<TITLE: Plant Club
ACADEMIC DOMAIN: natural sciences
DISCIPLINE: biology
EVENT TYPE: seminar discussion
FILE ID: USEMD270
NOTES: seminar includes presentations USEMP12A/C, USEMP12B not transcribed

RECORDING DURATION: 11 min 57 sec

RECORDING DATE: 11.4.2007

NUMBER OF PARTICIPANTS: 34

NUMBER OF SPEAKERS: 11

S1: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: senior staff; GENDER: male; AGE: 51-over

S2: NATIVE-SPEAKER STATUS: Portuguese; ACADEMIC ROLE: junior staff; GENDER: female; AGE: 31-50

S3: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: senior staff; GENDER: male; AGE: unknown

S4: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: research student; GENDER: female; AGE: 24-30

S5: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: unknown; GENDER: male; AGE: unknown

S6: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: junior staff; GENDER: male; AGE: unknown

S7: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: senior staff; GENDER: female; AGE: 31-50

S8: NATIVE-SPEAKER STATUS: German; ACADEMIC ROLE: junior staff; GENDER: male; AGE: 24-30

S9: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: senior staff; GENDER: male; AGE: 31-50

S10: NATIVE-SPEAKER STATUS: Finnish; ACADEMIC ROLE: junior staff; GENDER: male; AGE: 31-50

S11: NATIVE-SPEAKER STATUS: French; ACADEMIC ROLE: junior staff; GENDER: female; AGE: 24-30

SS: several simultaneous speakers>


<S1> okay i think we can start now so er it is er , plant club of april and er our first speaker will be <NAME S2> from <NAME S6>'s group and the title you see on the on there so please <NAME S2> </S1>

<PRESENTATION USEMP12A by S2>

<S1> so thanks <NAME S2> so there's plenty of time for [questions] </S1>
<S2> [yeah] i was a little bit @fast@ </S2>
<S1> so please <NAME S3> </S1>
<S3> did you generally associate any phenotypes of the roots to the changed GFP pattern or were they sort of normal </S3>
<S2> many so , it it's the thing so in this one i i have a a lot of (lost) of GFP and actually they look quite normal and i think it's probably some of the (xx) in GFP itself , yeah so er it's the thing so o- of all these mutants or putative mutants that i'm picking up are those that really (territorise) into phenotypes but i'm still in this process and er of course i will pick the ones that have the most interesting phenotypes </S2>
<S1> going on with <NAME S3>'s er question so in those in those strange mutants did you look at some other genes which should have the same expression pattern as this </S1>
<S2> as the H- AHP-6 </S2>
<S1> yeah </S1>
<S2> @er@ as far as i know there er <S1> [or is that the unique] </S1> [is there any @@] <S1> [@@] </S1> [oh @@] with with these two so specific in this protoxylem poles it it's quite unique i would say , there are many just or in the pericycle <S1> [mhm-hm] </S1> [or] in old vascular bundle i think there are a lot in (xx) but no (xx) <S1> mhm </S1> no @mhm@ </S2>
<S1> okay . <COUGH> any other <P:05> <S2> mhm [(that's) @@] </S2> [we-] well thank you , let's then move on to , <NAME S4> move on from <P:06> plant biology or sorry applied biology @@ and gerbera so please <S4> [yes] </S4> [do you need] the l- all the lights off <NAME S4> or </S1>
<S3> no </S3>
<S4> er i think this is quite okay </S4>
<S1> i'll put put this off then </S1>
<S4> yeah </S4>

<PRESENTATION USEMP12B by S4, NOT TRANSCRIBED>

<S1> so thanks <NAME S4> questions yeah </S1>
<S4> yeah </S4>
<S5> it seems like there's (been) a (xx) promoter (again) might play a (xx) (defects) so do you have some more specific promoters you could (direct) expression in the flowers early </S5>
<S4> yes we have thought about it of course some petal specific in- inflorescence specific but but we haven't yet started and as you know it takes a long time to produce transgenic gerberas so we have only these basic basic lines now available and some some RNA-I lines coming up in a month or so , so </S4>
<S1> yeah <NAME S6> </S1>
<S6> there are these (cresting) mutations and (if) you shown shown now that in your <COUGH> lines there was no difference in the expression <S4> mhm-hm </S4> but i think you should also sequence the gene in those lines because i mean there could be <S4> [mhm-hm] </S4> [a mutation] <S1> mhm-hm </S1> and that i mean that would be the critical test </S6>
<S4> yeah i know that's what that was the first check but it wasn't that promising @@ the first check , yeah and also that we don't have this crossing population it's quite hard to start to to (VCR) these crests that we have but but we've been thinking about that </S4>
<P:06>
<S1> somebody yeah [(go ahead)] </S1>
<S6> [i have] another so so you showed that in in there in the distant ray flowers there were in in there was an early phase or expression that was only in ray flowers <S4> [mhm] </S4> [and later] you showed expression in both in rays and discs <S4> yes </S4> so have you been looking at the in in in situ hybridisation whether there is actually could be a specific pattern in some of these two phases would be actually i mean you showed me the point that there is no er regional specificity in the during (xx) but how about <S4> [mhm] </S4> [this] very early (xx) <S4> mhm </S4> have you covered all the stages </S6>
<S4> no i haven't this is what i said as my future plans this is really the next stage [that next] </S4>
<S7> [er but they] have to be repeated </S7>
<S4> yes @@ this has to be repeated well it has been repeated but again repeated and take some other development and now that we know what we want to say we want to like cover it really carefully and do proper like not proper but yeah more specific in situs yes </S4>
<S1> how close is your gene to the antirrhinum cycloidea </S1>
<S4> it's quite close quite similar but but @@ it would be nice to have a a proper allogenic analysis and we are working on <S1> [mhm-hm] </S1> [that] but mhm i've done [only just] </S4>
<S1> [but okay in] antirrhinum if you mutate that you get a dramatic phenotype right </S1>
<S4> mhm n- er no you need also to mutate the dichotoma </S4>
<S1> okay so you [need two mutations] <S4> [like gen-] yes </S4> to get the <S4> [yes] </S4> [phenotype] okay i see </S1>
<S4> mhm-hm <S1> mhm </S1> but you do get a phenotype with only cycloidea mutated but it's not it's not totally radially <S1> [okay] </S1> [symmetrical] but it's a bit changed </S4>
<S1> have you considered trying that gene in gerbera </S1>
<S4> mhm we've done that <S1> and </S1> and er it didn't have an effect <S1> mhm-hm </S1> not not at least not mhm at the pe- at the flower symmetry <S1> [so you] </S1> [and we] also we have also gene from senecio <S1> [mhm-hm] </S1> [cycloidea gene] from senecio and and one line but only one line showed really interesting phenotype with these petals that have tubular shape <S1> mhm-hm </S1> and we are actually working on that as well but it's a pity that it's a single line it didn't er repeat </S4>
<S1> something else , no okay thanks , now let's go to , our third speaker of today <NAME S8> from plant biology will tell us about irregulators in er . in cell death so please </S1>

<PRESENTATION USEMP12C by S8>

<S1> thanks <NAME S8> questions or comments , <NAME S6> </S1>
<S6> so y- you added this kind of power er grid aspect in this so <S8> yeah </S8> so you showed some evidence that there was something wrong with the stigma but how about the pollen so ha- have you made reciprocal crosses [with pollen on (xx)] </S6>
<S8> [erm i have i have samples] and tubes i have to go through microscope </S8>
<S6> but have you done crosses RCD-5 pollen to the normal </S6>
<S8> yes yes <S6> okay </S6> these samples are in my tubes i have to go through microscope [and look at them] </S8>
<S6> [ah okay] okay </S6>
<S8> we are very curious about those but i haven't had time to look at them yet , but honestly i don't think they will show much different there why we didn't initia- initially notice erm that there was the sequent that RCD-5 is reduced is when we (xx) seeds we take a couple of plants and put it together in seed bag and then we rub them and take out the seed we could have like thirty per cent less seed or even forty per cent you might not always notice un- unless you look really carefully but here we really counted individual seeds in my experiment i only counted individual seed (leaves) i counted the seed but i also erm er looked at the embryos and this basically gave the same result </S8>
<P:05>
<S10> but crosses didn't work in either direction </S10>
<S8> crosses didn't work in either direction so erm RCD-5 m- , mo- most likely has also a function in pollen , but that's why we do the reciprocal crosses for staining embryo 'cause we have to have a look at that . but it it goes it goes together with the ideas that RCD-5 might be some some please stay alive signal </S8>
<S1> <NAME S3> </S1>
<S3> and you're suspecting that it's outside of the cell </S3>
<S8> yeah so the tobacco people have not hundred per cent convincing data but pretty good evidence that it's that it's er secreted </S8>
<S3> <NAME S9> is not here i guess </S3>
<S1> yeah he's [(also)] </S1>
<S8> [i i] saw him today </S8>
<S3> okay </S3>
<S8> oh </S8>
<S3> oh there you are <SS> @@ </SS> i mean i was just about to refer to your work with peroxidasis showing that it's apoplastic </S3>
<S9> yes are you sure about the signals because that in- that (xx) with the apoplastic , (xx) because the signal sequences are not that clear in plants </S9>
<S8> well that's at least what what i got from the prediction programmes but if you want to have a look at that i'd be happy to show you </S8>
<SS> @@ </SS>
<S3> but anyway anyway <NAME S9>'s point was that y- you put the retention signal in the in in the GFD fusion and then it was E-R and then your latest experiment was without the retention signal but then the cells started to make cell wall and it got out and stuck to the cell wall <S9> yes </S9> the [signal] </S3>
<S9> [(i got it)] in , [(in my study)] </S9>
<S3> [and the problem] with apoplastic is that y- you lose it you don't see it anywhere , basically </S3>
<S9> yeah but if <S3> [so that's why] </S3> [you have whole] plants or whole tissue [(xx)] </S9>
<S8> [that was that was] that was <S3> [yeah] </S3> [onion er] onion peels in the compartment and i think what what could yo- could correct me if i'm wrong but what you're doing is y- you use protoplasts </S8>
<S9> well we we use protoplasts [yes] </S9>
<S8> [yeah] and there you lose it into the into the medium </S8>
<S3> but after three days the protoplast have a cell wall <S8> mhm </S8> and what i understood th- then you <S9> [yeah then you see it] </S9> [see it in the cell wall] if you don't have an attention signal <S8> [mhm] </S8> [there] of course with the onion peel the cell wall was <S8> yeah </S8> stained </S3>
<S8> yeah (it was) yeah yes that it was the first pictures i did i did basically on the weekend and er we have to have to figure out now where this goes <S3> yeah </S3> i mean i'm i'm just as happy with cell walls i'm i'm happy with any loca- location localisation as long as i know where it is </S8>
<S3> i mean cell wall is in the apoplast i guess </S3>
<S8> yeah </S8>
<S1> okay </S1>
<S11> your antibody is er an anti-nuc- er antibody </S11>
<S8> there was anti- (nuclear) </S8>
<S11> yeah so do you think it could be possible for you to have a specific antibody for RCD-5 </S11>
<S8> yes but since i have two bands there on the gel we're not actually quite sure what that means erm that could either be just an artefact from the over-expression or from the tag or it could also mean that the protein is chopped up and then if we make the antibody against the wrong domain we might not see anything despite despite we have to stop there so we have to be a bit careful about where we direct an antibody against </S8>
<S11> yeah because a good way to see if it's apoplastic or not would be electronic microscopy <S8> yeah </S8> but then you have to have a [a very good antibody] </S11>
<S8> [have to have a specific antibody] </S8>
<S11> yeah </S11>
<S8> but you ca- you could also use t- use the tag but yeah there's also problems with that but you're right but we just have to really think carefully about where we put the antibody [against] </S8>
<S11> [mhm] </S11>
<S1> okay any further if not then let's thanks all the speakers again and er </S1>
<APPLAUSE>
