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Experiment Name:

Comparison of wild-type and cell death mutant of barley plants containing Mla6 powdery mildew resistance gene

Accession No:

BB46

Microarray:

Barley1

Visibility:

public

Experiment Type:

Experiment Factor(s):

genotype: •Mla6 •11542
pathogen isolates: •5874 •K1
time: •0 •8 •16 •20 •24 •32

Quality Control:

Treatment summary:

genotype	pathogen isolates	time	# replicates
Mla6	5874	0	3
Mla6	5874	8	3
Mla6	5874	16	3
Mla6	5874	20	3
Mla6	5874	24	3
Mla6	5874	32	3
Mla6	K1	0	3
Mla6	K1	8	3
Mla6	K1	16	3
Mla6	K1	20	3
Mla6	K1	24	3
Mla6	K1	32	3
11542	5874	0	3
11542	5874	8	3
11542	5874	16	3
11542	5874	20	3
11542	5874	24	3
11542	5874	32	3
11542	K1	0	3
11542	K1	8	3
11542	K1	16	3
11542	K1	20	3
11542	K1	24	3
11542	K1	32	3

Total hybridizations: 72

Description:

Time-course expression profiles of Bgh challenged barley cultivar C.I. 16151 (harboring the Mla6 powdery mildew resistance allele) and its fast-neutron-derived "Bgh-induced tip cell death1" mutant, bcd1, were compared using the 22K Barley1 GeneChip.

Planting, stage of seedlings, harvesting, and experimental design were part of a larger experiment described by Caldo et al. (2004). PLEXdb BB4.

Experiment Design:
C.I. 16151 (wildtype) and bcd1 (mutant) were planted in separate 20 x 30-cm flats using sterilized potting soil. Each experimental flat consisted of six rows of 15 seedlings, with rows randomly assigned to one of six harvest time points (0, 8, 16, 20, 24, and 32 hai). Seedlings grown to the 1st leaf stage with 2nd leaf unfolded were inoculated with a high density of fresh conidiospores (84 +/- 19 spores/mm2). Groups of flats were placed at 18C (8-hour darkness, 16-hour light) in separate controlled growth chambers corresponding to the Bgh isolates. Rows of plants were harvested at each assigned time points and snap frozen in liquid nitrogen. The entire experiment was repeated three times in a standard split-split-plot design with 72 experimental units (2 genotypes x 2 pathogen isolates x 6 time points x 3 replications).

Treatment Description:
The samples constituted pairwise combinations of the the cultivar C.I. 16151(containing the Mla6 resistance allele), and its fast-neutron-derived "Bgh-induced tip cell death1" mutant, bcd1 with the two Bgh (Blumeria graminis f. sp. hordei) isolates, 5874 (AvrMla6, AvrMla1) and K1 (AvrMla13, AvrMla1).

For each replication, individual genotypes were planted in separate 20 x 30 cm flats using sterilized potting soil. Each experimental flat consisted of six rows of 15 seedlings, with rows randomly assigned to one of six harvest times (0, 8, 16, 20, 24, and 32 hai). Seedlings were grown to the 2nd-leaf stage with 1st leaf unfolded, and inoculation was performed at 4 PM Central Standard Time by tipping the flats at 45oC and dusting the plants with a high density of fresh conidiospores [84 +/- 19 spores/mm2]. This procedure was repeated from the opposite angle to ensure that a high proportion of the cells are in contact with the fungus. This conidial density per unit leaf area routinely results in greater than 50% of epidermal cells that are successfully infected. Groups of flats were placed at 18oC (8 hours darkness, 16 hours light, 8 hours darkness) in separate controlled growth chambers corresponding to the Bgh isolate. Rows of plants were harvested at their assigned harvest times and flash-frozen in liquid nitrogen.

This experiment has been submitted at NCBI GEO via PLEXdb.

Publication:

'Transcript-Based Cloning of RRP46, a Regulator of rRNA Processing and R Gene-Independent Cell Death in Barley-Powdery Mildew Interactions.', Xi L, Moscou MJ, Meng Y, Xu W, Caldo RA, Shaver M, Nettleton D, Wise RP.
The Plant Cell 21:3280-3295 (2009)
pubmed: 19861556
'Interaction-Dependent Gene Expression in Mla-Specified Response to Barley Powdery Mildew', Rico A. Caldo, Dan Nettleton and Roger P. Wise
Plant Cell. 16:2514-2528 (2004)
pubmed: 15319481

Created:

2006-07-18 20:59:24

Last Update:

2008-01-24 14:33:14

GEO Accession

GSE14930

Submitter:

Roger P Wise

Experimenter:

Name:	Roger P Wise
Institution:	Iowa State University
Head of Laboratory:	Roger P Wise
email(s):	rpwise@iastate.edu
Homepage:


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