Rice Science ›› 2022, Vol. 29 ›› Issue (3): 268-276.DOI: 10.1016/j.rsci.2021.09.004
• Research Paper • Previous Articles Next Articles
Wang Chenjiaozi, Zhao Mei, Shu Canwei, Zhou Erxun()
Received:
2021-07-12
Accepted:
2021-09-17
Online:
2022-05-28
Published:
2022-03-10
Contact:
Zhou Erxun
Wang Chenjiaozi, Zhao Mei, Shu Canwei, Zhou Erxun. Three Genes Related to Trehalose Metabolism Affect Sclerotial Development of Rhizoctonia solani AG-1 IA, Causal Agent of Rice Sheath Blight[J]. Rice Science, 2022, 29(3): 268-276.
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Fig. 1. Gene relative expression, Western blot and enzyme activities of Rstre, Rstps1 and Rstpp in different developmental stages of sclerotia in Rhizoctonia solani AG-1 IA. A, Relative expression levels of Rstre, Rstps1 and Rstpp at different sclerotial development stages. Samples were collected at seven different time points (36, 48, 60, 72, 120, 168 and 336 h of strain culture) during the sclerotial development. Data were normalized to transcripts encoding GAPDH, and shown as Mean ± SE (n = 3). B, Western blot of RSTPS1 and RSTPP at different sclerotial development stages. Anti- GAPDH was used as an internal control. C, Enzyme activities of RSTRE, RSTPS1 and RSTPP at different sclerotial development stages. Data are Mean ± SE (n = 3).
Fig. 2. Functional verification of Rstre (A), Rstps1 (B) and Rstpp (C) in Pichia pastoris GS115. pPIC9K-Rstre, pPIC9K-Rstps1 and pPIC9K-Rstpp are shown in red lines, and GS115 strain containing pPIC9K vector shown in blue line was used as a control. Solvent peak, glucose peak, trehalose peak and maltose peak are represented by blue, red, green and black arrows, respectively. nRIU is the refraction coefficient unit of differential refractive detector.
Fig. 3. Reactive oxygen species (ROS) contents in Pichia pastoris GS115 transformants of Rstre (A), Rstps1 (B) and Rstpp (C). ROS measurements were performed by the fluorescence assay. Mycelia were stained with 2’,7’-dichloro-dihydro-fluorescein diacetate for 20 min and visualized under a fluorescence microscope at 405 nm. Data are Mean ± SE (n = 3). * and ** indicate significant differences at P < 0.05 and P < 0.01 by the Duncan’s multiple range test.
Fig. 4. Effects of fermentation broths of Pichia pastoris GS115 transformants of Rstre, Rstps1 and Rstpp on the amounts of sclerotia. A?D, Morphology of sclerotia on potato dextrose agar plate containing GS115 fermentation broth (A), GS115-Rstre yeast fermentation broth (B), GS115-Rstps1 yeast fermentation broth (C) and GS115-Rstpp yeast fermentation broth (D). E, Effect of fermentation broth of yeast inverters on sclerotia of R. solani. Sclerotial di?erentiation (SDIF) was defined as percent of dry weight of sclerotia (in respect to 100% SDIF of the control) per dish. Data are Mean ± SE (n = 3). * indicates significant difference at P < 0.05 by the Duncan's multiple range test.
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