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The top ten Twitter jokes about Findus and horse meat
Fresh on the back of the horse burger scandal, frozen meal producer Findus have taken their beef lasagnes off the shelves because they were found to contain traces off horse meat. Food officials have revealed that some of the ready meals contained '100 per cent' horse meat and shoppers have been told not to eat them.
Twitter users haven't been slow out of the gates and have taken a strong hold with some tasty one-liners.
I should be on a Stable Diet but I think I might give this #Findus meal a miss! #YFB pic.twitter.com/uLTwTcUK
— Chubby Brown (@RoyChubbsBrown) February 8, 2013
It was probably a good thing that they stopped sponsoring this race… #findus pic.twitter.com/h6Jzbbo2
— bettingpro (@bettingpro) February 8, 2013
What's the fuss? For years we've been told that Ready Meals contain too much Salt and Shergar. #findus
— Graeme Garden (@GraemeGarden1) February 8, 2013
Had #Findus beef lasagne for the mane course at lunch and now I've got the trots…
— Martyn Ziegler (@martynziegler) February 7, 2013
'Food beyond compare, food beyond belief, mix it in a mincer and pretend it's beef'. #Findus #dear #LesMis
— westendproducer (@westendproducer) February 7, 2013
#Findus website: "using only the best ingredients and a generous pinch of imagination in our recipes." I'd £10 at 5/1 on Imagination at Ayr
— Stewart Weir (@sweirz) February 7, 2013
So now Findus have withdrawn their products because of Horse meat contamination.. Shame..I really liked their Spaghetti Bologneighs. #findus
— The Carpenters Arms (@thecarpentersw1) February 8, 2013
Doctor says I should watch what I eat, so I've got tickets to the Grand National. #Hiyooo #findus
— Andrew Trendell (@AndrewTrendell) February 8, 2013
#Findus confirms its products will not be available as of tomorrow as they are running the 1:20 at Chepstow, 2:30 at Aintree & 3:20 at York
— Andy Cawthray (@ChickenStreet) February 8, 2013
I'm not sure I see the fuss about this #Findus story, quite Frankely.
— Paddy Power (@paddypower) February 8, 2013


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Tales of the Jedi [abridged - audio]
by Tom Veitch
No current Talk conversations about this book.
This is a small collection of short stories about the Jedi Knights in the age before the movies. These are stories of adventure of young jedi facing their first conflicts. I listened to the audio version of the book.
Unfortunately, the dialog is pretty bad. The characterization of the young Jedi is weak and poorly written. The author explains thing to the reader by using ignorance, often in the jedi, who do know know some of the basics in how the force works. I suspect the reader knows far more than the young jedi.
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Full cast audio drama adapted from the Dark Horse Comics series written by Tom Veitch.
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The Mysterious And Haunting Beauty Of Antelope Canyon, Arizona
Formed over hundreds of years of water running through sandstone, Antelope Canyon is both a sacred site for the Navajo and a favorite destination for tourists from all over the world. Antelope Canyon is the most-visited and most-photographed slot canyon in the American Southwest. It is located on Navajo land near Page, Arizona. Antelope Canyon includes two separate, photogenic slot canyon sections, referred to individually as Upper Antelope Canyon or The Crack; and Lower Antelope Canyon or The ... View More


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Forum indexCanon EOS 7D / 10D - 80D TalkChange forum
60D upgrade anytime soon?
Started May 10, 2012 | Discussions thread
DonaldHaines • New Member • Posts: 3
Re: 60D upgrade anytime soon?
In reply to joelroffman • May 11, 2012
I have a 60D and love it..... but mostly because of the glass I hang off of it. I have an 18-200 lens that is good for those times when you can only bring a single lens with you, but when you compare the quality of pictures from that lens to that from the 70-200 you realize that there is no substitute for good glass. Something in the 400mm range really brings the wildlife closer and might be a better idea than upgrading the body...
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Washouts should not be intimidating, especially if you approach them from the correct angle. Team USA Head Coach Rod Ross explains how you still can use your strike ball to pick up a washout by just moving your feet to the angle that is most advantageous to cover the spare.Watch Now >>
Some of the most common bowling injuries involve the shoulder. USBC Hall of Famer Lynda Barnes discusses how she overcame her shoulder injury in order to continue her career on the lanes. Watch as she goes over the exercises that got her back into shape. She also offers her insight on returning to the sport… Read more »Watch Now >>
Let Team USA Sports Performance Specialist Nick Bohanan show you how all bowlers can benefit from strength exercises. Learn how core strengthening involves your abs, hips and back muscles and how performing these exercises will improve your posture and leverage. By performing the exercises recommended by Bohanan, you will be able to enhance your strength… Read more »Watch Now >>
The bowling release is one of the most crucial aspects of the game. Team USA Head Coach Rod Ross talks about some of the ways you can practice your release from the comfort of your own home. Watch Rod and USBC Gold Coach Bryan O'Keefe demonstrate how a softball and football can be used to… Read more »Watch Now >>
Certain spares can be attacked multiple ways, and finding the way that is most comfortable for you to consistently repeat is a key to bigger scores. USBC Hall of Famer Carolyn Dorin-Ballard and Team USA Head Coach Rod Ross demonstrate options for converting the 2-8 spare combination.Watch Now >>
USBC Director Carolyn Dorin-Ballard and USBC Gold Coach Bill Monce teach you the differences between bowling grips and slugs. Find out what they are used for and learn the benefits of using bowling grips and/or bowling slugs. Also, learn more about the thumb hole and see what options you have regarding thumb inserts. Use these… Read more »Watch Now >>
Team USA Head Coach Rod Ross and Assistant Head Coach Kim Terrell-Kearney teach you how to create a more consistent approach to the line. By using two strips of tape to lay out a guideline to your approach, this drill will help you draw the ball back in a straight line rather than pulling it… Read more »Watch Now >>
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"Last year we came up a little short," Bolden said. "We went 4-5 and I really don't know what my stats were. We lost in the wild card game.
Bolden is more than a quarterback. He is like an extension of his coaching staff on the field and helps the others around him by the way that he goes out and plays.
"I have good ability and I am a leader on the field. I can make the other players around me better with the way that I play.
"I feel like I can throw the ball well and take off and run as well. I feel like I can do everything at the quarterback position."
Bolden had only been on one trip at the time of this interview, but would like to take some more as new offers seem to keep pouring in for him.
"I have only taken a trip to Northwestern for their junior day and that's it. I would like to take a few more and it just needs to line up with my schedule.
"I would like to go and see all of the schools that are contacting me and that have offered. I would like to know more about all of them."
Nebraska is one of many teams that have tendered an offer to Bolden. He isn't commenting on who he might be favoring and is saying that everything is open as of right now.
"I don't have a short list that I am favoring. I would say that I am considering all of the schools that I have offers from as of right now.
"At this point, I don't really have a specific opinion about any team. I am wide open. Everyone is in the same boat as of right now with me."


text:
Musictoday still sucks, but...
Now that the S&A DVD release has been postponed till June -- I called Musictoday again and asked them to cancel just the CD from my order -- telling them that when I placed my order, I did not expect to have to wait till June to get the CD. THE ACTUALLY AGREED TO DO IT. I talked to a guy who called himself "Bentley" -- I didn't have to argue with him or anything.
He said I should get a credit on my credit card in about 72 hours.
We'll see if they follow through.


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Wednesday, 5-Sep-2007 12:00
leicester.. down memory lane
after leaving irwin college back in 1998, never once hv i gone back to the place where i did my two-years of a levels.. bukan taknak, just that x berkesempatan je.. funny considering it is just a few mins away..
today, finally got my chance to go down the memory lane a bit.. lawak jugak la.. teringat zaman muda2 dulu.. i went back to the college, altho, it has been sold to montessori n no longer known as irwin college. cikgu2 sorang pun xde dah. went back to my host families' houses.. they're no longer there too.. wished i had kept in touch with them.. gi visit tmpt lepak, tmpt mkn, jln2 kat the shires mall..
hmm.. kalu bukan pasal attend interview.. dunno la bila nak sampai sini
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text: As an independent potter you have the freedom of creating and developing your own work and own style by renting out View's studio space on a monthly basis. (You must be an experienced potter to register. If you are new to our studio space contact Workshop Coordinator, Lauren Fix, for more information on how to become an Independent Potter).


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May 01, 2014
No Speech Can Fix This
President Obama has been on a foreign tour. How's it going on the world stage?
Probably not so well when New York Times columnist Maureen Dowd is accusing him of "whining" and "whiffing."
Dowd's eviscerating assessment of Obama's foreign policy is timely.
"It doesn't feel like leadership," she said yesterday. "It doesn't feel like you're in command of your world."
Why does this matter right now?
>>> Read More
|Did the White House Change the Benghazi Talking Points?
|What's Oprah Got to Do With Opportunity? Mike Lee Knows.
Quick Hits
- You won't hear about this record-breaking project in the mainstream news.
- This famous comedian doesn't think it's very funny that Common Core math makes his kids cry.
- Robin Hood was wrong: Taking income from the rich doesn't help the poor.
- Will taxpayers recoup the estimated $305 million spent on Oregon's failed Obamacare exchange?
- Charles Krauthammer challenges the mainstream media to cover this—but doesn't think they will.


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A recently described moth species has been named in honour of the new US president, Donald Trump.
The moth, described as Neopalpa donaldtrumpi, is found in Baja California, Mexico, as well as in southern California and Arizona.
The adult moth differs from the closely-related Neopalpa neonata by its distinctively silky head of white-blonde scales and its comparatively small genitalia.
Evolutionary biologist Vazrick Nazari, who described the new species, states in the paper: "The specific epithet is selected because of the resemblance of the scales on the frons (head) of the moth to Mr Trump's hairstyle."
DNA analysis backs up the observed physical differences, with analysed barcodes of N neonata and N donaldtrumpi illustrating 4.9-5.1 per cent divergence.
Adults fly throughout the year, probably in more than one generation. Studied specimens were collected in every month of the year in almost even numbers. The life history and host plant of N donaldtrumpi remain unknown.
Nazari added: "The discovery of this distinct micro-moth in the densely populated and otherwise zoologically well-studied southern California underscores the importance of conservation of the fragile habitats that still contain undescribed and threatened species, and highlights the paucity of interest in species-level taxonomy of smaller faunal elements in North America. By naming this species after the 45th President of the United States, I hope to bring some public attention to, and interest in, the importance of alpha-taxonomy in better understanding the neglected micro-fauna component of the North American biodiversity."
Nazari V. 2017. Review of Neopalpa Povolný, 1998 with description of a new species from California and Baja California, Mexico (Lepidoptera, Gelechiidae). ZooKeys 646: 79-94. DOI: https://doi.org/10.3897/zookeys.646.11411
Related articles
The information in this article was believed correct at the time of writing. BirdGuides accepts no responsibility for errors, or for any consequences of acting on information in the article. The opinions expressed are those of the author(s) and are not necessarily shared by BirdGuides Ltd.


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Tocopherols are natural antioxidants with both in vivo (vitamin E) and in vitro activity. Sunflower seeds contain predominantly alpha-tocopherol (>90% of total tocopherols), with maximum vitamin E effect but lower in vitro antioxidant action than other tocopherol forms such as gamma-tocopherol. Sunflower germplasm with stable high levels of gamma-tocopherol (>85%) has been developed. The trait is controlled by recessive alleles at a single locus Tph2 underlying a gamma-tocopherol methyltransferase (gamma-TMT). Additionally, unstable expression of increased gamma-tocopherol content in the range from 5 to 85% has been reported. The objective of this research was to determine the genetic basis of unstable expression of high gamma-tocopherol content in sunflower seeds.
Male sterile plants of nuclear male sterile line nmsT2100, with stable high gamma-tocopherol content, were crossed with plants of line IAST-1, with stable high gamma-tocopherol content but derived from a population that exhibited unstable expression of the trait. F2 seeds showed continuous segregation for gamma-tocopherol content from 1.0 to 99.7%. Gamma-tocopherol content in F2 plants (average of 24 individual F3 seeds) segregated from 59.4 to 99.4%. A genetic linkage map comprising 17 linkage groups (LGs) was constructed from this population using 109 SSR and 20 INDEL marker loci, including INDEL markers for tocopherol biosynthesis genes. QTL analysis revealed a major QTL on LG 8 that corresponded to the gamma-TMT Tph2 locus, which suggested that high gamma-tocopherol lines nmsT2100 and IAST-1 possess different alleles at this locus. Modifying genes were identified at LGs 1, 9, 14 and 16, corresponding in most cases with gamma-TMT duplicated loci.
Unstable expression of high gamma-tocopherol content is produced by the effect of modifying genes on tph2a allele at the gamma-TMT Tph2 gene. This allele is present in line IAST-1 and is different to allele tph2 present in line nmsT2100, which is not affected by modifying genes. No sequence differences at the gamma-TMT gene were found associated to allelic unstability. Our results suggested that modifying genes are mostly epistatically interacting gamma-TMT duplicated loci.
Tocopherols are the main antioxidants present in seed oils. They form a family of four fat-soluble compounds with vitamin E activity named alpha-, beta-, gamma-, and delta tocopherol. Tocopherols contain a 6-chromanol ring structure methylated to varying degrees at positions 5, 7, and 8, and an isoprenoid-derived C16 saturated side chain at position 2. The four tocopherols differ by the number and positions of the methyl groups on the 6-chromanol ring. Alpha-tocopherol is trimethylated, beta- and gamma-tocopherol are dimethylated, and delta-tocopherol is monomethylated [1].
Tocopherols are important antioxidants operating both in vivo, protecting cells from oxidative damage, as well as in vitro, protecting oils and oil-based products from oxidation [2]. The relative in vivo and in vitro antioxidant properties of the specific tocopherols is determined by their chemical structure. The relative biological activity of the tocopherols is estimated as 100% for alpha-tocopherol, 15 to 27% for beta-tocopherol, 3 to 20% for gamma-tocopherol, and 0.3 to 2% for delta-tocopherol [3]. However, there is a widespread confusion concerning their relative potency in vitro[4], though it is generally accepted that alpha-tocopherol shows better antioxidant activity in fats and oils at lower concentrations, but at higher concentrations gamma-tocopherol is a more active antioxidant [5]. Studies conducted in sunflower seed oil, in which alpha-tocopherol accounts for more than >90% of the total tocopherols, concluded that substitution of alpha-tocopherol by gamma-tocopherol has a positive impact on the stability of the oil [6-9].
Four sunflower germplasms named LG-17, T2100, IAST-1, and IAST-540 in which alpha-tocopherol in the seeds was almost completely replaced by gamma-tocopherol have been developed [6,10,11]. Gamma-tocopherol in the novel germplasm accounts for more than 85% of the total seed tocopherols, compared with more than 90% alpha-tocopherol in conventional sunflower seeds. The increased gamma-tocopherol levels are produced by recessive alleles at the Tph2 locus [6,12,13], which encodes a gamma-tocopherol methyltransferase (gamma-TMT) enzyme [14]. This enzyme catalyzes the methylation of delta- and gamma-tocopherol to yield beta- and alpha-tocopherol, respectively [15]. Gamma-TMT mutation in sunflower disrupts the synthesis of alpha-tocopherol and causes the accumulation of gamma-tocopherol [14]. In a detailed sequence analysis of the gamma-TMT gene in sunflower, Hass et al. [14] identified two gamma-TMT paralogs (gamma-TMT-1 and gamma-TMT-2) and five different haplotypes (haplotypes 4 and 5 corresponding to paralogs 1 and 2, respectively). Both gamma-TMT paralogs 1 and 2 cosegregated with Tph2 and were mapped to linkage group (LG) 8 of the sunflower linkage map. Even though the Tph2 mutation reduced or disrupted the expression of the two paralogs in developing sunflower seeds, none of the DNA polymorphisms found within the gamma-TMT Tph2 gene were associated with the high gamma-tocopherol phenotype [14]. The authors suggested that the Tph2 mutation must be very tightly linked to the gamma-TMT locus on LG 8 and may disrupt regulatory sequences.
Phenotypic studies of Demurin et al. [16] and García-Moreno et al. [13] concluded that the four high gamma-tocopherol lines LG-17, T2100, IAST-1, and IAST-540 possess the same allele at Tph2, as no transgressive segregations were observed in crosses involving the four lines. Lines LG-17, T2100 and IAST-540 were isolated from germplasm or M1 mutant plants that segregated for high alpha-tocopherol (>90%) and high gamma-tocopherol (>85%), with no intermediate levels of both tocopherol forms being observed [6,10-12]. Conversely, IAST-1 was isolated from an M2 plant that exhibited large variation for alpha- and gamma-tocopherol levels, with gamma-tocopherol content in M3 seeds showing a continuous variation from zero to 84.6% and M4 seeds from selected M3 plants showing a variation from 60.4 to 97.4% gamma-tocopherol, which was uniformly high (>90%) in M5 seeds from selected M4 plants [11]. García-Moreno et al. [17] suggested that the intermediate gamma-tocopherol levels observed during the isolation of IAST-1 might be produced by the presence of modifying genes that determined unstable expression of mutated alleles at Tph2 locus. Modifying genes have been found to influence important traits in sunflower such as high oleic acid content [18] or broomrape resistance [19].
In this study, the genetic analysis of a population that showed segregation from low to high gamma-tocopherol values obtained from the cross between the two high gamma-tocopherol lines IAST-1 and nmsT2100 has been carried out. In the course of this analysis, we identified two different alleles at the gamma-TMT Tph2 locus at LG 8. The allele tph2 was present in line nmsT2100, whereas the allele tph2a was identified in line IAST-1. Additionally, we found four modifying genes at LGs 1, 9, 14 and 16 that in most cases corresponded to duplicated gamma-TMT loci. Modifying genes influenced the expression of tph2a alleles, but did not affect tph2 alleles.
Seeds of sunflower lines nmsT2100 (fertile plants) and IAST-1 showed uniformly high gamma-tocopherol content, from 91.2 to 99.8% in nmsT2100 and from 92.3 to 99.4% in IAST-1. F1 seeds from the cross between nmsT2100 and IAST-1 had also high gamma-tocopherol content, from 92.6 to 97.2%. However, large segregation for gamma-tocopherol content was observed in F2 seeds from some F1 plants. Particularly, the analysis of 192F2 seeds from an F1 plant derived from an F1 seed with 95.5% gamma-tocopherol revealed a continuous large segregation for the trait, from 1.0 to 99.7%, with no discrete phenotypic classes being observed and a non-normal distribution (p<0.0001; Kolmogorov-Smirnov test) (Figure (Figure1).1). Variation for gamma-tocopherol content was smaller at the F2 plant generation (average of 24 individual F3 seeds), which segregated from 59.4 to 99.4% (Figure (Figure2).2). However, examination of variation of individual F3 seeds within each F2 plant showed that minimum gamma-tocopherol content in individual F3 seeds from the different F3 families ranged from 0.0 to 98.4%, whereas maximum gamma-tocopherol content was in all cases above 94%. No discrete classes could be distinguished for minimum gamma-tocopherol content in the F2:3 population (data not shown).
One hundred and sixty three out of 437 (37.3%) simple sequence repeat (SSR) and insertion-deletion (INDEL) markers were polymorphic in the screening of the high gamma-tocopherol lines nmsT2100 and IAST-1 and four randomly selected F2 individuals from their cross. A set of 128 high quality, evenly spaced, and preferably co-dominant markers were genotyped in the F2 population. Final linkage map for QTL analyses comprised 129 marker loci (109 SSR and 20 INDEL) grouped on 17 linkage groups (Table (Table1).1). The total genetic distance covered by these marker loci was 792.4cM, with an average marker interval of 7.0cM (Table (Table1).1). Linkage groups contained between 3 and 16 marker loci, with 99.6% of the mapped genome being within 20cM to the nearest marker. No marker loci had significantly distorted segregation ratios (P<0.001).
INDEL markers for the gamma-TMT gene described by Hass et al. [14] were not polymorphic between nmsT2100 and IAST-1 or showed polymorphism in secondary loci. Therefore, new primer combinations based on the full-lenght sequence of the gamma-TMT gene in nmsT2100 and IAST-1 (see below) were designed and tested. A primer combination (gamma-TMT-F1/F2/R24) based on the forward primers F1 and F2 placed upstream and dowstream, respectively, of the T39 transcription initiation site [14], and a reverse primer R24 complementary to the DNA sequence in exon 2 amplified a primary gamma-TMT locus that was polymorphic between IAST-1 and nmsT2100 (Figure (Figure3).3). The polymorphic band was about 1460bp in IAST-1 and 1420bp in nmsT2100 (Figure (Figure3).3). When screened against the individuals from the F2 population, the gamma-TMT-F1/F2/R24 locus was co-dominantly mapped to LG 8 at the previously described position of the gamma-TMT Tph2 gene [14]. Other INDEL markers for the gamma-TMT gene were also tested. The F9/R24 gamma-TMT INDEL marker showed three loci in the nmsT2100 and IAST-1 parental lines (Figure (Figure4).4). A primary locus (gamma-TMT-F9/R24a) corresponding to a band of about 1200bp was not polymorphic with this primer combination (Figure (Figure4).4). This locus, however, was demostrated to co-segregate with Tph2 and was co-dominantly mapped to LG 8 in the population CAS-12 x IAST-540, also segregating for gamma-tocopherol content [13]. A second locus (gamma-TMT-F9/R24b) that showed a dominant polymorphism, with a band of about 750bp present in nmsT2100 and absent in IAST-1, was mapped to LG 16 (Figure (Figure4).4). The locus was also mapped to LG 16 in the CAS-12 x IAST-540 population. Finally, a third locus (gamma-TMT-F9/R24c) that also showed a dominant polymorphism, with a band of 375bp absent in nmsT2100 and present in IAST-1, was mapped to LG 1 (Figure (Figure4).4). In addition to the gamma-TMT-F9/R24a, b and c loci, a fourth gamma-TMT locus was identified in populations other than nmsT2100 x IAST-1. This locus was named gamma-TMT-F9/R24d and was mapped to LG 14 in populations CAS-12 x IAST-540 [13] and IAST-413 x HA-89 (Del Moral L, unpublished data) (Figure (Figure4).4). Finally, the locus MT-2 of the 2-methyl-6-phytyl-1,4-benzoquinone/2-methyl-6-solanyl-1,4-benzoquinone methyltransferase (MPBQ/MSBQ-MT) gene was mapped to LG 4 using the F24/R25 INDEL marker [20]. None of the other markers for tocopherol biosynthesis genes tested were polymorphic between nmsT2100 and IAST-1.
One-factor analysis of variance revealed that the gamma-TMT-F1/F2/R24 locus on LG 8 was underlying a major QTL affecting gamma-tocopherol content. This locus had a significant effect on gamma-tocopherol content of the F2 and the F3 generations (Table (Table2)2) and corresponded to the gamma-TMT Tph2 gene also mapped to the same position on LG 8 by Hass et al. [14]. The mean and the standard deviation for gamma-tocopherol content in the F2 seed generation of plants homozygous for the IAST-1 allele were 52.3±29.6%, whereas those for plants homozygous for the nmsT2100 allele were 98.9±0.8% (Table (Table2).2). These results suggested the existence to two different alleles at the gamma-TMT Tph2 locus in lines nmsT2100 and IAST-1. The allele at IAST-1 was unstable, producing in homozygous condition a broad distribution ranging from 0.97 to 89.02% in the F2 seed generation, whereas the allele at nmsT2100 was highly stable and resulted in gamma-tocopherol values above 97%. Similar results were observed in the F3 seed generation (Table (Table2).2). It is interesting to note that only three F2 plants homozygous for the nmsT2100 allele produced sufficient number of F3 seeds for tocopherol analyses, compared to 26F2 plants homozygous for the IAST-1 allele (Table (Table2).2). This was probably caused by a close linkage between Tph2 and Ms11 loci at LG 8, the latter responsible for male sterility in the nuclear male sterile (NMS) line P21 [21] from which Ms11 was introgressed to nmsT2100.
Composite interval mapping analyses confirmed the existence of a QTL with a main effect centered on the gamma-TMT-F1/F2/R24 locus on LG 8 (Table (Table33 and Figure Figure5).5). This QTL was named Tph2-gamma-TMT and explained 41.2% and 44.4% of the F2 and F3 gamma-tocopherol phenotypic variance, respectively (Table (Table3).3). A second QTL peak on LG 8 16cM apart from Tph2-Gamma-TMT was identified only in the QTL analysis of the F3 data (Figure (Figure5).5). This adjacent peak might be a ghost QTL resulting from the distorted F3 data in this region and was not taken into consideration to protect against type I errors in declaring QTLs.
Besides Tph2-Gamma-TMT, four other QTL with a moderate effect on gamma-tocopherol content were observed at the F2 level. These QTL were located at LG 1 (GamT1.1), 9 (GamT9.1), 14 (GamT14.1), and 16 (GamT16.1). They individually accounted for 2.3-4.1% of trait variation (Table (Table33 and Figure Figure5).5). The QTL on LG 9, 14 and 16 were also detected in the F3 analyses at practically identical positions (Table (Table33 and Figure Figure5).5). The model with all the QTL explained 44.4% and 61.2% of the phenotypic variance for gamma-tocopherol content in the F2 and the F3 generations, respectively. The QTL peaks of GamT1.1 and GamT16.1 were centered on the gamma-TMT loci mapped at these linkage groups (gamma-TMT-F9/R24c in LG1 and gamma-TMT-F9/R24b in LG 16) (Figure (Figure5).5). Additionally, the QTL peak of GamT14.1 was likely to correspond to the gamma-TMT-F9/R24d locus mapped to LG 14 in populations CAS-12 x IAST-540 [13] and IAST-413 x HA-89 (Del Moral L, unpublished data), located 1.5cM from the ZVG64 marker locus at this LG.
Analysis for epistasis in the F2 generation through two-way ANOVA revealed significant interactions for gamma-tocopherol content involving the Tph2-gamma-TMT marker locus on LG 8 and markers on LG 1, LG 14 and LG 16. The most significant interactions involved the ORS185 marker locus on LG 14 (F=10.5, P<0.0001), the gamma-TMT_F9R24b locus on LG 16 (F=4.22, P=0.017), and the gamma-TMT_F9R24c locus on LG 1 (F=3.12, P=0.048), which were associated to the GamT14.1, GamT16.1, and GamT1.1 QTL, respectively. Significant interactions involving the Tph2-Gamma-TMT QTL on LG 8, the gamT14.1 on LG 14, and the gamT16.1 QTL on LG 16 were also detected in the composite interval mapping analyses, with the percentage of explained phenotypic variance increasing from 44.4% to 52.4% in the F2 and from 61.2% to 71.3% in the F3 when epistatic interactions were included in the multiple-locus model (Table (Table3).3). Genotypic means for F2 gamma-tocopherol content in allelic combinations of the epistatically interacting marker loci were calculated to detail these interactions. Since ORS185 and gamma-TMT_F9R24b marker loci were dominant, genotypic means were computed using close co-dominant markers, ORS578 on LG 14 and ORS126 on LG 16 (Table (Table4).4). The results showed that QTL regions on LG 14 and LG 16 only had a significant phenotypic effect when the IAST-1 allele was present at the Tph2-gamma-TMT locus on LG 8 (gamma-TMT-F1/F2/R24). Thus, no significant differences were observed between different genotypes at ORS578 on LG 14 or ORS126 on LG 16 in presence of the nmsT2100 allele at Tph2. However, when the Tph2 locus was homozygous for the IAST-1 allele, the presence of nmsT2100 alleles at ORS578 or ORS126 resulted in a drastic reduction of gamma-tocopherol content as compared to the presence of IAST-1 alleles (Table (Table44).
Several loci amplified by the gamma-TMT F9/R24 INDEL marker in IAST-1 and nmsT2100 (Figure (Figure4)4) were cloned and sequenced. A band of about 1200bp from the non-polymorphic locus a and a band of about 375bp from the dominant locus c that mapped to LG 1 were sequenced from IAST-1. A band of about 1200bp from the non-polymorphic locus a and a band of about 750bp from the dominant locus b that mapped to LG 16 were sequenced from nmsT2100. The locus d that mapped to LG 14 was sequenced from lines IAST-540 and HA-89.
The locus a fragment isolated from nmsT2100 harboured alleles from both gamma-TMT paralogs, which showed showed 100% sequence identity to gamma-TMT haplotype 4 (paralog 1) and 5 (paralog 2) from Hass et al. [14]. For IAST-1, the locus a sequence showed 100% sequence identity to gamma-TMT haplotype 4. The locus b consensus sequence was 785bp long and showed a significant homology to gamma-TMT haplotypes 2 and 3 (GenBank accessions nos. DQ229829 and DQ229830, 3e−58), with 94% maximum sequence identity spanning 61% of the locus b fragment coverage. The consensus sequence for locus c (388bp long) showed a significant homology to gamma-TMT haplotypes 4 and 5 (GenBank accessions nos. DQ229831 to DQ229834, 4e-14), with 86% maximum sequence identity spanning 19% of the locus c fragment coverage. The sequences of locus d from IAST-540 (405bp) and HA-89 (406bp) were very similar, with a 96% of sequence identity between them. The consensus sequence obtained from this alignment showed significant homology to gamma-TMT haplotypes 1, 3, 4 and 5 (GenBank accession nos. DQ229828, DQ229830, and DQ229831 and DQ229834; 8e−18), with 89% maximum sequence identity spanning 18% of the fragment coverage.
Nucleotide sequences from IAST-1 locus a, nmsT2100 locus a, nmsT2100 locus b, IAST-1 locus c, HA-89 locus d, and IAST-540 locus d were aligned. The sequence alignment tree (Figure (Figure6)6) revealed great similarity between loci a and b (85 to 87% of sequence identity) and between loci c and d (92–93% of sequence identity). Both groups were more distantly related, with 56 to 58% of sequence identity between locus a and the cluster of loci c and d.
Agarose gels from PCR products obtained with the F1 and R92 primers flanking the gamma-TMT gene mapped to LG 8 [14] revealed two bands in nmsT2100 and IAST-1 lines that were identified as the two gamma-TMT paralogs 1 and 2 described by Hass et al. [14]. Consensus nucleotide sequences for IAST-1 gamma-TMT paralog 1 (4126bp) and paralog 2 (4280bp) were identical to gamma-TMT haplotype 4 (GenBank accessions nos. DQ229831 and DQ229832) and 5 (GenBank accessions nos. DQ229833 to DQ229834), respectively, from Hass et al. [14] ( Additional file 1: Figure S1). Consensus nucleotide sequence for nmsT2100 gamma-TMT paralog 1 (4126bp) was identical to gamma-TMT haplotype 4 [14] ( Additional file 1: Figure S1). Consensus nucleotide sequence for nmsT2100 gamma-TMT paralog 2 (4281bp) was almost identical to gamma-TMT haplotype 5 [14], with the exception of five SNPs, one in the 5´UTR (G to A), one in intron 1 (C to T), and three in intron 4 (A to T) ( Additional file 1: Figure S1).
The results of this research suggest that high gamma-tocopherol lines IAST-1 and nmsT2100 possess different mutated alleles at the gamma-TMT Tph2 locus on LG 8. The gamma-TMT catalyses the methylation step from gamma-tocopherol to alpha-tocopherol in sunflower seeds [14]. Mutated tph2 alleles disrupt the activity of gamma-TMT, which results in accumulation of gamma-tocopherol [14]. The allele at nmsT2100 (tph2) is stable and not affected by modifying genes, whereas the allele at IAST-1 (tph2a) is unstable and affected by modifying genes. Putative modifying genes located at LG 1, 9, 14 and 16 were identified in the present research, being the effect of those on LG 1, 14 and 16 highly significant in the epistatic interaction with the Tph2 locus on LG 8. Studies on expression of Tph2 mutations in plant tissues other than seeds also pointed to differences between T2100 and IAST-1 lines, as the latter showed lower gamma-tocopherol content in leaves, roots, and pollen [22].
High gamma-tocopherol lines T2100 and IAST-1 were isolated following different strategies. T2100 derived from an open-pollinated cultivar that showed variation for high gamma-tocopherol content (>85%) at the single-seed level [9]. Genetic study of the trait in T2100 seeds indicated that the trait was controlled by recessive alleles at a single locus Tph2 in such a way that Tph2Tph2 and Tph2tph2 genotypes produced low levels of gamma-tocopherol (<5%) and tph2tph2 genotypes showed high levels of gamma-tocopherol (>85%), with no intermediate levels being observed [12]. On the contrary, IAST-1 derived from a mutagenesis program in which an M2 seed with intermediate gamma-tocopherol content (19.2%) was identified. The trait showed continuous variation for gamma-tocopherol levels (zero to 84.5%) in the M3 generation, which allowed selection of genotypes with stable high gamma-tocopherol content in the M4 generation [11]. A comparative genetic study between both lines concluded that they shared the same allele at Tph2, as no transgressive segregation was observed in the F1 and F2 generations from crosses between them [13]. However, the evaluation of several F2 populations from crosses of IAST-1 with T2100 and the conventional sunflower line HA89 showed that intermediate gamma-tocopherol values (5 to 85%) occurred in some F2 populations, whereas other F2 populations showed uniformly high gamma-tocopherol content (IAST-1 x T2100) or segregated into well-defined low and high gamma-tocopherol classes (IAST-1 x HA89) [17]. The results of this research suggest that intermediate gamma-tocopherol levels are produced by the effect of modifying genes on tph2a alleles from IAST-1. We hypothesize that modifying genes were present in the mutagenized population from which IAST-1 was developed, where selection against negative alleles at modifying genes led to the isolation of IAST-1, and also in other lines such as HA89 and T2100. Modifying genes have no phenotypic effect in absence of alteration in the major gene [23]. In this research, it was also found that modifying genes have no phenotypic effect on the mutated allele at T2100 (tph2). Accordingly, they are expected to be segregating in HA89 and T2100 lines, which produced different segregation patterns in crosses with IAST-1 [17]. The occurrence of different segregation patterns depending on allelic configuration of modifying genes has been previously reported in sunflower for high oleic acid content [24,25] and broomrape resistance [19].
Full length genomic gamma-TMT sequences corresponding to the Tph2 locus on LG 8 have been obtained in this study for both IAST-1 and nmsT2100. Both lines have two gamma-TMT paralogs. Paralog 1 is identical between the two lines and also to the gamma-TMT haplotype 4 from Hass et al. [14], which included sequences from both high and low gamma-tocopherol lines. Paralog 2 from IAST-1 is identical to gamma-TMT haplotype 5 from Hass et al. [14], which also included sequences from both high and low gamma-tocopherol lines, while paralog 2 from nmsT2100 carries slight SNP differences. Consequently, we have not found any sequence difference in the gamma-TMT gene from IAST-1 that would explain why the tph2a allele present in this line is unstable and affected by modifying genes. Previous research did not identify sequence differences in the gamma-TMT gene related to the high gamma-tocopherol phenotype [14]. The authors found identical gamma-TMT paralog 1 and 2 sequences in both high and low gamma-tocopherol lines and reported that mutation leading to the high gamma-tocopherol trait in the sunflower material derived from the high gamma-tocopherol line LG-17 [6] may disrupt regulatory sequences of the gamma-TMT gene. Relating the nmsT2100 line, the slight sequence differences in gamma-TMT paralog 2 were changes to those nucleotides also present in paralog 1 sequences from IAST-1 and nmsT2100 (2 SNPs in the 5´UTR region and in intron 1) or found in a repetitive sequence within an intron (3 SNPs in intron 4) and were not likely to be involved in determining a more stable gamma-TMT allele.
Modifying genes affecting important traits for sunflower breeding such as high oleic acid content [24,25] and broomrape resistance [19] have been reported. The discovery of modifying genes affecting high gamma-tocopherol content confirms that the occurrence of modifying genes is not a rare phenomenon in sunflower genetics. Thus far there was no indication on the nature and mode of action of modifying genes. The results of the present research suggest that Tph2 modifying genes are in most cases additional gamma-TMT loci duplicated in the sunflower genome. This was confirmed for modifying genes at LG 1 and 16, whereas there is also indication that the modifying gene at LG 14 might be an additional copy of a gamma-TMT. Duplicated gamma-TMT loci have also been found in safflower [26] and rapeseed [27]. In fact, the two gamma-TMT loci from safflower (one of them co-segregating with the safflower Tph2 gene determining high gamma-tocopherol content in this crop) were identified by using the F9/R24 primer combination from the sunflower gamma-TMT [26], the same one used in this study to map different gamma-TMT loci. These results suggested that the F9 and R24 primer sequences are within a highly conserved region of the gamma-TMT gene. In sunflower, Hass et al. [14] mapped, in addition to gamma-TMT-Tph2 on LG 8, another gamma-TMT locus on LG 16, although the authors considered this locus a randomly amplified polymorphic DNA (RAPD). In this study, the locus on LG 16 (gamma-TMT locus b) showed significant homology to the sunflower gamma-TMT gene and underlay a gamma-tocopherol QTL. Our results indicated that interaction between duplicated gamma-TMT loci revert the high gamma-tocopherol phenotype to intermediate-low gamma-tocopherol values. This effect has also been reported for modifying genes affecting the high oleic acid phenotype in sunflower, where modifying genes suppressed the effect of the Ol-FAD2-1 allele that is essential for accumulating high oleic acid content [18,28]. Also, epistatically interacting duplicated MPBQ/MSBQ-MT genes that modify beta-tocopherol levels have been reported in sunflower [20]. It is well known that duplicate gene pairs can form negative epistasis due to their overlapping functions [29] and that suppression of a mutant phenotype can be altered by gene dosage [30]. However, additional biochemical, functional, and sequence analyses are required to determine the mode of action of duplicated gamma-TMT loci in sunflower.
From a breeding perspective, the existence of several modifying genes influencing high gamma-tocopherol content should not be a constraint for the development of cultivars with seeds rich in gamma-tocopherol, as this research revealed that modifying genes do not alter the phenotypic expression of the mutated allele tph2 present at nmsT2100. On the other hand, the influence of modifying genes on expressivity of mutated tph2a alleles opens up the possibility of selection for balanced levels of alpha- and gamma-tocopherol content, provided that stable combinations of modifying genes can be obtained. The development of mid oleic acid hybrids of sunflower was based on genetic stocks carrying modifying genes that limit the expression of high oleic acid content produced by mutated alleles at Ol locus (>80%) to the mid-range level (55–75%) [31]. In this sense, determination of allelic diversity at gamma-TMT loci underlying gamma-tocopherol modifying genes present in hybrid parental lines might be a useful tool to predict the presence of adequate allelic combinations giving rise to the desired levels of gamma-tocopherol content. Further studies should elucidate whether the other two gamma-tocopherol sources developed thus far, IAST-540 and LG-17, possess stable or unstable alleles at Tph2. Demurin et al. [6] reported differences in the expressivity of the recessive homozygotes of the Tph2 gene from LG-17 in different genetic backgrounds, which might be indicative of allelic instability.
The results of this research suggest that high gamma-tocopherol in sunflower lines IAST-1 and nmsT2100 is determined by different mutated alleles at the gamma-TMT Tph2 locus on LG 8. The allele at nmsT2100 is not affected by modifying genes, whereas the allele at IAST-1 is unstable and affected by modifying genes. Putative modifying genes have been located at LG 1, 9, 14 and 16, being the effect of those on LG 1, 14 and 16 highly significant in their epistatic interaction with the Tph2 locus on LG 8. Finally, our results suggest that modifying genes correspond to gamma-TMT loci duplicated in the sunflower genome. Phenotypic effects of modifying genes altering the expression of important genes in crop plants are known [23], but in most cases the genetic basis for modification remains unclear. In this study, we shed light into the mode of action and nature of modifier genes in sunflower, and suggest the relevance of duplicated loci affecting the expression of seed quality specific mutations.
The study included the sunflower lines nmsT2100 and IAST-1, both with high gamma-tocopherol content (>85%). nmsT2100 is a NMS line developed by introgressing monogenic recessive NMS from line P21 [32] into high gamma-tocopherol line T2100 [10]. IAST-1 was isolated in the course of a chemical mutagenesis program on seeds of an accession of 'Peredovik' [11]. Twenty-four half seeds of nmsT2100 and IAST-1 were nondestructively analyzed for tocopherol profile as described below, germinated and planted in pots under open air conditions in the spring of 2005. NMS plants of nmsT2100 were pollinated with pollen of IAST-1 plants. Half seeds of the parents as well as F1 half seeds were analysed for tocopherol profile. F1 and parent half seeds were sown in March 2006 and the corresponding plants were grown in pots under open air conditions. F1 plants were bagged before flowering to obtain the F2 generation. F2 seeds from several F1 plants were analysed for seed tocopherol profile. In most cases, F2 seeds had uniformly high gamma-tocopherol content (>90%). However, some F1 plants showed segregation for gamma-tocopherol content at the F2 seed level, which indicated the expression of modifier genes. One population of 192F2 seeds from a single F1 plant that showed large segregation for gamma-tocopherol content was selected for the molecular analyses. F2 half seeds were germinated and the corresponding plants were grown in pots under open air conditions in 2007. F2 plants were bagged before flowering to produce the F3 seed. Five fully expanded leaves from each F2 plant (135F2 plants) were cut, frozen at −80°C, lyophilised and ground to a fine powder in a laboratory mill. DNA was isolated from ground leaf tissue from each F2 plant and from five plants of nmsT2100 and IAST-1 parental lines as described in Berry et al. [33]. Twenty four F3 seeds from each F2 plant that produced sufficient amount of seeds were analysed for tocopherol profile.
Two additional F2 populations were also used in this study with the objective of mapping tocopherol biosynthesis loci. One population, described in García-Moreno et al. [13], derived from a cross between CAS-12, with wild-type tocopherol profile mainly made up of alpha-tocopherol, and IAST-540, with high gamma-tocopherol content. The other population derived from a cross between lines IAST-413 and HA-89, both with wild-type tocopherol profile mainly made up of alpha-tocopherol, though IAST-413 is characterized by increased total tocopherol content [34].
The analysis of tocopherol profile was made for all analyzed generations and populations in half seeds. The half-seed technique is a common technique used in sunflower breeding consisting in cutting a small seed piece from the seed part distal to the embryo, which is used for nondestructive analysis of seed quality traits, as the remaining seed containing the embryo can be germinated after the corresponding analysis [35]. Individual half seeds were analysed for tocopherol profile following the method of Goffman et al. [36]. Half seeds were placed into 10-ml tubes with 2ml iso-octane. The half seeds were then crushed with a stainless steel rod as fine as possible. The samples were stirred and extracted overnight at room temperature in darkness (extraction time about 16h). After extraction, the samples were stirred again, centrifuged, and filtered. Twenty-five μl of the extract were analysed by HPLC using a fluorescence detector at 295nm excitation and 330nm emission and iso-octane/tert-butylmethylether (94:6) as eluent at an isocratic flow rate of 1mlmin-1. Chromatographic separation of the tocopherols was performed on a LiChrospher 100 diol column (250mm x 2mm I.D.) with 5-μm spherical particles, connected to a silica guard column (LiChrospher Si 60, 5mm x 4mm I.D.). The peak areas of the individual tocopherols were corrected according to their previously calculated response factors: alpha-tocopherol=1.0; beta-tocopherol=1.80; gamma-tocopherol=1.85; delta-tocopherol=2.30.
A complete linkage map for the nmsT2100 x IAST-1 population was constructed to scan the genome for modifier genes affecting the expression of the Tph2 gene. For this, the parental lines IAST-1 and nmsT2100 were initially screened for polymorphisms in two replicate samples together with four F2 individuals, using a genome-wide framework of 95 sunflower SSRs [37]. A preliminary genetic linkage map from this population was constructed. A set of INDEL markers [38], identified by ZVG prefixes, and an additional set of SSR markers mapped by Tang et al. [39] and Yu et al. [38], identified by ORS and CRT prefixes, were additionally screened for polymorphisms between these parental lines to complete the linkage map. INDEL markers for the tocopherol biosynthesis genes gamma-TMT, MPBQ/MSBQ-MT and tocopherol cyclase described by Hass et al. [14] and Tang et al. [20] were also screened for polymorphisms between nmsT2100 and IAST-1. PCRs for SSRs analyses were performed as described by Pérez-Vich et al. [40]. INDEL analyses were carried out following Yu et al. [38] and Hass et al. [14]. SSR and INDEL amplification products were separated on 3% (w/v) Metaphor® (BMA, Rockland, ME, USA) and 1.5% agarose gels, respectively, in 1x TBE buffer with ethidium bromide incorporated in the gel. SSR and INDEL markers revealing polymorphisms were then genotyped in the nmsT2100 x IAST-1F2 population, following the protocols mentioned above.
Chi-square statistics were computed on each genotyped locus to detect deviations from the expected Mendelian ratios for codominant (1:2:1) or dominant (3:1) markers. The nmsT2100 x IAST-1 linkage map was constructed using the software MAPMAKER/EXP version 3.0b (Whitehead Institute, Cambridge, MA, USA) [41]. Two-point analysis was used to identify linkage groups (LGs) at a LOD score of 3 and a maximum recombination frequency of 0.40. Three-point and multi-point analyses were used to determine the order and interval distances between the markers at each LG. The Haldane mapping function was used to compute the map distances in centiMorgans (cM) from the recombination fractions. Multiple loci detected by a single maker were coded with the marker name plus the suffix a, b, c, or d to indicate each duplicate locus. Linkage group maps were drawn using the MapChart software [42].
Genetic analysis of modifier genes was performed in several stages. In the first stage, the significance of each marker's association with the phenotypic trait [gamma-tocopherol content at the F2 seed and F2 plant (average value of 24F3 seeds per F2 plant) generations] was determined by one-way analysis of variance (ANOVA) using the statistical package SPSS Statistics v. 19, with marker genotypes being classes. In this analysis, we identified an unexpected macromutation on LG 8 at the Tph2 locus. The effects of the macromutations, if ignored, could dramatically reduce the power for identifying other genes or QTL affecting the studied trait.
In a second stage, composite interval mapping (CIM) [43,44] was used to scan the genome for QTL affecting gamma-tocopherol content, in order to strengthen and corroborate the results of the analyses of variance, evaluate the existence of additional QTL, and estimate the interaction and global effect of all the detected QTL. Computations were carried out using the software PLABQTL Version 1.1 [45]. The phenotypic data consisted on gamma-tocopherol content in the F2 seed and F2 plant generations. Additional analyses were carried out by using other parameters calculated from the F3 seed data such as the minimum, maximum and the standard deviation of gamma-tocopherol content in F3 seeds per F2 plant, and the number of F3 seeds within each F2 plant with less than 90% of gamma-tocopherol content. Since these analyses gave similar results to those obtained with the mean gamma-tocopherol value per F3 family, only results based on the F3 mean value are shown. Analyses were made initially with the "first" statement to check the database for errors and outliers. Next, simple interval mapping (SIM) was carried out for an initial scan and detection of QTL with main effects. Finally, CIM was performed with markers closest to the main QTL as co-factors. Genome-wide threshold values (α=0.05) for declaring the presence of QTL were estimated from 1000 permutations of each phenotypic trait [46]. The thresholds of the LOD score (and their 0.95 confidence intervals) were 2.57 (2.48–2.70) and 2.65 (2.58–2.76) for gamma-tocopherol content in the F2 and the F3 generations, respectively. Estimates of QTL positions were obtained at the point where the LOD score reaches its maximum in the region under consideration. One-LOD support limits for the position of each QTL were also calculated [47]. The proportion of phenotypic variance explained by each individual QTL was calculated as the square of the partial correlation coefficient (R2). Estimates of the additive (ai) and dominance (di) effects, as defined by Falconer [48], for the ith putative QTL, the total LOD score, as well as the total proportion of the phenotypic variance explained by all QTL, were obtained by fitting a multiple regression model including all putative QTL for the respective trait simultaneously [47]. The occurrence of QTL x QTL interactions was tested by adding digenic epistatic effects to the model.
QTL software such as PLABQTL estimate epistatic interactions among previously identified QTL. Since modifying genes are defined as genes having no known effect except to intensify or diminish the expression of a major gene [23], their effect as individual loci and subsequently their interaction with major loci may be undetectable with this type of analyses. In consequence, two-way interactions between the Tph2 major locus and all the marker loci genotyped in this study were also tested. Two-way interactions were analyzed at a significance threshold of P≤0.05 by analysis of variance using the general linear model (GLM) of SPSS Statistics v. 19. Statistical significance of differences for gamma-tocopherol content in different genotypes combining two marker loci were also computed using Duncan's multiple range test. The significant epistatic interaction terms were combined with those of the previously identified QTL in multiple locus models using the "seq" statement of PLABQTL.
In the course of the genetic analyses of the nmsT2100 x IAST-1 population, we identified different gamma-TMT loci amplified with INDEL marker gamma-TMT-F9/R24 associated to gamma-tocopherol QTL. In order to confirm their nature, these loci were sequenced as follows. F9/R24 INDEL fragments amplified from the IAST-1 and nmsT2100 parental lines were separated on a 1.5% agarose gel, excised and purified by means of the QIAquick gel extraction kit (Qiagen GmbH, Hilden, Germany). The purified fragments were ligated into the T/A vector (pCR2.1) and the recombinants were transformed to TOP10 Chemically Competent E. coli using the TOPO-TA cloning kit (Invitrogen, San Diego, CA, USA) as described by the manufacturer. Five recombinant bacterial colonies (white) per isolated band were picked from the plate containing ampicillin and X-gal as selective media and cultured overnight at 37°C. Plasmids were extracted and purified using QIAprep Spin Miniprep Kit (Qiagen GmbH, Hilden, Germany). PCR with M13 forward and reverse vector primers and F9 and R24 primers, and restriction enzyme digestion was performed to confirm the presence and size of the insert. Sequencing in both forward and reverse orientations of the cloned fragments (two clones per locus) was performed at GATC Biotechnology (Konstanz, Germany) using the M13 forward and reverse sequencing primers. Sequence analysis was conducted with the aid of the software Vector NTI Advance 10.3.0 (Invitrogen, San Diego, CA, USA).
Full-length gamma-TMT genomic DNA sequences were isolated from the high gamma-tocopherol lines nmsT2100 and IAST-1 by long distance PCR using primers developed at the 5´end (forward primer gamma-TMT-F1 from Hass et al. [14]) and the 3´end (reverse primer R92: TAATTCCTTGGGATGCCATT) of the sunflower gamma-TMT gene (GenBank accessions nos. DQ229828 to DQ229834). AccuPrime High Fidelity Taq DNA Polymerase (Invitrogen Life Technologies, Carlsbad, CA, USA) was used for PCR amplification in three individuals of nmsT2100 and five of IAST-1 as described by the manufacturer. The amplified products from each individual were separated on 1.5% agarose gels, showing in both lines two bands of a size higher than 4kb that corresponded to the two gamma-TMT paralogs described by Hass et al. [14]. The upper (paralog 2) and the lower (paralog 1) bands were independently purified in each individual and cloned using the TOPO-TA cloning kit (Invitrogen Life Technologies, Carlsbad, CA, USA) as described above, with the exception that twenty recombinant bacterial colonies (white) per isolated band were picked from the plate. Restriction enzyme digestion was performed to confirm the presence of the insert and the restriction patterns characteristic for each gamma-TMT paralog. Sequencing in both forward and reverse orientations of the cloned fragments (a total of 10 clones for IAST-1 paralog 1, 13 clones for IAST-1 paralog 2, 5 clones for nmsT2100 paralog 1, and 6 clones for nmsT2100 paralog 2) was performed at GATC Biotechnology (Konstanz, Germany) using the universal M13 forward and reverse primers and internal primers (Gamma-TMT-F9, F27, F67, R10, R24, R35 and R78 from Hass et al. [14], and primers from Table Table5)5) designed at 500 to 1000bp intervals from the sunflower gamma-TMT gene. Sequence analysis was conducted using Vector NTI Advance 10.3.0 (Invitrogen, San Diego, CA, USA). A consensus sequence for IAST-1 gamma-TMT paralog 1, IAST-1 gamma-TMT paralog 2, nmsT2100 gamma-TMT paralog 1, and nmsT2100 gamma-TMT paralog 2 was made from the analysis of 10, 13, 5, and 6 sequenced clones, respectively. Changes in the nucleotide sequence were only included in the consensus sequence when they were conserved among the different clones and individuals sequenced from each line.
Gamma-TMT: gamma-tocopherol methyltransferase; INDEL: insertion-deletion; LG: linkage group; MPBQ/MSBQ-MT: 2-methyl-6-phytyl-1,4-benzoquinone/2-methyl-6-solanyl-1,4-benzoquinone methyltransferase; NMS: nuclear male sterile; SSR: simple sequence repeat.
MJG-M carried out genetic map construction and all other molecular analyses. JMF-M and LV selected the sunflower lines, crossed them, produced and studied the phenotypic data, and participated in the design of the study. BP-V conceived and designed the study and supervised molecular analyses and interpretation of results. All authors contributed to the manuscript preparation, and read and approved the final manuscript.
Figure S1. Gamma-TMT sequence alignment. Sunflower gamma-tocopherol methyltransferase genomic DNA sequence alignment for the high gamma-tocopherol lines IAST-1 and nmsT2100 as well as genbank sequences from gamma-TMT haplotypes 1 (DQ DQ229828), 2 (DQ DQ229829), 3 (DQ DQ229830), 4 (DQ DQ229831 and DQ DQ229832), and 5 (DQ DQ229833 and DQ DQ229834) and the cDNA gamma-TMT EF495161 sequence.
The research was funded by the Spanish Ministry of Science and Innovation and the European Union FEDER (research project AGL2007-62834) and Dow Agrosciences LLC. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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David Lewis
David Lewis (Production) will be a second year student at TCU this fall where he is pursuing his B.F.A. in Theatre with an emphasis in Production. At TCU, he has assisted with Theatre TCU's productions of Tigers Be Still (Set Build), Guys & Dolls (Properties and Set Build), Brighton Beach Memoirs (Set Build), We Need a Little Christmas (Set Build), Anton in Show Business (Set Run and Build), and Three Sisters (Set Build).


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Jasmine Seafood Restaurant
4609 Convoy Street, Suite A
San Diego, CA 92111
(858) 268-0888
Open 365 days a year
Mon to Fri 10am to 10pm
Sat and Sun 9:30 to 10pm
Jasmine is centrally located in Kearny Mesa, San Diego surrounded by the 805, 163 and 52 freeways on the corner of Balboa Avenue and Dagget Street.
There is free parking to the North and East of Jasmine outside of the parking structure. There is ample neighborhood street parking and a large free parking lot at the corner of Dagget Street and Ruffner Road available on nights, weekends and holidays.
Jasmine Express
Fast, hot meals and Dim Sum
To-Go Orders before 6pm
(858) 268-0888
Please make your reservation over the phone.
*Reservations on weekends from 9:30am to 3pm are reserved for parties greater than 10.
For assistance with planning a wedding, party, event or meeting at Jasmine, e-mail us at email@example.com
or call us at: 858-268-0888.
General Inquiries


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Tips for dining on a budget
With restaurant week here we are helping you stretch your dollar so you don't overspend on food.
Sponsored by:
With restaurant week here we are helping you stretch your dollar so you don't overspend on food.
Ryan Kristafer brings you in the kitchen at Mario Batali's Tarry Lodge to show you the specials for New Haven Restaurant Week.
Members from the East Haven Rotary Club stopped by to give us a preview of their fundraising event this week, Anginette Wars.
Meteorologist Kevin Arnone took on the Small Space Workout Challenge this morning when GYMGUYZ stopped by the studio.
A recall of the emergency anti-allergy medicine EpiPen is expanding to the U.S. and other markets in North America, Europe, Asia and South A…
Happy National Grilled Cheese Day! This morning The Whey Station stopped by to cook up a unique recipe for a classic favorite.
Today is April 1st and that means its April Fools Day; Founder of PrettyWellness.com, Caryn Sullivan, stopped by our studio to talk about we…
News8's Ryan Kristafer travels out of this world to the Connecticut Science Center to celebrate the "NASA Space Day" exhibit.
Stretch Your Dollar// We are in the first week of April and that means we are also looking for the best deals in the new month.
The FBI has released several new pictures from the Pentagon on September 11, 2001.
Bills, bills, bills. They can cause headaches and inspire hit songs, but if you want to lower your bills it's time to pick up the phone and …
News 8 went back into our archives to find some classic footage of the first UConn National Championship team, and we've compiled them here …
Are you a foodie? Do you love to eat out?
Finally it feels like spring outside…sunshine and some warmth so let's head into the nursery and find out if there are some things that we…


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New Delhi, Dec 12: Chinese smartphone maker Xiaomi has suspended its sales in India following directions of the Delhi High Court, restraining sales of its products.
However, following the order passed by the Delhi High Court, "we have been forced to suspend sales until further notice. As a law abiding company, we are investigating the matter carefully and assessing our legal options", he said.
Recently, the Delhi High Court in an interim order has restrained Xiaomi as well as Flipkart from selling in India handsets of the Chinese mobile maker that run on the technology patented by Ericsson.
"Our sincere apologies to all Indian Mi fans! Please rest assured that we're doing all we can to revert the situation.
We have greatly enjoyed our journey with you in India over the last 5 months and we firmly intend to continue it!", he said in the post.


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First Stage Home
Our facility and staff are ready to help you embark on the road to recovery.
The friendly counsellors at LIFE Recovery Home understand the difficult choices and issues faced by woman bound by substance additions. Confidential consultation is available to these individuals every weekday during regular working hours. Many draw from their own addiction and recovery experiences.
LIFE Recovery is ready to help willing individuals embark on the road to recovery. In order to be accepted into the LIFE Recovery Program, prospective residents must meet the following criteria:
- Be a female, 19 years of age or older
- Have substance abuse issues
- Complete a TB test and ensure all other medical issues are under control
- Possess a desire to overcome and be set free from substance addictions
- Commit to a minimum of 3 months in treatment at LIFE Recovery Home
- Be willing to participate fully and comply with program requirements
- Be open to growing spiritually in Christ as you gain freedom from addictive lifestyles
- Make payment arrangements for treatment prior to entry
Program Activities
Programs at LIFE Recovery Home consist of both individual and group therapy.
Residents must meet with a counsellor on a regular basis to monitor progress and readiness for discharge. Residents must also complete a curriculum of educational material, including written assignments, both individually and in small groups. Residents must also attend Alcoholics Anonymous (AA) meetings and other 12-Step meetings on a daily basis.
Each woman at LIFE Recovery Home is treated as a child of God, with care and respect, as they choose to pursue a life free from addiction.
Program Topics
- Feelings "check-in"
- Self-esteem assessment
- Education about chemical dependency, addiction, boundaries, co-dependency and relapse prevention
- Big Book study
- 12-Steps study
- Workshops on communication skills, healthy relationships, goal-setting, planning and problem-solving skills
- Opportunities for life skills development and employment
- Devotions
- Art therapy
- Music therapy
- Weekly guest speakers
- Sing-along time
- Physical exercise
Christian 12-Step Program
The 12 Steps as applied by Alcoholics Anonymous (AA) have been drawn from the heart of the Scriptures. The Christian 12-Step Program is an integration of these foundational truths as they relate to chemically-addicted and emotionally-wounded women.
By applying Biblical principles, LIFE Recovery residents begin to grow spiritually and gain the opportunity to be free from addictive, compulsive and dysfunctional behaviours. This freedom gives way to peace, joy and most importantly, a stronger personal relationship with God and others.
The 12 Steps (as adapted from the original AA Steps):
- We admitted we were powerless over alcohol and that our lives had become unmanageable.
- We came to believe that a power greater than ourselves could restore us to sanity.
- We made a decision to turn our will and our lives over to the care of God.
- We made a searching and fearless moral inventory of us.
- We admitted to God, to ourselves, and to another human being, the exact nature of our wrongs.
- We were entirely ready to have God remove all these defects of character.
- We humbly asked Him to remove all our shortcomings.
- We made a list of all persons we had harmed and became willing to make amends to them all.
- We made direct amends to such people whenever possible, except when to do so would harm them or others.
- We continued to take personal inventory and when we were wrong promptly admitted it.
- We sought through prayer and meditation to improve our conscious contact with God, praying only for knowledge of His will for us and power to carry that out.
- We, having had a spiritual experience as the result of these steps, tried to carry this message to others, and practice these principles in all our affairs.


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Product #: 006-02828-99-0
Additional Information
- gorgeous glow balancing face wash gives you a deep and thorough cleanse, balancing your skin beautifully with priobotics and moisturizing organic omega-rich oils natural exfoliants, mild astringents, and tea tree extract will help keep hormonal breakouts at bay
- green tea will keep skin bouncy and strong
- pregnancy safe
- soap free
- 3.4 fl oz
We ship orders within the Continental US, Alaska, Hawaii & its territories. Shipping charges are flat rate based on service level. Find more shipping details.
We are not able to ship products to International addresses from Motherhood.com
Please visit MotherhoodCanada.ca to order and ship to Canadian addresses.
If for any reason you are not satisfied with your purchase, you can return it by mail or take it to any Motherhood store.
Gorgeous Glow Cleanser By Mama Mio is rated out of 5 by 1.
Rated 5 out of 5 by BoyMom2017 from Love this - definitely recommend! This has been the only thing to help keep any breakouts on my face at bay during my pregnancy. It's gentle, smells clean without being overpowering and is an overall great value.
Date published: 2016-10-14


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The two men will share a reported purse of £15million when they fight for the World Boxing Council and International Boxing Federation belts in Las Vegas on Saturday 17 November.
Promoter Don King said: "This is one of the most trying fights I've ever put together."
Rahman won the titles when he knocked out Lewis five months ago.
- More about:
- Boxing
- Las Vegas
- World Boxing Council


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Singapore Food Dictionary
Constantly on the lookout for the best places to indulge ourselves in our favourite foods, Singaporeans always have a diverse range of recommendations for every kind of cuisine. Must-try Singaporean foods include Hainanese Chicken Rice, Chilli Crab, Katong Laksa and the Singapore Sling. Other signature dishes in Singapore include Nasi Lemak, Bak Kut Teh, Kaya Toast and Roti Prata. Spend some time indulging your stomach in Singaporean classics during your stay here. Browse through the list we have compiled in our Food Dictionary and start your culinary adventure here!
|Satay is a Malay dish of marinated, skewered meats grilled over a wood or charcoal fire and served with a peanut dipping sauce. Skewered meat usually consist of sliced chicken, goat, mutton, beef or pork. Satay is also typically accompanied with chopped onion, cucumber and ketupat (Malay rice cakes).
|At night, the street next to Lau Pa Sat Festival Market is closed off and converted into a lively night market where people congregate for mouth-watering satay.
|Singapore Sling
|Singapore's official cocktail invented by Ngiam Tong Boon. The Singapore Sling is made from gin, Cherry Heering, Bénédictine, and fresh pineapple juice from Sarawak.
|Request for a shaken version instead of blended to experience a more genuine experience. To get the Singapore Sling that is closest to the original, visit Raffles Hotel's Long Bar.
|Steamboat is a Chinese-style hotpot eaten communally. Raw meats such as chicken, pork, beef, liver, meatballs, fish, squid, and vegetables are served for patrons to cook in the hot boiling soup. Some places even divide the steamboat pot into two halves so that patrons can have the option of having two different soups. Other steamboat eateries even come with a grill surrounding the boiling soup for patrons to grill their food.
|Tau Suan
|Dao Suean
|Meaning 'split green bean soup', this is a dessert that can be found in hawker centres and markets all over Singapore. The beans are known as moong dal, which are actually green in colour when husked and yellow when dehusked. This explains why the dessert is obviously yellow in colour despite its name. Served hot, it is usually accompanied by a few small chunks of you tiao (fried dough).
|Teh Tarik
|tay tah-rik
|Teh tarik (Malay for 'pulled tea') is a tea served with condensed milk (specifically, a Singaporean brand called "Carnation") and sugar. The concoction is then poured from one mug to another from a great height thus giving the illusion that the tea is being pulled. This technique enables the tea to froth which enhances its flavour. Although generally served hot, the teh tarik can also be served with ice, without milk or sugar upon request.
|Although the teh tarik is widely available in every hawker stall across town, some of the best teh tariks are concocted by Indian stallholders with their artfully crafted 'pulling' techniques.
|Turtle Soup
|Turtle soup is Chinese delicacy made from the flesh of the turtle. Fresh (soft-shell) turtle meat is brewed in a mixture of Chinese herbs till the herbs' essence are fully immersed in the turtle meat. Turtle meat is highly sought for its dense collection of collagen. Its fats has also been said to enhance blood circulation.
According to Chinese folklore, turtle eggs are supposed to be aphrodisiacs and in specialty shops, these are also served in the soup.
|Wanton Mee
|wun-tun mi
|No, there is nothing immoral or lewd about this dish. Wanton Mee is a Chinese noodle dish served with dumplings (wantons), barbecued pork slices (char siew) and leafy vegetables. The dish can be served dry or in a soup. Fried wantons (wantons deep fried in oil) are sometimes served instead of the usual boiled wantons.
|Yong Tau Foo
|yong tow foo
|Yong Tau Fu is an assortment of pre-cooked ingredients mostly made from a soya bean curd base (tau hu, tau kwa) and vegetables such as kangkong and mushrooms. The customer grabs a tong and picks the various ingredients (fish balls, meat balls, crab sticks, bittergourds, eggplant, mushrooms, cuttlefish, lettuce, ladies fingers and chilis) he would like from the shelves before passing it to the vendor to reheat them. Customers can also request for rice, thick yellow noodles or bee hoon (vermicelli) to go with their soup. Sauces such as vinegar, chili sauce and brown sweet bean sauce are usually available at the side for customers to use as dips.
|You Tiao
|yoh tiow
|You Tiao is a long, golden-brown, deep fried strip of dough and is usually eaten for breakfast. Conventionally, You Tiaos are lightly salted and made so they can be torn lengthwise in two. You Tiaos are normally eaten as an accompaniment to rice congee or soy milk.
|The You Tiao has a very interesting story behind it that dates back to the story of Yue Fei during the Southern Song dynasty. Yue Fei was a patriotic and well-respected General who fought many wars to protect the kingdom. The Prime Minister of the time, Qin Hui, resented him out of thirst for power. Together with his wife, Qin Hui fabricated charges which resulted in Yue Fei's death. The general public despised Qin Hui for killing their beloved war hero. Out of anger, a baker invented the You Tiao which resembles two people being twisted together to signify Qin Hui and his wife. This fried bread was named "You Zha Gui", meaning deep-fried ghost. Patrons who ate the "You Zha Gui" did so as a sign that they had burnt Qin Hui and his wife in hot oil and eaten them up.


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It has been 10 YEARS since ISLAGA (Indiana Society of Lampwork & Art Glass Artists) was formed in July of 2003. Contributions of time, knowledge, fellowship and dedication have made this milestone possible. Many have formed deep friendships over the years, and have grown professionally with their Art. It's been an exciting journey, and one we hope will continue for many years!
You are invited to join us in celebrating ISLAGA's 10th Anniversary celebration and The Hoosier Glass Trail, ISLAGAPALOOZA:
Complex Circle City
Indy Flame Art Glass
1125 Brookside
Ave. Suite C16
You are invited to join us in celebrating ISLAGA's 10th Anniversary celebration and The Hoosier Glass Trail, ISLAGAPALOOZA:
Indy Flame Art Glass
(Go inside the YELLOW door and Follow the signs inside the building to Indy Flame Art Studios)
Time: 10:00 to 2:00 pm with Studio open til 4:00 pm
Events include:
- Lampwork Demonstrations
- Intro to Lampwork sessions (FREE 1 hour mini classes at 10:00 AM and 3:00 PM)
- Refreshments
- Bead exhibit and sale
Meet old and New Friends - If you are a bead or jewelry artist, please bring YOUR work and publications to show and tell!
Free gift to the 1st 100 people! Please join us - it would be wonderful to see you there!
The Indiana Glass Trail is making a video for the Department of Tourism and all of us at Indy Flame Art are very honored to be part of it!
Thanks so much to Cassie Tamm of Indiana Glass Trail for coordinating and arranging this, and for all that she does for Indiana glass artists.


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A group of Acholi leaders led by Aruu County MP, Odonga Otto have come out to bash Besigye's wife Winnie Byanyima for allegedly supporting a murderer.
Of all people, Winnie Byanyima, the wife of Besigye to become the one to stand surety for Kanyamunyu! It is all over, the political lines have been blocked." Otto said.
The leaders are spitting venom purely because Winnie came in support of businessman Matthew Kanyamunyu who is accused of murdering Kenneth Akena.
The High Court in Kampala commenced the hearing of a bail application by businessman Mathew Kanyamunyu and his two co-accused persons where a group of eight sureties including Winnie Byanyima were presented before court to plead for Kanyamunyu's bail.
The anger among the Acholi leaders is so bad that they have even warned Besigye against going to the Acholi region without an armoured vehicle because anything can happen to him.


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Low credit interest, high overdraft charges and big fees on spending abroad means banking in the UK is far from free, according to a survey by consumer magazine Which?
Although many customers believe they are benefiting from free banking, research by Which? into the real cost of current accounts has uncovered a system where charging methods are deceptively complex, making it virtually impossible for consumers to compare costs.
The study found that while most current accounts do not charge a monthly free, high overdraft charges combined with low credit interest mean consumers end up paying more than they bargained for.
The biggest confusion concerned overdrafts, where many disparities were found between accounts. The annual cost of an authorised overdraft of £200 for six days a month indirectly takes £185 out of your account at First Trust, whereas the same amount at Coventry BS is not subject to charges.
The cost of unauthorised overdrafts was even higher. Santander was the most expensive for a regular unauthorised overdraft where the account holder goes overdrawn through payment of two direct debits each month, incurring costs of £95 a month or £1,140 over a year. The second most expensive bank for the same unauthorised overdraft was Clydesdale/Yorkshire Bank, which charged £950 over the year, while the Co-operative Bank was the cheapest, costing £51 over the year.
But even customers who stay out of the red do not get off scot free. One short trip overseas a year can set you back as much as £57 simply by using your direct debit card to make two £100 withdrawals and two £50 direct card payments if you bank with Norwich & Peterborough, while Lloyds TSB and Ulster Bank charge £56.
Which? said that while current accounts offer free facilities such as machine withdrawals and direct debits, low interest rates all but negate the positive effects of these benefits.
As current account costs are not levied upfront, customers do not feel the need to hunt around for better deals and changing accounts is seen as an inconvenience.
Which? chief executive, Peter Vicary-Smith, said: "The complicated ways banks present charges makes it difficult for people to work out whether they're getting a good deal. If you regularly go into the red or want a better return on your money, then the figures speak for themselves – it's worth making the switch to a more suitable account."
The organisation looked at 29 current accounts and found 14 do not offer credit interest and few match or beat the bank base rate of 0.5%. Which? based its calculations on the cost of in-credit banking, a four-day break overseas and credit interest on a balance of £1,500.


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Winegrowers Supplies - Automatic counter-pressure (Isobaric) bottle fillers + crown cappers
A counter-pressure filler is necessary for bottling already carbonated drinks, such as beer, Sparkling cider or wine.
The drink will already be stored under pressure, so there is no need for a pump.
To avoid excess foaming the temperature of the liquid (when inside the filler's header tank) should be between - 3 °C and + 4 °C.
C-C: 6 bottle Isobaric filler and integrated crown capper: 47,945.00 Euros
The photograph shows an in-feed table, 6-gripper bottle rinser, Isobaric 6 bottle filler with double pre-evacuation, and integrated crown capper, plus collecting table. A video of it in use: http://www.youtube.com/watch?v=6fetnfWH_bg
Automatic bottle rinser: not essential, you can use a semi-automatic rinser and place bottles on the in-feed table.
In-feed and collecting table: well worthwhile
Sensors on the filler conveyor, In and Out: 1,650 Euros
Filler with fittings for Cip cleaning: 1,100 Euros
Different Fill level: 340 Euros
Equipment for additional bottle size:
Kit of false bottles for cleaning: 1,500 Euros
Upgrade to the crown capper for additional format: 4,250 Euros
Tank for CIP cleaning (including pump, heating elements and control panel): 6,420 Euros
Kompakt: system ISO:-
Isobaric 9 bottle filler: 56,220 Euros
Pre-evacuation of the air (or double pre-evacuation): 4,500 Euros
Automatic fill level: 3,600 Euros
Kit of false bottles for CIP (cleaning in place): 2,665 Euros
False bottles and CIP (cleaning in place) are essential for isobaric fillers. The Kit of false (dummy) bottles includes everything needed on the filler for CIP, including dummy bottles, also the hoses on the filler's tank and control panel on the filler. You cannot easily open the tank of an isobaric filler for cleaning. Therefore CIP is essential.
Isobaric 12 bottle filler: 64,095 Euros
Pre-evacuation of the air (or double pre-evacuation): 6,090 Euros
Automatic fill level: 4,290 Euros
Kit of false bottles for cleaning: 3,550 Euros
Isobaric 16 bottle filler: 80,250 Euros
Pre-evacuation of the air (or double pre-evacuation): 7,490 Euros
Automatic fill level: 5,125 Euros
Kit of false bottles for cleaning: 5,140 Euros
Photograph of Isobaric bottle filler and crown capper:-
Photograph of bottle rinser, Isobaric bottle filler and crown capper:-
Tank for CIP cleaning (including pump, heating elements and control panel): 6,420.00 Euros
The CIP cleaning tank is a SS tank with heating elements, float, pump, and is controlled from the control panel of the filler. From the control panel you can start the automatic cleaning with re-circulation etc.
Additional bottle size: 1,340 Euros
Pressure gauge in the filler's tank: 320 Euros
Fully automatic integrated crown capper: 17,280 Euros
CO2 injection on the capper: 300 Euros
Electronic height adjustment: 1,480 Euros
Option for Mignon bottles - less than 160 mm tall: 2,465 Euros
Additional bottle size on the crown capper: 320 Euros
Prices shown are exclusive of Vat.
Delivery usually will need to be charged at cost.


text: Team Gasse has recently announced that we will be unable to participate in the Key West World Championships this year. This has been a great year of racing and hate we are going to miss the big event. We wish everyone good luck and we will see you on the race course in 2014!


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North Cornwall Region | 6,735 Yards, Par 72, Parkland | On-site Accommodation
Bude & North Cornwall is a traditional seaside links situated in the centre of Bude. The natural all weather fairways and greens are rarely unplayable making this a year round must visit!
Jaw-dropping views over St Austell Bay, beautifully kept greens and a mix of parkland and links golf, Carlyon Bay is a truly unique venue!
Described as "one of the finest 18 hole golf courses in Cornwall and the South West", this wonderful parkland course is situated in the Bodmin Valley.
The course is really well-designed and always presented in good condition. The front nine is quite open and the back nine, as well as being more protected is very scenic.
Copyright 2016 by GolfSouthWest.co.uk | Site by WebCreationUK.


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This set consists of a cushion, bedsheet, and a quilt. The quilt features hand-block prints of cute owls. The patterns are hand carved into wood and then stamped onto the fabric with skilled precision. The cushion features a big version of the owl motif on one side. The bedsheet has a polka dot inspired design. Made from pure cotton fabric, this set also makes for a great baby shower gift. Available in blue.
© Copyright 2017 Fabindia.com. All Rights Reserved.


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All Saints Church, Wheatacre
Sometimes, I come across random churches by accident. When trying to find Aldeby church, we went down all these country roads which were leading towards the Waveney River Centre, getting a bit lost we decided to give up. Surrounded by flatland fields, and marsh land, in the distance I spotted this little gem, beaming in… Continue reading All Saints Church, Wheatacre


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Awards season is in full swing and red carpet stocks are in short supply. First the US rolled it out for the Globes and Grammys – then came the Baftas at the Royal Opera House, while the Brits hit the O2 tonight. But don't breathe out just yet; the Oscars and Oliviers are still to come.
I love perusing the post-party fashion coverage with the best of you. But are these shiny affairs anything more than backslaps for the already rich and famous, laid on and voted for by industry insiders to provide marketing fodder for posters and promotion?
At a time when the arts sector is struggling most at the bottom, couldn't all that money be better spent on grass roots rather than glasses of champagne?
Or do awards ceremonies serve as an important showcase to investors, not to mention inspiring a new generation of artists? More than 70,000 people voted in this year's Whatonstage.com awards, the only major theatre awards decided entirely by audiences – is this the way forward?
Do awards ceremonies make a difference to you and your organisation? Have you say by voting in our poll and adding a comment or two below.


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Feed contaminated by salmonella bacteria is a familiar and costly problem for the animal feed industry all over the world. Some types of salmonella have succeeded in establishing themselves in feed and fish meal factories and have persisted there for several years because it has proved impossible to eradicate them.
In her doctoral thesis, Lene Karine Vestby therefore studied why it is so difficult to get rid of salmonella once they have managed to establish themselves in Norwegian feed and fish meal factories. She discovered that salmonella bacteria efficient at forming biofilm (bacteria coating) survived for longer in the factories than those that had a reduced ability to form this coating. The ability to survive in factories therefore appears to be connected with the ability to form a biofilm and it would seem that removing biofilm is a necessary step towards eradicating salmonella from the factories.
In a biofilm, bacteria are well protected by a slime (matrix) which they produce themselves. Vestby has studied the effect of the nine most frequently used disinfectants in the Norwegian animal feed industry and found that the efficiency of the disinfectants is substantially reduced if the salmonella has managed to form a biofilm. The effect of the majority of the disinfectants was then no longer satisfactory, but a product containing 70% ethanol was the most efficient, followed by one called Virkon S.
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Washington, Jan. 16 (ANI): In a bid to support Tony Blair's role of Middle East peace envoy, a whopping 600,000 pounds are being spent annually, but the former British Prime Minister has not spoken to a Foreign Office minister since October.
It was revealed during parliamentary questions that 600,000 pounds a year is being spent to support Blair's office, as four Whitehall officials are working for him full-time, whom he selected personally, and is given public money for office accommodation and IT.
"Everyone knows that the post of Middle East Envoy was created simply to pamper Tony Blair's ego and wasn't a serious appointment. What we didn't know, however, was that this pointless ego trip was to be heavily funded by the British taxpayer, at the whopping cost of over half a million a year," TheTelegraph quoted Norman Baker, the Liberal Democrat MP for Lewes, as saying.
"How could the man who helped invade Iraq possibly be seen as an honest broker in the Middle East? After two years, he has barely set foot in the Palestinian territories, preferring it seems to hole up in his luxurious hotel suite," he added.
As soon as Blair left Downing Street in 2007 he was given the high-profile diplomatic post of Representative of the Quartet - the UN, EU, US and Russia - in the Middle East.
It was also revealed that Blair's last meeting with British Foreign Secretary David Miliband was on 14 October 2009, and he has never met ministers at the Department for International Development, who provide much of the support for his position as Office of the Quartet Representative.
"The fact that the Foreign Secretary has not even met Tony Blair for three months and that meetings only happen rarely in any case suggests that there is very little return for the taxpayer for the vast amounts of public money being committed," Baker asked.
"The British government should end their funding for this pointless post now. The former Prime Minster, whose apparent willingness to engage almost any cause if there is money attached, is becoming a national embarrassment for this country," he added. (ANI)


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Beautiful Dove Dish
This is a beautiful handmade dove dish perfect for a minimalist dining experience. I just love the simplicity of this piece. I think I might have to buy it! Look at this artist, MayLuk's other handcrafted work for sale!
Posted by Amy Bird at 10:03 AM
Labels: dinnerware, dove, dove dish
1 comment:
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No products
Amount payable includes 7% GST.
Check out
Cold Steel Knives Singapore
Product Code 92BKKB
Our new improved Japanese Training Swords are closely modeled after our world-famous Warrior series swords. Featuring an intricately detailed imitation cord wrap grip - molded in high impact polypropylene - they offer superior grip and comfort as well as more realistic training potential for the modern Martial Artist.
Product Code 92BKKAZ
Product Code 95BUSK
For the money, the Bushman® knives might be the strongest survival/outdoor knives in the world!
Product Code 91SPB
As the saying goes, "the pen is mightier than the sword." Indeed, Cold Steel makes some mighty strong swords, but this little pen makes for some clever competition. It's the biggest, baddest, permanent marker around!
Product Code 62KGCM
The Kukri's best attributes and enormous potential as a weapon. We've made different variations of the Kukri over the years, some with longer blades, and some with shorter, but never, until now, have we had a version you could carry in your pocket as a folder.
Product Code 62KGC
A good Kukri, with its broad heavy duty blade, is a true power - able to cut, chop, slice and shear through even the toughest targets. An outstanding multi-purpose tool that can put in exceptional work, as well as serve as a truly devastating weapon during wartime or in a last ditch self-defense situation.
Product Code 30ULH
A design collaboration between custom knife-maker Andrew Demko and Lynn C Thompson, the Ultimate Hunter represents the culmination of almost 3 years of design evolution.
Product Code 14AKVG
Our AK47 series folding knives have set the standard for hard use tactical folders. A firm favorite with military personnel and special units, these rugged, dependable folding knives have seen use all over the world and in several conflict zones, and have been in our line up in various iterations for almost a decade.
Product Code 27TMCS
Now With U.S. Made Carpenter CTS® XHP Alloy Blade Steel Featuring DLC Coating* Our Recon I series continues to set a standard for Tactical Folding Knives that's hard to beat.
Product Code 27TLCS
Product Code 20LT
Tough enough to do the job and light enough to be handy, Cold Steel's Tuff-Lite folders are there when you need them. They're perfectly sized to slip into a pocket, clip to your waistband, or wear around your neck on a lanyard.
Product Code 92SF
The Special Forces Shovel is modeled after the original Soviet Spetznaz shovel.
Product Code 92R13RT
Cold Steel's Recon Tanto Rubber Training Knife is made of a 7" Santoprene Rubber blade. This 11 ¾" knife is soft enough to prevent the likelihood of most bodily injury, and perfect for Solo Practicing, Training Drills, Demonstrations, and Airsoft and Paintball.
Product Code 58TPCS
Product Code 29TMC
Product Code 27TDSRY
Product Code 27TDSG
Product Code 92R39LSF
Product Code 27TLSH
Product Code 20T
Product Code 27TLS
Product Code 27TDS
Product Code 39LSFD
Product Code 39LSFT
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