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Journals
Journal of Nematology
Volume 52 (2020): Issue 1 (January 2020)
Open Access
Root-knot nematodes demonstrate temporal variation in host penetration
Shova Mishra
Shova Mishra
and
Peter DiGennaro
Peter DiGennaro
| Apr 13, 2020
Journal of Nematology
Volume 52 (2020): Issue 1 (January 2020)
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Published Online:
Apr 13, 2020
Page range:
1 - 8
Received:
Feb 07, 2020
DOI:
https://doi.org/10.21307/jofnem-2020-037
Keywords
Root-knot nematodes
,
Temporal variation
,
Circadian rhythms
,
Bioassays
,
Nematode biology
© 2020 Shova Mishra et al., published by Sciendo.
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Figure 1:
Host plants, Medicago truncatula were inoculated with RKN at six different time points; three times points during the light period and three time points in the dark period. In total, 24 hr post-inoculation, root samples were collected at respective time points to check the temporal variation in penetration. All infective juveniles were exposed to host roots for a 24 hr period before quantification.
Figure 2:
Sampling timepoints for normal (A) and inverted (B) light/dark cycles. To exclude the direct effect of light as a contributing factor to diurnal changes in penetration, we inverted light/dark cycles for equal period of light and dark as in normal condition. Inoculation was done at 2 hr interval for three time points at dark and three time points at light under both conditions. Root samples were collected at 24 hr post-inoculation at respective time points.
Figure 3:
RKN displays temporal variation in penetration with the highest penetration at night, 3:00 a.m. M. truncatula was inoculated with M incognita (A) and M. hapla (B) at six time points, three times points light and three timepoints dark. In total, 24-hr post-inoculation root samples were collected at respective time points. Bars represent standard error and columns with different letters are significantly different (p < 0.05). Data shown in each figure represent three independent experiments with four replications per experiment.
Figure 4:
M. truncatula plants maintained under normal 16 hr day/8 hr night conditions were inoculated with M. incognita at six time points. One set of plants was kept in normal light condition (A) and another set of plants were immediately transferred to an inverted light condition (B). In total, 24 hr post-inoculation root samples were collected at respective time points and nematode inside root was quantified. Bars represent standard error and columns with different letters are significantly different (p < 0.05). Data shown in each figure are from three experiments with five replications per experiment.
Figure 5:
RKN do not exhibit temporal difference in penetration when plants are entrained under constant light conditions. Three weeks old M. truncatula plants were kept in constant light for one week. Plants were inoculated with M. incognita at six time points. In total, 24 hr post-inoculation root samples were collected at respective time points and nematode inside root was counted. Bars represent standard error. No timepoint was significantly different (p > 0.05). Data shown in each figure are from two experiments with four replications in each experiment.
Figure 6:
Percent egg hatch is not influenced by light and do not show temporal differences at different time points. Freshly harvested RKN eggs were observed for hatching over a period of 68 hr. Hatched juveniles were counted, and percent hatch was calculated. To examine influence of light we compared percent egg hatch between constant light and constant dark timepoints and to assess any temporal differences, percent egg hatch between consecutive time points under constant dark and constant light were measured. Bars represent standard error and means (n =12) are not significantly different (p > 0.05) according to paired t-test.
Figure 7:
Light does not have influence on RKN juvenile mobility; 300 freshly hatched J2 were placed in a modified Baermann funnel under constant light or constant dark conditions. After 4 hr, the number of J2 that passed through sieve were counted. Bars represent standard error and means (n =12) are not significantly different (p > 0.05) according to paired t-test.
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