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StKU80, a component in the NHEJ repair pathway, is involved in mycelial morphogenesis, conidiation, appressorium development, and oxidative stress reactions in Exserohilum turcicum
GONG Xiao-dong, LIU Yu-wei, BI Huan-huan, YANG Xiao-rong, HAN Jian-min, DONG Jin-gao, GU Shou-qin
2021, 20 (1): 147-158.   DOI: 10.1016/S2095-3119(20)63233-6
Abstract133)      PDF in ScienceDirect      
Homologous recombination (HR) and nonhomologous end joining (NHEJ) are considered the two main double-strand break (DSB) repair approaches in eukaryotes.  Inhibiting the activities of the key component in NHEJ commonly enhances the efficiency of targeted gene knockouts or affects growth and development in higher eukaryotes.  However, little is known about the roles of the NHEJ pathway in foliar pathogens.  Here we identified a gene designated StKU80, which encodes a putative DNA end-binding protein homologous to yeast Ku80, in the foliar pathogen Exserohilum turcicum.  Conserved domain analysis showed that the typical domains VWA, Ku78 and Ku-PK-bind are usually present in Ku70/80 proteins in eukaryotes and are also present in StKu80.  Phylogenetic analysis indicated that StKu80 is most closely related to Ku80 (XP_001802136.1) from Parastagonospora nodorum, followed by Ku80 (AGF90044.1) from Monascus ruber.  Furthermore, the gene knockout mutants ΔStKU80-1 and ΔStKU80-2 were obtained.  These mutants displayed longer septas, thinner cell walls, smaller amounts of substances on cell wall surfaces, and more mitochondria per cell than the wild-type (WT) strain but similar HT-toxin activity.  The mutants did not produce conidia and mature appressoria.  On the other hand, the mutants were highly sensitive to H2O2, but not to ultraviolet radiation.  In summary, the StKU80 plays devious roles in regulating the development of E.?turcicum.
 
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MAP kinase gene STK1 is required for hyphal, conidial, and appressorial development, toxin biosynthesis, pathogenicity, and hypertonic stress response in the plant pathogenic fungus Setosphaeria turcica
LI Po, GONG Xiao-dong, JIA Hui, FAN Yong-shan, ZHANG Yun-feng, CAO Zhi-yan, HAO Zhi-min, HAN Jian-min, GU Shou-qin, DONG Jin-gao
2016, 15 (12): 2786-2794.   DOI: 10.1016/S2095-3119(16)61472-7
Abstract1188)      PDF in ScienceDirect      
The mitogen-activated protein kinase (MAPK), a key signal transduction component in the MAPK cascade pathway, regulates a variety of physiological activities in eukaryotes.  However, little is known of the role MAPK plays in phytopathogenic fungi.  In this research, we cloned the MAPK gene STK1 from the northern corn leaf blight pathogen Setosphaeria turcica and found that the gene shared high homology with the high osmolality glycerol (HOG) MAPK gene HOG1 of Saccharomyces cerevisiae.  In addition, gene knockout technology was employed to investigate the function of STK1.  Gene knockout mutants (KOs) were found to have altered hyphae morphology and no conidiogenesis, though they did show similar radial growth rate compared to the wild-type strain (WT).  Furthermore, microscope observations indicated that STK1 KOs did not form normal appressoria at 48 h post-inoculation on a hydrophobic surface.  STK1 KOs had reduced virulence, a significantly altered Helminthosporium turcicum (HT)-toxin composition, and diminished pathogenicity on the leaves of susceptible inbred corn OH43.  Mycelium morphology appeared to be significantly swollen and the radial growth rates of STK1 KOs declined in comparison with WT under high osmotic stress.  These results suggested that STK1 affects the hyphae development, conidiogenesis, and pathogenicity of S. turcica by regulating appressorium development and HT-toxin biosynthesis.  Moreover, the gene appears to be involved in the hypertonic stress response in S. turcica.
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Stk2, a Mitogen-Activated Protein Kinase from Setosphaeria turcica, Specifically Complements the Functions of the Fus3 and Kss1 of Saccharomyces cerevisiae in Filamentation, Invasive Growth, and Mating Behavior
GU Shou-qin, YANG Yang, LI Po, ZHANG Chang-zhi, FAN Yu, ZHANG Xiao-yu, TIAN Lan
2013, 12 (12): 2209-2216.   DOI: 10.1016/S2095-3119(13)60296-8
Abstract2429)      PDF in ScienceDirect      
Setosphaeria turcica, an essential phytopathogenic fungus, is the primary cause of serious yield losses in corn; however,
its pathogenic mechanism is poorly understood. We cloned STK2, a newly discovered mitogen-activated protein kinase gene with a deduced amino acid sequence that is 96% identical to MAK2 from Phaeosphaeria nodorum, 56% identical to
KSS1 and 57% identical to FUS3 from Saccharomyces cerevisiae. To deduce Stk2 function in S. turcica and to identify the
genetic relationship between STK2 and KSS1/FUS3 from S. cerevisiae, a restructured vector containing the open reading
frame of STK2 was transformed into a fus3/kss1 double deletion mutant of S. cerevisiae. The results show that the STK2
complementary strain clearly formed pseudohyphae and ascospores, and the strain grew on the surface of the medium after
rinsing with sterile water and the characteristics of the complementary strain was the same as the wild-type strain. Moreover,
STK2 complemented the function of KSS1 in filamentation and invasive growth, as well as the mating behavior of FUS3 in
S. cerevisiae, however, its exact functions in S. turcica will be studied in the future research.
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