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Apache Tomcat

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    Clone of potato Rieske FeS gene and its expression analysis under drought stress
    YE He-jun, YE Chong-ze, YU Jing, ZHAO Rui-ying, GUO Zhi-ping
    HUBEI AGRICULTURAL SCIENCES    2025, 64 (3): 176-181.   DOI: 10.14088/j.cnki.issn0439-8114.2025.03.028
    Abstract28)      PDF (6337KB)(1)       Save
    The potato (Solanum tuberosum) was used as the research object. The cDNA sequence of the Rieske FeS gene of the potato was cloned by RT-PCR technology, and bioinformatics analysis was performed on the gene. The expression of the Rieske FeS gene under simulated drought stress (0, 12, 24 h) was detected by fluorescence quantitative PCR. The results showed that the full-length cDNA sequence of the potato Rieske FeS gene was 693 bp, containing one open reading frame and encoding 230 amino acids. The molecular weight of the potato Rieske FeS protein was 24.27 ku, with a theoretical isoelectric point of 8.20;the Rieske FeS protein contained the CytB6-F_Fe-S domain (59~97 aa), transmembrane region (72~94 aa), and Rieske domain (113~203 aa);the secondary structure of potato Rieske FeS protein had the highest proportion of irregular curls, followed by β-sheets, and the lowest proportion was α - helices. The phylogenetic tree analysis showed that the potato Rieske FeS protein was most closely related to the tobacco Rieske FeS protein. The qPCR results showed that the Rieske FeS gene of the potato was expressed in all tested tissues, with the highest relative expression level of Rieske FeS gene in leaves, followed by stems and roots, and the lowest relative expression level in tubers. After PEG drought stress, the relative expression level of Rieske FeS gene in leaves showed a decreasing trend followed by an increasing trend, and the relative expression level at 12 hours was significantly lower than that at 0 hours (control group). The relative expression level of Rieske FeS gene in stem segments was significantly upregulated after 12 hours of simulated drought treatment; the relative expression level of Rieske FeS gene was significantly upregulated in tubers under simulated drought stress for 24 hours; there was no significant difference in the relative expression levels of Rieske FeS gene among the drought treatments in the roots. The potato Rieske FeS gene was closely related to drought stress.
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    Establishment and optimization of Agrobacterium mediated genetic transformation system for Magnaporthe oryzae
    LI Shuang, ZHANG Shu-mei, TIAN Yuan, PAN Yu, LIU Wei, YAN Geng-xuan
    HUBEI AGRICULTURAL SCIENCES    2025, 64 (3): 182-189.   DOI: 10.14088/j.cnki.issn0439-8114.2025.03.029
    Abstract24)      PDF (7188KB)(1)       Save
    Magnaporthe oryzae was used as the experimental material. The GFP gene was obtained through PCR, and a pBarg-GFP-BAR-Amp recombinant vector was constructed. The Magnaporthe oryzae strain containing the GFP gene with stable inheritance was obtained using Agrobacterium-mediated transformation. The transformation conditions were optimized using single factor and response surface methodology. The optimal transformation conditions for the Magnaporthe oryzae genetic transformation system were an OD600 nm of 0.38 (Agrobacterium concentration), a co-culture temperature of 28.2 ℃, a co-culture time of 2.12 days, and a transformation efficiency of 280.00×10-5. PCR identification and fluorescence microscopy observation revealed that the GFP gene of the transformant Magnaporthe oryzae could be expressed normally, with no significant difference in pathogenicity compared to the wild-type Magnaporthe oryzae.
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