Podcasts about nicotiana

With (sort of) special appearances by Tobin Bell, Rosalind Franklin, and a 19th-century Dutch scientist whose name I certainly did not get right. Music by James Milor from Pixabay Information provided by: Tobacco (Nicotiana tabacum L.) - A model system for tissue culture interventions and genetic engineering by Thumballi R. Ganapathi, et al. (2004) [Abstract] In vitro transformation of cultured cells from Nicotiana tabacum by Agrobacterium tumefaciens by L. Marton, et al. (1979) [Abstract] https://www.pmiscience.com/en/smoke-free/nicotine/tobacco-plant-research/ On the historical significance of Beijerinck and his contagium vivum fluidum for modern virology by Neeraja Sankaran (2018) Therapeutic potential and phytoremediation capabilities of the tobacco plant: Advancements through genetic engineering and cultivation techniques by Nidhi Selwal, et al. (2023) https://doi.org/10.1016/j.bcab.2023.102845 PLANT vs. PATHOGEN: Enlisting Tobacco in the Fight Against Anthrax by Graeme Stemp-Morlock (2006) https://doi.org/10.1289%2Fehp.114-a364 “Tobacco Research and Its Relevance to Science, Medicine and Industry” by T.C. Tso (2006) DOI: 10.2478/cttr-2013-0824 Phytochemicals derived from Nicotiana tabacum L. plant contribute to pharmaceutical development by Wenji Zhang, et al. (2024) https://doi.org/10.3389%2Ffphar.2024.1372456

I actually planned to have two parts this time. And I hope this goes without saying, but I'll say it anyway: this podcast is for entertainment and educational purposes, not medical advice. Please, please do not attempt using tobacco for any of the historical medical purposes mentioned in this episode just because I said it's been done before. Not everything that has been done should be repeated. Music by James Milor from Pixabay Information provided by: Medicinal uses of tobacco in history by Anne Charlton (2004) https://doi.org/10.1258%2Fjrsm.97.6.292 Nicotiana tabacum (tobacco) by Marianna Jennifer Datiles & Pedro Acevedo-Rodriguez (2014) https://doi.org/10.1079/cabicompendium.36326 https://encyclopediavirginia.org/entries/tobacco-in-colonial-virginia/ https://www.worldhistory.org/article/1677/a-brief-history-of-tobacco-in-the-americas/ https://www.nps.gov/jame/learn/historyculture/tobacco-the-early-history-of-a-new-world-crop.htm https://gobotany.nativeplanttrust.org/species/nicotiana/tabacum/ https://plants.ces.ncsu.edu/plants/nicotiana-tabacum/ https://herbaria.plants.ox.ac.uk/bol/plants400/Profiles/MN/Nicotiana Tobacco smoking: the leading cause of preventable disease worldwide by Jonathan M. Samet (2013) https://doi.org/10.1016/j.thorsurg.2013.01.009 https://www.st-group.com/about-us/our-tobacco/history-of-tobacco/ https://tropical.theferns.info/viewtropical.php?id=Nicotiana+tabacum

Welcome to the Instant Trivia podcast episode 1147, where we ask the best trivia on the Internet. Round 1. Category: Hymns And Spirituals 1: Title that precedes "How sweet the sound, that saved a wretch like me". "Amazing Grace". 2: It's where "I'm goin' to lay down my burden", and "my sword and shield". "down by the riverside". 3: A hymn and anthem, "Lift" this "and sing, till earth and heaven ring". every voice. 4: "We shall come rejoicing, bringing in" these. the sheaves. 5: Asking for comfort and guidance, do this, "fast falls the eventide", "in life, in death, O Lord" do this. abide with me. Round 2. Category: Presidents And Baseball 1: On April 19, 1909 Taft attended a home game of this team and probably had a hot dog or 9. the Washington Senators. 2: Saying it was in the national interest, he kept baseball going during World War II. FDR. 3: It's the only team that has a U.S. president on its roster of former owners. the Texas Rangers. 4: One of his first jobs was re-creating Cubs games over the radio in Iowa. Ronald Reagan. 5: Before his Army years, he played baseball under an assumed name in the Kansas minor leagues. (Dwight) Eisenhower. Round 3. Category: Abbreviated States 1: This state that acts as a conjunction between Nevada and Washington has an abbreviation that is a conjunction. Oregon. 2: The abbreviation of this state is also an abbreviation for the largest city in California. Louisiana. 3: Show me that the abbreviation for this state means the habits of a predictable criminal. Missouri. 4: When abbreviated before the number 47, this state becomes an assault weapon. Alaska. 5: State whose abbreviation is also a cabinet department that was formed in 1989. Virginia. Round 4. Category: American Food And Drink 1: Maryland and Mississippi both "got" this as their state drink or beverage. milk. 2: This type of bread made from a starter has been a San Francisco specialty since the gold rush days. sourdough. 3: Packets of this fruity soft drink were first sold through the mail in 1927 for 10 cents apiece. Kool-Aid. 4: Some recipes for this sweet treat call for beets instead of food coloring. red velvet cake. 5: They're the 2 beverages that are combined to make an Arnold Palmer. iced tea and lemonade. Round 5. Category: Top Seeds 1: As the picture proves, you don't need the luck of the Irish for this plant; just scatter the seed about. clover. 2: This plant of genus Nicotiana has tiny seeds that use light to germinate so they can grow after top seeding. tobacco. 3: As Porky Pig's gal knows, seeds of these flowers should stay on top of the soil as they need light to germinate. petunia. 4: The seeds of this edible plant, whether Butterhead or Burgundy Boston, are planted on top. lettuce. 5: Just place the seeds of this flower on top of your soil and they'll be sprouting in no time. snapdragons. Thanks for listening! Come back tomorrow for more exciting trivia!Special thanks to https://blog.feedspot.com/trivia_podcasts/ AI Voices used

Isn't every plant great in a group? Well, the answer is no. Some plants are too vigorous in their growth habits to share the stage, while others are better if put on a pedestal all their own (i.e. the focal point plants of the landscape). Today's episode we talk about plants that are great in masses—that is to say—in groups of three or more. We have options for shade, choices for sun, and selections for those in-between exposures situations. We'll also feature some great plants that we've seen grouped to perfection in gardens featured in Fine Gardening. And you don't have to be a millionaire to group plants. Many of our suggestions are easily divided after just a year or two, providing you with multiple plants for the price of just one. Expert guest:  Daniel Robarts is a horticulturist at Coastal Maine Botanical Gardens in Boothbay, Maine. Danielle's Plants 'Kitten Around' catmint (Nepeta faassenii 'Kitten Around', Zones 3-8) ‘All Gold' Japanese forest grass (Hakonechloa macra 'All Gold', Zones 5-9) 'Kobold' liatris (Liatris spicata 'Kobold', Zones 3-8) Smooth hydrangea (Hydrangea arborescens and cvs., Zones 3-9)   Carol's Plants ‘Galaxy Blue' agapanthus (Agapanthus ‘Galaxy Blue', Zones 6-10) Feather Falls™ sedge (Carex 'ET CRX01', Zones 5-9) Firefinch™ coneflower (Echinacea purpurea 'G0052Y', Zones 4-8) Flowering tobacco (Nicotiana sylvestris, Zones 10-11)   Expert's Plants ‘Millenium' allium (Allium ‘Millenium', Zones 5-8) 'Brunette' bugbane (Actaea simplex 'Brunette', Zones 3-8) 'Starry Starry Night' hibiscus (Hibiscus 'Starry Starry Night', Zones 4-9)

Welcome to the Instant Trivia podcast episode 939, where we ask the best trivia on the Internet. Round 1. Category: boy story 1: What, were you raised by wolves? well, if you're this "Jungle Book" boy, then yes, you were. Mowgli. 2: Parzival is the avatar of a kid with an extremely good handle on '80s pop culture in this novel. Ready Player One. 3: The name "Pip" is mentioned 3 times in the first 2 sentences of this novel--we get it, the guy's name is Pip. Great Expectations. 4: R.J. Palacio said this bestseller about Auggie Pullman and his unique face was "a meditation on kindness". Wonder. 5: It's Nick Hornby's novel concerning a 12-year-old lad and a man in his 30s and how the 2 bond. About a Boy. Round 2. Category: of the tiger 1: The Jersey boys who play sports for this university are the Tigers. Princeton. 2: First graders work to achieve tiger rank in this organization. the Cub Scouts. 3: Here's the story of a dog named Tiger who was a family pet on this TV show. The Brady Bunch. 4: "Tyger tyger, burning bright, in the forests of the night", begins a poem in his 1794 collection "Songs of Experience". (William) Blake. 5: The largest carnivorous marsupial of modern times was the now-extinct thylacine, also known as this alliterative tiger. Tasmanian. Round 3. Category: logo a-gogo 1: The dynamic ribbon device (AKA "the wave") is part of the logo of this soft drink. Coca-Cola. 2: Its red and white bullseye logo was born in 1962 along with this retailer's brand name. Target. 3: This purveyor of roast beef and other sandwiches wanted to be called Big Tex, so the stylized cowboy hat logo makes more sense. Arby's. 4: Larry, the white-haired gent on the label of steel-cut and other products of this brand, got a subtle makeover in 2012. Quaker Oats. 5: A black prancing horse on a canary yellow background, the color of Modena, is the logo of this Italian car. Ferrari. Round 4. Category: time for "t" 1: These small pincers are used to remove splinters or hair. tweezers. 2: It's a newspaper concentrating on lurid news or whose pages are about half the standard size. a tabloid. 3: It's a set of kettledrums that's used in an orchestra. a timpani. 4: This plant belongs to the genus Nicotiana. tobacco. 5: It's the 11-letter word for an excessively fast heartbeat. tachycardia. Round 5. Category: russian literature 1: The last words spoken in this novel are "Hurrah for Karamazov!". The Brothers Karamazov. 2: In this last Chekov play, the character Trofimov says, "All Russia is our orchard". The Cherry Orchard. 3: This 1957 Pasternak novel was finally published in the USSR in 1987. Doctor Zhivago. 4: His 1863 novel "The Cossacks" grew out of his service in the Russian Army in the Caucasus. Tolstoy. 5: He was expelled from the Soviet Writer's Union in 1969; in 1970 he won the Nobel Prize for Literature. Solzhenitsyn. Thanks for listening! Come back tomorrow for more exciting trivia! Special thanks to https://blog.feedspot.com/trivia_podcasts/

RNA Interference, known as RNAi, is a biological process that leads to the silencing of gene expression.  A lot of plant viruses are RNA viruses including grapevine leafroll-associated virus and grapevine red blotch virus. Yen-Wen Kuo, Assistant Project Scientist in the Department of Plant Pathology at the University of California, Davis is researching ways to induce RNAi in grapevines to target virus. Growers may have heard of double-stranded RNA sprays which are intended to initiate RNAi. The challenge has been that double-stranded RNA breaks down quickly in the elements. The Kou lab is working to improve this process and look for alternatives that will have little impact on the ecology. Resources: 71: New Techniques to Detect Grapevine Leafroll Disease 131: Virus Detection in Grapevines Abstract: Development of Agrobacterium tumefaciens Infiltration of Infectious Clones of Grapevine Geminivirus A Directly into Greenhouse-Grown Grapevine and Nicotiana benthamiana Plants Kuo Laboratory – Plant Virology Maher Al Rwahnih, Foundation plant services RNA-Based Vaccination of Plants for Control of Viruses Yen-wen Kuo's Google Scholar page Vineyard Team Programs: Juan Nevarez Memorial Scholarship - Donate SIP Certified – Show your care for the people and planet   Sustainable Ag Expo – The premiere winegrowing event of the year Sustainable Winegrowing Education On-Demand (Western SARE) – Sign Up! Vineyard Team – Become a Member Get More Subscribe wherever you listen so you never miss an episode on the latest science and research with the Sustainable Winegrowing Podcast. Since 1994, Vineyard Team has been your resource for workshops and field demonstrations, research, and events dedicated to the stewardship of our natural resources. Learn more at www.vineyardteam.org.   Transcript Craig Macmillan  0:00  Our guest today is Yen-Wen Kuo. And she is Assistant Professor in the Department of Plant Pathology at UC Davis. I'm Craig Macmillan, your host, and I'm very excited to have Dr. Koh here with us today. Welcome.   Yen-Wen Kuo  0:11  Thank you for having me.   Craig Macmillan  0:13  So you've been doing some interesting work the lab on interference RNA, and also how it affects plant viruses and possibly insects in the future. Can you explain for those of us that did not take genetics like we were supposed to in college, what interference RNA is and how it works?   Yen-Wen Kuo  0:29  Sure. So RNA interference is a biological process in which certain types of RNA RNAs can trigger RNA interference. And then once it's triggered, it will produce specifics more RNAs, that can regulate gene expression, by degrading or binding to the target RNAs containing a homologous sequence containing a similar sequence of those small RNAs. So this is a general concept of RNA interference, we also call it RNAi is very complicated the whole process. And there are different pathways and mechanisms included in the RNA interference. RNAi is a primary and effective antiviral defense in plants, but also found in some fungi and insects and lower eukaryotes. And because of all these different mechanisms, scientists and researchers, they they work on different aspects of this mechanism for either plants or animals. And they're also looking for different potential and better ways to use RNAi for different applications.   Craig Macmillan  1:45  So if I understand correctly, you have cell and there is DNA in that cell, and there's genes that code for certain things. And so the RNA is was transmitting or was carrying information from that's encoded with the gene out into the world to do something, is that a fair explanation?   Yen-Wen Kuo  2:05  So the genome there in plants or animals and human is their DNA genomes is DNA, and then the DNA will transcribed into RNA. And those RNA, some of the messenger RNAs can translate into proteins. So it's a how the central dogma from DNA makes RNA and then RNA makes protein. In the old days, we thought that oh, the protein is the important things because the protein can have different functional, different functions in different ways to to regulate everything in the body or in different organisms. But then afterwards, we found that actually RNAs they have many different forms and they can function at the RNA level. So it can interfere with gene expressions and many different things.   Craig Macmillan  3:03  And how does this apply to plant viruses because you've done some really exciting work with Gemini viruses, I believe with grapevine virus a Tell me a little bit about that work and how that works.   Yen-Wen Kuo  3:15  A lot of plant viruses, they are RNA viruses, a lot of those devastating viruses in grapevines, for example, grapevine leaf roll associated virus or grapevine red blotch virus they. So grapevine leaf roll associated viruses and RNA virus and grapevine red blotch is DNA virus. So there are different types of viruses. And so my work is trying to use different viruses making them into viral vectors to induce RNAi in Grapevine plants, to target those important viruses causing diseases in the field for the grapevines. And because so for example, when the viruses they are infecting plants, they will trigger RNAi in the plant, so that plants can protect themselves from virus infection. And because of that, we're trying to develop viral vectors can trigger RNA interference to target those viruses that's causing diseases. The work I have on the grapevine Gemini virus A that GGVA is to either develop the virus into viral vectors to target RNA virus first. So that's the initial plan for us to use. GGVA the grapevine Gemini virus A target grapevine leaf roll associated viruses. So before we eventually target that virus, we have to do a lot of different tests. We need to know if the clones the constructs or DNA constructs we have of this, GGVA can actually affect Gravelines plants, so we have to do that. And then we want to see if we can develop it into viral vector to carry the sequence we want them to express in grapevines to do the work we want them to do. So then we use it to target genes in the plants to see if they can silence the genes in the plants. So then we did that, we found that yes, we can use that viral vector to silence genes in plants. And then now we try to see that if we can use this viral vector to target other RNA viruses, or other grapevine RNA viruses, because we are actually at the same time developing different viral vectors, and one of them is GBA, is grapevine virus, a another's name, it can be very confusing. GGVA is a DNA virus. GVA is an RNA virus totally different to viruses. So since we have both viruses in the lab, so first, we try to prove the concept. We use the GGVA, the DNA virus, to target the GBA wild type virus, to see if we can see any effects. The GBA infection viral titers in the infected grapevines. So this is what we're working on right now. And so eventually, we want to use this viral vector, and potentially other viral vectors to to target grapevine leaf roll associated virus. And maybe we can use it to target mealybugs too.   Craig Macmillan  6:35  How are these vectors introduced to the plant?   Yen-Wen Kuo  6:38  We modify from the previous reports how people try to deliver those constructs the plasmids into grapevines. Most of the experiments or the assays, from before, they needed to have grapevine plants grown from in vitro, on media or from embryos. But that's really a lot of work. And it will be harder to have applications in the field. So then we develop vacuuming filtration method that we can directly vacuum infiltrate those plasmids that those DNA construct plasmids directly into the greenhouse grown grapevine plants. So those plants are propagated from the cuttings and then those plants, they are usually maybe 12 to 19 inches high above the soil when we infiltrated those plasmids into those grow vine plants. So this is an we got pretty good results, we successfully introduced those DNA constructs into the grapevine plans and those constructs can be infectious and initiate the whole the virus replicate in the grapevine.   Craig Macmillan  7:50  So is this something that can be done in a nursery then with new plants? And basically, they then would come with the vector or is it something you could do in the field?   Yen-Wen Kuo  7:57  Yes, I think the plan is that we can introduce those plasmas in the nursery in greenhouse plants before we plant them into the field. So then the plants that's planted into the field, they can have this viral vector to protect the plants from specific viruses.   Craig Macmillan  8:18  Got it. That's really neat. That's a great idea. And it's pretty cool. So that's fantastic. And in the work that you're doing so far, it sounds really exciting. And it sounds like the direction that you're kind of going in the future is with leaf roll virus that you mentioned. And then also, interaction with mealybugs you mentioned. Can you tell me more about that? What's that work all about?   Yen-Wen Kuo  8:39  Because this virus does GGVA and other viral vectors we're working on to a lot of viruses infecting grape vines, their phloem limited virus, so this GGVA is also phloem limited, meaning that the virus is can only infect the tissues around or in the phloem  is restricted. It doesn't go to like mesophyll cells or epidermal cells in infected plants, because mealybugs they feed on phloems. So we think if they can pick up those RNA interference signals, may be those RNA interference signals those small RNAs can target mealybugs too. So we can choose different target sequences in mealybugs. Hopefully you can see some effects for many bucks to to prevent that from transmitting viruses or have lethal effects for mealybugs. That's the plan. Hopefully we can do that. But we have to do tests to see how the efficacy and everything though it can have mealybugs, because there are previously they are different studies they use RNAi on insects, and many people prove that they can see some effects. We hope that the viral vector approach can also use for really apply this into the field for grapevine plants.   Craig Macmillan  10:00  What kind of index on insects are we talking about?   Yen-Wen Kuo  10:03  Depends on what target genes or sequences we choose. For my first choice, I would like to have a target that can prevent the transmission of the virus by mealybug, that will be my choice. I'm not sure if it's good to kill the insects, if it's going to affect the ecology too much. So if we can make the mealybug not transmitting the virus or other diseases, I think there will be a very good first step if we can see a lower transmission rate. And and then we can see if we need to adjust from there.   Craig Macmillan  10:40  That is amazing. And we haven't, yeah, the little bit of research that I did we have we do have proof of concept basically on this in other cropping systems. Is that right?   Yen-Wen Kuo  10:55  Yes,   Craig Macmillan  10:55  Can you tell me a little bit more about that, because that might give us some some vision of where we might go in the vineyard industry.   Yen-Wen Kuo  11:01  So, the RNAi applications, people are already trying to do some of those works. So, one example is that before people can spray double stranded RNA into the field. So, let me talk a little bit about the introduction of why using double stranded RNA. So, there are different types of RNAs that can induce RNA interference, certain types, one of them is double stranded RNA, either double stranded RNA or the single stranded RNA, they can form into a secondary structure in folding into a structure like a hairpin RNA, those are found to be able to induce RNA interference. And there's also other things like artificial micro RNAs, there are different types of RNAs that can induce RNAi and most convenient ways to make double stranded RNA. And people have been synthesizing the double stranded RNA or using bacteria to produce those double stranded RNA and then spraying to the field to get some protection for the plants. It worked at some level, but it's just not stable enough. Although double stranded RNA is more stable compared to single stranded RNA, steroids and RNA can be degraded in the field with the sun and everything the whole environment it can be degraded, people started to look for ways like bio clay to protect the RNA, and then so, they can spray in the field. So, the RNA can last longer and cause the effects. So, those double stranded RNAs can be absorbed by the insects, they can pick up from the surface of the plant or the plant can absorb those double stranded RNA into the plants. So, those are different ways and people started to see some effects on that, but still, we have to improve those different methods delivering double stranded RNA or other types of RNA to induce RNA interference in the plant. So, they are different different approaches. So, one of that is now we are trying using virus to introduce the RNAi to induce the RNAi in the plants. So, people are trying different ways to deliver those specific RNAs to induce RNAi to target specific diseases, sometimes not just viral diseases, that they will try to target fungal disease or something else and insects. This is what many different groups they are trying to do also previously, another way is to try to make transgenic plants. So if we can make plants to express those RNAs that can induce RNAi targeting to specific diseases, then you don't need to really use any tool to the deliver because the transgenic plants itself can produce those RNAs doing to induce RNAi plants. So that's also another way that people are trying to do we call that host induced gene silencing HIGS, and the virus induced gene silencing is the way my group is working on and we call it VIGs vigs. So there are different ways that which we would use to introduce those RNAs to induce RNAi in the plants.   Craig Macmillan  14:31  And right now you are at the greenhouse stage, if I understand correctly.   Yen-Wen Kuo  14:35  Yes.   Craig Macmillan  14:36  Have you introduced mealybug into your experiments into your work yet?   Yen-Wen Kuo  14:40  Not yet. We are just working on targeting grapevine virus first to see the effects. So where we have to continue monitoring those tested plants to see if the effects can last long, and the efficacy and how good they can be. So now we're at four for five months, so it's still we can see the targeted virus is being suppressed in a very, very low titer. So GVA can cause some symptoms in the grapevine plants when they see the plans are infected. But we have to peel off the bark to see the symptoms, we want to see that after targeting to the GBA virus, we saw that the viral titer is very low, if we can see that, also, the symptoms is not there anymore, is now like wild type, when when the virus was infecting in the plants alone, if we can see the difference, we don't even see the symptoms there will be really great. And this part, hopefully I can collaborate with the collaborators, Maher, he's run the foundation plan services, he can help my group on this, to see that how good the effects can be using this GGVA viral vector. So after that, if we can successfully target two different viruses, then we will start to work to change the target sequence in this viral vector to target mealybugs. So that's after the virus work.   Craig Macmillan  16:12  Yeah, well, that's very exciting. This is a really fascinating idea, and obviously is still relatively new. And I think it's really great that you and everybody else is working on this sounds like there's tremendous potential, and I hope that you folks continue on are able to continue on, is there one thing really related to this topic, you would tell growers one thing that you would advise them or you would educate them with?   Yen-Wen Kuo  16:34  I understand that there could be some concerns and maybe doubts, questioning RNAi applications in the field, because before, they already probably heard about the spray of double stranded RNA or other methods, and they saw some effects but not stable enough. So they may have some concerns or doubts, I think many scientists are trying different delivery methods that can be applied efficiently in the field. And we will do different types of tests and trials to make sure we work on any potential issues of this technology before applying them in the field and try not to affect the whole ecology or anything in the field too. And obviously, the current approaches we have are not enough to keep certain grapevine diseases, at low enough incidence. So we have to explore more potential control approaches before those diseases get worse, and adjust the ways to manage those different grapevine diseases with this changing environment. And I think hopefully, we can all work together to achieve this same goal. And I understand this is something new, I hope everyone can keep an open mind and willing to work with us to do different trials and see if we can improve different approaches to control different diseases.   Craig Macmillan  17:58  Well, I hope so too. grape growers are very creative. And they're always looking for solutions to their problems that very much fit what you're describing. And it sounds to me, this could be another tool in the IPM toolbox that may not be the single solution may not be a silver bullet. But it sounds very exciting that it may play a very important role to improve the efficacy of other techniques we have, which is great. Where can people find out more about you?   Yen-Wen Kuo  18:22  So because I will, setting up my lab, so hopefully I can have a lab website soon. I don't have accounts at Twitter or Instagram.   Craig Macmillan  18:34  Neither do I.   Yen-Wen Kuo  18:36  I don't use social media a lot. So my email that people can reach me through the email. And hopefully, when this is up or in your podcast, I will have my lab website set up so people can find us our work, my lab website.   Craig Macmillan  18:53  And we will have links and everything else that we can find posted on the episode page at the Vineyard Team podcast website. I want to thank you for being on the program. This was really, really interesting and is a kind of a view into the future of what's possible. Yeah. Our guest today was Dr. Yen-Wen Kuo. She is with the Department of Plant Pathology at the University of California Davis. And I want to thank you for being on the podcast.   Yen-Wen Kuo  19:20  Thank you for having me on the show. I really appreciate this opportunity to talk about research to explain some details about our work to the course and hopefully, I answer some questions that growers might have. I look forward to in the future maybe collaborating with different people to make this thing to work.   Nearly Perfect Transcription by https://otter.ai

Discover the monochromatic and sophisticated beauty of Nicotiana ‘Lime Green' blooms, as well as how to grow them with BBC Gardeners' World Magazine commissioning editor Adam Duxbury. Get free nicotiana seeds with our May issue, buy it here: https://bit.ly/BuyGWMagSowalong Learn more about your ad choices. Visit podcastchoices.com/adchoices

Based on the research of Dr. Alan Sandstrom in his book Corn is Our Blood, Yara and Pythia discuss the ritual practices of the Nahua people of Mexico and how you can incorporate aspects of their philosophy into your own practice. They also discuss how to make your own kombucha along with information and the uses of Nicotiana rustica.

Green plants? Well of course! What else would our topic be for a St. Patrick's Day episode? Today we're talking about plants that are stunners despite being “just green.” Turns out that these selections of perennials, annuals, trees, and shrubs are some of the most textural and eye-catching plants you can grow. And, we'll remind listeners right at the top of the show—green comes in tons of different shades. So make way for some chartreuse, emerald, and lime colored options that are just what your garden needs. We can't guarantee planting these gems will attract leprechauns but having them in your beds will make you feel like you've scored a pot of gold. Expert guest: Mark Dwyer is garden manager for the Edgerton Hospital Healing Garden and operates Landscape Prescriptions by MD in Wisconsin. Danielle's Plants Irish moss (Sagina subulata, Zones 4-8) Beesia (Beesia deltophylla,Zones 6-9) ‘Soft Caress' mahonia (Mahonia eurybracteata 'Soft Caress', Zones 6-11) Hacquetia (Hacquetia epipactis syn. Sanicula epipactis, Zones 5-7)   Carol's Plants Langsdorff's tobacco (Nicotiana langsdorffii, annual) Maidenhair fern (Adiantum pedatum, Zones 3-8) Lady's mantle (Alchemilla mollis, Zones 3-8) 'Cupressina' Norway spruce (Picea abies 'Cupressina', Zones 3-8)   Expert's Plants ‘Virdis' and ‘Green Tails' love-lies-bleeding (Amaranthus caudatus 'Green Tails' and 'Viridis', annual) 'Act Green' and 'Spring Green' cockscomb (Celosia cristata 'Act Green' and 'Spring Green', annual) 'Sunday Green' and 'Sylphid' plume celosia (Celosia plumosa 'Sunday Green' and 'Sylphid', annual) 'Green Ball' dianthus (Dianthus 'Green Ball', annual)  Bells-of-Ireland (Moluccella laevis, annual) 'Queen Lime' zinnia (Zinnia elegans 'Queen Lime', annual) 'August Forest' gloriosa daisy (Rudbeckia hirta 'August Forest', annual) ‘Green Jewel' coneflower (Echinacea purpurea 'Green Jewel', Zones 4-9) ‘Green Envy' coneflower (Echinacea purpurea 'Green Envy', Zones 4-9) ‘Limelight' panicle hydrangea (Hydrangea paniculata 'Limelight', Zones 3-9) Little Lime ™ panicle hydrangea (Hydrangea paniculata 'Bulk', Zones 3-9) Little Lime Punch ™ panicle hydrangea (Hydrangea paniculata 'SMNHPH', Zones 3-9)

Welcome to the farm to fire podcast. I'm Ash, a certified tobacconist and cigar connoisseur; and I'm here to share knowledge on The Art Of Tobacco from the planting of the seed all the way to the experience of smoking. Join me for a smoke while learning the full cigar sommelier teachings and tips. Today we are talking about Nicotiana Tabacum (nuh-kow-shee-ann-uh taba-come) Nicotiana tabacum, or cultivated tobacco, is an annually grown plant of the Nicotiana genus..

 Over the next couple of episodes we are going to be dipping into a different group of life than we usually do, the plants. Mike has been getting back into carnivorous plant rearing and wants to share all he knows about the world of plants that eat bugs. How the tables can turn!    Darwin's drawings of the leaves and tentacles of a sundew (Drosera rotundifolia), Figures 1, 4, and 5 from "Insectivorous Plants" (1897), in the public domain.  Aphids and other small insects caught in the sticky trichomes of Nicotiana insecticida, a newly described species of tobacco from Australia. Photograph by Maarten Christenhusz, Figure 1 in Chase & Lambkin (2021).   Phylogeny angiosperm plants with carnivorous taxa indicated by numbered circles. Illustration by Andreas Fleischmann, in Fleischmann et al. (2017) "Evolution of carnivory in angiosperms" in Ellison & Adamec (eds) "Carnivorous Plants: Physiology, ecology, and evolution".A wetland in Loyalsock State Forest, Lycoming County, Pennsylvania that is home to temperate sundews. Photography by Nicholas_T via Flickr, used under a CC BY 2.0 license.  Tropical swamp in New Caledonia, habitat for Drosera neocaledonica. Photograph by  Boaz Ng via Flickr, used under a CC BY-NC-ND 2.0 license.Utricularia jamesoniana growing as an epiphyte on a tree. Photography by Dr. Alexey Yakovlev via Flickr, used under a CC BY-SA 2.0 licenseUtricularia corunta growing as a dense matt in an aquatic habitat. Photograph by peupleloup via Flickr, used under a CC BY-SA 2.0 license.Catopsis berteroniana in the Florida Everglades growing as an epiphyte in the upper branches of mangrove trees. Photograph by  Scott Zona via Flickr, used under a CC BY-NC 2.0 license.Albany pitcher plant (Cephalotus follicularis) in culture. Photograph by Lucas Arrrrgh via Flickr, used under a CC BY-NC-ND 2.0 license.Wild Nepenthes mirabilis growing in Hong Kong. Photograph by  Boaz Ng via Flickr, used under a CC BY-NC-ND 2.0 license.Nepenthes albomarginata growing from on a cliff side above a beach. This species has white trichomes around the rim of the pitcher that are attractive to foraging termites. Photograph by Bernard DUPONT via Flickr, used under a CC BY-SA 2.0 license.Nepenthes ampularia are a species that have adapted away from carnivory and instead capture leaves that fall from the canopy. Photograph by CIFOR via Flickr, used under a CC BY-NC-ND 2.0 license.A spoon-leaved sundew, Drosera spatulata. This species has a circumboreal distribution with an isolated population also found in the highlands of Borneo. Photograph by  Boaz Ng via Flickr, used under a CC BY-NC-ND 2.0 license.Forked-leaf sundews (Drosera binata) in the wild. Photograph by Doug Beckers via Flickr, used under a CC BY-SA 2.0 license. Oblong-leaved sundews (Drosera intermedia), showing the semi-aquatic habitat of this species. Photograph by Ashley Basil via Flickr, used under a CC BY 2.0 license.Vining sundew showing the round sticky traps along the vine and flowers. Photograph by Jean and Fred Hort via Flickr, used under a CC BY 2.0 license.Wild Venus fly trap in a natural environment. Photograph by NC Wetlands via Flickr, in the public domain.Venus fly traps being sold commercially. Photograph by Mike Mozart via Flickr, used under a CC BY 2.0 license.A waterwheel plant (Aldrovanda vesiculosa), which have snap traps and are closely related to Venus fly traps. This species is at risk in their native range but have been introduced into North America, where they may be invasive. This specimen was photographed at Fort AP Hill in New York, USA. Photograph by the U.S. Government, in the public domain.Individual Aldrovanda nodes showing the whorl of leaves and snap traps. Photograph by David Short via Flickr, used under a CC BY 2.0 license.A young dewy pine (Drosophyllum lusitanicum) grown in culture. While this sticky-leaved plant may look like a sundew, they are only distantly related to one another. Photograph by incidencematrix via Flickr, used under a CC BY 2.0 license.Abundant prey captured by a dewy pine. Photograph by incidencematrix via Flickr, used under a CC BY 2.0 license.Young Triphyophylum plants showing the characteristic wavy primary leaves. Photograph by  Carel Jongkind via iNaturalist, used under a CC BY-NC 4.0 license.Secondary carnivorous leaves of Triphyophylum. Photograph by Lotus-Salvinia.de via Flickr.Tertiary leaves of Triphyophylum, note the the apical hooks. Photograph by  Carel Jongkind via iNaturalist, used under a CC BY-NC 4.0 license. Questions? Comments? Follow the show on Twitter @Arthro_PodshowFollow the hosts on Twitter @bugmanjon, @JodyBugsmeUNL, and @MSkvarla36Get the show through Apple Podcast, Stitcher, Spotify, or your favorite podcatching app!If you can spare a moment, we appreciate when you subscribe to the show on those apps or when you take time to leave a review!Subscribe to our feed on Feedburner!  This episode is freely available on archive.org and is licensed under Creative Commons: By Attribution 3.0. http://creativecommons.org/licenses/by/3.0/

Gabi SchaffnerDirekter Link zur Audiodatei

As of February 12th 2022, it's been 113 years since Charles Robert Darwin was born. As such, we're back with another Darwin Day episode! This time around, we turn our attention to His Majesty's Ship Beagle, which Darwin would famously sail on. However, most people probably don't know a lot about the ship besides this fact. So, let's explore the Beagle's past and its possible connection to the present... Sources for this episode: 1) Burchett, M. (1996), Oceanography and Marine Biology: Water Movements and Oceanic Circulation Patterns. In: Waller, G. (ed.), Dando, M. and Burchett, M. (principal contributors) (1996), SeaLife: A Complete Guide to the Marine Environment. Pica Press: Smithsonian Institution Press. 2) Darwin, C. R. (1945), The Voyage of the Beagle. The Temple Press Letchworth: J. M. Dent & Sons Ltd. 3) Dubowsky, N., and Dubowsky, S. M. (1994), The final mission of HMS Beagle: clarifying the historical record. BJHS 27: 105-111. 4) The Editors, Encyclopaedia Britannica (2014), Arafura Sea (online) [Accessed 09/02/2022]. 5) Goodin, M. M., Zaitlin, D., Naidu, R. A. and Lommel, S. A. (2008), Nicotiana benthamiana: Its History and Future as a Model for Plant–Pathogen Interactions. Molecular Plant-Microbe Interactions 21(8): 1015-1026. 6) Thomson, K. S., Encyclopaedia Britannica (2019), Beagle (online) [Accessed 03/02/2022]. 7) Author unknown, National Geographic (date unknown), HMS Beagle: Darwin's trip around the World (online) [Accessed 05/02/2022]. 8) Author unknown, Royal Museums Greenwich (date unknown), HMS Beagle: Discover more about the ship that took Darwin around the world (online) [Accessed 03/02/2022]. 9) Author unknown, Wikipedia (date unknown), Beagle Channel (online) [Accessed 07/02/2022]. 10) Author unknown, Wikipedia (date unknown), Beagle Gulf (online) [Accessed 09/02/2022]. 11) Author unknown, Wikipedia (date unknown), HMS Beagle (online) [Accessed 03/02/2022]. 12) Author unknown, Wikipedia (date unknown), Robert FitzRoy (online) [Accessed 07/02/2022]. Darwin Day: Author unknown, Galapagos Conservation Trust (2015), Darwin Day Comic Strip (online) [Accessed 06/02/2022].

Researchers at the Council for Scientific and Industrial Research (CSIR) briefed the media on Friday on some of the organisation's research, development and innovation (RDI) efforts aimed at addressing the HIV/AIDS and Tuberculosis (TB) epidemics in South Africa. CSIR researcher Dr Advaita Singh shed light on an innovative plant-based, highly potent anti-HIV antibodies research initiative. “The CSIR's plant-based anti-HIV antibodies research is aimed at developing a cost-effective pre prophylaxis vaccine which will passively immunise against HIV/AIDS, pre and post exposure. This initiative was inspired by the desire to empower young girls and women to protect themselves against the virus as studies have indicated that women are theoretically, at higher risk of HIV acquisition,” said Singh. Working with the National Institute of Communicable Diseases (NICD), the CSIR is developing broadly neutralising antibodies against HIV-1, the most widespread human immunodeficiency virus, for the prevention and treatment of HIV/AIDS. CSIR scientists and their research partners were able to demonstrate the ability to engineer tobacco plants to produce unique glycosylation and rare sulfation post-translational modifications, a result that is not usually seen in plants. Glycosylation is a biochemical process during which a glycan – a large carbohydrate molecule – attaches to a protein, a lipid, or another organic molecule. The glycoengineered Nicotiana benthamiana (a relative of the tobacco plant) expressed two broadly neutralising antibodies named CAP256-VRC26 08 and CAP256-VRC26 09. These two antibodies were originally isolated by collaboration Aids research centre CAPRISA and the NICD from a South African HIV positive patient during a trial conducted by CAPRISA. The outcome showed that the antibodies from the CAP256 lineage were the most active, with high potency and broad specificity. “We still have a number of steps to get through, as the research process commonly requires, before this technology is available in the market, however, the CSIR and its partners intends on making it accessible to patients through an industry partner once it is ready,” added Singh. Another technology presented by the CSIR at the media briefing aims to provide rural areas with a diagnostic assay that will test for HIV and TB simultaneously using one device in the comfort of the patient's home. CSIR lab technician Kanyane Malatji provided insight into a multiplex HIV and TB point-of-care diagnostic test being developed for use in remote and rural areas. “The effective management of patients infected with HIV and TB is restricted by separate diagnoses. The situation is exacerbated in remote areas where patients must either travel long distances to reach a healthcare facility or wait a long time before obtaining their TB diagnostic results. Therefore, through our multiplex HIV and TB point-of-care diagnostic assay, we hope to mitigate these challenges with the objective of improving treatment outcomes for patients in rural areas,” said Malatji. The locally developed diagnostic test uses a solid surface that couples the HIV and TB antigens and makes use of a locally produced fluorescence detector. “The benefits of this technology are that it offers a cost-effective solution with a low turnaround time of approximately 2 hours. Additionally, no laboratory infrastructure is required, the device is easy to use and can be performed at point-of-care without highly trained personnel,” added Malatji. The CSIR's multiplex HIV and TB point-of-care diagnostic assay is funded by government and the plant-based anti-HIV antibodies research work is funded by the Department of Science and Innovation and the South African Medical Research Council.

A estas alturas de la pandemia, a muchos de nosotros nos han hecho un test de antígenos o de anticuerpos. Estas pruebas diagnósticas necesitan utilizar componentes del virus que hay que generar y las plantas son posibles fábricas de las proteínas del coronavirus, por supuesto modificadas adecuadamente mediante técnicas de biotecnología vegetal. Eso es lo que ha hecho nuestro invitado, Fernando Ponz Ascaso, y su equipo del INIA junto con otros grupos de investigación. La planta utilizada se llama Nicotiana benthamiana, es de origen australiano y ha sido modificada para que genere una proteína del coronavirus SARS-Cov-2, la proteína N. Esta no es la proteína utilizada habitualmente en los test actuales pero tiene, entre otras, la ventaja de que podría distinguir entre las personas que han sido vacunadas de las que sí han sido infectadas por el virus.

A estas alturas de la pandemia, a muchos de nosotros nos han hecho un test de antígenos o de anticuerpos. Estas pruebas diagnósticas necesitan utilizar componentes del virus que hay que generar y las plantas son posibles fábricas de las proteínas del coronavirus, por supuesto modificadas adecuadamente mediante técnicas de biotecnología vegetal. Eso es lo que ha hecho nuestro invitado, Fernando Ponz Ascaso, y su equipo del INIA junto con otros grupos de investigación. La planta utilizada se llama Nicotiana benthamiana, es de origen australiano y ha sido modificada para que genere una proteína del coronavirus SARS-Cov-2, la proteína N. Esta no es la proteína utilizada habitualmente en los test actuales pero tiene, entre otras, la ventaja de que podría distinguir entre las personas que han sido vacunadas de las que sí han sido infectadas por el virus.

Today in botanical history, we celebrate the 4th Earl of Chesterfield, an English botanist and a Patron Saint of gardeners. We'll hear an excerpt from a book by Tim Robbins featuring September in Louisiana. We Grow That Garden Library™ with a book that inspires us to make plants feel right at home in our homes. And then we'll wrap things up with a milestone moment in the history of Australia - the stunning loss of the Garden Palace that happened on this day 139 years ago today.   Subscribe Apple | Google | Spotify | Stitcher | iHeart To listen to the show while you're at home, just ask Alexa or Google to “Play the latest episode of The Daily Gardener Podcast.” And she will. It's just that easy.   The Daily Gardener Friday Newsletter Sign up for the FREE Friday Newsletter featuring: A personal update from me Garden-related items for your calendar The Grow That Garden Library™ featured books for the week Gardener gift ideas Garden-inspired recipes Exclusive updates regarding the show Plus, each week, one lucky subscriber wins a book from the Grow That Garden Library™ bookshelf.   Gardener Greetings Send your garden pics, stories, birthday wishes, and so forth to Jennifer@theDailyGardener.org   Facebook Group If you'd like to check out my curated news articles and original blog posts for yourself, you're in luck. I share all of it with the Listener Community in the Free Facebook Group - The Daily Gardener Community. So, there's no need to take notes or search for links. The next time you're on Facebook, search for Daily Gardener Community, where you'd search for a friend... and request to join. I'd love to meet you in the group.   Curated News 2022 Garden Trends Report: From Crisis to Innovation | Garden Media Group    Important Events September 22, 1694  Birth of Philip Dormer Stanhope, 4th Earl of Chesterfield, English statesman and writer. He's remembered for his letters to his son and other notable people of his day. He once advised his son, l recommend you to take care of the minutes, for hours will take care of themselves… Yale University has Chesterfield's note containing the words to On a Lady Stung By a Bee.   To heal a wound a bee had made  Upon my Chloe's face, It's honey to the part she laid, And bade me kiss the place. Pleased, I obeyed, and from the wound Suck'd both the sweet and smart ; The honey on my lips I found, The sting within my heart.   September 22, 1800  Birth of George Bentham, English botanist, writer, and teacher. He was going to be an attorney but pursued botany after living in the country. His thinking was preserved in a diary, which he kept for over fifty years. George once wrote, I decided that my means were sufficient to enable me to devote myself to botany, a determination which I never…. [had] any reason to [regret]. George's longest professional friendship was with the botanist John Stuart Mills who had lived with the Bentham family as a teenager. A pragmatist, George finished his Flora of the British Islands by writing every morning before breakfast. He purposely used simple language so that his book could reach a wider audience. George wanted everyone to see fundamental differences in plants. The useful way he classified plants laid the foundation for modern taxonomy. Later in his career, George co-authored the three-volume Genera Plantarum with Sir Joseph Hooker. The "Bentham & Hooker system" was widely used and made plant classification easier. George also worked with Ferdinand Mueller to create an impressive nineteen-volume flora of Australia. In 1830, George discovered Opal Basil (purple) which is prized for its flavor and color. But the plant George is most associated with is an Australian sister plant to tobacco, Nicotiana benthamiana. The plant was named in his honor and is used to create vaccines for the Ebola virus and the coronavirus. George died two weeks shy of his 84th birthday.   September 22nd   Today is the Feast Day of Phocas the Gardener, a Turkish innkeeper and gardener who lived during the third century. A protector of persecuted Christians, Phocas grew crops in his garden to help feed the poor. His garden aided him in living his most-remembered virtues: hospitality and generosity. When Roman soldiers arrived in his village, Phocas offered them lodging and a homemade meal using the bounty of his garden. As they talked, Phocas realized they had come for him. While the soldiers slept, Phocas went out to the garden to dig his own grave and pray for the soldiers. In the morning, Phocas revealed his identity to the soldiers who reluctantly killed him. Although gardening can be a solitary activity, Phocas illustrated how gardens create connection and community. Phocas is the Patron Saint of flower and ornamental gardens,  farmers, field hands, and market gardeners.   Unearthed Words Louisiana in September was like an obscene phone call from nature. The air--moist, sultry, secretive, and far from fresh--felt as if it were being exhaled into one's face. Sometimes it even sounded like heavy breathing. Honeysuckle, swamp flowers, magnolia, and the mystery smell of the river scented the atmosphere, amplifying the intrusion of organic sleaze. It was aphrodisiac and repressive, soft and violent at the same time. In New Orleans, in the French Quarter, miles from the barking lungs of alligators, the air maintained this quality of breath, although here it acquired a tinge of metallic halitosis, due to fumes expelled by tourist buses, trucks delivering Dixie beer, and, on Decatur Street, a mass-transit motor coach named Desire. ― Tom Robbins, Jitterbug Perfume   Grow That Garden Library Wild Interiors by Hilton Carter This book came out in 2020, and the subtitle is Beautiful plants in beautiful spaces. And this book has one of my favorite covers ever! So hats off to the book designer who came up with that incredible cover. Hilton is a plant stylist, a plant whisperer, and a plant coach, and all of that comes into play in this inspiring book of home interiors that are full of life, style, balance, health, and of course, plants. Carter is a master of greenery, and his approach to creating a welcoming room is making your plants feel right at home. Carter uses his book to take us on a tour of a dozen different homes that all feature their own unique ways of incorporating plants into their interiors and design. Each space is thoughtfully laid out, super comfortable, and beautiful. This book is 224 pages of plants at home in the home - and what a welcome addition for each of us to make. Lots of plant styling inspo in this book! You can get a copy of Wild Interiors by Hilton Carter and support the show using the Amazon Link in today's Show Notes for around $17   Today's Botanic Spark Reviving the little botanic spark in your heart September 22, 1882  On this day, at 5:40 am, the iconic Garden Palace in the Royal Botanic Garden in Sydney was destroyed in a fire that consumed the entire fourteen-hectare structure in forty minutes. The flames could be seen for twenty miles. Modeled after the Crystal Palace but constructed primarily with timber, The Garden Palace was built at a record pace and completed in just over eight months for the Sydney International Exhibition in 1879. It dominated Sydney's skyline for only three years. In its glory, a statue of the Queen stood beneath the palace dome made of thirty-six stained-glass windows. After the Exhibition closed, the Garden Palace was unfortunately used to store important records (including the 1881 census) and countless irreplaceable Indigenous artifacts. The cause of the fire has never been established. At the time of the fire, a French artist named Lucien Henry captured the fire on canvas. His assistant, George Hippolyte Aurousseau, recalled the moment in a 1912 edition of the Technical Gazette: Mister Henry went out onto the balcony and watched until the Great Dome toppled in; it was then early morning; he went back to his studio procured a canvas, sat down, and painted the whole scene in a most realistic manner, showing the fig trees in the Domain, the flames rising through the towers, the dome falling in and the reflected light of the flames all around. Today the Pioneer Memorial Garden rests on the site where the dome would have been. Built in 1938, the garden commemorated the 150th anniversary of European settlement in Australia.   Thanks for listening to The Daily Gardener. And remember: "For a happy, healthy life, garden every day."

With lots of questions to answer, Thordis and Alan Gray (East Ruston Old Vicarage) host a Postbag edition of the podcast, plus plenty of seed-sowing tips and Alpine trough advice with a focus on special plants that you could try. If you want to ask a question of the team you can email hello@getgardeningnow.co.uk, contact them on social media or comment on the Get Gardening YouTube channel. PLANT LIST Lunaria annua 'Chedglow' Eschscholzia californica Papaver dubium 'Albiflorum'  Papaver commutatum 'Ladybird' Papaver rhoeas 'Amazing Grey' Saxifraga burseriana   Gentiana asclepiadea   Calceolaria paralia 'Lemon Drops' Calceolaria Herbeohybrida Group 'Sunset Orange' Aloe aristata Clematis 'Pixie' Narcissus cantabricus Betula nana Forsythia ovata 'Paulina' Saxifraga longifolia 'Tumbling Waters'   Viola odorata 'King of Violets' Nicotiana x hybrida 'Tinkerbell' Fritillaria meleagris Geum 'Mai Tai'  Geum 'Mrs J Bradshaw' Geum 'Lady Stratheden' Geum 'Coral Tempest' Geum 'Totally Tangerine' Chrysanthemum 'Emperor of China'   Chrysanthemum 'Tula Improved'

An informal chat about what I've been up to in the garden, where I discuss methods of sowing seeds, planting up herbaceous borders and potting up dahlias. While I am sowing lots so different varieties of Nicotiana or Tobacco plants

Autor: Gabi Schaffner Sendung: Wurfsendung - Podcast Hören bis: ..

Este episodio es especial pues nos adentramos al chismecito sobre el trabajo galardonado con el Premio Nobel de Química 2020. Acompáñanos mientras Mel nos explica el porqué las tijeras genéticas serán una de las herramientas más importantes en un futuro no muy lejano. Manda tus correcciones a cienciaenchanclas@gmail.com Instagram: @cienciachanclas https://www.instagram.com/cienciachanclas/ Facebook: https://www.facebook.com/cienciachanclas Twitter: @CienciaChanclas https://twitter.com/CienciaChanclas YoutTube: https://youtube.com/channel/UCaBT5jpjwLs6elG7tE8CGQg Referencias: Mojica, F.J., Díez-Villaseñor, C., García-Martínez, J. et al. Intervening Sequences of Regularly Spaced Prokaryotic Repeats Derive from Foreign Genetic Elements. J Mol Evol 60, 174–182 (2005). https://link.springer.com/article/10.1007/s00239-004-0046-3 Mojica, F., Ferrer, C., Juez, G. and Rodríguez‐Valera, F. (1995), Long stretches of short tandem repeats are present in the largest replicons of the Archaea Haloferax mediterranei and Haloferax volcanii and could be involved in replicon partitioning. Molecular Microbiology, 17: 85-93. https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1365-2958.1995.mmi_17010085.x Press release: The Nobel Prize in Chemistry 2020. NobelPrize.org. Nobel Media AB 2020. Sun. 1 Nov 2020. https://www.nobelprize.org/prizes/chemistry/2020/press-release/ Doudna, J. A., & Charpentier, E. (2014). The new frontier of genome engineering with CRISPR-Cas9. Science, 346(6213), 1258096. https://science.sciencemag.org/content/346/6213/1258096.abstract?casa_token=TjyvAlzAs9cAAAAA:4d53SmLKJqrpZC9hIxcvx6EP72ipKy3jgdVqxmDYLPvHTm0EQ8BjoemEV-w_rvDcsTTn1LHAH3dd2i2y Li, J., Norville, J., Aach, J. et al. Multiplex and homologous recombination–mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nat Biotechnol 31, 688–691 (2013). https://www.nature.com/articles/nbt.2654

Gardening podcast nicotiana. Penny goes a bit zen over nicotiana success. A have-a-go gardener, join Penny Haslam as she heads outdoors to explore, learn and grow. My Garden Podcast is produced by Bit Famous Ltd. https://gardenpodcast.co.uk/ https://twitter.com/podcast_garden

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.30.249292v1?rss=1 Authors: Yin, Q., Zhang, J., Wang, S., Chen, J., Gao, H., Guo, C., Ma, L., Sun, L., Chen, S., Liu, A. Abstract: As a group of the most important phytohormone, auxin homeostasis is regulated in a complex manner. Generally, auxin conjugations especially IAA glucosides are dominant on high auxin level conditions. Former terminal glucosylation researches mainly focus on O-position, while IAA-N-glucoside or IAA-Asp-N-glucoside has been neglected since their found in 2001. In our study, IAA-Asp-N-glucoside was firstly found specifically abundant (as high as 4.13 mg/g) in ginkgo seeds of 58 cultivars from Ginkgo Resource Nursery built in 1990. Furthermore, a novel N-glucosyltransferase GbNGT1, which could catalyze IAA-Asp and IAA to form their corresponding N-glucoside, was identified through differential transcriptome analysis and in vitro enzymatic test. The enzyme was demonstrated to possess specific catalyze capacity toward the N-position of IAA-amino acid or IAA among 52 substrates, and was typical of acid tolerance, metal ion independence and high temperature sensitivity. Docking and site-directed mutagenesis of this enzyme confirmed that E15G mutant could almost abolish enzyme catalytic activity towards IAA-Asp and IAA in vitro and in vivo. The IAA modification of GbNGT1 and GbGH3.5 was verified by transient expression assay in Nicotiana benthamiana. In conclusion, our results complement the terminal metabolic pathway of auxin, and the specific catalytic function of GbNGT1 towards IAA-amino acid provide a new way to biosynthesis indole-amide compounds. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.29.225110v1?rss=1 Authors: Sun, G., Putkaradze, N., Bohnacker, S., Joncyk, R., Fida, T., Hoffmann, T., Bernhardt, R., Härtl, K., Schwab, W. Abstract: C13-apocarotenoids (norisoprenoids) are carotenoid-derived oxidation products, which perform important physiological functions in plants. Although their biosynthetic pathways have been extensively studied, their metabolism including glycosylation remains elusive. Candidate uridine-diphosphate glycosyltransferase genes (UGTs) were selected for their high transcript abundance in comparison with other UGTs in vegetative tissues of Nicotiana benthamiana and Mentha x piperita, as these tissues are rich sources of apocarotenoid glucosides. Hydroxylated C13-apocarotenol substrates were produced by P450-catalyzed biotransformation and microbial/plant enzyme systems were established for the synthesis of glycosides. Natural substrates were identified by physiological aglycone libraries prepared from isolated plant glycosides. In total, we identified six UGTs that catalyze the unprecedented glucosylation of C13-apocarotenols, where glucose is bound either to the cyclohexene ring or butane side chain. MpUGT86C10 is a superior novel enzyme that catalyzes the glucosylation of allelopathic 3-hydroxy--damascone, 3-oxo--ionol, 3-oxo-7,8-dihydro--ionol (Blumenol C) and 3-hydroxy-7,8-dihydro-{beta}-ionol, while a germination test demonstrated the higher phytotoxic potential of a norisoprenoid glucoside in comparison to its aglycone. Glycosylation of C13-apocarotenoids has several functions in plants, including increased allelopathic activity of the aglycone, facilitating exudation by roots and allowing symbiosis with arbuscular mycorrhizal fungi. The results enable in-depth analyses of the roles of glycosylated norisoprenoid allelochemicals, the physiological functions of apocarotenoids during arbuscular mycorrhizal colonization and the associated maintenance of carotenoid homeostasis. Copy rights belong to original authors. Visit the link for more info

Dee and Carol discuss flowering tobacco (Nicotiana sp.), vegetables that are easy to grow and hard to grow plus a new app to help all gardeners grow vegetables, and a few of their favorite old gardening books.  Here are some helpful links:Flowers:  The lime-green Nicotiana langsdorffii, with a narrow bell-shaped flower is hard to find seeds for but Louis the Plant Geek also grows this flower.Nan Ondra from Hayefield has an entire post on Nicotianas she's loved. She also has a cross between Nicotiana langsdorffii and N. alata ‘Lime Green' called ‘Ondra's Green Mix' which she may have in her  Etsy shop now.  Botanical Interests also sells a flowering tobacco called Indian Peace Pipe.Veggies:Information on a new app for growing vegetables is at From Seed to Spoon.Dirt:A few favorite old books include:Theme Gardens by Barbara Damrosch Mrs. Greenthumbs: How I Turned a Boring Yard into a Glorious Garden and How You Can, Too by Cassandra Danz (1993) Mrs. Greenthumbs Plows Ahead: Five Steps to the Drop-Dead Gorgeous Garden of Your Dreams by Cassandra Danz (1998)Email us at TheGardenangelists@gmail.com and look for us on Instagram, Facebook, Twitter and now Pinterest.For more info on Carol, visit her website.For more info on Dee, visit her website.We also invite you to join The Gardenangelists Garden Club on Facebook.(Some links are affiliate links. If you make a purchase after clicking on an affiliate link, we may receive a tiny commission. It does not affect the price you pay!)

Today we’re talking about something near and dear to every hobbit’s heart, the history and art of smoking pipe-weed. Obviously I want to be clear that this is something to be enjoyed in moderation by adults, and I know for many smoking can become an addiction so I just want to note that we aren’t endorsing illicit use of pipe tobacco and if you are prone to addiction or are struggling with addiction, this might not be a good episode for you. But because it’s such an important aspect of Hobbit history and culture, I did think it would be fun to have a chat about it. Concerning Pipe-Weed, from the Prologue to The Lord of the Rings“There is another astonishing thing about Hobbits of old that must be mentioned, an astonishing habit: they imbibed or inhaled, through pipes of clay or wood, the smoke of the burning leaves of a herb, which they called pipe-weed or leaf, a variety probably of Nicotiana. A great deal of mystery surrounds the origin of this peculiar custom, or ‘art’ as the Hobbits preferred to call it. All that could be discovered about it in antiquity was put together by Meriadoc Brandybuck (later Master of Buckland), and since he and the tobacco of the Southfarthing play a part in the history that follows, his remarks in the introduction of this Herblore of the Shire may be quoted…” - Concerning Pipe-weedThe smoking of pipe-weed, writes Merry Brandybuck, “is the one art that we can certainly claim to be our own invention.” Today we’re chatting with one of our patreon supporters, Timothy Martin! If you’d like to support Tea with Tolkien, you can learn more about our mission and the rewards here. Highlights from this Week’s Conversation:Pipe-weed obviously means a lot to Hobbit culture as a whole, so I wonder if Tolkien loved it as much as they did? Do you know what kind of tobacco he preferred? Tolkien refers to the smoking of pipe-weed as an “art” in the prologue to The Lord of the Rings. I feel like this ties in so well with the idea of Enchantment, the magic of the elves, which Tolkien also describes as an art. In what ways do you see it as an art-form?You mentioned when we were talking before that you have built sort of an online community for Catholics who also enjoy smoking a pipe. What’s that been like and what fruit have you seen from that community? What was the inspiration for starting this community?The Catholic Pipe Smoker’s Den Facebook Group

Professor Peter Waterhouse and the wonder plant Nicotiana benthamiana.

Professor Peter Waterhouse and the wonder plant Nicotiana benthamiana.

The TWiV team reviews the first FDA approved gene therapy, accidental exposure to poliovirus type 2 in a manufacturing plant, and production of a candidate poliovirus vaccine in plants. Hosts: Vincent Racaniello, Dickson Despommier, Alan Dove, Rich Condit, and Kathy Spindler Become a patron of TWiV! Links for this episode ASM Conference on Viral Manipulation of Nuclear Processes ASM Public Outreach Fellowship Kymriah approved (PennMed) CAR T cells (NCBI) Cost of Kymriah (NYTimes) Accidental exposure to poliovirus type 2 (Eurosurveill) GAPIII (WHO) Poliovaccine candidate in plants (Nat Commun) Vertical vaccine farm (TWiV 47) All picornaviruses, all the time (TWiV 425) Image credit Letters read on TWiV 459 This episode is brought to you by the Defense Threat Reduction Agency. Part of the U.S. Department of Defense, the Agency’s Chemical and Biological Technologies Department hosts the 2017 Chemical and Biological Defense Science & Technology Conference to exchange information on the latest and most dynamic developments for countering chemical and biological weapons of mass destruction. Find out more at http://www.cbdstconference.com Weekly Science Picks Kathy - Cassini photos Dickson - Caliber Biotherapeutics Alan - Grav Rich - Google street view of the International Space Station (article) Vincent - the bioinformatics chat Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Today we decipher an ancient message. It is the Feast Day of St. Nicotiana, and in the garden, there is almost lavender. Be Safe, and Remember: You Are Not Alone.

In this episode we talk about two lovely self-seeding annuals, being worried that a plant problem will spread, and growing random acts of kindness.

Tue, 22 May 2012 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/14494/ https://edoc.ub.uni-muenchen.de/14494/1/Stotz_Mathias_M.pdf Stotz, Mathias Michael ddc:540, ddc:500, Fakultät für Chemie und Pharmazie

Plastid transformation is a well-established tool for biotechnology as it allows highexpression levels of proteins and as it provides biological transgene containment because of maternal inheritance of cytoplasmic genes in most crops. In basic research the function and regulation of plastid encoded genes can be further studied with the help of plastid transformation. This study is focused on the three most critical steps to achieve plastid transformation in Arabidopsis thaliana and Rapid-Cycling Brassica rapa (RCBr), the establishment of a regeneration protocol, the construction of species specific vectors and the use of different transformation protocols. First, to allow the generation of fertile plants out of transformed cells, a reliable regeneration protocol was established. Fertile plants of Arabidopsis thaliana were obtained with high regeneration efficiencies from cotyledons, seed derived calli and protoplasts. In RCBr, fertile plants could be regenerated from tissue culture of hypocotyls. For introduction of genes into plastids, appropriate vectors with different selection and visual markers were constructed. These ensure site specific integration of desired sequences and enable selection of transformed cell lines. In this study, species specific vectors for Nicotiana tabacum, Arabidopsis thaliana and RCBr were designed and cloned, using aminoglycoside resistance markers such as the aadA gene (confers resistance to spectinomycin and streptinomycin), the nptII or the aphA6 gene (conferring resistance to kanamycin). To allow the selection through the herbicides phosphinothricin or bialaphos, the bar gene was employed. In addition, fluorescence markers such as GFP, DsRed and AmCyan were taken as visual markers. After the establishment of a regeneration protocol and the construction of specific vectors, two different protocols for plastid transformation, the particle gun- or PEG (Polyethylene glycol)-mediated transformation method, were applied. However, no transformed plant in Arabidopsis thaliana and RCBr could be recovered so far. That the transformation protocol does work in principal could be shown by the use of Nicotiana tabacum. Plastids of Nicotiana tabacum were successfully transformed using both transformation methods and AmCyan as new visual marker. Out of these experiments, fertile homoplasmic tobacco plants could be obtained as was shown by Southern blot analysis and reciprocal crosses. Thus, the functionality of the used transgene expression cassettes was clearly proven.

Für die Entwicklung einer extern regulierbaren Expression plastidärer Fremdgene wurde das bakterielle Induktionssystem des quorum sensing in Nicotiana tabacum (Tabak) getestet. Die Komponenten zur Kontrolle der Expression eines Reporterproteins, β-Glucuronidase (GUS), wurden von Vibrio fischeri entnommen und an die Expressionsmaschinerie der Plastiden adaptiert. Um die Induzierbarkeit der GUS-Expression zu testen, wurden Blattstücke der generierten stabilen Plastomtransformanten mit N-3-(Oxohexanoyl)-L-Homoserin-Lacton (VfHSL) bzw. Ethanol als Kontrolle inkubiert. Die Expression von GUS fand bereits im nicht induzierten Zustand statt, konnte jedoch durch die Auswahl geeigneter Promotorelemente auf eine Konzentration von 0,0003 % vom gesamtlöslichen Protein erheblich gesenkt werden. Mit dem Induktor VfHSL konnte hingegen keine signifikante Steigerung der GUS-Expression erzielt werden. Mögliche Ursachen dafür werden anhand der Literatur diskutiert. Weiterhin wurde in dieser Arbeit ein transientes Expressionssystem aufgebaut, um Vektoren auf ihre Funktionalität zu überprüfen. Mittels Polyethylenglykol (PEG) wurden dafür Tabak-Protoplasten mit einem konstitutiven GUS-Expressionsvektor transient transformiert und reproduzierbar eine signifikante GUS-Aktivität im Zellextrakt gemessen. Die plastidäre GUS-Konzentration ließ sich indessen nicht ermitteln - Ursachen und Probleme werden diskutiert. Insgesamt lassen die Ergebnisse jedoch darauf schließen, dass der Großteil der GUS-Expression im Kern/ Cytosol stattfand, während die Expressionsrate in Plastiden für einen Nachweis zu gering war.

Jasmonate sind Phytohormone mit vielfältiger Wirkung in Entwicklung und Stressmanagement der Pflanzen. Über die Perzeption und Transduktion der Jasmonatsignale ist bisher kaum etwas bekannt. Unter Verwendung des synthetischen Jasmonat-Analogons 6-Azido-1-oxoindanoyl(14C)isoleucinmethylester (IndAz(14C)IleMe) als Radioligand wurde eine spezifische Bindestelle in Sojabohne (Glycine max) biochemisch charakterisiert, in der Erwartung, eine Bindestelle für Jasmonate zu beschreiben. Die IndAz(14C)IleMe-Bindung erwies sich als spezifisch, saturierbar und reversibel. Da es sich aber um eine niedrigaffine Bindestelle handelt und die Affinität verschiedener Jasmonate und synthetischer Indanoyl-Isoleucin-Konjugate nicht mit deren biologischer Aktivität in Sojabohne korreliert, dürfte es sich bei der IndAz(14C)IleMe-Bindestelle nicht um einen Jasmonatrezeptor handeln. Sowohl bei Jasmonaten als auch bei Indanoyl-Isoleucin-Konjugaten wurden Methylester gegenüber den entsprechenden freien Säuren bevorzugt gebunden. Ein Enzym, das den Liganden umsetzt, scheint nicht vorzuliegen, da die IndAz(14C)IleMe-Bindung kein pH-Optimum aufwies und keine Umsetzung des Liganden beobachtet wurde. Mit fortschreitendem Alter der Pflanze nahm die Bindungsaktivität zu. Die IndAz(14C)IleMe-Bindestelle kommt in verschiedenen höheren Pflanzenarten vor, war hauptsächlich in der Wurzel nachweisbar und wurde in der Zellwand lokalisiert. Da die Bindestelle weder mit Salzen noch mit Detergenzien extrahiert werden konnte, gegenüber Proteinase K, DTT, Periodat, Lipase, Cellulase, Hemicellulase, Pectinase und Pectolyase resistent und zu 50 % hitzestabil war, wird vermutet, dass ein in der Zellwand fest verankertes Protein vorliegt. Zu den intrazellulären Signalvermittlern von Pflanzen gehört Calcium, nicht nur im Cytosol, sondern auch im Zellkern. In transgenen Nicotiana tabacum BY-2-Zellen wurden mit Hilfe des Photoproteins Aequorin erstmals jasmonatinduzierte Änderungen der Calciumkonzentration in beiden Kompartimenten gezeigt. Auch ein Vertreter aus der Gruppe der Phytoprostane, Phytoprostan B1 Typ II, löste Calciumantworten in Cytosol und Zellkern aus. JA und OPDA induzierten unterschiedliche Calciumsignaturen, die sich jeweils aus einer cytosolischen Calciumantwort gefolgt von einem Calciumsignal im Zellkern zusammensetzten. Die Unterschiede in Form, Höhe und Kinetik der einzelnen Antworten lassen auf zwei verschiedene Signaltransduktionswege bei JA und OPDA schließen. MeJA war in beiden Kompartimenten inaktiv und demonstriert dadurch, dass MeJA nicht immer, wie häufig angenommen wird, wie JA wirkt. Durch das Isoleucin-Konjugat der JA (JA-Ile) wurde eine dritte Calciumsignatur ausgelöst, die sich von der JA-induzierten Calciumsignatur durch das Fehlen der JA-ähnlichen cytosolischen Calciumantwort unterscheidet. Dieser Befund lässt vermuten, dass die Unterscheidung von JA- und JA-Ile-Signalen möglicherweise auf Ebene des Calciums stattfindet. Eine Struktur-Aktivitätsanalyse mit Indanoyl-Isoleucin-Konjugaten bestätigte, dass die Konjugation mit Isoleucin zur Veränderung der Calciumsignatur führt. Die unkonjugierte 1-Oxoindan-4-carbonsäure (Ind) verhielt sich wie JA, das Konjugat Ind-Ile wie JA-Ile. Ferner wurde gezeigt, dass für die Induktion der Calciumantworten eine freie, negativ geladene Carboxylgruppe unerlässlich ist. Neben MeJA erwiesen sich JA-IleMe, Ind-IleMe und 3-(Nitro-methyl)-2-((Z)-pent-2-enyl)cyclopentanon als inaktiv. 6-substituierte Indanoyl-Isoleucin-Konjugate zeichnen sich durch verstärkte biologische Aktivität aus. Tatsächlich verlieh der Ethyl-Substituent dem IndEt-IleMe calciuminduzierende Aktivität im Zellkern. Bei den freien Säuren Ind-Ile und IndEt-Ile wurde aber keine Aktivitätssteigerung durch Substitution festgestellt. Die Untersuchung der Expression einiger JA-responsiver Gene zeigte, dass unter den Versuchsbedingungen, die die Induktion von Calciumantworten ermöglichten, keine jasmonatinduzierte Genexpression erfolgte. Sollten die beschriebenen Calciumsignale die Expression bestimmter Gene vermitteln, ist eine ausgewählte Gruppe von Genen zu erwarten, deren Expression eventuell einen besonderen physiologischen Zustand der Zellen erfordert.

Die Pflanzenzelle enthält ein integriertes, kompartimentiertes genetisches System, mit den Subgenomen im Zellkern, in den Mitochondrien und den Plastiden, das aus Endocytobioseereignissen mit prokaryotischen Zellen hervorgegangen ist. Im Laufe der Evolution der eukaryotischen Zelle wurden die genetischen Potentiale der symbiontischen Partnerzellen vermischt. Dabei ging ein Teil genetischer Information verloren, ein anderer wurde aus den Organellen in den Kern transferiert, und außerdem wurde neue Information hinzugewonnen. Dies ging einher mit der Einbettung von Mitochondrien und Plastiden in die Signaltransduktionsketten und Regelkreise der Wirtszelle. Heute interagieren die Subgenome auf vielen Ebenen; ihre Expression wird in der Pflanzenzelle koordiniert in Raum, Zeit und Quantität reguliert. Die Interdependenz der Subgenome hatte ihre Koevolution zur Folge, so daß die genetischen Kompartimente der Zelle nicht mehr ohne weiteres zwischen Arten ausgetauscht werden können. Kombinationen von artfremden Organellen können zu Entwicklungsstörungen führen, wie sie sowohl von "kompartimentellen" (Genom/Plastom-) Hybriden als auch von Cybriden beschrieben worden sind (Bastardbleichheit, Bastardscheckung). In dieser Arbeit wurden reziproke Cybriden der Arten Atropa belladonna und Nicotiana tabacum auf molekulare Determinanten von Genom/Plastom-Inkompatibilität untersucht. Die Cybriden sind je nach Kombination elterlicher Organellen entweder albinotisch [Kern von Atropa, Plastide vom Tabak; Ab(Nt)-Cybride] oder gleichen dem Wildtyp [Kern von Tabak; Plastide von Atropa, Nt(Ab)-Cybride]. 1. Als Voraussetzung für einen Sequenzvergleich der plastidären Chromosomen beider Solanaceen-Arten wurde das Plastidenchromosom von Atropa komplett sequenziert. Der Vergleich der (Atropa)-Sequenz mit der bekannten des Chromosoms aus dem Tabak und anschließende molekularbiologische Untersuchungen führten zur Identifizierung von zwei potenziellen Ursachen für die Defekte im albinotischen Material. 2. Die Ab(Nt)-Cybride zeigt eine gestörte Akkumulation von Transkripten für eine Reihe von Operonen. Das resultierende aberrante Transkriptmuster ähnelte verblüffend dem von Tabakpflanzen mit Defizienz der plastidenkodierten RNA-Polymerase (PEP). Möglicherweise ist in der Cybride die Interaktion des PEP-Apoenzyms mit einem oder mehreren der kernkodierten Sigmafaktoren gestört. Tatsächlich unterscheiden sich die für eine Untereinheit der PEP kodierenden (plastidären) rpoC2-Gene von Tabak und Atropa durch eine Insertion/Deletion an einer Stelle im Molekül, die mit Sigmafaktoren interagieren kann. Transformation der Plastiden der Ab(Nt)-Cybride mit dem rpoC2-Gen aus Tabak führte in der Tat zu einer partiellen Reversion zum WT und macht Transkriptionsdefekte als eines von offenbar mehreren Determinanten für die Genom/Plastom-Inkompatibilität in Artbastarden wahrscheinlich. 3. Neben der Transkription ist im albinotischen Material auch die RNA-Edierung gestört. Die plastidären Editotypen beider Solanaceen ähneln einander, doch gibt es für beide Arten spezifische Edierungsstellen. Von den fünf tabakspezifischen Stellen in der Ab(Nt)-Cybride werden vier nicht ediert. Offensichtlich besitzt der Atropa-Kern nicht die notwendigen Kernfaktoren zur Prozessierung dieser Stellen. Da Edierung generell hochkonservierte und funktionell wichtige Aminosäurepositionen betrifft, trägt der Ausfall der Edierung sehr wahrscheinlich ebenfalls zum beobachteten Defekt in der Plastidenentwicklung bei. 4. Auf der anderen Seite werden die Stellen der grünen Nt(Ab)-Cybride, bemerkenswerterweise auch Atropa-spezifische, heterolog vom Tabakkern ediert. Der erstmalige Befund von heterologem Edieren stellte sich als Folge der Allotetraploidie von Tabak heraus. Untersuchungen dieser Stellen in den diploiden Eltern des allotetraploiden Tabaks, N. tomentosiformis als Nachkomme des Vaters und N. sylvestris als Nachkomme der Mutter, zeigten, daß der Tabak die Fähigkeit zur heterologen Edierung von Atropa-spezifischen Stellen wohl vom Vater ererbt hat. Dies wurde auch durch einen transplastomischen Ansatz bestätigt. In diesen Experimenten wurde die intronnahe ndhA-Edierungsstelle aus Spinat, die es auch in N. tomentosiformis gibt, nicht aber in N. sylvestris, in Tabak über ballistische Transformation eingebracht. 5. Über Konstruktionen, die entweder der gespleißen oder ungespleißten ndhA-mRNA inklusive der Edierungsstelle entsprachen, konnte gezeigt werden, daß die Edierung an dieser Stelle immer erst nach dem Spleißen erfolgt. Dies ist der erste Nachweis einer strikten kinetischen Verknüpfung von RNA-Edierung mit einem anderen mRNA-Reifungsschritt in Plastiden. Er zeigt an, daß das ndhA-Intron phylogenetisch älter als die ndhA-Edierungsstelle ist. Mechanistische Implikationen dieses Befundes werden diskutiert.

Plastid chromosomes from the variety of plant species contain several conserved open reading frames of unknown function, which most probably represent functional genes. The primary aim of this thesis was the analysis of the role of two such ORFs, designated ycfs or hypothetical chloroplast reading frames, namely ycf9 (ORF62) and ycf10 (ORF229, cemA). Both were analyzed in Nicotiana tabacum (tobacco) via their inactivation using biolistic plastid transformation. A new experimental protocol, based on pulsed-field gel electrophoresis (PFGE), was established to reliably assess the homoplastomic state of transformed plants. 1. Functional analysis of the ycf9 gene product: The inactivation of ycf9 in N. tabacum as well as in Chlamydomonas reinhardtii yielded a homoplastomic mutant phenotype after several rounds of regeneration under selective pressure. The mutant plants grew photoautotrophically, but displayed two clear phenotypes, a light-sensitive one, increasing with the light intensity, and a dwarf phenotype under low-light combined with temperatures below 20°C. The ycf9 gene product was exclusively located in PSII core complexes. This localization was based on the isolation of protein complexes released from thylakoids by controlled, partial lysis, followed by sucrose density gradient centrifugation or 2D gel electrophoresis. This finding revised data of the literature. Biochemical analysis indicated an involvement of the protein in the interaction of the light harvesting antenna II complex (LHCII) with PSII cores. In particular, PSII-LHCII supercomplexes could no longer be isolated from transplastomic tobacco plants. Furthermore, the minor chlorophyll a/b-binding proteins CP26, and to a lesser extent CP29, were substantially reduced under most growth conditions analyzed, in both, tobacco and photoautotrophically grown Chlamydomonas mutants (Swiatek et al. 2001). The gene was therefore renamed psbZ. The ∆psbZ-related alterations in the supramolecular organization of PSII complexes were accompanied by considerable modification in (i) the phosphorylation pattern of PSII subunits, (ii) the rate of deepoxydation of xanthophylls, and (iii) the kinetics and amplitude of non-photochemical quenching. The proposed position of PsbZ in close proximity to CP43 enables the protein to interact with PSII cores to elicit an adaptation process in response to excess light excitation. The molecular mechanism underlying this energy dissipation process remains to be investigated. 2. Functional analysis of the ycf10 gene product: Biolistic plastid transformation was also used to inactivate the ycf10 reading frame in tobacco. After several rounds of regeneration under selective pressure, homoplastomic plants were obtained. Northern analysis uncovered co-transcription of ycf10 within the psaI-ycf4-ycf10-petA gene cluster, with at least two promotor regions upstream of the psaI gene. The mutant plants grew photoautotrophically and developed dark green leaves with numerous pale green to white regions, the latter devoid of photosynthetic activity. The loss of ycf10 did not affect photosynthetic activity, as indicated by unaltered chlorophyll fluorescence. The tobacco ycf10 gene product was localized in the chloroplast inner envelope membrane. Neither protein composition of stroma or thylakoid fractions, nor the stability of the photosynthetic protein complexes were affected in the mutant plants. In contrast, CO2- dependent oxygen evolution was strongly reduced, with a maximum rate of Ci-dependent photosynthesis being approximately 50% lower than in wild-type plants. Two explanations can account for the observed phenomenon: (i) de-regulation of carbonconcentrating mechanisms in transformed cells, or (ii) an indirect effect on CO2-uptake in ∆ycf10 plants. 3. Pulsed-field gel electrophoresis is an ideal tool to verify the homoplastomic state of transformed plants: To enhance the sensitivity of detection of heteroplastomic states, and to distinguish between plastome-located wild-type segments in transplastomic material and promiscuous DNA, a new approach was developed. Customary Southern and PCR techniques are not sensitive enough or not discriminating the latter alternatives, respectively. Pulsed-field gel electrophoresis allows to isolate virtually contamination-free plastid DNA. Plastid DNA isolated this way lacked traces of nuclear and mitochondrial DNA at a detection level of 50 DNA molecules. This excludes that gene-specific PCR amplification products originate from promiscuous nuclear or mitochondrial gene copies. Therefore, PFGE appears to be an ideal tool to investigate the homoplastomic state of transformed plants, especially when combined with radiolabeled probes and Southern techniques.