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Current status of laurel wilt research - Shared screen with speaker view
Andrew Johnson
36:41
Add questions here!
Jiri Hulcr
41:44
Question for Jeff Rollins: when you say that R. aquacate is "associated with avocado", does it mean native to the Americas?
Matthew Kasson
43:01
And to follow up on Jiri's question: What beetle host is R. aquacate isolated from?
Jiri Hulcr
44:07
Isn't R. quercivora from a different clade? I.e., independenlty evolved?
Luis Mendez
44:37
knowing that these sulfur enzymes are in the infection process, would they make good targets for quelling? Is it conserved to Raffaella spp.?
Jason Smith
44:40
aguacate was isolated from a dying avocado years ago, but was shown to be non-pathogenic…
Jason Smith
45:06
I don’t think its ever been found in any beetles?
Romina Gazis-Seregina
46:43
That’s aguacate
Denita Hadziabdic Guerry
47:54
What are the time points used for presented experiment
Jason Smith
57:08
Did you look for the pathogen in leaves?
Carlos de la Torre
57:49
Have you check this mobility in avocado?
Luis Mendez
57:57
What promoter is GFP under? Great work!
Christina Burns
58:01
Josh, were root sections studied?
Michael Wingfield
58:01
Josh- could you say a bit about swamp bay- as related to red bay.
rjoseph
58:08
For the mean estimated CFU experiments, how do you estimate across the entire tree? Are you assuming that there's an even distribution of the fungi?
Jason Smith
58:40
Agreed. Beautiful study.
You Li
58:58
how long after inoculation the swampbay die?
Patricia Merced Manosalva
59:12
Are you planing in testing this GFP strain in avocado with different vascular system architectures? for example narrow as Mexican races and Wide as West Indian
Guest
59:20
Interesting Josh!
DanV
01:07:10
Could I see the first slide again?
DanV
01:09:00
What is the MM supplemented with that you get growth on?
rjoseph
01:09:13
Is the expression of cas9 transient because the promoter is inducible?
DanV
01:09:26
2nd slide
Chase Mayers
01:16:11
What kind of symbiosis mechanisms with the beetles might be tested with these methods?
Jiri Hulcr
01:17:06
Ross - a big problem in studying these symbioses is contamination in beetle-fungus colonies. Can we use your method to introduce an antibiotic resistance and rear the symbiosis on antibiotic-amended media?
Jason Smith
01:17:11
Is there an advantage for visualization of signal in beetles vs. host wrt reporter?
Jiri Hulcr
01:17:57
Can you even see it fluorescing inside a beetle?
Luis Mendez
01:20:10
Using the gfp model, could this be used to visualize fungal “infection” in the mycangium? (not a beetle expert) or is the fungus just sitting on the exoskeleton?
Jiri Hulcr
01:27:01
Ever seen volvulus or bispinatus colonizing living trees?
Wilma Nel
01:31:26
Hi Daniel, very interesting presentation. Do you think these other beetles are actively cultivating Raffaelea lauricola in their galleries or is it possible that they have accidentally acquired inoculum of R. lauricola and it is out competing their primary symbionts?
Jiri Hulcr
01:32:08
Wilma - that's an excellent question for Octavio!
Wilma Nel
01:33:28
Thanks Jiri, let's pose the question to him and hear what he says
rjoseph
01:33:30
Cool work!
Chase Mayers
01:34:12
Does the competitive binary experiment include subsequent transmission tests (which enters and persists in the mycangia)?
You Li
01:34:24
If the yeast symbiont could keep the beetle live in the test?
Chase Mayers
01:36:12
(Not a question just a comment) Awesome! It would be hugely useful to let it put both symbionts in mycangium and see which transfers, perhaps through several generations
rjoseph
01:38:37
How do you go about disinfecting pupae prior to transferring to rearing tubes?
rjoseph
01:41:07
Thank you!
Denita Hadziabdic Guerry
01:41:50
Are you pooling your samples?
Chase Mayers
01:42:04
How did you avoid infamous ITS amplification problems in Raffaelea - were these ITS2 primers?
Chase Mayers
01:43:54
It looks likeITS7 would indeed avoid it - awesome that makes sense. Thanks!
Matt Kasson
01:46:25
Are you looking at northern spicebush in and around diseased sassafras?
Luis Mendez
01:47:19
Don’t move firewood!
Jiri Hulcr
01:47:28
:)
Caterina Villari
01:48:21
Was the CHK pcr performed directly on wood samples, or on fungal cultures?
Jason Smith
01:48:34
Did you have any that seemed positive that you couldn’t confirm>
Jason Smith
01:48:34
?
Jeffrey Hamilton
01:52:05
Do you have Xylosandrus compactus in Tennessee?
Jiri Hulcr
01:56:23
Who, or which agency, is monitoring for LW in Texas, and how actively?
Jason Smith
01:56:56
Texas Forest Service…Shane Harrington
William Klingeman III
01:57:16
X. compactus does occur in TN but is very infrequently collected and likely patchy in distribution
Andrew Johnson
01:57:18
X glabratus has also been found on other host such as oaks. Are there other ambrosia beetles found in irrigated urban settings in the south area of texas?
Jason Smith
01:59:34
Has sassafras been found to be impacted in TX?
Jason Smith
02:04:43
We have tested susceptibility of a few Lauraceae native to Mexico..P. podadenia, etc., would that information help with your modeling? (P. podadenia is highly susceptible for example)
Demian Gomez Damiano
02:10:16
Any data on what would happen if you look even southern? Like South America?
Shannon Lynch
02:11:26
thanks Andres! super cool!
Matt Kasson
02:16:49
was this all done in orchards or were you outplanting genotypes in disease epicenters?
Denita Hadziabdic Guerry
02:19:08
What inoculation method worked the best?
Jiri Hulcr
02:19:17
Scary!
Robin Choudhury
02:19:52
Is the leaf accumulation of R. lauricola the reason for reduced abscission or the change in hyperspectral imaging?
rjoseph
02:19:54
Dr. Rollins suggested that the R. lauricola genome my contain genes coding for effectors that can elicit host defenses. Do you think that tolerant species might have genetic differences related to defense responses, and comparative genetic studies between tolerant and susceptible cultivars might uncover these differences?
Patricia Merced Manosalva
02:20:17
How reproducible and consistent are the symptoms using leaf infection assays
Patricia Merced Manosalva
02:20:41
For example if you inoculate 100 leaf you have 100% of infection
Pedro Parra Giraldo
02:21:08
How far away from the the inoculation point was the fungus recovered in the leaves.
Michael Wingfield
02:21:44
Jason- your thoughts of leaves being infected is intriguing. Does R. l spread in borer frass. Have you looked at frass as a source of inoculum on leaves?
Matt Kasson
02:29:15
Did you test it on the new Raffaelea Jason mentioned they are recovering from Indiana?
Denita Hadziabdic Guerry
02:29:16
How many other species were tested? or any other closely related fungi as well?
Jason Smith
02:29:16
Mike: Never looked at frass.
Demian Gomez Damiano
02:30:29
Have you tried this with the beetles?
Matt Kasson
02:30:30
How cheap is equipment?
Denita Hadziabdic Guerry
02:30:38
What about false positive samples?
Carlos de la Torre
02:30:55
When do you think is going to be available for growers to use
Jason Smith
02:31:08
Pedro: The inoculation point was done at 10 cm above soil line at two leaves (one on each leaf on opposite sides of stem)…then recovered from leaves at apex of stems on potted trees in greenhouse.
Guest
02:31:15
Is the equipment wildly available?
Karan Chahal
02:31:23
Does that 50 conidia alone as pure propagules or mixed with wood/frass and then detected?
Guest
02:31:29
Widely *
Karan Chahal
02:32:02
I was talking about sensitivity
Jeffrey Hamilton
02:32:42
The 50 conidia was a spore suspension
Jason Smith
02:32:45
Patricia: Not 100%. Varied depending on method and whether field or greenhouse. We are currently doing a larger experiment to determine reproducibility during different conditions.
rjoseph
02:33:00
Sorry if I missed it, but what is the sample prep for the assay like?
Caterina Villari
02:35:10
@Demian, Yes, and it works perfectly on insects as well
Demian Gomez Damiano
02:35:22
Thanks!
Pedro Parra Giraldo
02:35:43
Thank you Jason
Patricia Merced Manosalva
02:35:53
Thank you Jason
Xavier Martini
02:38:47
Did you treat only the trees next by pre-symptomatic ones?
Denita Hadziabdic Guerry
02:38:53
So non-scientific question. Who takes the pups home after their hard work in the field? :)
DanV
02:39:40
Awesome Julian, pre-symptomatic, how long after infection are they able to detect? Have you checked the CFU threshold of detection?
Chase Mayers
02:39:44
Could some of the detected metabolites be the same strong fruity ester aroma that Raffaelea produce in culture?
rjoseph
02:40:19
Super cool work!
José L Olivares
02:40:48
Amazing work!
Demian Gomez Damiano
02:41:59
Have this been tried in ports of entry? Like containers?
Julian Mendel
02:43:19
DavV we did studies with pure culture and determined the canines could detect as low as 50 spores. In terms of how long after infection, we are unsure, our studies show they can detect as early as 40 days before you see the wilt.
Julian Mendel
02:44:02
Chase: we saw alot of interesting compounds. One thing we really want to explore more is the fact that in RL cultures we found plant hormones associated with defense
Julian Mendel
02:44:47
Damien Gomez Damiano: We have not, would definitely work, it has been done for a number of other non laurel odors.
Caterina Villari
02:47:28
In our study we had issues with CHK cross reacting with host tissues. Have you find similar results?
Jason Smith
02:50:26
Using only one SSR may be risky since it is always possible that it may amplify from something else. Two were originally chosen because it greatly decreases chances of random amplification.
Jason Smith
02:50:46
Should a new marker be developed?
Diana Sanchez-Rangel
02:59:37
Hi, have you search for some molecular markers in the trees exposed to low doses?
Chase Mayers
02:59:55
Could this explain the "resistance" in native population in Asia - they are vaccinated?
Patricia Merced Manosalva
03:00:45
Romina which cultivars you have looked so far?
Caterina Villari
03:01:13
Have you noticed any reduction of growth in "vaccinated" plants?
Jason Smith
03:02:00
what defines a “low dose”?
Dave Anderson
03:02:28
What response did you see in the "unvacinated" trees?
Caterina Villari
03:04:14
Thank you!!
Matt Kasson
03:04:23
We've seen stunted growth as the only outward symptom in Verticillium tolerant trees
Ben Held
03:04:56
Interesting work! Have you considered timing of inoculation?. For example when actively growing and earlywood is present disease can develop more quickly than when latewood is present.
Michael Wingfield
03:05:45
Dear Jiri and colleagues. I am afraid that I need to leave the meeting to join a IUFRO discussion. This has been a fantastic symposium- thanks for organising and thanks to all the speakers and participants. Take care- keep in touch. Mike AND apologies to the last speakers!
Romina Gazis-Seregina
03:06:25
Patricia: we looked at Lula, Doni on Waldin, Waldin on Waldin
Romina Gazis-Seregina
03:06:40
We will look into Simmonds/waldin and hopefully Mexican race ones!
Romina Gazis-Seregina
03:08:14
Jason, we define low dose as 600 CFUs but we are planning to scale up this experiment and test 100-1,000
Daniel Carrillo
03:09:14
How are the beetles exposed to the insecticides?
JR
03:09:46
HI Romina!!! We have multiple seedlings used as rootstocks from the Mexican race. Please let me know if you would be interested to test them
Romina Gazis-Seregina
03:10:01
Dave: unvaccinated trees showed symptoms of diseases - but we are trying now to standardize our experimental plants (in terms of cultivar combination, size, branching patterns, canopy size etc) so we can have even symptom progress
Xavier Martini
03:10:06
These insecticides will be systemic?
Daniel Carrillo
03:11:02
how do you envision using this insecticides in the field?
José L Olivares
03:13:50
Hi, the experimental results suggest that this compound is systemic.
Jason Smith
03:15:47
Hi Romina. In redbay 100 spores will cause lethal wilt. It will be interesting to see what happens over time with those. It may be that the pathogen will increase in titer and wilt will eventually happen. But I could be wrong. We used 1500 spores with avocado and caused lethal wilt. See the paper published by Hughes et al on inoculum conc. and laurel wilt in swamp bay (Forest Pathology, 2014?).
José L Olivares
03:15:54
To Daniel Carrillo, we are thinking to nano encapsulate our compound and administer directly to the tree. Any suggestion is very welcome.
Jiri Hulcr
03:16:23
When you lop off the tree canopy, doesn't the stump become much more attractive to the beetles?
Demian Gomez Damiano
03:17:19
How recommendations and mitigation tactics change for home landscapes? What are the current recommendations?
Romina Gazis-Seregina
03:27:42
Hi Jason: I agree it will be interesting to follow these trees through time. I believe there is tons of variation on how different avocado cultivar react to infection - in terms on cultivar genotype, age and graft combinations. We are trying to standardize our experimental plants, but use graft combinations representative of the Florida industry (so we will settle for Doni/Waldin or Simmonds/Lula) and then replicate the experiments. Maybe “more tolerant” trees suppress the multiplication of the fungus so the titer goes up slower or there is something going on not he host side of the equation…
Jason Smith
03:28:44
Romina, good points. Host tolerance and reaction to the pathogen early on may be significant.
Salvador Ochoa
03:32:40
Excellent symposium. Thanks to the organizers and all the speakers.
Simon
03:32:53
tHANKS everyone, VERY INTERESTING TALKS
Simon
03:33:06
Ignore the caps :)