contains a zinc finger protein conserved domain that recognizes and binds the 5'-(A/T) GATA (A/G)-3' motif to regulate the transcription and expression of multiple functional genes in eukaryotes. It is closely related to the growth and development of pathogenic fungi and the utilization of nitrogen sources. In order to explore the function of the GATA transcription factor family of Setosphaeria turcica, the Softberry database (http://linux1.softberry.com/berry.phtm) was employed to carry out genetic structure analysis, prediction of conserved domains and promoters; qRT-PCR was used to detect the effects of nitrogen source concentration levels and species on gene transcription levels of family members. The results showed that there were 7 members in the GATA transcription factor family of S. turcica. Compared with KNO3 as the nitrogen source, in the medium with NH4Cl as the only nitrogen source, the growth rate of hyphae was accelerated and the conidia production was reduced by about 15 times, and the transcription levels of all GATA transcription factors were increased by 2 to 4 times. Compared with the normal Richard's medium, when KNO3 content was 0, the growth rate of hyphae was increased and the yield of conidia was reduced by about 10 times. At the same time, the expression of all GATA transcription factors was increased by 20 to 40 times, indicating that GATA transcription factor expressions was highly induced under the completely restricted nitrogen source. The above results preliminarily clarified the response of GATA transcription factors to nitrogen metabolism, revealed the availability of nitrogen and the regulatory role of GATA transcription factors in the development of S. turcica, and provide new ideas for the control of northern corn leaf blight.
[1] 周晓罡, 姚春馨, 丁玉梅, 等. 2012. 氮源受限条件下植物病原真菌氮调控基因表达特性[J]. 遗传, 34(7): 848-856. (Zhou X G, Yao C X, Ding Y M, et al.2012. Progress on nitrogen regulation gene expression of plant pathogenic fungi under nitrogen starvation[J]. Hereditas, 34(7): 848-856.) [2] Bi F, Ment D, Luria N, et al.2017. Mutation of AREA affects growth, sporulation, nitrogen regulation, and pathogenicity in Colletotrichum gloeosporioides[J]. Fungal Genetics and Biology, 99: 29-39. [3] Brown S H, Yarden O, Gollop N, et al.2008. Differential protein expression in Colletotrichum acutatum: Changes associated with reactive oxygen species and nitrogen starvation implicated in pathogenicity on strawberry[J]. Molecular Plant Pathology, 9(2): 171. [4] Caretti G, Salsi V, Vecchi C, et al.2003. Dynamic recruitment of NF-Y and hats on cell-cycle prometers[J]. Journal of Biological Chemistry, 278(33): 30435-30440. [5] Coffman J A, Rai R, Loprete D M, et al.1997. Cross regulation of four GATA factors that control nitrogen catabolic gene expression in Saccharomyces cerevisiae[J]. Journal of Bacteriology, 179(11): 3416-3429. [6] Crespo J L, Daicho K, Ushimaru T, et al.2001. The GATA transcription factors GLN3 and GAT1 link TOR to salt stress in Saccharomyces cerevisiae[J]. Journal of Biological Chemistry, 276(37): 34441-34444. [7] Dangl J L.2007. Nibbling at the plant cell nucleus[J]. Science, 315(5815): 1088-1089. [8] Divon H H, Ziv C, Davydov O, et al.2006. The global nitrogen regulator, FNR1, regulates fungal nutrition-genes and fitness during Fusarium oxysporum pathogenesis[J]. Molecular Plant Pathology, 7(6): 485-497. [9] Donofrio N M, Oh Y, Lundy R, et al.2006. Global gene expression during nitrogen starvation in the rice blast fungus, Magnaporthe grisea[J]. Fungal Genetics and Biology, 43(9): 605-617. [10] Gimeno C J, Ljungdahl P O, Styles C A, et al.1992. Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: Regulation by starvation and RAS[J]. Cell, 68(6): 1077-1090. [11] Guan F, Pan Y, Li J, et al.2017. A GATA-type transcription factor AcAREB for nitrogen metabolism is involved in regulation of cephalosporin biosynthesis in Acremonium chrysogenum[J]. Science China Life Sciences, 60(9): 958-967. [12] Han K H, Han K Y, Yu J H, et al.2001. The nsdD gene encodes a putative GATA-type transcription factor necessary for sexual development of Aspergillus nidulans[J]. Molecular Microbiology, 41(2): 299-309. [13] Han X, Qiu M, Wang B, et al.2016. Functional analysis of the nitrogen metabolite repression regulator gene mrA in Aspergillus flavus[J]. Frontiers in Microbiology, 7: 1794. [14] Hoe K L, Won M S, Chung K S, et al.1998. Molecular cloning of Gaf1, a Schizosaccharomyces pombe GATA factor, which can function as a transcriptional activator[J]. Gene, 215(2): 319-328. [15] Kudla B, Caddick M X, Langdon T, et al.1990. The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger[J]. EMBO Journal, 9(5): 1355-1364. [16] Li J, Pan Y, Liu G.2013. Disruption of the nitrogen regulatory gene AcareA in Acremonium chrysogenum leads to reduction of cephalosporin production and repression of nitrogen metabolism[J]. Fungal Genetic Biology, 61: 69-79. [17] Lowry J A, Atchley W R.2000. Molecular evolution of the GATA family of transcription factors: Conservation within the DNA-Binding domain[J]. Journal of Molecular Evolution, 50(2): 103-115. [18] Margarita M G, Richard A. Wilson.2015. GATA-Dependent glutaminolysis drives appressorium formation in Magnaporthe oryzae by suppressing TOR Inhibition of cAMP/PKA signaling[J]. PLoS Pathogens, 11(4): e1004851. [19] Michielse C B, Pfannmüller A, Macios M, et al.2014. The interplay between the GATA transcription factors AreA, the global nitrogen regulator and AreB in Fusarium fujikuroi[J]. Molecular Microbiology, 91(3): 472-493. [20] Pfannmüller A, Leufken J, Studt L, et al.2017. Comparative transcriptome and proteome analysis reveals a global impact of the nitrogen regulators AreA and AreB on secondary metabolism in Fusarium fujikuroi[J]. PLOS ONE, 12(4): e0176194. [21] Punt P J, Strauss J, Smit R, et al.1995. The intergenic region between the divergently transcribed niiA and niaD genes of Aspergillus nidulans contains multiple NirA binding sites which act bidirectionally[J]. Molecular and Cellular Biology, 15(10): 5688-5699. [22] Quispe C F.2011. GATA-Family transcription factors in Magnaporthe oryzae[D]. Thesis for M.S., University of Ne‐braska-Lincoln, Supervisor: Wilson R A. pp,c3-16. [23] Smith D G, Garcia-Pedrajas M D, Gold S E, et al.2003. Isolation and characterization from pathogenic fungi of genes encoding ammonium permeases and their roles in dimorphism[J]. Molecular Microbiology, 50(1): 259. [24] Soundararajan S, Jedd G, Li X, et al.2004. Woronin body function in Magnaporthe grisea is essential for efficient pathogenesis and for survival during nitrogen starvation stress[J]. Plant Cell, 16(6): 1564-1574. [25] Wickes B L, Mayorga M E, Edman U, et al.1996. Dimorphism and haploid fruiting in Cryptococcus neoformans: Association with the alpha-mating type[J]. Proceedings of the National Academy of Sciences of the USA, 93(14): 7327. [26] Wilson R A, Arst H N.1998. Mutational analysis of AREA, a transcriptional activator mediating nitrogen metabolite repression in Aspergillus nidulans and a member of the "streetwise" GATA family of transcription factors[J]. Microbiology and Molecular Biology Reviews, 62(3): 586-596. [27] Xiao X, Marzluf G A.1996. Identification of the native NIT2 major nitrogen regulatory protein in nuclear extracts of Neurospora crassa[J]. Genetica, 97(2): 153-163. [28] Zwicker J, Gross C, Lucibello F C, et al.1995. Cell cycle regulation of cdc25C transcription is mediated by the periodic repression of the glutamine-rich activators NF-Y and Sp1[J]. Nucleic Acids Research, 23(19): 3822-3830.
