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Date (2010)	Time	Observations Made on One Inflorescence Bagged to Exclude Insect Visitation, Accession 694 Plant 1
27 Aug	3 pm, 7:30 pm	Three flowers having unopened corollas were tagged at 3 pm: tags 28 (red), 29 (green) and 30
28 Aug	7 am, 7: 30 am, 8 am, 8:30 am, 9 am	Flowers 28 (left in photo in next row) and 29 (top right in photo) - corolla longer than wide and corolla still closed. Flower 30 (middle of photo) - corolla is also longer than wide, still closed, and it is evident that abscission is starting (corolla is nearly in its original place but close inspection reveals that it is beginning to abscise as shown in photo in the next row).
28 Aug	Corollas are whitish; the calyces they are in are green. The three corollas shown have never been open.   The middle corolla has begun abscision (look closely where it meets the calyx).
28 Aug	9:30 am, 10:00, 10:30, 11, 11:30, noon, 12:30	Flower 30 - corolla, having never been open, detached but remains largely in place such that one strong gust of wind would allow it to completely drop off
28 Aug	1 pm, 1:30, 2:00, 2:30	Flower 30 - Corolla is 1 mm lower, in other words closer to dropping completely
28 Aug	3:00 pm	Flower 30 - corolla is withering, changing color
28 Aug	3:30 pm	The corollas of flowers 28 and 30, having never been open, abscised and dropped completely off; opening of these corollas revealed anthers had dechisced prior to corolla drop [same 4pm, 5 pm, 5:30 corolla of flower 29 is still on and is still closed]
29 Aug	8 am, 7 pm	Flower 29 - corolla is still closed, shriveling/senescing in place. Pollinator exclusion bag is still present.
30 Aug	7 am, 4 pm	Flower 29 - corolla senesced in place.
31 Aug	8 am	Ovaries of all three flowers are expanding. Another flower of the same inflorescence, not tagged, is abscising its never-been-open corolla.
3 Sept	8:30 am	Ovaries of all three flowers are expanding. Another flower of the same inflorescence, not tagged, is abscising its never-been-open corolla.
9,15 Sept	7:30 am	Ovaries of all three flowers are expanding
end Oct		28, 29 &30 all resulted in ripe fruits
	Summary	Three flowers that did not open produced ripe fruit; these flowers were inside of a pollinator exclusion bag when they flowered. Cleistogamy of all three tagged flowers.

Date (2010)	Time	Observations Made on One Inflorescence Bagged to Exclude Insect Visitation, Accession 694 Plant 1
20 Aug	10:30 am, 2 pm	Three flowers having never-been-open corollas were tagged, tags 16 (red), 17 (green), 18 (white), tagging and bagging by Alison Cameron and Thomas Mione. No change noted at 2 pm.
21 Aug	8 am	Flowers 16 & 18 - corolla closed & rather flat-topped, wider than long
	8 am	Flower 17 - corolla abscising from calyx without ever having been open, photo in next row
	Tag 17 hangs from a flower that is dropping a corolla that has never been open
	11:30	Flowers 16, 17 and 18, no change
	4 pm	Flower 16, same. Flower 17's corolla fell completely off. Flower 18's corolla might be senescing
22 Aug	7 am	Flower 16's corolla is missing, must have abscised and fallen. Flower 17 is same as yesterday, missing its corolla. Flower 18's corolla looks like it is senescing.
	2 pm	Flowers 16 and 17 - same. The corolla of flower 18 dropped off, abscised.
23 Aug	9 am, 3 pm	Same as yesterday. There are no open flowers on this inflorescence.
24 Aug	8:30 am	No change
25 Aug	8:30 am	Ovary of fl 17 is expanding. Ovaries of fls 16 and 18 are not expanding.
26, 27, 29 Aug	1 pm, 8:30 am	The ovaries of flowers 16 and 17 are both expanding
31 Aug	8 am	Immature fruits within same inflorescence removed (these from flowers that were not tagged) so I can more easily see fruits developing from flowers 16 &17.
3 Sept	8:30 am	16 & 17 are now immature fruits. Flower 18 aborted with no ovary expansion. Another flower of the same infl, not tagged, absiced its corolla; carefully opening the corolla revealed dehisced anthers.
9 Sept	7:30 am	16 & 17 - fruits are expanding
15 Sept	3 pm	16 & 17 - Immature fruits
end Oct		16 & 17 both resulted in ripe fruits
	Summary	Cleistogamy!

Date (2010)	Time	Observations Made on One Inflorescence Bagged to Exclude Insect Visitation, Accession 694 Plant 1
18 Aug	morning	Shown in the next row of this table is a flower bud (under a ruler) that is aborting its corolla before the corolla opened, corolla clearly longer than wide. Other flowers on the same plant are wide open (chasmogamous) at the same time.
18 Aug	Same time of day as previous row in this table 4 mm long ruler rests on calyx; corolla is lighter and lower
19 Aug	7:45 am	Flower 12. Corolla that had never been open dropped off. Corolla was inspected and contained stamens having dehisced anthers. Tulle bag was removed for inspection of stigma with a 14X hand lens; revealed pollen, lots of pollen, all over stigma, and then bag was replaced.
	4:45 pm	Tulle bag removed, Tag 13 added to a flower that had never been open but looked like it might open soon. Tulle bag was replaced.
20 Aug	6:45 am	Flower 12 same as yesterday, pedicel-calyx-style unit present Flower 13 Corolla that had never been open absiced but resting on style
	10 am	Flower 13. Inspection of cor reveals that cor has never been open and contains anthers that all dehisced.
	3 pm	No change
21 Aug	8 am	No change
22 Aug	7 am	No change to tagged fls. Another fl (not tagged) within same tulle bag is dropping a cor that looks like it has never been open.
	2 pm	One wide open (chasmogamous) fl in tulle bag (a flower that was not tagged).
23 Aug	9 am	Pollinator exclusion (tulle) bag removed. Ovaries of flowers 12 and 13 are expanding. A flower that was not tagged for this study is wide open (chasmogamous).
	3 pm	Two flowers on this inflorescence are now wide open, chasmogamous, flowers not tagged for this study
24 Aug	8 am	Two immature fruits, still small, were removed (with scissors) from this inflorescence; these were never tagged and their removal will allow me to more easily see flowers 12 and 13 now showing expanding ovaries
25, 26, 27, 29 Aug	8:30 am, 1pm, 8:30 am, 8 am	The ovaries of flowers 12 & 13 have expanded into green immature fruits
31 Aug	8 am	Flowers 12 & 13 - ovaries continue to expand. Immature fruits within same inflorescence removed (these from flowers that were not tagged) so I can more easily see fruits developing from flower 12 & 13.
3 Sept	8:30 am	12 & 13 continuing to expand, fruits getting larger. One flower of this inflorescence, not tagged, will open in a few days.
9 Sept	7:30 am	12 & 13 continuing to expand, fruits getting larger. One flower of this inflorescence, not tagged, will open in a few days.
15 Sept	3 pm	12 & 13 are both immature fruits
end Oct		12 & 13 both resulted in ripe fruits
	Summary	Two flowers that abscised unopened corollas produced ripe fruit!

All of the following observations were made on plants of accession 694.

Observations on a flower. Year 2000.
14 Sept. 7:30 am—I manually opened a flower that (I am certain) had never opened before. All 5 anthers had already dehisced! I marked the tip of the sepal with white out.
8:30am—distal portion of petals reflexed. 11:30—Nectar where androecium meets corolla between stamens.
4:30—flower now a very shallow bowl; thus flower closing . Tips of corolla lobes still reflexed.
15 Sept. When I got home from work at about 4pm--flower looked closed and finished.

Comment of September 2002—this flower must have been on the plant I was growing under lights in the spare bedroom in 2000.
19 Sept. 6:00am—sepal marked with a very small amount of whiteout. 6pm—Flower open. 8pm—half open. 10pm—closed (not all wrinkled as if senescing).
20 Sept. 5:50 am—corolla has dropped.
Summary: Flower lasted only one day.

Comment of September 2002—this flower must have been on the plant I was growing under lights in the spare bedroom in 2000.
20 Sept. 11:00 pm. Corolla shut, has never opened, very small drop of whiteout to mark flower.
21 Sept. 6:52am still shut. 2pm—Flower open wide with filaments angling out and style curved. 6:40pm—flower closed. 9:44pm—corolla dropped.
Summary: Flower lasted one day.

Comment of September 2002—this flower must have been on the plant I was growing under lights in the spare bedroom in 2000.
21 Sept. Corolla shut, looks very much like it has never opened. I put two small drops of whiteout on branch by flower.
22 Sept. 5:30am—closed. 3pm—wide open & somewhat reflexed. 5:45 pm corolla still open but not open all the way (see original hand-writen notes for sketch).
23 Sept. 5:30am—corolla has dropped.
Summary: Flower Lasted One Day.

Observations on a flower, year 2000.
23 Oct. 7am—Closed flower (bud) marked with white tape. 8:30am—closed. 12:00—closed. 1:30pm—closed. 3:00pm—closed.
24 Oct. 6:20am—Open and anthers have dehisced, filaments have elongated; style curved. 1:30—same.
25 Oct. 7:00am—Flower finished.
Summary: No protogyny. Flower lasted one day.

Observations on a flower. Year 2001.
3 Sept. A flower was not yet open but has a large looking bud (large compared to other accessions). I carefully forced open the corolla and the anthers had dehisced before the corolla opened.
4 Sept. 4:00 pm--This flower’s corolla had dropped.
Summary: I am not sure if this corolla would have opened on its own, or would have dropped off unopened.

Observations on a flower. Year 2001.
4 Sept. A flower having a corolla that has never opened was manually opened, and all anthers had dehisced before the corolla opened.
5 Sept. 3:30--Corolla has dropped.
Summary: Flower lasted one day.

7 Sept. 2001. Two more flowers have anthers dehisced prior to the corolla opening for the first time. I gently manually opened the corolla before it had even opened, and found dehisced anthers. I looked the next day,
8 Sept., and the corollas of these two flowers had dropped. Does this mean that some flowers of 694 never have an open condition? Comment of October 2002—Sept. 2001 was the first time it occurred to me that some flowers might be self-pollinating in the bud with the corolla of these flowers never opening.

15 Aug. 2002, Thursday. I noticed a flower bud that was not open but was as large as a flower bud could be without the corolla opening. I carefully opened the corolla, and all 5 anthers had dehisced prior to my peek inside. This plant has been growing since May, so it is approximately three months old, and has been outdoors in a pot for most of the summer.

21 Aug. 2002. Same as 15 August; plant growing in pot.

23 Aug. 2002. Same as 15 August, but plant rooted in ground. I see pollen on the edge of the stigma. Did the pollen get on the stigma when I picked the flower, because of the mechanical disturbance created by my picking the flower?

5 Sept. 2002. Same as 15 August 2002, but plant rooted in ground. May be self-pollinating in bud, but pollen could have been touched to stigma when I manually (and carefully) popped open the corolla.

7 Sept. 2002. I noticed a flower bud, not open, but as large as a flower bud could be without opening. Without removing the bud from the plant I carefully popped open the corolla—it opened easily suggesting the bud would have opened today. With a 14X hand lens I noticed that a little bit of pollen got on the side of the stigma, not the top. Bud photographed, and is 9.5 to 10 mm in diameter. It is dark for the photo; there is no direct sun. Immediately after photographing I opened the corolla, and all 5 anthers has dehisced. 14X lens used and there is definitely pollen on the stigma, but the pollen may have gotten there when I gently pried open the corolla lobes.

14 Sept. 2002, Saturday, warm and sunny. Two corolla-androecium units were seen falling off of the plant growing in the ground at home; the corolla had never opened. I opened the corolla and found all 5 anthers had dehisced. One stigma has a little bit of pollen on its edge; the other stigma was covered with pollen.

15 Sept. 2002, Sunday. Same as 14 September 2002 but stigma half covered with pollen.

19 Sept. 2002, Thurs.– just to see if the method of D’Souza (in Stain Technology) would allow us to see Jaltomata pollen tubes, we manually self-pollinated flowers on 16 and 18 Sept., harvested, fixed, stained, and observed. We could not see the tubes in the style, but could see the pollen tubes in the stigma. Conclusion: this method should be abandoned. It is interesting that pollen stays on the stigma through all of the steps, this must be because the tube growing out of the pollen into the stigma holds it in place.

20 Sept. 2002, Fri. – changed day/night on plant growing in a pot in the hall to 12 hours day and 12 hours night. My thinking is that if the plant at home (rooted in the ground) is showing self-pollination inside the bud, but the plant under lights is not, then perhaps the day length is involved. I don’t have much confidence that shortening the day (increasing the night) will actually trigger self-pollination in bud, but we shall see.

20 Sept. 2002 – three flowers collected 2 days prior were used. These flowers had closed corollas and dehisced anthers (the corolla was not falling off when the flowers were harvested). Rich Th. and I used the technique exactly as described by D’Souza in Stain Technology, and saw pollen tubes penetrating the stigma (not style) of one flower out of three. The other two stigmas showed no pollen (and therefore no pollen tubes). Seeing the pollen tubes tells us that the process of gently popping open a closed corolla to see if the anthers had dehisced in the bud is not what caused the pollen to get on the stigma. In other words, in one of the three flowers the pollen must have gotten on the stigma before I opened the corolla to look inside! However, we can’t know from this, if I had collected the buds a day later, if we would have seen pollen tubes on the other two as well.

20 Sept. 2002 – within the last week we have started to remove developing fruits from all plants (two at home, one at work) to promote flowering. We are removing nearly all, but not all, developing fruits.

22 Sept. 2002. On the plant of J. darcyana growing in the ground at home, I have seen many unopened corollas (with all 5 anthers dehisced) abscise (begin falling off) this weekend. In contrast, the plant nearby that is growing in the pot has no unopened flowers that are falling off [but see notes of 27 Sept). These two plants are the same age, and have been growing within 20 feet of each other for months, and have received approximately the same amount of fertilizer. The plant in the pot is not as large, and has not produced as many fruits. Perhaps, when a plant of J. darcyana produces a certain critical mass of fruits, then the cleistogamy starts, but this does not seem like a good strategy? The weather has been gorgeous, warm sunny days, and nights that have been in the 40s and 50s.

23 Sept. 2002. Gregory Shenk, worked with a bottle of about 6 flowers that I labeled "694 flowers: unopened corolla (with attached androecium) fell off." I gave him the bottle when he visited my house to see the plant on Monday I believe 16 September. “Your pollen tubes glow like my white legs in the dark! I tried one pistil over the weekend at 24 hrs in 8N NaOH and 8hrs in fresh aniline blue w/ K3PO4. The style looked like a super highway with all of the pollen tubes. I also brought the Jaltomata to the UConn GH and will be watching it for self-pollinated-in-bud flowers.

24 Sept. The plant that has been inside Copernicus Hall all summer and early fall produces no flowers that drop off having never opened. I see several flowers open (chasmogamous flowers) today, but I don’t know if these self-pollinated in the bud.

24 Sept. On the plant at home in the ground, there is at least one open chasmogamous flower (low and kind of hidden). Today there is at least one flower with a withered corolla that never opened. I removed the withered corolla and found all anthers dehisced. I tagged this flower after seeing what is almost certainly some pollen on the stigma.

26 Sept. 2002. Greg Shenk wrote: If you haven't already committed your pistils to the NaOH, you may want to try some for a shorter duration. Worked with a bottle of about 6 flowers that were labeled "694 flowers (corolla with attached androecium) fell off w/out ever opening." What worked well for the first one seems to have oversoftened the other four. Applying too much pressure when I squash may be part of the problem. They all stained beautifully and all have pollen tubes growing in them but the oversoftened ones broke up a bit so the pollen tubes are interrupted at a few points. As data, they'll be fine and I'll get you pictures. As pictures for publication, only the one will be very good. Unless you want close-ups anyway; then they may all work.

27 Sept. 2002. Plant growing in pot at home all summer was moved to Copernicus. I then noticed a flower having a corolla that has never opened on it that was beginning to fall off. The corolla had detached, but had not yet dropped completely, so I removed the corolla with forceps, noted that the stigma was covered with self-pollen (a hand lens was used), and tagged the flower. No pollinator exclusion netting is needed because the plant is indoors in Copernicus. Is this flower just doing this because it is away from its normal climate and doing bizarre things in an unfamiliar climate?

28 Sept. 2002. I have been tagging flowers (that have an unopened corolla fall off) on plant growing in ground at home. Ovaries are expanding!

30 Sept. – 4 October. 2002
a) 30 Sept. 10:00. Two flowers manually self-pollinated
b) 2 Oct. 10:00 transferred to 70% ethanol
c) 3 Oct. 11:20am I put flowers in 4N NaOH, and noticed that one flower had lost its style.
d) 4 Oct. 55 min. in tap water, thoroughly rinsing, followed by 2 hours in stain. At least 20 pollen tubes were observed. Pollen tubes stained well. When the style split longitudinally (I believe because of pressure applied when pressing down on cover slip) the pollen tubes were much easier to observe.
Another flower, removed from the indoor plant when its corolla was senescing, was fixed in 70% EtOH for only about 3 hours, and then put in 4 NaOH at 2:15 on 3 October. On 4 October: 55 min. in tap water, thoroughly rinsing, followed by 2 hours in stain. OK pollen tubes, but not as good as longer fixation. Obviously, flower had self-pollinated without aid from a pollinator.
Yet another flower was immersed in the Lactic Acid fixative of the D’Souza (in Stain Technology) method for weeks, after being collected when the corolla (unopened) was in the process of falling off. Pollen tubes were observed in this style! This clinches it: the plant self-pollinates inside the bud.

7 Oct. 2002
We have numbered the three living plants of Jaltomata darcyana accession 694 (I have no other accessions of this species). All plants, including the one grown by Greg Shenk (unnumbered, perhaps we will refer to it as #4), are the same age; the seeds were planted in late April or early May 2002.
Plant #1 – Has been growing in Copernicus Hall all summer, on window sill mostly. Was under fluorescent grow bulbs for a while, but is back on the windowsill as of 4 October 2002. Has been in, and continues to be in an 8” pot.
Plant #2 – This plant grew all summer at home outdoors in a 10” pot and is still in a 10” pot; was moved to Copernicus on 27 Sept. (see notes of 27 Sept.). Now under 4 fluorescent lightTubes in Copernicus Hall.
Plant #3 – This plant grew rooted in the ground all summer at home. When it was moved to CopernHall 3-4 Oct. 2002, an potted it in a 14”pot. Now under 4 fluorescent lightTubes in Copernicus Hall.

Fertilizer
Starting today, 7 Oct. 2002, we will give plants one, two and three one ml of a fertilizer solution for every inch of pot. Thus, the 8” pot (plant #1) received 8 ml today. The solution is 3 level, loosely packed tablespoons of fertilizer per liter. I unfortunately do not know the chemical make-up of the fertilizer; it is the blue, powder fertilizer that has been up in the greenhouse for many years.
The plants grown at home during the summer of 2002 received an unmeasured amount of granular fertilizer most weekends, delivered to the soil immediately around the plant, over the roots. The amount varied depending on plant size, with larger plants receiving up to a tablespoon, and smaller plants receiving half this much.
Plants grown in pots received an unmeasured amount of slow release pellets, probably about a teaspoon, only once sometime during the summer. Perhaps we should apply again in the fall, because of the micronutrients.

7 Oct. 2002
I see about 5 open flowers on plant #2.
I saw a large bud that had never opened; there were very small slits where corolla lobes meet, so flower looked like it might open soon or the next day. All 5 anthers had dehisced, but no pollen on stigma. Flower was destructively sampled.

11 October 2002
One of the questions I had/have is/was, do regular (chasmogamous) flowers that don’t self-pollinate in the bud exist? The answer is yes. I can see absolutely no pollen on the stigma of the open flowers of plant 2, here in the pollinator-free environment of the hallway of Copernicus Hall.

16 Oct.
Plant #2 got covered with aphids (while growing under lights in the hall) and I had to put it out of its misery today.

Study of the Fall of 2002.

Flower was tagged when the corolla, having never opened, was observed to be in the process of abscising. Result: No fruit.
Flower 1 (tagged on 22 September, tag harvested 17 October).

Flower was tagged when the corolla, having never opened, was observed to be in the process of abscising. Fruit Was Observed to Develop for at least one month.
Flower 2
Flower 4 (ripe fruit harvested 15 Nov.)
Possibly Flower 5
Flower 6 (tagged 22 Sept., fruit ripening 5 Nov.)
Flower 7 (tagged 21 Sept., fruit beginning to ripen 28 October)
Flower 8 (ripe fruit harvested 15 Nov.)
Flower 10 (tagged 22 Sept., about to start ripening when plant died 15 Nov.)
Flower 11 (tagged 20 Sept., about to start ripening when plant died 15 Nov.)

Of the nine flowers that were tagged when the corolla, having never opened, was observed to be in the process of abscising, eight fruits formed (as defined by enlarging and remaining on the plant for at least one month). Only four of these eight fruits actually ripened, because the plant on which they were developing died.

Flower was tagged when I noticed that the corolla withered (but looked like it had never opened).
Fruit Was Observed to Develop for at least one month.
Flower 3.

Flower was tagged when I noticed that the corolla withered (but looked like it had never opened).
Result: No fruit.
Flower 9