Biology Reference
In-Depth Information
With the above procedure were identifi ed 25 from 78 variable
spots. This low rate of identifi cation is due to the lack of reported
sequences for carnation in the NCBInr. A specifi c species database
construction using results of large-scale sequencing of RNA and
DNA, as well as its corresponding annotation, is necessary in order
to development researches in plant species poorly studied at pro-
teomic level as carnation. Despite that, we have identifi ed some con-
stitutive proteins associated with the resistant phenotype, such as a
class III peroxidase and a NB-ARC resistance protein (Nucleotide
Binding domain shared by Apaf-1, certain R gene products, and
CED-4 fused to C-terminal leucine-rich repeats). Likewise, a differ-
ential protein accumulation associated to carbohydrate metabolism
reorganization was found in the resistant cultivar during pathogen
infection at root level. The next step will be focused to develop a
specifi c database for identifying those non-identifi ed remaining pro-
teins which are differentially accumulated in the resistant cultivar,
and fi nally elucidate the principal earliest resistant mechanisms acting
in carnation against
Fusarium oxysporum
f. sp.
dianthi
.
4
Notes
1. The most appropriate experimental approach should be
selected according to the type of pathogen and previous stud-
ies about the plant-pathogen interaction model. Considering
previous studies with
F. oxysporum
f. sp.
dianthi
, an in vivo
inoculation assay with the pathogen was chosen [
15
].
2. At least two cultivars differing in terms of resistance to the patho-
gen must be selected for the in vivo inoculation assay. A certain
number of plants must be used to obtain, at least, biological
triplicates for each treatment during each sampling [
16
].
3. Inoculation conditions and sampling timing must be selected
considering the nature of the pathogen and the type of dis-
ease. In this case, the previously reported conditions are fol-
lowed [
15
], but sampling is done at early times within the fi rst
4 days (0, 6, 12, 24, 48, and 96 post-inoculation hours). It is
important to consider the local or systemic nature of the host
plant response. A completely randomized experimental design
using treatments including no inoculated controls becomes a
suitable alternative for this type of studies in plants. In order to
do this, it is recommended to use the following treatments:
T1, Resistant Control; T2, Resistant Inoculated; T3,
Susceptible Control; T4, Susceptible Inoculated.
4. Non-lyophilized material can be used for extraction. In this
case, report the results as mg of protein per g of fresh vegetable
material, using amounts ten times larger than for lyophilized
material due to the high moisture level in vegetable samples
(around 90 %). Different vegetal material amounts (lyophilized
