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Gene regulatory networks of kiwifruit response to Pseudomonas syringae pv. actinidiae

ABSTRACT: To study the responses of kiwifruit to Pseudomonas syringae pv. actinidiae, one-year-old potted seeding A. c. var. deliciosa cultivar ‘Jinkui’ and the pandemic Pseudomonas syringae pv. actinidiae bacterial strain JF8 (CCTCC AB2018305) were used for this study. This bacterial strain was originally isolated from A. c. var. chinensis cultivar ‘Jinfeng’ and further characterized . Plants were maintained in an aseptic room, with 95% of relative humidity, have natural light and no further fertilization after their receiving from the nursery. For inoculation, the P. s.pv. actinidiae strain was streaked on nutrient-sucrose agar (NSA) and incubated at 25 °C for 48-h. Ten microliters of a bacterial suspension (1-2×107cfu/mL) prepared in sterile 0.85 % w NaCl were inoculated in the plants chosen for investigation. The bacterial suspension was sprayed to entirety tree. In parallel, control plants were treated in the same way with sterile 0.85 % w NaCl solution. The inoculated and control plants were randomly distributed in the room at 15 ± 3 °C. 24-h after inoculation, ‘Jinkui’ leaves were sampled from the infected and control plants for further analyses. Each sample consisted of the leaves of one tree. Three biological replicates were used for each line.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Actinidia Chinensis Var. Chinensis

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Liu Pu

LAB HEAD: Pu Liu

PROVIDER: PXD014787 | Pride | 2021-11-10

REPOSITORIES: Pride

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Project description:To study basal resistance we inoculated tobacco plants with Pseudomonas syringae pv. syringae 61 hrcC mutants (gift of A. Collmer). To compare the expression profiles of basal resistance to hypersensitive response, we used P. syringae pv. syringae 61. Gene activation triggered by the live compatible pathogen P. syringae pv. tabaci was compared to kanamycine-inactivated P. tabaci (10 min in 100 ug/ml KM, then washed 2x in H2O). Water-infiltrated control leaves were used. The aim of another set of experiments was to investigate the influence of various signal transduction pathways on basal resistance. Control plants were infiltrated with P. syringae pv syringae hrcC mutant bacteria. These were compared to treatments, in which the bacterial suspensions were supplemented with inhibitors that can block a specific type of signal transduction pathway. The inhibitors represented Ca2+ signaling, lipid derived signals generated by phospholipase C and A, protein phosphorilation (kinases) and protein degradation (degradation of transcription repressors or activators). The used inhibitors were LaCl3, neomycin, aristolochic acid, K252a and MG115 respectively. 2-2.5 months old tobacco plants (Nicotiana tabacum cv. Samsun nn) were grown in a greenhouse in soil with cca. 16/8 hr light/dark period at 20°C, then in a growth chamber 2 days before and during experiments, under identical conditions. A syringe was used for the injection of bacteria or chemical solutions into the tobacco leaves. At 6 or 48 hours post inoculation, leaf samples were taken, and frozen in liquid nitrogen. Every experiment was carried out in 3 independent replicates. Total RNA was extracted using the Quiagen RNeasy Plant Mini kit and the Quiagen Rnase-free Dnase Set, then re-purified and concentrated on Microcon-30 (Millipore) columns. The concentration and quality of isolated RNA was estimated by measuring its absorbency at 260 and 280 nm and running on 1% agarose gel. Keywords: Direct comparison

			Action	DRS
	F19FTSCCKF0661_J190530011_rep1.dat	Other
	Rawdata_1.tar.gz	Other
	Rawdata_2.tar.gz	Other
	Rawdata_3.tar.gz	Other
	Rawdata_4.tar.gz	Other

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Gene regulatory networks of kiwifruit response to Pseudomonas syringae pv. actinidiae

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