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WANTED: plants for okc plant swap: april 30
I have reserved the South Will Rogers pavillion ("the castle") for a plant swap on April 30 from 1-3PM. This is located at NW 30th and Portland in Oklahoma City.
This is what I am hoping to bring. Let me know if you would like to trade for them. I am just now getting my seeds planted, and I will make sure I have extra.
A few tomatoes: brandwine, San Marzano, Prize of the Trails, could also plant Red House Freestaning.
A few cucumbers: armenian and mideast prolific
Herbs: epazote, borage, basils: sweet, lemon, lime
Flowers: to be determined
I have
Clary Sage
Yellow Lavender
Lambs Ear
one Mexican Oregano
Variegated Mint Pineapple
I started some sweet 100 cherry tomatoes. But they are taking their sweet time.
Obedient Plant
A few Sedum's
One peony
some creeping phlox Purple.
I could dig up some crown vetch If some wants a starts
I have 2 aloe noid they stay on the small side and grow kind of fan shaped
one banana tree
I want
Tall Phlox
Sedum's I don't have
Candy stripe Creeping Phlox
Shasta Daisy
I would try about any thing I haven't tried yet
Well now, I've learned another important Oklahoma Gardening lesson.
The wind here not only comes sweeping down the plains, but it also burns young maters to death!
Notwithstanding, I should have some of the following for the swap:
Maters (all heirloom varieties):
Cherokee Purple
Cherokee Red
Reisenstraub Red Cherry
Galina Yellow Cherry
Wisconsin 55
and maybe a couple of Hellfucht.
Depending on what survives hardening off, I may have some:
Cayenne peppers (unnamed variety)
Buran sweet peppers (Polish heirloom)(only a couple)
Cubanelle sweet peppers.
If anyone is interested, I could pot up some Oregano, russian sage, Cinnamon Basil, spearmint (I think) or Thyme.
HPD: I am definetly interested in a Cherokee Red and a cayenne if any survive. I'm hoping the plants I put out today survive the wind also. What are looking for? Would you like lemon or lime basil?
All: I also have Autumn Joy Sedums. (WindSurfGirl: I'll put your name on a few since I didn't have enough last year)
I am looking for heirloom tomatoe Arkansas Traveler and the "Ugly" tomato.
Happy Gardening!
If I had known about this swap like in February, I could have started extra heirloom tomato plants specifically for the event; I started only what I wanted to grow for myself, darn it.
I do need to thin out my aloes and it looks like that's the only plant I can be definite about bringing. Depending on how planting goes, I may bring some Grand Bell bell peppers and other stuff that I may discover I have no room for after all. Too early to tell what's what at this point, tho.
What I'm gonna have to do is make a point to plan better for next year.
I took an inventory of some of my plants for the swap. I'll have the following heirloom tomatoes - Sioux, Porter, Arkansas Traveler, Mule Team, Mortgage Lifter, Boxcar Willie, Amish Paste and Galina.
I'll also have some hot peppers - Chilipeno, Salsa, and Concho. Plus a few heirloom sweet peppers - Corno Di Toro and Quatro D'asti Rosso.
I'll have some catnip, oregano, garlic chives and possibly some fennel. There may be more but we'll just have to see how much time I have to continue digging.....
Mrs Frodo,
I'll have to look to see if the wind spared any more of the Cherokee Reds. If so, you'll be first on the list.
I would love sedum if you have any extras.
WSG-Those Galina seeds you sent-what monstrous plants they produce!
I don't think I can bring any to the swap as they will be much too large for transplanting! Maybe a couple as potted plants?
I really had a problem with damping off and wind breakage, so I'm not sure how much I'll have to bring. I'll have some small cherokee purples and Brandywines, and whatever extras don't go in the garden (darn cutworms are really trying my patience!)
But I'll be there with HotDogs and 2 year olds!
HPD- Is the Cinnamon Basil still available?
PlanterMum- I have a few Shasta Daisies with your name on them. I got 2 pots of 2 plants each at my firt swap. I got them in October and by spring they had multipled (they are in a protected place).
Happy Gardening!
I have Hoya cuttings I could take. I have a collection of around 500! Any Hoya lovers in here that would be interested?
I have some freckles outdoor violets I could dig from outside. They went nuts last year re-seeding.
I could take some rose cuttings if anyone has luck rooting them?
I need to thin my iris's. They are a brownish purple with a violet edge.
My bf and I would be interested in sedums, dwarf or stay small pond plants, elephant ears and caladiums. Any takers?
In another post on this subject I also agree to supply the hot dog buns.,,,Debbie
Mrs Frodo,
I have a few very small starts of the cinamon basil, I'll bring those along with all my leftover seeds in case anyone is interested.
Does anyone have any Lavendar?
Every time I start some, they sprout, start to grow then disappear.
Its driving me nuts!
Since Im new to this forum I have a question concerning the plant swapping at the meet. Thus far no one has showed any interest to what I have to offer,,,which is fine because I understand Im new to outdoor gardening so havent anything spectacular to share yet,,haha. So do I bring the plants anyway? How is the swapping set up,,,,do we bargain prior to the 30th so we know what to dig/cut to bring or is there a grab table or how?
I really wouldnt want to cut the Hoya's if no one is interested nor dig the violets and iris to only bring them back home to replant. UGH! haha.
Please dont think Im trying to be difficult,,,thats not my intention. Im just wanting to understand how the swapping works.,,,,Debbie
Yes bring your plants anyway. Bargaining prior to the swap is fine, and fun, but you don't have to have any prior swaps set up. I usually post what I have so if someone just has to have some I can save one for them. I will bring lots of things no one has shown an interest in. Most things I dig are extra anyway and if I don't swap or give them away I'll take them home and give them to someone else.
In fact Debbie I'm interested in your hoya and violets but I don't know if I will have anything you are interested in. I have lots of pond plants but nothing dwarf or small. I do have an elephant ear plant I could bring you.
Last year we all put our plants on tables and just walked around looking. If I saw something I wanted I ask for it and then the person would go to my table and see if I had something they wanted. I gave alot of plants away that I did not get a trade for and was perfectly happy to share with everyone. When I got plants in trade I usually put them UNDER my table so everyone would know they were off limits.
Let me know if you have any more questions!
Thanks WindsurfGirl for the explaination on how the swapping works!
Do you have any Hoya's already? Please email me a list if you do. I may bring a couple mature plants also.
Ill get some violets ready this weekend if possible. If not I will during the week.,,,Debbie
mrsfrodo-I'd love to try some epazote! (Next month I'm doing a couple of cooking classes involving Mexican food, I know the instructor uses epazote.)
I don't have much to bring, but have a few extra ancho chile plants, and I don't know what else until I get some re-potting dome today.
Darn! I really wanted to go to the plant swap. I won't be able to make it.I am on a fire department and take collections of items for families who get burnt out of their house and I promised this weekend I would take the time to stop by a couple houses and pick up things they would like to donate.I don't want to break my promise since these people are being nice enough to donate things....Hmmm maybe they will donate a plant to me since I am missing the swap...LOL...Everyone take notes and let me know how it goes. *sniff* I really want to go.
I am new to OKC and plan on attending this weekend, but I only have rooted Jade plant cuttings to offer. Hopefully I'll have some takers!
See you this weekend,
Just FYI for those expecting lime basil or epazote. Since I was sick these got started late. They have sprouted, but have not had a growth spurt. I keep hoping for several days of warm weather to help them along, but that has been wishful. I will bring them for any taker. They will give us all a good laugh.
I have some sedums spoken for, but should have one or two extra. I have two extra Shasta Daisies, and a rooted trumpeter vine cutting. Lots of star of bethelehem bulbs. Time willing, pink oxalis, wild onions, and garden sage may be thrown in.
Anyone interested in a free red rose bush- and willing to dig and move- may take it.
If anyone has any of the following I would be interested:
Lily of the Valley, Cardinal Flower, tarragon, evenning primrose, red or white flowers for full shade, stella day lilies.........
See you Saturday Deb.
Happy Gardening!
Mrs. Frodo,
I can bring you some cardinal flower seeds and I can dig up some evening primrose plants for you if the kind you want is the showy pink evening primrose. Let me know. I also have 4 tomato plants sitting on my driveway with your name on them--from a post long, long ago: think the 4 are Yellow Pear, Arkansas Traveler, Ugly and Beefmaster.
I'll be bringing the rest of my excess tomato plants--mostly hybrids as I've given away most of the heirlooms already. They are quite large and want to be planted!
I do have some VERY TINY heirlooms started from seeds I got in a trade earlier this month. They are barely sprouted, but I might bring them if any heirloom tomato afficianados are looking for Aunt Ginny's Purple, Marianna's Peace, Cherokee Chocolate or maybe Earl's Faux (only 2 of the Earl's Faux have sprouted though, so I might part with one).
I'll be bringing lots of packs of seeds, especially tomato seeds for the seed bin, and some flower seeds as well.
In the plant area, I've been digging and thinning and will have at least the following (and maybe others depending on what I get done today and tomorrow):
Black-eyed Susan Vine
Four O'Clocks
Veronica--blue flowered
Pink & Purple Cleome from a mixed planting
Verbena Bonariensis (this is the tall one with little purple flower heads on the end of slender stalks; very heat tolerant and attracts butterflies like crazy)
I'm sure I'll come up with more but I'll have to get off the computer and go outside and get some work done!!!
I've never been to anything like this and I plant to come and check it out .I was wondering if anyboby has any Brugs for trading ? I have lots to trade and can not bring everthing please check my trade list and I also have some things not listed like bronze leaf fennel , clary sage , basil's ,chives ect ... and if you do not see it ask . I do not have everything listed .
OKieDeb: Can you describe the violets. I'm interested and will save you a sedum.
PlanterMunn- The mexican oregano sounds intriquing. Your Shasta has been dug and I am babying it.
Okie Dawn- Hmmmmm. Do you want my first born child? Thanks for keeping the tomatos for so long. My husband is looking forward to the Ugly tomato especially. The primrose offer is appreciated, but I am looking for the yellow evenning primrose. If the four-o-clocks and black-eyed susan vine are still available, I'm interested. Can you tell how big the verbena is and what color the flowers are? Lastly, what are you looking for?
Orchidtrader: Are the Stellas or the clematis available? What are you looking for?
FYI- I understand if it is too late for any of you to prepare these plants.
Happy Gardening! See you Saturday!
Mrs. Frodo,
I'll let you keep your first-born child! lol The tomato plants are rather large, and except for taking a beating from the wind a little now & then, and getting too dry once or twice, they've been fine. My driveway will be quite lonely once they leave our home for yours (grinning).
The four o'clocks and black-eyed susan vine are still available, and I'm bringing lots of each.
The verbena bonariensis forms a mound about one foot in diameter. Then very thin, tall stalks shoot up. At the top of the stalks you will have tiny cluster of lilac-colored flowers. The stalks also branch out some, and flowers form on each branch. The bees and butterflies fight over the flowers. If you deadhead by cutting the flowering stalks off near the ground, the plant will re-bloom over and over again. It is a very light and airy plant, amazingly tough and drought-resistant. Sometimes reseeds itself.
I've attached a link with a picture of verbena for your viewing pleasure.
And, what am I looking for? Just about any herb or veggie. Or flower. Or anything else. I'm not picky and I like trying new things, so anything you have to trade is fine. And, if you run out of things to trade, that's fine too.
Thank you so much for all the work you've done to organize the swap. I hope you know how much your efforts are appreciated!
The 2 verbena photos are at the bottom of the linked page.
Here is a link that might be useful: Verbena details/photo
A final note to all:
Bring what you have, don't be timid about bringing small seedlings, seeds, or even nothing at all if your just getting started. Believe me, you won't go home empty handed!
The purpose of this is to meet and greet our fellow OK gardeners and to share share share!
Everyone please note the start time is NOON. The orginal plan was for 1:00 but mrsfrodo managed to get the pavillon for a little bit longer time so it will be NOON to 3:00.
I spent some time potting up some more plants last night. And I'd like to also restate what HPD said - don't worry if you don't have lots of plants or even anything at all. Just come and meet everyone. I'm bringing so many plants I bet I'll have something to send home with you. This is called a swap but I think my favorite part is giving plants away. :)
I have a question.
I am bringing some rooted Jade plant cuttings. Currently I have them planted in a tray, they are not in individual pots and I don't have enough pots to plant them in. Is that OK? I figured I'd bring some small dixie cups with me and just "dig" them up and put them in the cup as they are taken. Is this acceptable?
Dixie cups will fit right in at the swap. As a matter of fact, I found a few from a previous swap, that may be recycled. Many of the plants I am bringing are in folgers cans. One good thing about having a swap at a park is any loose dirt can easily be swept out. Large scale potting is probably a bad idea, but what Deb is describing sounds fine.
Let me add my invitation along with WindSurfGirls and HotPepperDad to join us Saturday evenning if you are worried you don't have many plants. We all love to share are plants. You will be suprised what you will have to swap in a two years.


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A serious game helps us discover what our environment might be like in 50 years' time
What will our environment be like in 50 years' time and in how much does our behaviour influence our climate as well as our everyday lives? The priority for researchers at the Leibniz Centre for Agricultural Landscape Research (ZALF) e.V. and their cooperation partners such as DHI WASY GmbH is to inform citizens about the possible consequences of landscape use and to start a conversation with them.read more

subset: commoncrawl


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The Week in Photos for 3/30/2015
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Dubai, Australia, Japan, Fair Grounds, Gulfstream Park, Hawthorne, Oaklawn & More
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A glimpse into the legacy of Hall of Fame trainer H. Allen Jerkens. Allen Jerkens was elected to the Hall of Fame in 1975 at age 46, then the youngest trainer to be so honored.
Pioneering racetrack photographer Charles Cook served as an aerial reconnaissance photographer in World War I.
Oaklawn Park, Santa Anita, Gulfstream, Woodbine, Golden Gate Fields, Sunland & More
Tampa Bay Downs, Santa Anita, Aqueduct, Oaklawn Park, Gulfstream, Sunland & More.
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Bode Miller, Michael Phelps, Rashard Lewis, Wes Welker, Michael Owen & More
Australia, Gulfstream Park, Santa Anita, Charles Town, Turfway, Oaklawn & More
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Candy Ride continues the series of training offspring of sires, which can be found in the Jan. 17, 2015 issue of The Blood-Horse.
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As a companion to the January 10, 2015 issue of The Blood-Horse, we present a look at Tapit's 2014 successes on the track and at the sales.
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Highlights from the racing career of Japan's champion mare Gentildonna.


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Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
MicroRNA399 is a long-distance signal for the regulation of plant phosphate homeostasis
Article first published online: 6 NOV 2007
DOI: 10.1111/j.1365-313X.2007.03363.x
© 2007 The Authors
The Plant Journal
Volume 53, Issue 5, pages 731–738, March 2008
Additional Information
How to Cite
Pant, B. D., Buhtz, A., Kehr, J. and Scheible, W.-R. (2008), MicroRNA399 is a long-distance signal for the regulation of plant phosphate homeostasis. The Plant Journal, 53: 731–738. doi: 10.1111/j.1365-313X.2007.03363.x
Publication History
- Issue published online: 6 NOV 2007
- Article first published online: 6 NOV 2007
- Received 31 August 2007; revised 24 October 2007; accepted 30 October 2007.
- microRNA;
- phosphate homeostasis;
- long-distance signaling;
- nutrient;
- Arabidopsis;
- Brassica
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
The presence of microRNA species in plant phloem sap suggests potential signaling roles by long-distance regulation of gene expression. Proof for such a role for a phloem-mobile microRNA is lacking. Here we show that phosphate (Pi) starvation-induced microRNA399 (miR399) is present in the phloem sap of two diverse plant species, rapeseed and pumpkin, and levels are strongly and specifically increased in phloem sap during Pi deprivation. By performing micro-grafting experiments using Arabidopsis, we further show that chimeric plants constitutively over-expressing miR399 in the shoot accumulate mature miR399 species to very high levels in their wild-type roots, while corresponding primary transcripts are virtually absent in roots, demonstrating shoot-to-root transport. The chimeric plants exhibit (i) down-regulation of the miR399 target transcript (PHO2), which encodes a critical component for maintenance of Pi homeostasis, in the wild-type root, and (ii) Pi accumulation in the shoot, which is the phenotype of pho2 mutants, miR399 over-expressers or chimeric plants with a genetic knock-out of PHO2 in the root. Hence the transported miR399 molecules retain biological activity. This is a demonstration of systemic control of a biological process, i.e. maintenance of plant Pi homeostasis, by a phloem-mobile microRNA.
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
Inorganic phosphate (Pi) is one of the most limiting mineral nutrients for plant growth (Bieleski, 1973; Poirier and Bucher, 2002; Marschner, 1995). The levels in the soil are often low, much of the Pi is covalently or non-covalently bound, and mobility is restricted. Strategies of plants to cope with and to improve Pi acquisition during Pi limitation include increasing the root/shoot ratio (Lynch, 1995), alterations in root architecture (López-Bucio et al., 2002; Williamson et al., 2001), production of lateral roots into Pi-rich patches in the soil (Robinson, 1994), proliferation of long root hairs (Bates and Lynch, 2001), increasing high-affinity Pi uptake capacities (Mudge et al., 2002; Smith et al., 2003), establishing symbiotic interactions with mycorrhizal fungi (Harrison, 1999), production and root secretion of organic acids to solubilize Pi in the soil and apoplast (Raghothama, 1999), and induction of phosphatases (del Pozo et al., 1999; Li et al., 2002) and RNases (Bariola et al., 1994) that mobilize organically bound phosphate inside and outside the plant. Changes in metabolism allow re-mobilization of Pi in the plant, including a general decrease in the levels of P-containing intermediates and co-factors such as nucleotides (Zrenner et al., 2006), or replacement of phospholipids by sulfo- and galactolipids (Dörmann and Benning, 2002; Härtel et al., 2000).
Correct allocation of the available Pi in the plant, i.e. maintenance of Pi homeostasis by supplying growing organs and tissues such as young leaves and inflorescences with the spare resources taken up by roots, is also important for plant productivity and survival during Pi limitation. The existence of mutants with altered levels and distribution of Pi (pho1, pho2 and pup1; see Delhaize and Randall, 1995; Poirier et al., 1991; Trull and Deikman, 1998) suggests that plants regulate the long-distance transport and allocation of Pi between shoot and root. PHO1 encodes the founder member of a novel transporter family (Hamburger et al., 2002) and probably is involved in Pi transport into the xylem (Wang et al., 2004), and PHO2 encodes an E2 ubiquitin-conjugating enzyme (Aung et al., 2006; Bari et al., 2006). The PHO2 E2 conjugase is a major component for the maintenance of Pi homeostasis in Arabidopsis. pho2 mutants grown in Pi-replete conditions display continuous induction of a subset of phosphate starvation-induced (PSI) genes, including some encoding phosphate transporters (Aung et al., 2006; Bari et al., 2006), and accumulate Pi to high levels (three- to fivefold increased) in shoots but not roots (Bari et al., 2006; Delhaize and Randall, 1995).
PHO2 expression is post-transcriptionally regulated by microRNA399 (miR399). miR399 acts through transcript cleavage (Allen et al., 2005) and probably also translational repression (Bari et al., 2006). Consistently, over-expression of miR399 leads to high shoot Pi levels, thus phenocopying pho2 mutants (Aung et al., 2006; Bari et al., 2006; Fujii et al., 2005). The PHO2 gene harbors five binding sites complementary to miR399 in its 5′ UTR approximately 200–400 nucleotides upstream of the start codon, and miR399-dependent PHO2 transcript cleavage at these sites has been experimentally verified (Allen et al., 2005). Six miR399 species (a–f) that arise from five primary transcripts (PTs) have been identified in Arabidopsis (http://microrna.sanger.ac.uk). miR399d is by far the most prominent one during Pi limitation, based on the abundance of its primary transcript (Bari et al., 2006). The levels of mature miR399 and the five miR399 PTs are highly and specifically increased during Pi limitation (Bari et al., 2006; Fujii et al., 2005), suggesting inhibition of PHO2 under these conditions. Consistently, removal of the miR399 binding sites results in high and stable PHO2 transcript levels in Pi-limited plants (Fujii et al., 2005). Micrografting experiments revealed that a pho2 mutant root genotype is sufficient to result in shoot Pi accumulation (Bari et al., 2006). On the other hand, MIR399 genes, and especially MIR399d, display particularly strong induction in shoots (as compared to roots) during Pi limitation, with the shoot miR399d PT level exceeding the root PHO2 transcript level (Bari et al., 2006). This led us to hypothesize that miR399 itself might act as a long-distance Pi starvation signal in Arabidopsis.
Long-distance signaling in plants is known to be important for the regulation of several processes including leaf development, flowering, pathogen defense and resource allocation (Jaeger and Wigge, 2007; Khamis et al., 1990; Palauqui et al., 1997; Voinnet et al., 1998; for review, see Lough and Lucas, 2006). The phloem translocation stream of higher plants contains a multitude of small molecules and macromolecules, including proteins, mRNA and small RNAs (Lough and Lucas, 2006; Ruiz-Medrano et al., 2001). Several microRNAs have also been detected in the phloem sap of pumpkin (Yoo et al., 2004) and oilseed rape (Buhtz et al.). However, unlike siRNA or mRNA species (Haywood et al., 2005; Ruiz-Medrano et al., 1999; Voinnet et al., 1998; Yoo et al., 2004), microRNAs have not been functionally implicated in plant long-distance signaling. Here we present data that identify miR399 as a phloem-mobile long-distance signal for regulation of Pi homeostasis in Arabidopsis.
Results and discussion
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
Presence and Pi-specific induction of miR399 in phloem sap
To demonstrate the presence of miR399 and its Pi-status-dependent changes in phloem sap, we grew rapeseed (Brassica napus) and pumpkin (Cucurbita maxima) plants under abundant and limiting Pi conditions, and collected phloem sap for the determination of microRNA levels. Doing this type of experiment with Arabidopsis is impracticable because of (i) major difficulties associated with the collection of sufficiently large quantities of phloem sap from this small plant species, (ii) problems with RNase contamination and/or wounding-inducible RNase activity (Köck et al., 2004; LeBrasseur et al., 2002) in phloem sap exudates collected by decapitation or incision methods, making isolation and quantification of RNA impossible (A.B. and J.K., unpublished observations), and (iii) contamination of the sap with content (e.g. RNA) from other cells. Rapeseed plants are a useful alternative to Arabidopsis due to the close genetic relatedness of the two species, as well as their similar phloem sap composition, as judged by their protein profiles (J.K., unpublished data). In addition, phloem sap exudation from incised rapeseed inflorescence stems is quite strong, thus enabling rapid sampling and minimization of problems involving contaminating/induced RNase activity.
Mature miR399 was detected by quantitative real-time PCR and RNA gel blotting in rapeseed and pumpkin phloem sap (Figure 1), but miR399 precursors or primary transcripts were not detectable (results not shown). Mature miR399 levels increased very strongly in the phloem sap of Pi-limited plants (a difference of 7–10 quantitative real-time PCR CT value units, equivalent to a >100-fold change) (Figure 1a–c,e). The increase was even stronger and the abundance higher in phloem sap than in leaves, for example (Figure 1c), suggesting efficient transport of miR399 into the sap and/or strong induction of miR399 expression in phloem companion cells. The increase/accumulation of miR399 in phloem sap or other plant organs such as roots was also specific, as levels for other microRNA species, i.e. miR164 (Figure 1d) and miR172 (Figure 1f), did not change during Pi limitation. These results demonstrate that miR399 is present and highly abundant in the phloem, and indicate that the increase during Pi limitation is not related to decreased phloem mass flow. They also underscore the conserved Pi-responsiveness of miR399 in distantly related plant species (Bari et al., 2006; Chiou et al., 2006).
We further investigated the Pi specificity of this response by investigating miR399 levels in phloem sap of plants starved of sulfate. RNA gel blotting showed that mature miR399 abundance was unaffected by sulfate deprivation in phloem sap (Figure 1g) and leaves and roots (results not shown) of rapeseed plants. This is in accordance with previous results showing that miR399 primary transcripts are unaffected by limitation of sulfate, nitrogen or carbohydrates (Bari et al., 2006). It was also demonstrated that mature miR399 is unaffected by nitrogen (Bari et al., 2006) or potassium limitation (Fujii et al., 2005). Taken together, these results show that the responsiveness of miR399 and its primary transcripts is Pi-specific in all plant organs/samples, including phloem sap.
Micrografting reveals shoot-to-root transport and biological activity of shoot-derived miR399 in roots
We next investigated whether miR399 produced in the shoot can be detected and consequently alter expression of its target gene (PHO2) in the root. For this purpose, we used micro-grafting (Turnbull et al., 2002) to generate chimeric Arabidopsis plants that over-express miR399 in either shoots or roots. Six miR399 species (a–f) that arise from five primary transcripts have been identified in Arabidopsis. miR399d is the most prominent one during Pi limitation, based on the abundance of its primary transcript (Bari et al., 2006) (Figures 2 and 3). Transgenic plants strongly over-expressing miR399d (OX) under control of the 35S promoter were previously shown to have approximately 10-fold lower levels of the target PHO2 transcript (Bari et al., 2006), and to accumulate Pi in leaves to levels as high as in pho2 mutants or in micro-grafted chimeric plants with a pho2 root and a wild-type (WT) shoot scion (Bari et al., 2006).
As expected, the primary transcripts of miR399d (miR399d PT) and mature miR399 were strongly increased (threshold fluorescence was reached 12–13 cycles earlier during quantitative real-time PCR, equivalent to a >1000-fold increase) in root and shoot material from OX plants compared to the same material from WT plants (Figures 2 and 3 and Table S1). The levels of miR399d PT and mature miR399 in OX shoots matched the high levels found in Pi-starved WT shoots (Figure 2d,f), but the levels of the other miR399 primary transcripts (a, b, c, e/f) remained at barely detectable or undetectable levels, as in Pi-replete WT (see Table S1; note that in Figures 2 and 3 values for 40-deltaCT of less than approximately 30 correspond to transcript copy numbers of far less than 1 per cell in our quantitative real-time PCR system (Czechowski et al., 2004; also see Experimental Procedures)). In Figures 2 and 3, values for 40 −ΔCT of less than approximately 30 correspond to transcript copy numbers of much less than 1 per cell in our quantitative real-time PCR system (see Experimental procedures). The level of PHO2 transcript, measured using two quantitative real-time PCR primer pairs amplifying near the 3′ and 5′ ends of the PHO2 coding sequence (CDS), was approximately 10-fold decreased in OX plants (equivalent to 3 or 4 quantitative real-time PCR cycles) (Figure 2g,h and Table S1), and the leaf Pi level was four- to fivefold increased (Figure 4) as previously reported (Bari et al., 2006).
When shoot material from chimeras with an OX shoot and WT root (OX/WT) was investigated, it showed the same behavior as OX shoots of non-chimeric plants with respect to the levels of miR399 PTs, mature miR399 and PHO2 transcript (Figure 2). The root material of the same OX/WT chimeras exhibited WT-like miR399 PT levels (Figure 3a–e and Table S1), which were all below the expression level of some of the rarest transcription factor transcripts known (e.g. WUSCHEL, At2g17950), equivalent to an abundance of approximately 1 copy per 100 cells or less for each miR399 PT (see Czechowski et al., 2004). In contrast, the same WT roots had very high, OX-like, mature miR399 levels (Figure 3f), equivalent to a copy number of > 500 per cell, which is similar to that of one of the most abundant plant transcripts, UBQ10 (see Czechowski et al., 2005) and OX-like low PHO2 transcript level (Figure 3g,h). The shoot material had high Pi levels in the leaves, similar to those of genotypes in which PHO2 is inhibited in roots (Figure 4). We have previously shown by micro-grafting of pho2 and WT that decreased PHO2 in the shoot alone does not lead to increased Pi in the shoot (Bari et al., 2006).
A theoretical explanation for the virtually absent miR399 PTs and the simultaneous massive accumulation of mature miR399 in roots of OX/WT chimeras could be highly efficient cleavage of miR399 PTs into shorter fragments that escape oligo(dT)-primed reverse transcription and subsequent quantitative real-time PCR detection. Although there is no indication of highly efficient cleavage in roots of OX plants or WT/OX chimeras (i.e. the level of miR399 PTs mirrors the mature miR399 level), we analyzed whether there is an increase of corresponding fragments by reverse-transcribing total RNA with a primer binding to the 3′ region of the miR399d precursor sequence (Figure 5) (see Experimental procedures). However, there was no evidence for accumulation of such fragments in the roots of the OX/WT chimeras, as the level was as low as in Pi-replete WT. Similarly, the miR399 complementary duplex strand (miR399*) could not be detected in roots of OX/WT chimeras or roots of Pi-replete WT, but was clearly present in Pi-limited WT roots and miR399d PT over-expresser roots (results not shown). These results further suggest that the MIR399 genes are truly inactive in the roots of the OX/WT chimeras. The explanation for the massive accumulation of mature miR399 found in roots of OX/WT chimeras therefore must be transport of miR399 molecules from the OX scion to the WT root, where they exert their biological function, i.e. suppress PHO2. Co-localization of miR399 and the PHO2 target transcript in the root vascular cylinder has been reported previously (Aung et al., 2006), and further supports their interaction in vivo. The inactivity of the MIR399 genes in the roots of OX/WT chimeras also excludes the possibility that another phloem-mobile molecular species (e.g. a hormone or metabolite) acts as an intermediate signal to trigger miR399 synthesis.
A situation similar to that in the OX/WT chimera was also observed in Pi-deprived WT plants. During Pi starvation, levels of the miR399 PT rise in WT roots, but only to levels that are approximately 1% of the level of mature miR399 (Figure 3 and Figure S1a), while miR399 PT levels in WT shoots equal or exceed the mature miR399 level (Figure 2 and Figure S1a). This result indicates that miR399 phloem transport is relevant in the normal biological situation. In addition, given the relatively long half-life time of mature miR399 (approximately 12 h, Figure S1b) (Bari et al., 2006), we estimated that phloem transport rates are sufficiently high to achieve and maintain very high (i.e. PHO-inhibitory) levels of mature miR399 in roots that do not express miR399 PTs within a physiological response time of 2–3 h (see Appendix S1). In summary, Pi deprivation in the shoot can be rapidly signaled to the roots through miR399. Subsequent inhibition of PHO2 by miR399 in the root will lead to increased Pi uptake and transport to the shoot (Aung et al., 2006; Bari et al., 2006; Delhaize and Randall, 1995). This elegant mechanism is capable of maintaining shoot Pi homeostasis and optimizing shoot growth during Pi limitation.
No evidence for root-to-shoot transport of miR399
Reciprocal chimeras (WT/OX) that over-express miR399d PT and mature miR399 in roots were also investigated. The root material (Figure 3) resembled that of normal OX plants, while the shoot material (Figure 2) resembled WT material, with low miR399d PT and low mature miR399. This indicates that root-to-shoot transport of miR399, which would occur via the xylem, does not take place or occurs to a negligible extent. Unlike water or ions, the transport of larger molecules, including miR399, is not circulatory between shoot and root (Lough and Lucas, 2006). Accordingly, miR399 was not detectable in xylem sap from oilseed rape plants grown under normal or phosphate-deficient conditions (Buhtz et al., in press). The level of PHO2 transcript in the shoots of WT/OX chimeras was high, resembling Pi-replete WT shoots (Figure 2g,h). Leaf Pi levels were also high (Figure 4), resembling genotypes in which the level of PHO2 transcript is low in roots (Bari et al., 2006).
Taken together, these results pinpoint the mature miR399 as a phloem-mobile molecule that increases in concentration in phloem sap during Pi limitation, and that can be transported between plant organs and exert its biological role at its destination. Hence, miR399 fulfills the criteria for a long-distance signaling molecule. The existence of multiple microRNA species in phloem sap of diverse plant species (Buhtz et al., in press; Yoo et al., 2004) indicates that this might only be one example of systemic control of a biological process by a microRNA.
Despite the demonstration that miR399 is transported in the phloem conduit, and thereby plays a central role in the maintenance of whole-plant Pi homeostasis, several questions remain to be addressed with respect to the transport mechanism. These include the mechanism and proteins required for miR399 entry into and exit from the sieve tubes, and the requirement and nature of RNA binding proteins to allow miR399 phloem transport, such as the 27 kDa phloem small RNA binding protein CmPSRP1 (Yoo et al., 2004) or the 16 kDa phloem protein 16 (CmPP16) (Xoconostle-Cázares et al., 1999) in Cucurbita maxima. With respect to translocation in the phloem, the current data indicate that miR399, as already suggested for the whole miRNA population (Yoo et al., 2004), travels as a single-stranded molecule, as, in contrast to the miR399 sense strand, the level of the miR399* strand does not show any increase under phosphate starvation (Buhtz et al., in press). miR399 is efficiently translocated from Pi-replete OX shoots to Pi-replete WT roots. This suggests that the components of miR399 phloem uptake, translocation and exit are either constitutively present or induced during Pi limitation in a miR399-dependent manner.
Long-distance signaling in biological systems was put in place when multi-cellular organisms evolved. In this regard, it is interesting to note that miR399 and its target PHO2 gene are found in higher multi-cellular plants only, including rice, poplar and Medicago, but not in single-celled algae (Bari et al., 2006). The Pi-dependence of miR399 expression is strongly conserved between dicots and monocots, as is the target gene with respect to gene structure, encoded protein sequence, and the number and positions of miR399 binding sites in its 5′ UTR (Bari et al., 2006). This suggests co-evolution and conservation of the molecular components required for Pi homeostasis in higher multi-cellular plants. Long-distance transport of miR399 further cements this view.
Experimental procedures
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
Growth of rapeseed and pumpkin, and phloem sap sampling
Rapeseed (Brassica napus cv. Drakkar) and pumpkin (Cucurbita maxima) plants were grown in 18 cm pots containing a low-Pi soil mix in a phytotron at 25°C (day)/20°C (night), with a 16 h light period at an intensity of 550 μE. Plants were divided into two sets; one set was supplied with a full nutrient solution containing sufficient (3 mm) Pi, and the other one was supplied with the same nutrient solution free of Pi. Rapeseed plants were grown until flowering (7–8 weeks) and phloem sap was then collected by incising inflorescence stems (Giavalisco et al., 2006). Phloem sap from pumpkin plants was collected by stem incision 3 weeks after germination. After discarding the first droplets, the exuded phloem sap was collected into four volumes of ice-cold Trizol reagent (Invitrogen, http://www.invitrogen.com/).
Hydroponically grown rapeseed plants were germinated on filter paper for 1 week before transferring them to a hydroponics system containing nutrient medium (Buhtz et al., in press) for 7–8 weeks. Nutrient solutions were changed after 3 weeks, and subsequently renewed once a week. Phosphate and sulfur starvation were initiated after 3 weeks by changing to medium containing no phosphate or sulfur, respectively, and phloem sap was collected from inflorescence stems as described above.
Micrografting of Arabidopsis seedlings
Micrografting of 6-day-old seedlings grown on sterile vertical agar plates was performed as described previously (Turnbull et al., 2002). Seeds of the miR399d over-expressers, wild-type (ecotype Col-0) and pho2 mutant were surface-sterilized, and stratified at 4°C for 3 days. The seed were then laid onto sterile half-strength MS agar (0.7%) plates. The plates were kept vertically in constant light (approximately 120 μE) at 20°C for 3 days in an Arabidopsis growth chamber (Percival Scientific Inc., http://www.percival-scientific.com/), and then kept at 28°C under an 8 h photoperiod (60 μE) for another 3 days. The seedlings were grafted using silicon tubing (0.3 mm internal diameter) as the collar, and grafted seedlings were kept on identical sterile agar plates and under the same growth conditions for another 6–8 days until the graft junction had healed. Successfully grafted plantlets (approximately 14 days old) were transferred to a hydroponic culture system with half-strength nutrient solution containing 1.5 mm or no Pi (Scheible et al., 2004), and grown for another 3 weeks before harvesting of leaf and root material into liquid nitrogen. This material was used for analysis of gene expression and Pi content. Special care was taken to avoid any plants with adventitious roots.
Pi measurements
Pi levels were measured using a colorimetric micromethod (Itaya and Ui, 1966). Discs were taken from leaves of the same age using a 6 mm cork borer, and individually ground in 50 μl distilled water using plastic rods (Sarstedt, http://www.sarstedt.com) in 96-well plates (Greiner Bio-One, http://www.greinerbioone.com). Extracts were diluted with water to 125 μl, and then the microtiter plate was centrifuged at 3000 gfor 4 min. Aliquots (10 μl) of the supernatant were mixed with 15 μl water, and 100 μl 1 m HCl and 100 μl color reagent (one volume of 4.2% (NH4)6Mo7O24·H2O in 5 N HCl, three volumes of 0.2% malachite green dye in water) were added. The plate was incubated at room temperature for 15 min before adding 100 μl 1.5% Tween-20. The absorbance at 660 nm was measured another 15 min later. The Pi concentration in the samples was determined against a calibration curve.
RNA isolation, cDNA preparation, and quantitative real-time PCR
RNA isolation of total RNA from Brassica napus and Curcurbita maxima was achieved using Trizol reagent, and isolation of RNA from roots and shoots of micrografted Arabidopsis plants was performed using RNeasy mini kits (Qiagen; http://www.qiagen.com). cDNA preparation and quantitative real-time PCR were performed using the primer pairs previously described (Bari et al., 2006; Czechowski et al., 2004, 2005). cDNA synthesis for the detection of miR399d precursors/primary transcript fragments was primed using 5′- GGCAAATCTCCTTTGGCAGAG-3′ (RT2 in Figure 5). Typically, approximately 5–10 ng of cDNA, the amount produced from the RNA of several thousand cells (Czechowski et al., 2004), was used per assay, yielding a CT value of 16–17 for UBQ10 (see Table S1), which is equivalent to 1–2 million single-stranded UBQ10 template copies in the PCR reaction vessel.
For quantification of mature miR399 and miR164, total RNA was reverse-transcribed using stem-loop primers (5′-GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCGGGGCAAA-3′ and 5′-GTCGTATCCAGTGCAGGGTCCGAzGGTATTCGCACTGGAT ACGACCGCACG-3′, respectively) and MultiScribe reverse transcriptase (Applied Biosystems; http://www.appliedbiosystems.com/) (Chen et al., 2005). In brief, 1 μg total RNA was mixed with 1 μl 10 mm dNTPs, 1 μl 2.5 μm miRNA-specific stem-loop RT primer and RNase-free water to a final volume of 36.5 μl, heated to 65°C for 5 min, and then chilled on ice. Then 10 μl of 5x first-strand buffer, 2 μl 0.1 m DTT, 0.5 μl RNase inhibitor and 1 μl MultiScribe reverse transcriptase were added. The reaction mixture was incubated for 30 min at 16°C, followed by 30 min at 42°C and 5 min at 85°C, and then held at 4°C until quantitative real-time PCR amplification using primers miR399fwd (5′-CGACGTGCCAAAGGAGATTTG-3′), miR164fwd (5′-CACGTGGAGAAGCAG GGCA-3′) and miRrev (5′-CCAGTGCAGGGTCCGAGGT-3′). Rather than TaqMan probes (Chen et al., 2005), SYBR® Green (Applied Biosystems; http://www.appliedbiosystems.com) was used as the fluorescence dye for real-time monitoring of DNA amplification.
Primer efficiency (E) was determined for all amplicons using the freeware program LinRegPCR (Ramakers et al., 2003). The value did not change significantly between different cDNA samples, and was always higher than 0.80.
RNA gel blot analysis of microRNA species
Northern analysis was performed as described by Bari et al. (2006). Aliquots (20 μg) of total RNA were resolved on denaturing 17% polyacrylamide gels containing 7 m urea in 0.5x Tris-Borate/EDTA. RNA was blotted to positively charged nylon membranes (Biodyne B, Pall Europe Ltd; http://www.pall.com/) using a semi-dry transfer cell (Bio-Rad; http://www.bio-rad.com/) and auto-crosslinked at 0.12 mJ in a Stratalinker 1800 (Stratagene; http://www.stratagene.com/). The membrane was hybridized at 42°C with 32P end-labeled probes (5′-TTACAGGGCAAATCTCCTTTGGCA-3′ and 5′-ATGCAGCATCATCAAGATTCT-3′) complementary to miR399 and miR172, respectively, and visualized using a phosphorimager BAS-1800 II (Fuji Photo Film Co. Ltd; http://www.fujifilm.com/).
Materials not commercially available and used in the experiments reported will be made available for non-profit research on request.
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
We thank Professor Mark Stitt (Max-Planck Institute for Molecular Plant Physiology) for his comments on the manuscript, and Dr Tzyy-Jen Chiou (Academia Sinica, Taipei, Taiwan) for discussions on the results. The work was supported by the Max-Planck Society.
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
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Supporting Information
- Top of page
- Summary
- Introduction
- Results and discussion
- Experimental procedures
- Acknowledgements
- References
- Supporting Information
Figure S1. Expression levels of mature miR399 and miR399d primary transcripts in Arabidopsis wild-type during Pi limitation and re-addition.
Table S1. Quantitative real-time RT-PCR threshold cycle values.
Appendix S1. Estimation of the time required to establish high mature miR399 levels by shoot-to-root phloem transport in roots that do not express miR399 PTs.
Please note: Blackwell Publishing are not responsible for the content or functionality of any supplementary materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article.
|TPJ_3363_sm_Appendix S1.doc
|Supporting info item
|Supporting info item
|Supporting info item
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.


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Audio: Romney Proudly Touts Anti-LGBT Positions
October 26, 2012 by HRC staff
Post submitted by Dan Rafter, Former HRC Associate Director of Communications
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Atlatsa Resources Corporation announces its operating and financial results for the three months ended September 30, 2014. Read more...
8 October 2014
Bokoni Platinum Mine investigates fatal injury to a community member
Opencast mining operations suspended
Atlatsa Resources Corporation has suspended operations at its Klipfontein opencast mine at Bokoni Platinum Mine pending the outcome of an urgent and full investigation into the fatal injury of a Mosotsi community member, Mr Bernard Moropane. Read more...
29 August 2014
Atlatsa appoints Chief Financial Officer
Atlatsa Resources Corporation is pleased to announce the appointment of Ms Boipelo Lekubo as part of its executive management team as chief financial officer effective 1 September 2014. Read more...
20 August 2014
Wage agreement reached at the Bokoni Mine
Atlatsa Resources Corporation is pleased to announce that Bokoni Platinum Mines Limited has reached a two-year wage agreement with its recognised labour union bodies. Read more...
15 August 2014
Atlatsa talks to Amplats about status of Bokoni joint venture
[Business Day] - Atlatsa Resources is holding talks with Anglo American Platinum (Amplats) and the Department of Mineral Resources, about the continued participation of the world's largest platinum producer in the Bokoni project. Read more...
The latest company filings may be found on the SEDAR website: www.sedar.com
14 November 2014
Condensed consolidated interim financial statements three and nine months ended 30 September 2014 Download (PDF, 220KB)
Management Discussion and Analysis of Financial Condition and Results of Operations for the three and nine months ended September 30, 2014 Download (PDF, 671KB)
14 August 2014
Results presentation for the quarter ended 30 June 2014
Download (PDF – 1.8MB)
Fact sheet
28 May 2014
Atlatsa Resources Corporate Fact sheet
Download (PDF – 462KB)


text:
Latest U.N. Food and Agriculture Organization Stories
Experts at the UN Food and Agriculture Organization (FAO) said in a report on Monday that fish consumption has reached record levels and world stocks need to be urgently rebuilt.
According to the UN Food and Agriculture Organization (FAO), a joint food safety commission has set rules to cut the level of cancer-causing chemicals in food.
The global economic slowdown combined with high food prices is causing one in six of the world's population, more than one billion people, to go hungry, the UN Food and Agriculture Organization (FAO) said Friday.
FAO calls for review of biofuel policies, subsidies ROME, Oct. 7 (Xinhua) -- Biofuel policies and subsidies should be urgently reviewed to preserve the goal of world food security, protect poor farmers, promote broad-based rural development and ensure environmental sustainability, said a U.N.
The move toward biofuel proliferation in the West may need to be reconsidered, a United Nations report advised on Tuesday.
Scientists reported Wednesday that the major forces behind recent avian flu outbreaks in Vietnam and Thailand are people, ducks, and rice patties, and not the number of chickens. Although avian influenza has been closely associated with chickens in the past, the new study found the number of chickens to be a less important predictor.
One hundred and eight countries have agreed on an action plan to save endangered livestock breeds, the U.N. Food and Agriculture Organization said Friday.
The rate at which livestock breeds are disappearing is "alarming," a senior official at the U.N. Food and Agriculture Organization said Tuesday, warning that precious genes could be lost forever.
Ministers and experts from across West Africa met on Wednesday to draft a plan to contain the bird flu virus as the U.N. Food and Agriculture Organization (FAO) warned of a looming regional disaster.
- Forsooth! indeed! originally a parenthetical phrase used in repeating the words of another with more or less contempt or disdain.


text:
It's time to expose them.
Kill Facebook before it becomes an unstoppable agent of chaos, its own Mr. Smith.
It only took one courageous selfie to start a national movement.
Selfie sticks, taco breaks, and Patagonia jackets. What more could you want?
Mark Zuckerberg is throwing things at the wall and seeing what sticks.
The perverse Facebook cult of Germanwings co-pilot Andreas Lubitz.


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Hairy stud Rich Kelly jerks off in the shower
For this week's solo scene, High Performance Men introduces hairy stud Rich Kelly. While this is his very first scene for High Performance Men, Rich has filmed several hardcore scenes with Falcon Studios and Raging Stallion last year and he will probably appear in a hardcore scene for High Performance Men soon.
Rich loves the ocean, but he doesn't live anywhere near the ocean, so when he arrived in Florida for his shoot, he wanted to go to the beach straight away. There are some beautiful shots of Rich at the beach and in the sea. The video starts when Rich takes a shower to rinse off the sand and salt water. It doesn't take long before he starts playing with his gorgeous cock and he's rock hard in an instant.
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I'm an easy going guy.
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- Catholicism, but not too serious about it
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- Capricorn, but it doesn't matter
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Global warming, lunch, or your next vacation… it's all fair game.
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Loeb winner gets N.E. honor
David Lang, president of the Professional Fire Fighters, will receive the inaugural Citzen's Right to Know Award. Lang's group spent years seeking records from the Local Government Center regarding how insurance premiums were being used. The LGC fought the efforts of the center to prevent public scrutiny, with the New Hampshire Supreme Court ultimately agreeing with the firefighters' position, and the LGC was forced to open its books. Millions of dollars have been refunded as a result.
Also to be honored at the NEFAC luncheon here will be Hartford Courant reporter Don Stacom, who exposed misdeeds in a Connecticut police department, and the lifetime achievements of New England Cable News founder and GlobalPost co-founder Philip Balboni.
"NEFAC's mission goes on year-round," said coalition Executive Director Rosanna Cavanagh. "We have training programs for journalists on FOI (Freedom of Information) and media law, and we advocate for greater access to public records through friend of the court briefs and expert legislative testimony."
Likewise, the Nackey S. Loeb School of Communications Inc., headquartered in Manchester, offers many opportunities for journalists and the general public to expand their knowledge of the First Amendment.
Tickets to the event are available on NEFAC's website, www.Northeasteren.edu/firstamendmentcenter.
The awards luncheon is held in conjunction with the New England Newspaper & Press Association's convention and trade show at the Boston Park Plaza Hotel.
|NH Angle >> Human Interest
0 New Hampshire students build Habitat house in Valdosta
0 Manchester priest convicted of molestation continues long road to clear his name
0 Maine charity founder commits suicide amid fraud probe that found he spent $7K at NH liquor stores alone
0 Case of missing Newton girl still active after 35 years
0 Hasbro selects Portsmouth for new Monopoly board
0 Mitt Romney to fight Evander Holyfield May 15 in Salt Lake City
0 'Clueless' actress Stacey Dash cast as NH woman sentenced to death in Sudan for being Christian
0 Cheer championship passes from Sunapee to Newport through act of sportsmanship
0 HuffPo: Jersey girl transplant apologizes to New Hampshire for thinking were big into flatulence
0 �Million Dollar Contractor� cast caught in real-life gun drama involving NH gunman
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NH natives make finals of NESN video contest
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Joe McQuaid's Publisher's Notes: If you see leprechauns tomorrow, you might soon be seeing Jean Valjean
Joe McQuaid's Publisher's Notes: The 'professionals' may say Trump is unpolished, but voters may find him more than an apprentice


text: Adobe PDF Jobs 32 were found based on your criteria Post a job. It's free! Newest Jobs First Oldest Jobs First Most Employer Hours Least Employer Hours Most Dollars Paid Least Dollars Paid Highest Feedback Score Lowest Feedback Score 1 2 3 4 › » MindMeister Expert/Specialist Fixed-Price – Est. Budget: $5.00 Posted 9 hours, 58 minutes ago 1.The applicant must be creative. 2.Good at brainstorming 3.Good at excerpting keywords 4.Must be logical and organized 5.Can work under pressure administrative-support adobe-pdf data-entry internet-research transcription PDF creation from "Word for Mac 2011" file Fixed-Price – Est. Budget: $15.00 Posted 13 hours, 48 minutes ago Dear Sir or Madam, I am looking for someone who can create ONE PDF file out of my "Word for Mac 2011" file. An experienced PDF creator shouldn't need longer than 10 mins and therefore I pay a fixed price of 5 US dollars. 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We primarily use R to do analysis and generate graphics, so the template should be easy to integrate with R output (txt files, csv files, pdfs, etc ... adobe-illustrator adobe-pdf latex


text:
Company Overview of Tuck School of Business at Dartmouth
Company Overview
Tuck School of Business at Dartmouth is an educational institution.
100 Tuck Hall
Hanover, NH 03755
United States
Key Executives for Tuck School of Business at Dartmouth
Tuck School of Business at Dartmouth does not have any Key Executives recorded.
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To contact Tuck School of Business at Dartmouth, please visit www.tuck.dartmouth.edu/. Company data is provided by Capital IQ. Please use this form to report any data issues.
Our data partners will research the update request and update the information on this page if necessary. Research and follow-up could take several weeks. If you have questions, you can contact them at email@example.com.


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Pollinators, Painting & Planting
- Food & Drink
- Whole Foods Market, Jacksonville FL
Health Starts Here Shopping Healthy on a Budget Store Tour
Whole Foods Market Jacksonville: Events
Thursday, March 20, 2014 from 6:00 PM to 7:00 PM (EDT)
Our own Healthy Eating Specialist, Heather, teaches you to follow the Four Pillars of Healthy Eating without breaking the bank! Learn to navigate the aisles to find the healthiest and most budget-friendly foods Whole Foods Market has to offer. Registration includes a chance to win a $25 Whole Foods Market gift card (must be present to win)!
Tour is limited to 10 participants, and registration is required. Please reserve your spot by Wednesday, March 19th.
*Interested in a private tour for yourself or a group? Contact firstname.lastname@example.com
Join millions of people on Eventbrite.
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text:
Stock Photo:
A dog and young boy with Down Syndrome together on a sofa.
- Image ID: 9453139
- Release Information: Signed model release filed with Shutterstock, Inc
- Copyright: Tomasz Markowski
High quality images curated from over 150 countries.
Over 40 million photos, vectors, illustrations, and more.
Simple and flat pricing by image or by month.
Over 230,000 images downloaded every day.


text:
Luke Johnson
I am Washington Correspondent for Radio Free Europe/Radio Liberty.
Previously, I was Politics Reporter at the Huffington Post in Washington, D.C.
Proficient in Russian, I returned to Moscow in December 2012 as part of the Russia-U.S. Journalist Exchange organized by the Knight Foundation and the International Center for Journalists.
I worked as a reporter covering campaigns and elections for The American Independent during the 2010 election cycle, part of the network of sites that included The Washington Independent.
I have appeared on MSNBC, NewsChannel 8, Current TV and BBC Radio, and am a frequent guest on The Bill Press Show.
I have lived in Washington since 2010. I grew up in the Boston suburbs and graduated from Claremont McKenna College in Southern California.
I can be reached at LUKEGJOHNSON (AT) GMAIL (DOT) COM


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Premium car manufacturers are likely to "lead the way in introducing connectivity and autonomous technologies", a UK study has found. 27 Mar 2015
The £100 million pledged by the UK chancellor earlier this week for new research into driverless cars shows the government is seeking to "lead the world in relation to new technologies", an expert has said. 23 Mar 2015
Automotive manufacturers selling vehicles in China are subject to strict rules governing product safety and must follow procedures set by a regulator when undertaking recalls. Manufacturers face financial losses, regulatory fines and significant reputational...
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This guide is based on UK law. It was last updated in March 2009. Sometimes a company infringes another company's rights without realising it – so intellectual property laws distinguish deliberate...
The UK's biggest car manufacturers have re-directed £1 billion of spending from overseas suppliers to Britain-based suppliers in the past three years, according to a new report. 16 Mar 2015
The UK government needs to develop a strategy to help the UK make the most of technological advancements in the transport sector, a committee of MPs has said. 09 Mar 2015
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Registered charity number 1151287 Wansford Cambridgeshire
We rescue unwanted dogs and puppies, and rehome to new
loving homes.
Thousands of unwanted dogs, strays and pound dogs have found their way through our kennels during the 30 years we have
been rescuing dogs.
You are very welcome to visit our kennels by appointment
Registered Charity Number 1151287
Have you ever considered offering a Staffordshire Bull terrier a forever home. The answer maybe "no" it maybe "yes" but before you decide please see our
and reasons you may be surprised by for considering a Staffordshire Bull Terrier
Welcome to our website and we hope you find the perfect companion for youself. Before you start your search please do take a minute to read about our appeal to find loving new homes for some of our overlooked residents. At Animal Helpline we try to help any breed who needs our help space permitting, for this reason a majority of the dogs we do take into our care are the ones who may wait longer for a new home.
A large percentage of people looking to help a rescue dog will want a Spaniel a Labrador a small dog or a Collie type. The plain crossbreed the Staffordshire Bull terrier the large dog will all be bottom of most peoples list or not even make that list.
This appeal is a push for that dog, the one people will pass and not even give a second glance to, the one who if you saw a picture of you would quickly pass over, the one who may not have the looks but the nature that would just be perfect for you.
Would you take A PEEK at some of these dogs waiting patiently
Help us find loving new homes for our dogs.Please print out and display our
These can be dispalyed in a variety of locations such as pet stores ,vets, dog training venues, popular dog walking areas, in your car.
Do you shop at Pets at home?
Do you have a VIP card?
Do you know you can help the rescue raise funds by registering your card to Animal Helpline?
Was your new years resolution to get fit? Well we can help. If you enjoy walking and have a passion for dogs come and join our dog walking club and get fit in the process.
We are looking for volunteers to walk our dogs during the week. Please ring 07849 637862 or 01780783331
Fun Agility/ obedience training and socializing held at the rescue. These are FUN Sessions – Focussing on bonding with your dog. We welcome all dogs .
Cost is £5 and is organised by Lynn & Liz – All the fee goes to AH to support our rescue dogs.
To find out more : Call Liz on : 07999 145311 or firstname.lastname@example.com
Do come along for a great day.
See our FUNDRAISING PAGE and let us fill your diary with dates of fun events planned through the year.
Happy endings are stories of some of our dogs now happily rehomed and celebrations of Gotcha days (day you adopted your canine chum) anyone who has adopted a dog from us or wants to celebrate their dog's Gotcha day send us a email and we can add to these pages. We love hearing how our dogs are doing.
Our dedicated fundraising team work tirelessly beavering away organizing all sorts of events to raise much needed funds for the rescue.
If you have a few hours spare would you consider joining this dedicated merry team.
To join our fundraising team for more information please contact Lynn on
Alot of people are not able to adopt or volunteer but you can still make a dog's wish come true by sending a gift via our
Dog Food:
Blankets & bedding - especially blankets/ throws/ bedspreads which we are very low on.
Spare leads, collars etc. new or used - especially slip leads.
Thank you to all the wonderful items already donated.
Please meet our beautiful 'Taz' a male Staffordshire Bull terrier. who we are desperately trying to find a foster home for. This sweet natured lad suddenly, unexpectedly lost some of the function intermittently of his back legs and our vet has advised it is believed to be neurological issues. The long term prognosis is currently unknown and it maybe that this will be terminal, if this is the case we want for any time Taz has left to be in a loving home. This lad has all our volunteers wrapped round his paws they are all devastated for Taz, he is such a bidding, friendly boy. A foster home preferably in or around Peterborough for access to our vets or a willingness to travel to our vets. If you think you can help please ring Roz on 07849 637862 or 01780783331 for more information.


text:
Sunday, October 26 2014
'The Best American Mystery Stories 2014' Delivers a Short Sharp Stab Into Many Different Lives
These stories pulsate with the everyday wrongs and trivial annoyances and tragedies with which each and every one of us is only too familiar.
Sunday, October 12 2014
In 'The Beggar and the Hare', Beggars Can Be Choosers
What really happens when you hit rock bottom?
Thursday, September 11 2014
Various Artists: XL Records - Pay Close Attention
A concise, pure and punchy pop history lesson.
Monday, June 16 2014
Radio Moscow: Magical Dirt
Ry Cooder + Raw Rhythms = Radio Moscow.
Friday, June 6 2014
Pharmakos: Nude
One cold, sensual slab of synth rock.
Marvin Gaye: The Soul Legend [5.Jun.14]
Moby: Almost Home - Live at the Fonda, L.A. [4.Jun.14]
The Trouble with Templeton: Rookie [29.May.14]
Paolo Nutini: Iron Sky EP [22.May.14]


text: Movies 'Cloverfield' Release Will Be Test of Online Hype January 11, 2008 Cloverfield, the highly anticipated new film produced by the creator of such TV shows as Alias and Lost, is out in theaters next week. For months, Internet speculation about the movie has been rampant. Listen Loading… Download Playlist


text:
Apparently The 5th Element Is Glass
These glasses have the flute and stem but are missing the stand. That's because you're supposed to stick them in the stand when not in use. The sand you ask? Yes, they were designed for hot Swedish summers at the beach.
Designer: Sebastian Kjersen & KayJay Design


text:
Wall Street Systems Boosts Revenues, Cuts Costs by Migrating to Red Hat Enterprise Linux
Free registration required
Executive Summary
Wall Street Systems helps financial institutions as well as enterprises throughout a variety of other industries improve their workflows and increase control of corporate treasury. They wanted to create new revenue streams by responding to customer demands for treasury and financial management software available as a service 24/7. Cut costs and improve reliability and scalability of internal systems. Ensure market competitiveness by reducing the infrastructure investment required by customers. For this they chose Red Hat. Wall Street Systems offers Red Hat Enterprise Linux based versions of all its flagship financial applications. Additionally, it runs Red Hat Enterprise Linux Advanced Platform running on Intel Xeon based servers internally for all development and Quality Assurance (QA) activities.
- Format: PDF
- Size: 415.05 KB


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- Shopping Bag ( 0 items )
From the Publisher"Klein is obviously an admirer, but also strives for balance....the definitive Moose story."
–Toronto Star
Author Biography: Jeff Z. Klein is the author of Mario Lemieux (1995), and the hockey fan's manifesto The Death of Hockey (1998) and co-author of The Coolest Guys on Ice (1996) and The Hockey Compendium (1987, 2001). Though a life-long Sabres fan, the peak moment of his life occurred on June 14th, 1994, when he drank champagne out of the Stanley Cup in the dressing room of Messier's victorious Rangers.
Posted December 2, 2003
Anyone who cares about hockey will be fascinated by this insightful biography of a living legend. Klein writes beautifully and really knows his stuff.Was this review helpful? Yes NoThank you for your feedback. Report this reviewThank you, this review has been flagged.
Posted July 22, 2010
No text was provided for this review.


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Art and Crafts
Michael Jackson Ink Rendering Fine Art Print
The Life and Times of Elvis Presley New York Times Compilation
Paul McCartney Limited Edition Fine Art Print
Love Me Tender Dog House Musical Figurine
Play It Again Elvis Jukebox Radio
Precious Moments I'll Never Let You Go Elvis Fan Figurine
Elvis Sonic Musical Wallet
Billboard Magazine Hot 100 50th Anniversary Songbook
Elvis Presley Pop Art Limited Edition
John Lennon Oil Painting Fine Art Print
John Lennon Oil Painting Print
Jimi Hendrix Pop Art Limited Edition Print
Michael Jackson Pop Art Print
Beatles Pop Art Limited Edition Art Print
Carrie Underwood Oil Painting Print
Elvis Presley GI Blues Bear
Handmade Elvis Impersonator Wine Caddy
The Beatles Yellow Submarine Poster
Grateful Dead Dancing Bears Tie Dye T-Shirt
Beatles Let It Be Black and White Latte Mug
The Best Of Kiss Bass Guitar Tab Songbook
The Beatles: It was 50 Years Ago Today Deluxe Book and DVD Set
Punk Rock Fun Time Activity Book
The Beatles Hollywood Bowl Poster


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ESPN.com "In a landmark decision, a federal judge ruled Friday that the NCAA is in violation of the nation's antitrust laws by restricting the compensation that major college football and men's basketball student-athletes can receive for use of their names, images and likenesses. U.S. District Court..." August 08
CBSSports.com "Bowling Green has fired head coach Louis Orr, the school announced Tuesday afternoon. Orr's contract was up after seven seasons, and the Falcons decided not to renew it. "We want to thank Coach Orr for his dedication to BGSU and his leadership in developing men of character in the Falcon..." March 12
College Basketball Talk "Bowling Green will not be renewing the contract of head coach Louis Orr, the school announced on Tuesday. Orr had coached the Falcons for seven seasons, finishing with a 101-121 record and just one season with a .500 record or better. BGSU was 12-20 this past season." March 11
Akron Beacon Journal "It was only two weeks ago that Devareaux Manley beat the final buzzer with a falling-away 3-pointer to allow Kent State to steal a win against Bowling Green, which held 13-point leads at two points in that game. Coming off a loss to Buffalo, Kent State (15-12, 6-8) again draws Bowling Green..." February 26
Akron Beacon Journal "Kent State was down and out more than once against Bowling Green on Wednesday night. The Golden Flashes trailed 13-0 to open the game and were behind another 13 points (40-27) not long into the second half. But twice Kent State fought back, and a team desperate for a win finally downed Bowling..." February 14
Akron Beacon Journal "Forward Quincy Diggs answered a game-tying shot with one of his own to lead the University of Akron men's basketball team to a last-minute victory over Bowling Green State University 65-63 at the Stroh Center on Sunday. No one could blame the Zips (16-7, 8-2 in Mid-American Conference) and..." February 11


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Books Worth Reading
Books Worth Reading
- 5 Pins
Cinnamon Swirl Cake | simplykierste.com
Buttercream Frosting
Cinnamon Swirl Cake with Buttercream Frosting
Cinnamon Swirl Cake with Simply Kierste
Biscuits and Gravy Casserole...I think I may have died and gone to heaven.
Breakfast Casseroles, Biscuit And Gravy Casserole, Lemon Sugar, Super Delish, Casseroles Recipe, Gravy And Biscuits, Biscuits And Gravy, Quick Breakfast Casserole, Gravy Casseroles
Biscuits and Gravy Casserole. I usually make a breakfast casserole at Christmas or biscuits and gravy. Now I can make both. Still gonna do homemade biscuits though.
Biscuits and Gravy Casserole - Lemon Sugar
Biscuits and Gravy #Casserole recipe. Lord.
Quick and easy, and super delish! Biscuit and Gravy Casserole
Biscuits and Gravy Casserole (but going to add the seasonings I use in my regular sausage gravy and biscuits to spice it up a bit)
Biscuits and Gravy Casserole - Lemon Sugar
Hunger Games Mockingjay Peeta, Katniss And Peeta Love, Hunger Games Feels, Katniss Peeta, Peeta And Katniss Love, Katniss And Peeta Mockingjay, Hunger Games Trilogy, Hunger Games Peeta And Katniss, Hunger Games Katniss And Peeta
Excuse while I go and cry out my soul... The Hunger Games Trilogy, Catching Fire
Marcelle 14kt White Gold Cushion FB Moissanite and by RosadosBox; FUTURE HUSBAND: PICK THIS STYLE RING.
Moissanite Engagement Ring, Cushion Cut Ring, Diamond Cut, Cushion Cut Engagement Ring, Simple Wedding Ring, Thin Band Engagement Ring, Cushion Engagement Ring, Princess Cut Wedding Ring, Cushion Cut Wedding Ring
Cushion cut engagement ring
Cushion cut ring
oh lordie