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The hotel is clean and nicely furnished.There is plenty of hot water and thick towels.
The staff are helpful if asked advice but otherwise not very friendly.
The breakfast is not good.Terrible coffee and mediocre food.
Review collected in partnership with this hotel
I would like to recommend everyone to take the opportunity to stay at the Hotel Romantic as it is well run and the staff are friendly and efficient and I would like to say that I was impressed with the refurbishment.
Would definately come back to this hotel again.
Food was very good, located on the sea front it was close to bars, restaurants and shops. Although the hotel didnt have a pool, we could use the one locally just a short walk away. Rooms were spotless and spacious. Bathroom never had a bath just a really good powerful shower which...
Stayed here as a party of 8 aged between 44 and 64. Hotel is nicely decorated, staff friendly enough but we found that there was no atmosphere. Couples were just sitting looking at each other drinking wine not even talking to each other let alone us! At breakfast it was like being in a OAP's home, maybe it was the...
This was our second year at the a Romantic. Last year we had a wonderful time in a somewhat traditional Spanish hotel. This year after a major refurb we went back. Hotel has been tastefully updated. The food we thought was not up to the standard as last year, the staff although the same we're not as friendly. There was...
Staff lovely, very clean and in a great position opposite the beach. Rooms were fine but view on our side of the hotel was a blank wall. Breakfast was good. We went half board and the evening meals were generally OK.
We have visited this hotel for many years enjoying its simplicity.
However it has had a major upgrade which I am sure will appeal to younger visitors .(Who do not appear to need shelves in the bathroom ,because there are none ).
The rooms look lovely and the beds are comfortable. There is now a TV
which gets some major...
Just got back from our holiday at romantic hotel.The refurbishment truly makes this hotel much better.Our room 101 was bright and very clean with a lovely balacony and sea views.We had booked a standard double room, which was the only room available at the time of booking, when we came to check in we were offered the option to upgrade...
It was our first visit to the Romantic hotel. We read a lot of mixed reviews about this hotel,some good and some bad.We didn't know what to expect,but apart from the evening meal we really enjoyed our holiday.The hotel is only two stars and you get what you pay for.We would go back as the hotel is very clean, friendly...
We have been visiting this hotel annually for the last 26 years and have seen a few changes but none as radical as this. Overall the management have done a good job in the transformation, but I do wonder how long the paintwork is going to last, as it appears to be wearing off already. We liked the new look...
Is the hotel, suitable for a wheelchair user who is partly mobile. ie can the lifts be accessed without any steps. Thanks Ade
Thanks for information on fridge hire.How can l organise a fridge for hire? Can l do so before my holiday,or do l have to order on arrival at hotel,Regards Carolyn.
We are a couple, a grandmother and a 10 year old grandson wishing to stay at the hotel. Is that a problem?
Hello are there hair dryers in the rooms please???
Hi, we are hoping to stay in the at the start of July, and are wondering if all the rooms and facilities will be renovated by then?
See all questions


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If Occupy Wall Street had a national plan to flood the IRS with illegal claims for tax-exempt status and some IRS clerks had run a search for "occupy," they would probably get a bonus and the story would be Occupy's "criminal conspiracy." But if the search is for "tea party," you have a different "scandal," as William Boardman notes.
Obama Administration
A New Front in War on Wikileaks
With Private Bradley Manning's leak trial about to start and with Wikileaks founder Julian Assange still holed up in the Ecuador Embassy in London, "We Steal Secrets," a new big-budget documentary purports to explain the controversy but has more the look of a hit job, says Danny Schechter.
Ecocide and the Soul of a Nation
At the front of the political madness enveloping the United States are anti-government, anti-science extremists who reject evidence of global warming and block any response to this existential threat. But the disconnect between environmental destruction and today's humanity goes deeper, says Phil Rockstroh.
America's Long History of Bloodletting
Americans like to think of themselves as peace-loving, but their history belies that self-image. From the genocidal wars against Native Americans through the current multi-front "war on terror," the United States has been fighting and killing for most of its history, as Lawrence Davidson notes.
Bad Mind-Reading on Iran
Propagandists often speculate about the evil intentions of some rival state and then bait anyone who suggests that the other side is just looking out for its own interests or harboring its own fears. This propaganda technique has been honed into a fine art form regarding Iran, as ex-CIA analyst Paul R. Pillar describes.
The Folly of 'Self-Regulation'
The recent loss of human life in Bangladesh sweatshops and the 2008 Wall Street meltdown that devastated the world's economy should demonstrate that relying on corporate executives to "self-regulate" is a deadly and dangerous way to protect the broader society, as Michael Winship explains.
New Risks from 9/11 War Act
Presidents have been stretching their commander-in-chief powers since Thomas Jefferson dispatched the Navy to make war on the Barbary pirates. But Congress risks a perpetual war of presidential choice if it carelessly rewrites the 9/11 force-authorization act, warns Independent Institute's Ivan Eland.
'42' and 44
Exclusive: Most Americans now celebrate the courage of Jackie Robinson in facing down racism in 1947 as the first black Major League baseball player in the modern era. But there has been remarkably little appreciation for the bravery of Barack Obama as he has served as the nation's first black president, writes Robert Parry.
Doubting Obama's Resolve to Do Right
Exclusive: In his counterterrorism speech, President Obama ruminated about the moral and legal dilemma of balancing the safety of the American people against the use of targeted killings abroad. But Obama's handwringing did not sit well with some critics including ex-CIA analyst Ray McGovern.
Hezbollah's Risky Syrian Gambit
The sectarian rifts, which were opened by George W. Bush's invasion of Iraq, continue to tear apart the Middle East, now involving Syria and Lebanon. Hezbollah, the Lebanese Shiite militia, has plunged into Syria to fight Sunni-led rebels, as ex-CIA analyst Paul R. Pillar notes.

subset: commoncrawl


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First Day 5K
Haaaappy Newww Yeeearrrr! Its time to make some serious improvements.
Hmm, where to start? The case of the mysteriously missing shoe? Check prices on steam-cleaner rentals? Figure out who that trans-gendered monkey with an eye patch belongs to? Sounds like youve got one heck of a resolution list.
Lucky for you, theres a New Years Day 5K around the red holes of Papago Park to get your year off on the good foot. This particular event boasts a 5K run or a 1K fun run/walk. The start location is ramadas nine and ten, right next to the Phoenix Zoo.
Hey, maybe theyll recognize that monkey!
Sat., Jan. 1, 9 a.m., 2011
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Red Rock Ranch & Farms
Concho, AZ 85924
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This Orange Standard V-neck solid scrub top features a tonal screen print at the neck and pocket. A waistband, elastic back and front darts make for a flattering fit. There are two front patch pockets. The left pocket has a second layer with an angled opening. There are side slits in this poly/cotton top.
I like the color and the design but the size was smaller then a large should be. I had to cut out the elastic band that was in back just so I could get it on..Maybe a Xlarge would be a fit for someone ordering a large.
nice look, just a smidgeon too tight, if you need a lot of room to move, which I do, go a size larger.


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Photo by Tina Rupp
Leave a Review
Followed recipe exactly. Simple. Delicious. Gone in a minute. Teenagers were on the attack, trying to finish them off before the adults could get their fill.flag if inappropriate
Delicious - I tried the method of oven-baking the shrimp the last few minutes the toasts were in the oven, and also pan-frying as called for in the recipe. The pan fried looked better, browner; and it took longer in the oven than I expected, maybe 6 min. I think I would stick with the recipe but I was hoping the oven version would work because it would be easier. I was planning to serve these as hors d'oeuvres but I think they're a little awkward to eat as a finger food; better as a plated appetizer.flag if inappropriate
I used one shrimp per toast and garnished with chopped fennel fronds instead of capers. Took as a appetizer to a dinner party. Everyone loved them. The rouille was delicious. Can't wait to try it with crabcakes.flag if inappropriate
Simple and deliciousflag if inappropriate
Great recipe, simple to make. Prefer it to be made with home made mayonnaise w hich we did and it came out more flavorful. Served up as an amuse bouche on Christmas day. enjoy!flag if inappropriate
Easy and delish! I also only used one shrimp per toast, which was plenty for an h-d. It kind of lacks in presentation, but the addition of capers helps it along.flag if inappropriate
This was such a hit with my guests They just gobbled them up. The left over sauce will be great with crab cakes too. I look forward to serving this many more times!flag if inappropriate
Made this twice to bring to parties. I prepped everthing in advance. Found that to simplify preparation in a crowded kitchen, cook toasts as described but for about 7-8 minutes, then distribute the shrimp on the toasts and bake a few minutes longer just until shrimp is done. Excellent reviews from the guests - Everyone was in line to be sure they didn't get left out!flag if inappropriate
grilled the shrimp and they were all eaten before they reached the toast!flag if inappropriate
Made this for friends one night... and they loved it! Highly recommended for appetizers, delicious!!!!flag if inappropriate


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In the midst of a blowout loss to the Philadelphia Phillies on Sunday afternoon, Cubs manager Joe Maddon wanted to do something to lighten the mood in the clubhouse.
This was supposed to be the easy part of the Cubs' schedule.
ATLANTA — While other playoff contenders scramble to acquire relief pitching before the July 31 nonwaiver trade deadline, the Cubs are on track to add a reinforcement to their bullpen by as
With the trade deadline just a few weeks away, the Cubs admit they have to address their starting pitching depth.
Joe Maddon decided to focus on all the positive things the Cubs did on a chilly Wednesday night against the first-place St. Louis Cardinals.


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Banks crack down on interest-only mortgage deals
Borrowers hoping to cut their mortgage repayments by switching to interest-only deals may find such a move is frowned on by banks and building societies.
Mortgage companies are asking for bigger guarantees from anyone who wants to take out interest-only loans or switch their existing home loans from repayment to interest- only, particularly if they have no means of paying off their capital.
Some of the most popular mortgage companies lead the crackdown. Abbey demands borrowers have at least a 50 per cent deposit if they want to go interest-only.
And Barclays and Co-op Bank limit the maximum interest-only part of the mortgage to 75 per cent. Any borrowing in excess of this must be repayment.
Crack down: Banks are making it more difficult to get interest-only mortgages
Since the beginning of the year, City watchdog the Financial Services Authority (FSA) has warned banks and building societies they should be able to demonstrate that they are not lending too much money to customers who cannot afford to repay it.
A spokesman for Barclays said: 'There has been significant pressure from the FSA for us to demonstrate that we are lending responsibly.'
Borrowers with Co-op can still get a loan with just a 10 per cent deposit, provided the interest-only part is no greater than 75 per cent of the property value.
With a repayment mortgage, the monthly home-loan bill will pay some interest and some capital off the money borrowed.
An interest-only mortgage does precisely what the name suggests: pays back only the interest on your loan. You would need to find another way to pay off the capital owed.
Interest-only mortgages are attractive to those on tight budgets because monthly repayments are lower.
On a typical £150,000 loan at 6 per cent over 25 years, an interest-only monthly bill is £750, while for repayment it is £966.
However, at the end of 25 years the debt would be completely cleared with the repayment mortgage, but you would still owe £150,000 on the interest-only.
Over the past two years, the percentage of home movers and first-time buyers opting for interest-only without specifying how it will be repaid has steadily increased.
In the first three months of this year, up to 28 per cent of home movers opted for an interest-only loan and had no proof of how they were going to repay the money. In 2006, this figure was 23 per cent.
Last year, the percentage of first-time buyers who took out an interest-only loan with no means of repaying was 22 per cent, but it has steadily fallen since the credit crunch began as banks get tough with the handful of new buyers still entering the market.
By July the figure had dropped to 13 per cent.
A common strategy from borrowers on a tight budget is to take half of their mortgage interest-only and half repayment.
This would give repayments in our example of £858 a month - £483 towards the repayment and £375 towards the interest.
Of course, after 25 years you would still need to pay back £75,000 capital.
The falling housing market - latest reports from Nationwide BS show the average property value has fallen by around £20,000 in 12 months - has meant lenders are particularly concerned about those who show no means of repaying the capital on their loan.
This is because these homeowners will have an asset that is falling in value without them chipping away at the debt.
David Hollingworth, from brokers London & Country, says: 'Lenders are going to be more cautious in a falling market.
'They are reviewing their criteria in a changing market to protect themselves and the borrower.
'Borrowers can expect some tougher questions with regard to how they expect to repay their loans.'
If you really are struggling to keep up with your mortgage payments moving on to interest-only is preferable to falling behind with your bills.
However, be disciplined. Set a deadline to move back on to repayment as soon as you can.
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|Bruins name Doug Jarvis assistant coach
|08.04.10 at 3:25 pm ET
The Bruins announced Wednesday that they have named Doug Jarvis as an assistant coach. The move rounds out a group that also consists of head coach Claude Julien as well as assistants Doug Houda and Geoff Ward. Craig Ramsay left the Bruins earlier in the offseason to become the head coach of the Atlanta Thrashers.
Jarvis most recently coached with the Canadiens from 2005 to 2009 and worked under Julien during the 2005-06 season. Prior to that, he served as a coach for the North Stars/Stars from 1988-2002. After leaving Dallas, he became head coach of the Hamilton Bulldogs (the Canadiens' AHL affiliate) for two seasons.
As a player, Jarvis holds the current record for the longest games played streak with 964. He played 13 seasons in the NHL and won Stanley Cups with the Canadiens in each of his first four seasons. He totaled 403 points in his professional career.
|Bruins choose to skip Game 7 morning skate
|05.14.10 at 11:43 am ET
On the morning of the biggest game of the season, Bruins coach Claude Julien decided to let his players have the morning off as opposed to coming to TD Garden for a morning skate prior to Game 7 against the Flyers on Friday night. Julien said that since the Bruins had a late night in Philadelphia on Wednesday and practiced at Ristuccia Arena in Wilmington on Thursday, that it was appropriate to give his players the day to rest.
"First thing that I think you have to remember is that we had a late game the other night and didn't get in until the morning. We practiced yesterday and did all the things that we needed to do to prepare for tonight. So, give our guys an opportunity to get some rest, simple as that," Julien said. "We have done it before, sometimes in back-to-backs. Stay home and come to the rink. That was our idea behind this is that our guys need to rest and basically we have played these guys or we are going to play these guys seven times in a row. There is not much we don't already know about the other team. So, come ready to play."
It is the first time of the playoffs that the Bruins have completely skipped the morning skate through they have had optional skates and days off between games where only players receiving treatment have been required to report.
"Both teams are going to be reay," Julien said. "Anytime you play a Game 7 where the winner moves on you have to be ready. For us it is a matter of not bogging them down with what has happened the last few games but focus on what we have to do tonight. The bottom line is, if you win tonight you move on and that is forgotten. But that is what we have got to do here is stay focused and stay prepared and I think that our guys are up to the task."
As per usual, Julien declined to give any indication of what his roster would look like on Friday night but there was speculation after practice on Thursday that rookie center/forward Brad Marchand might get into the game after dressing in a grey practice sweater, normally reserved for the second line, as opposed to a red sweater that indicates the checking line and "fifth" line.
"Big players have to come up big in big games but at the same time I have also seen unsung heroes do that same thing," Julien said in response to how Zdeno Chara could play on Friday. "In those kind of games you go in there and hope that somebody will make a difference and I don't care if it is your best players or your role players. I just care that somebody comes up big and does something good for us and that is what you try to challenge your whole team to do. If you challenge only your key players then you are telling your other players you don't care what they do. You want everybody to be difference makers, or go in with that intent anyway."
|Julien chooses not to discipline his team
|05.13.10 at 4:27 am ET
PHILADELPHIA — At least publicly, Bruins coach Claude Julien failed to lash out at his team for their 2-1 loss to the Flyers that has forced a seventh and deciding game in the series.
Nor did Julien take issue with Daniel Paille for an elbow penalty that the coach thought – at the very least – was questionable, leading to a Flyers 5-on-3 advantage in the second, and eventually a 4-on-3 power play goal for the Flyers.
'It's important, but again not to criticize. It puts us down five on three when they called the elbow," Julien said. "If you look at the replay he doesn't even touch him. There's a space between his arm and the guys face but he puts his head back and the [referee] calls it. Do you blame your players for that? I don't think so, I think we have to stay on the puck.
"There's no doubt about that, but I don't know that we were overly undisciplined. We were the other night [Game 5] and it ended up costing us. Tonight, we had some power plays and we weren't able to capitalize. We had some opportunities as well, a couple tough penalties, but other than that I thought we were pretty disciplined.'
Paille's penalty was made even worse because Marc Savard was already serving a penalty. Then to finish the second period, Blake Wheeler takes an even worse penalty for holding with just under 27 seconds left in the period.
Can Julien sense tenseness in his players?
'It's a situation where the winner of the game moves on," the coach said. "Tonight, in the first three minutes of the game they really took it to us, but after that I thought we settled in and played hard. We didn't probably get enough scoring chances although we had the puck in our end for quite a bit. In the third, we got more scoring chances and hit a few posts; the pucks just weren't going in for us tonight. I'm not going to criticize my players' effort.
"I thought we were ready, but somehow we have to find a way to score goals. They had about 30 blocked shots tonight so we shot about 61 shots at the net and 30 of them got blocked so they did a great job at fronting our shots. You have to give them credit for doing that.'
Milan Lucic did finally score in the final 60 seconds of the game, snapping a 134-minute drought for the team. Julien can only hope that momentum carries into Game 7 on home ice.
"It just gave us an opportunity to be in the game and you hope you can go get him a goal soon after," Julien said of the Lucic goal. "Unfortunately, it was a little too late and we weren't able to get that last one, but these are things that we're going to have to find ways to score hopefully earlier and get more scoring chances earlier in the game instead of the third period tonight. If we can do what we did in the third, I think our chances are good.
'You really wish that line would get rewarded with some goals with the work they put in there. They work so hard, they make good things happen but unfortunately they haven't been rewarded with the goals and that's the unfortunate part. I guess everyone on the bench was routing for [Trent Whitfield] to score that goal because he's been a good soldier for us, he's been working hard and waiting for his turn to get in there and did a great job to spring himself loose. That would have been a big goal for us.'
|B's getting defensive about tonight
|05.12.10 at 1:24 pm ET
PHILADELPHIA — While Zdeno Chara wears the 'C' as captain, Mark Stuart's return has been a steadying influence in the Bruins dressing room.
He missed the last two weeks of the regular season with an infection following surgery on his pinkie. And then he missed the opening round against Buffalo and the first three games against the Flyers. When he finally returned in Game 4 of this series against the Flyers, a quiet leader had returned to the Bruins dressing room.
That leadership was on full display on Wednesday morning when asked if he and his fellow defensemen need to get more offensive pressure on just-returning Flyers goalie Michael Leighton after getting just two shots on net in Monday's 4-0 loss.
"Obviously, we want to get as many shots as we can," Stuart said. "We didn't do a great job of that last game. Tonight's a new night here. If we can just forget about last game and quit talking about it and go play tonight."
"Anybody who's been out for a long time, it just doesn't come back overnight," Julien added. "What you need to see is a guy getting better and better and I think Mark was better in the second game than he was in the first and hopefully he'll be better in the third than he was in the second so it's like every player that comes back, you hope that they get better as they move on here.
"Mark, to me, was a much better player than he was in the first and hopefully, that continues."
Just as obvious to the Bruins is the mission at hand tonight. They are hoping to avoid the pressure of a do-or-die Game 7 back in Boston on Friday night by beat the Philadelphia Flyers tonight at the Wachovia Center. The Bruins now lead the series, 3-2, after dropping the last two games while the Flyers have gained momentum with star Simon Gagne back in the lineup.
Stuart is not the only defenseman looking to pick up his game after not getting a shot on net. Matt Hunwick was also blanked on the shot chart.
"I think we just need to relax a little bit more and not hold the stick too tight and just play how we played the first three games," Hunwick said. "We battled the first three games. They were tight, they were close but we found ways to win. I think we can look back on those efforts and we should do a little bit better tonight."
As for the Bruins getting together for Kumbaya, Unbuntu or any other team-building the night before Game 6, Julien said there was no need.
"I don't think there was anything special [Tuesday] night. We didn't do a team dinner but I think our guys, in order to have success, we can't be tense," Julien said following this morning's optional pre-game skate at the Wachovia Center. "Being relaxed is maybe not even the right word but in the right frame of mind and hopefully that's what we show tonight when we start the game."
The Flyers can become the first NHL team since 1976 to force a Game 7 after being down 3-0 with a win on home ice tonight and are looking to become the fourth major pro sports team to win a series after being in a 3-0 hole.
'We haven't talked about it much,' admitted Flyers coach Peter Laviolette Wednesday. 'Certainly everybody understands the difficult challenge that an 0-3 series presents. That's part of the story you write through the course of the playoffs if you're able to get through it, it's part of the story. Our guys are well aware of it. Certainly they know not many teams get an opportunity at something like this. Any time you can put your name to a positive mark in history like that, it certainly would be a good thing.'
For the Flyers, head coach Peter Laviolette indicated that forward Claude Giroux would be ready for Game 6 after being held out of the third period on Monday after a hit to the boards from Steve Begin.
|Taking the edge off the Bruins
|05.11.10 at 2:30 pm ET
WILMINGTON — Somebody needs to take a little bit of an edge off these Bruins.
Captain Zdeno Chara made a half-admission after Game 5 on Monday night that the team may have been a touch nervous heading into what could have been a series-clinching victory.
I don't know if we were maybe a little bit nervous. It's hard to explain and really find words for it so for sure we didn't play with the composure we were playing with," Chara said Monday. "Maybe it wasn't nervous, it was just'¦ we couldn't make those plays we normally do, strong plays with the puck, plays that we are normally doing and all of the sudden it was tough for us to make those plays."
In the grand world of hockey cliches, this is what is called "clutching the stick." The Bruins need someone, be it Johnny Boychuk and his eccentric antics, Shawn Thornton and his smile and his wife's cooking or Claude Julien putting "Who Framed Roger Rabbit" on repeat on the team plane.
"Everybody can keep it loose and there is no reason to tense up and grip the sticks too tight," Boychuk said. "We know what we have to do and just go out there and do it. There are times to keep loose and times to focus and we know that and that is what we have been trying to do."
Thornton was of the opinion that, heading into Game 5, the team was relatively loose and had a good energy level. For the most part the Bruins tend to be a loose team. Chara and Patrice Bergeron are serious with the media and on the ice but there are moments when you catch them joking around with the guys. Thornton thinks that everybody on the team has a role to play in taking the edge off. He would not name specific characters for fear of being labeled the jokester by the coaching staff.
"We have got a few guys who like to keep things loose. It wasn't too tense today [Tuesday," Thornton said. "We did a pretty good job of forgetting about losses and forgetting about wins and moving on. We learned some things today and move on to the next one. There is nothing you can do. There was only seven on the ice but before the game too, there was a lot of energy. I don't know. We definitely didn't play the game we wanted to but honestly I thought going into it that we felt pretty good."
Coach Claude Julien agreed that everybody on the teams plays their part in keeping the room loose and said that, when it really come down to it, winning is what puts a smiles on everyone's face.
"We all have a part to do in that. I am telling you right now that we have too put yesterday aside and learn from it," Julien said. "That is what the players have to do and so do the coaches. You know, we have to take the same approach as a group and that is what we have done here. We have to focus as a group and do what we need to do tomorrow and hopefully those are good things and that we can come back with smiles on our faces."
|Bergeron the stick that stirs Bruins offense
|05.10.10 at 12:30 pm ET
For all the talk about Miroslav Satan, his hands and his legions or Mark Recchi and how he is the remarkable ageless one during these 2010 Stanley Cup playoffs, it is Patrice Bergeron who actually leads the Bruins in points this postseason.
Bergeron has four goals and seven assists through the first 11 games of the playoffs and he has been a difference-maker on both sides of the blue line. One has to wonder, though, if a guy like Bergeron, known especially to be a great defensive forward who is strong on the faceoff, purposely started to ramp up his offensive production. It seems in the nature of a guy like Bergeron, quiet yet with a developed sense of responsibility, to take it upon himself to create more offense for a team that struggled to light the lamp throughout the year.
"If you play defensively sound and it starts for a good offense. You know, I have always done that and it has been going well," Bergeron said. "I think right now, I don't think I am doing anything different, it is just going in. Obviously we needed it in the playoffs and everyone wants to chip in anyway possible and you know, right now, I am just happy that it is going the way it and and I just want it to keep on going."
Bergeron leads the team in playoff shots at 33 (two more than Michael Ryder, four more Satan and six more than the nearest defensemen, Zdeno Chara and Johnny Boychuk at 27). He is second in assists (Dennis Wideman leads with nine) and has been dominant on the faceoff dot against the likes of Flyers captain Mike Richards or Sabres center Derek Roy. He posted his first sub-50 percent faceoff game of the playoffs on Friday in Game 4 but his numbers in the circle have been closer to 60 and 70 percent for most games this postseason.
"I think in Game 4 we didn't do as good on the faceoff that we would have liked to so, it is huge to get the puck back and work with the puck and play a puck possession game and we have done a great job of that," Bergeron said. "So, obviously we want to start with the puck more often to start with the puck as much as we can."
It is not like Bergeron has all of a sudden flipped a switch in terms of offensive efficiency. He led the Bruins in point at 52 this year, which is not a lot in consideration with the NHL points leaders but still not a paltry sum. But, as the Bruins offense has come awake during the playoffs, either by good fortune that was not present during the regular season or increased efforts by guys like Recchi and Satan in the offensive zone, Bergeron by been the swizzle in the Bruins coffee.
Bruins coach Claude Julien knows that it is the responsibility of his players to play a good two-way game, such as the standard is Bergeron. The center was chosen for the Canadian Olympic team because of his defensive prowess and responsibility. For the Bruins, that approach has also turned into points on the scoreboard.
"We expect everybody to play a good two-way game. We always encourage our guys to be proactive offensively and we want them to be responsible defensively that is what you want in a well rounded team," Juliens said. "That is what we have encouraged all year, whether is has happened or not, the way we like it, that is a different story. But, to have those guys do that it just makes us a better hockey team and certainly we encourage our guys to be more proactive."
|Sunday notes: Pressure? What pressure for Game 5?
|05.09.10 at 3:18 pm ET
WILMINGTON — Pressure?
There has been a lot of talk in this Eastern Conference semifinals series about where the pressure lays. Flyers coach Peter Laviolette said that the pressure is on the Bruins before Game 4, Mark Recchi said afterward that he does not see where Laviolette gets that notion. Really though, we are talking about pressure. It is like talking about "character" — some esoteric notion that you know exists and it effects how a team plays and is perceived but it cannot be quantitated or examined until well after the point of pressure and high anxiety has passed.
"I feel that every game there has got to be a sense of urgency and that is the way we have approached it," coach Claude Julien said. "Some people call it pressure some people call it something else. You put the pressure on yourself to do well because we want to do well. I think pressure is something that, if you handle well, is a great thing to have on your side. If you can't handle it well it is certainly something that can be detrimental to your team."
Part of the reason the Bruins may have been playing so well through the playoffs is that their definition of "pressure" may have worn off. The ultimate embarrassment for Boston would have been to not qualify for the playoffs at all a season after having the best record in the Eastern Conference and starting the year as one of the Stanley Cup favorites. In that regard there was more pressure through the end of March into April than there has been during the playoffs.
"Well, we have been better for quite a while. We did it when we got into the playoffs. This is a better team and you move on from there," Julien said. "There was a sense of urgency or whatever you want to call it before the playoffs started so, we have gone through that and are adjusting to it right now. I find we are very focused team right now. We just have to keep that in the right direction and for us, everybody game is a must-win. So, no matter what, every game is a must win, we take that approach and it has served us well."
After making the tournament and getting out of the first round, the fear of embarrassment or failure, which might be a good definition of pressure, has not been present. They are able to go out and play hard and have fun while working hard. It is the playoffs, it is supposed to be fun because, after all, hockey is just a game.
"You can't play this game and not have fun. You guys can't do your job and not enjoy it. Otherwise, might as well changed your job right?" Julien said. "It is the same thing for players. You have to go out and love this time of year. There's a bunch of teams right now watching us play that would love to be where we are and that is fun. We have to take that approach and we have taken that approach. We have come into the dressing room after a period either down a couple of goals or tied or whatever and say 'guys, lets just go out there and win this game and have fun doing it.' And the guys have taken that approach and it has worked well for us."
Notes: The full compliment of healthy Bruins skaters were present at Ristuccia with Adam McQuaid the only player who might have been a possibility missing. He is still out with a "lower-body injury" and remains doubtful for Monday's Game 5. It is not likely that McQuaid would play either as Mark Stuart has come back to the lineup and, after a poor Game 4, feels that he will be able to get back up to mental and physical speed in his second playoff game of the year.
"Yeah, it was a little different actually, I felt like I was crashing the party," Stuart said. "I thought my emotion level would be there because of the playoffs and it definitely was because of the situation and the intensity is way up and everything is faster. I think I will be up to speed tomorrow."
Dennis Seidenberg skated on the Ristuccia ice after the rest of the team had completed its practice and was worked out by trainer John Whitesides. Seidenberg has been on the ice for two days in a row now as he battles back from a lacerated tendon suffered in Toronto on April 3. He had a hard cast taken off the left forearm last Monday to reveal a long, horizontal scar five inches up from his wrist. He is not expected to be back until at least eight weeks after the surgery but Julien said that it has been encouraging to gets guys back into the lineup even as big performers (Marco Sturm, David Krejci) have hit the infirmary.
"Any time you see that kind of thing around your team it is a positive," Julien said. "We have been hit this week with some big injuries but then you look at the other side and you see some other guys start to come around. So, hopefully we continue to win hockey games to give those guys and opportunity to come back."


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Genetic systems involving multiple X chromosomes have arisen repeatedly in sexually reproducing animals. Tiger beetles (Cicindelidae) exhibit a phylogenetically ancient multiple-X system typically consisting of 2–4 X chromosomes and a single Y. Because recombination rates are suppressed in sex chromosomes, changes in their numbers and movement of genes between sex chromosomes and autosomes, could have important consequences for gene evolution and rates of speciation induced by these rearrangements. However, it remains unclear how frequent these rearrangements are and which genes are affected.
Karyotype analyses were performed for a total of 26 North American species in the highly diverse genus Cicindela, tallying the number of X chromosomes and autosomes during mitosis and meiosis. The chromosomal location of the ribosomal rRNA gene cluster (rDNA) was used as an easily scored marker for genic turnover between sex chromosomes or autosomes. The findings were assessed in the light of a recent phylogenetic analysis of the group. While autosome numbers remained constant throughout the lineage, sex chromosome numbers varied. The predominant karyotype was n = 9+X1X2X3Y which was also inferred to be the ancestral state, with several changes to X1X2Y and X1X2X3X4Y confined to phylogenetically isolated species. The total (haploid) numbers of rDNA clusters varied between two, three, and six (in one exceptional case), and clusters were localized either on the autosomes, the sex chromosomes, or both. Transitions in rDNA localization and in numbers of rDNA clusters varied independently of each other, and also independently of changes in sex chromosome numbers.
Changes of X chromosome numbers and transposition of the rDNA locus (and presumably other genes) between autosomes and sex chromosomes in Cicindela occur frequently, and are likely to be the result of fusions or fissions between X chromosomes, rather than between sex chromosomes and autosomes. Yet, translocations between sex chromosomes and autosomes appear to be common, as indicated by the patterns of rDNA localization. Rearranged karyotypes involving multiple sex chromosomes would reduce recombination, and hybrid dysgenesis selects against polymorphic populations. Hence, the high frequency of these rearrangements could be a cause of the great species diversity in Cicindela.
Chromosomal rearrangements have been described in many groups of animals, distinguishing even very closely related species and populations [1-4]. As rearrangements would change the position of genes, generating new linkage groups that may lead to an increase of genetic differentiation between populations, they are expected to restrict reproductive compatibility and eventually promote speciation. Chromosomal rearrangements produce unbalanced gametes which have been thought to be the primary effect of rearrangements in speciation because of the diminished fecundity or viability of the heterozygotes [5]. However, this notion of 'hybrid-dysfunction' as a major cause of karyotype-mediated speciation may not be an effective isolation mechanism [1,6,7]. More recently, it is being realized that the primary effect of chromosomal rearrangements is the reduced recombination between genomes, favoring 'suppressed-recombination' scenarios for the evolution of reproductive isolation following chromosomal rearrangements [1,6,8].
Recombination rates vary widely depending on the genomic position and the linkage with genes under selection, affecting the tempo and mode of gene evolution [e.g. [7,9,10]]. Specifically, recombination is greatly reduced in sex-chromosomal linkage groups, because of the lack of a homologous chromosome in the heterogametic sex, or it is prevented altogether where sex chromosomes are achiasmatic in one of the sexes. In addition, due to the hemizygosity of the X chromosome (or the W chromosome in ZW systems with heterogametic females), recessive mutations associated with the sex chromosomes are exposed to selection in the heterogametic sex [11]. Hence, as [12] pointed out, the origination of new X chromosomes leads to fixation of a great deal of heterozygosity (while new Y chromosomes lead to the rapid degradation of gene content; see [13]). As the result of reduced recombination, genetic barriers to gene flow may arise rapidly between populations which are fixed for sex chromosomal variants.
Lineages with multiple-X chromosome systems may be especially strongly affected by these evolutionary regimes because a great proportion of their total genome is captured in the sex chromosomes. The large size of these X chromosomes and their limited heterochromatin content in karyotypic analyses suggest that they contain a large proportion of genes. Multiple-X chromosomes occur widely in vertebrates and invertebrates, for example in monotremes (Platypus) [14], howler monkeys (Aloutta) [15], fishes [16], gastropod molluscs [17,18], spiders [19], Lepidoptera [20] and several groups of Coleoptera [21-24]. Among these, the widely conserved XXY system in araneomorph spiders [19] and the ancestrally XXXY system in Cicindelidae (tiger beetles) are the most ancient multiple-X system known [25]. Novel X chromosomes of this kind originate readily as a consequence of karyotypic rearrangements involving a sex chromosome and an autosomal pair, producing chiasmatic sex chromosomes. For example, a fusion of the X chromosome with an autosome will produce an expanded X chromosome while the remaining sister chromosome will become an additional sex chromosome. In XY systems, this will produce XXY, whereas in the case of an X0 karyotype, this fusion will produce an enlarged X chromosome and the homolog will act as Y chromosome (producing a neo-XY system). An alternative pathway is that rare chromosomal rearrangements with the autosomes increase the size of the X chromosomes, followed by their dissociation to produce multiple X chromosomes [12]. In all cases, the consequence is that genes are shifted from an autosomal to a sex chromosomal position, and hence alter the evolutionary regime experienced by these genes.
However, the effects of autosomal-heterosomal rearrangements on the evolution of gene regions and recombination-suppressed speciation depend on the evolutionary dynamics and genomic extent of such rearrangements. An unknown factor is what is the frequency of autosomal-heterosomal changes, and if particularly gene regions would be affected repeatedly in the evolution of a lineage. While several studies have assessed these phenomena in populations and closely related species to investigate the microevolutionary consequences of rearrangements [1], only a few studies have addressed these questions on a macroevolutionary (between-species) scale [26]. Here, we conducted a survey of X chromosome numbers and rDNA localization on a representative sample of North American tiger beetles in the genus Cicindela. This group exhibits a conspicuous, non-chiasmatic multiple-X chromosome system consisting of 2 to 4 X chromosomes, plus a single Y, and between 9 to 11 pairs of autosomes. The multiple-X system in Cicindelidae is apparently ancient and is present widely in this families except for a set of basal lineages exhibiting an XY system [25]. Despite the presumed antiquity of the system, secondary reversion to simple XY or X0 genetic systems have occurred, but are rare [25,27]. In addition to the karyotypic variation in sex chromosome numbers, species of Cicindela differ in the number and localization of rDNA loci which include either two or four pairs of clusters and which are localised on either the autosomes or heterosomes [23,28,29].
The Cicindela of the North American continent constitute at least four distinct radiations, each of which with an estimated origin of several million years ago. Based on a phylogenetic tree available at the species level for each of these four lineages [30], the character transitions in sex chromosome number and switches in the position of rDNA between sex chromosomes and autosomes can be analyzed. Information on the tempo and mode of rearrangements in this genetic system that retains a large proportion of the physically observed karyotype in multiple X chromosomes, will provide the backdrop for assessing the effect of autosomal-heterosomal turnover on the evolution of coding regions and the origination of species.
X chromosome numbers and rDNA localization
Karyotype analysis and chromosomal localization of rDNA clusters was carried out in 26 North American species, sampled to cover the main phylogenetic groups of the continent [30]. Examples of karyotype and FISH visualization are shown in Fig. 1, and the results for all species are summarized in Table 1. Autosome numbers were constant for all species tested (n = 9). Heterosome numbers were variable, but most species exhibited an X1X2X3Y karyotype, which is also the most common type in Cicindela from the Palearctic, Australia and India, and presumably represents the ancestral state for the entire lineage that also includes other genera [25]. We also found the X1X2X3Y genetic system in several species which in the older literature were reported to exhibit a single X chromosome (C. pimeriana [31]; C. punctulata, C. purpurea, and C. repanda [32]; or X1X2Y and n = 9 for Cicindela repanda, C. tranquebarica, C. scutellaris and C. sexguttata [32], but some of the X chromosomes may have been overlooked in these studies. Our analysis found four species each showing X1X2X3X4Y (C. lemniscata, C. marutha, C. nigrocoerulea and C. marginata) or X1X2Y (C. sedecimpunctata, C. ocellata, C. rugatilis, C. nebuligera) (Table 1). For the autosomes, pairs of homologous chromosomes in meiosis could be paired easily by their size. The autosomal karyotype in all species had many features in common, even where heterosome numbers differed. For example, the largest pair was metacentric and clearly distinguishable from the rest. The second and third pairs were also recognizable as metacentric whereas the fourth was submetacentric. From the third pair to the ninth the size decreased gradually (not shown).
Table 1. Karyotypic data of North American species of the genus Cicindela
Figure 1. Fluorescence in situ hybridization of tiger beetle chromosomes with a ribosomal probe. A. Cicindela punctulata, n = 9 + XXXY, early prophase I nucleus showing two rDNA loci, one located on the Y and another located on one of the four X chromosomes. B. C. scutellaris, diakinesis with n = 9 + XXXY showing two rDNA loci located on an autosomal pair. C. C. repanda, diakinesis with n = 9 + XXXY showing two rDNA loci located on an autosomal pair D. C. tranquebarica, diakinesis with n = 9 + XXXY showing three rDNA loci, two located on an autosomal pair and one located on one of the 3 X chromosomes. E. C. ocellata, metaphase I with n = 9 + XXY showing two rDNA loci located on an autosomal pair and one on the X chromosome. F. C. marginata, spermatogonial metaphase with 2n = 23 chromosomes showing six rDNA loci, one located on each of the four X chromosomes and two loci located on the Y chromosome. The white arrows indicate the heterosomal loci, the empty arrows indicate the autosomal loci. The arrowheads point to the sex chromosomes which are condensed in Fig. 1D forming the sex vesicle, and are recognizable individually forming the rosette-like structure in Figs. 1A, B, C, and E.
We found substantial variation in the number and localization of rDNA clusters (Table 1; Fig. 1). Three types of rDNA constellations were evident, including: localization on the autosomes (one autosomal pair) in about half of the species; localization on the heterosomes (one of the X chromosomes and the Y) found in six species; and localization on both (one autosomal pair plus heterosomal copies located on one of the X chromosome) in the remaining five species. An additional type was found in C. marginata where mitotic metaphases showed six rDNA stains on five chromosomes (Figure 1F), with a single chromosome carrying rDNA clusters on both arms. We identified this chromosome as the Y, due to its morphology and orientation on first meiotic plates. In this image the fluorescent signal was located exclusively on the sex vesicle. Whereas all rDNA clusters present in the genome can be detected with FISH, silver staining on spermatogonial cells only shows those genes that are actively transcribed during spermatogenesis. Results from FISH and silver staining were similar generally, indicating that most rDNA clusters were active, but in species with rDNA clusters found on both heterosomes and autosomes, expression was mostly limited to the heterosomal locus, indicating the functional importance of the sex-chromosome linked copies.
Character reconstruction and covariation
Parsimony reconstruction (accelerated transformation) on the mtDNA tree inferred the X1X2X3Y system as the ancestral state, with several independent derivations of the X1X2X3X4Y and X1X2Y state (Fig. 2). Whereas the species exhibiting four X chromosomes were found in distant parts of the tree, all species with two X chromosomes were limited to the subgenus Cicindelidia. The results were consistent with a repeated transition from X1X2X3Y to either of the two derived states, but not in the reverse direction. The transitions to the X1X2Y and X1X2X3X4Y states were entirely limited to the terminal branches of the tree, and there was no direct transition between these two character states. The localization of rDNA was similarly variable phylogenetically, however the species with exclusively autosomal rDNA (labelled AA in Fig. 2) were largely limited to two clades, the subgenus Cicindela (sensu stricto) and the 'red abdomen' group of Cicindelidia. All other groups exhibited rDNA on the heterosomes, but without phylogenetic conservation of the two main states (autosomal pair plus one X, labelled AAX in Figure 2; or autosomal pair plus one X and Y, labelled XY).
Figure 2. Phylogenetic distribution of character states in sex chromosome number and rDNA localization. Character optimization using parsimony was conducted separately for the number of sex chromosomes (2, 3, or 4 X chromosomes) and rDNA localization (localization on autosomes only, on the autosomes and X chromosomes, and on the heterosomes only). The tree is from Vogler et al. (2005) representing approx. 75% of the Nearctic species of Cicindela (s.l.) and species names are preceded by an abbreviated subgenus label (CDIA, Cicindelidia; CELA, Cicindela; TRIB, Tribonia; PACH, Pachydela; HABR, Habroscelimorpha; CYLI, Cylindera; ELLI, Ellipsoptera; BRAS, Brasiella). Several major groupings established in the previous phylogenetic analysis are indicated on the tree. Asterisks above the branches indicate nodes with good support (Bremer Support ≥3 and Bootstrap values ≥60%). Karyotype information is presented by symbols at the tips for all taxa represented in the current study. Different shading of branches indicates the character optimization for rDNA localization. Due to the incomplete taxon sampling it was not possible to assign the character changes to a precise branch deep in the tree, indicated by thin lines.
Variation in X chromosome numbers and rDNA localization was largely independent. For example, ten sister species pairs can be assessed for covariation between rDNA positional changes and differences in X chromosome numbers on the tree of Figure 2. Between the sister species a character change in X chromosome number was observed six times, a change in rDNA localization was observed three times, and three species pairs were invariable. Among those sister pairs that were variable for one parameter, a transition in the other parameter is inferred only in two cases, and leading to different states. This demonstrated a broad lack of co-variation indicating that the chromosomal rearrangements producing variation in both karyotypic features were not connected.
Phylogenetic character reconstruction at the species level is useful to test for trait changes associated with speciation events [33]. The incompatibility of rearranged chromosomes could be an important factor leading to reproductive isolation and speciation, in particular where such changes are linked to the sex chromosomes. The genus Cicindela is very species rich, which is usually ascribed to ecological shifts in habitat associations [34,35] and a tendency for the isolation of local populations [36,37]. However, karyotypic features have not previously been considered to affect species diversification in this group. Karyotype analyses in Cicindela are technically challenging, due to the small size of chromosomes and the absence of the polytene chromosomes common in Diptera, plus the difficulties of obtaining meiotic cells (where multiple sex chromosomes can be accurately identified) except during a short season of the year. Yet, our sampling was sufficiently dense for a comparative analysis. We found that the multiple sex chromosomes provide an evolutionarily dynamic system affected by repeated gains and losses of X chromosomes, and repeated shifts in the localization of the rDNA loci between autosomes and heterosomes, with potential consequences for speciation.
Mechanistically, these shifts in X chromosome numbers do not appear to involve reciprocal rearrangements in the autosomes, as the latter remain invariable, arguing against a mechanism for the origination and loss of X chromosomes through fusions with autosomes. The independence of autosome and heterosome numbers has previously been established in lineages of Cicindela in the Palearctic, India and Australia, although in those cases changes affected mainly the autosome numbers while heterosomes were invariable for the modal X1X2X3Y system [38,39]. The absence of autosomal-heterosomal fusions and fissions was also supported by the fact that we did not observe any physical associations of a chiasmatic type between autosomes and heterosomes during meiosis which would be required for such rearrangements. Instead, in all cases studied here the multiple X chromosomes and the Y were connected at the telomeric ends during pairing in male meiosis and were visible as a conspicuous multivalent where (Fig. 1), as had already been described for the Palearctic species C. hybrida [40].
The lack of chiasmatic associations between heterosomes and autosomes also argues against a role of fusions and fissions to be responsible for the positional changes of the rDNA clusters. The number and position of rDNA loci can readily be altered by Robertsonian changes (fissions or fusions) via terminal nucleolus organizer regions (NOR) usually containing the rDNA clusters. Two chromosomes each carrying a terminal NOR may either fuse to produce an interstitial NOR, or a single chromosome with an interstitial NOR may undergo fissions resulting in two chromosomes with terminal NORs. However, except perhaps for the case of C. marginata with six rDNA loci, we did not see the expected changes in number and position of rDNA loci, as changes in chromosome number and rDNA loci were independent. Therefore neither fissions nor fusions at NORs are supported, unless one invokes additional rearrangements such as pericentric inversion to accommodate the predominantly mediocentric chromosomes in Cicindela. Furthermore, considering the achiasmatic nature of heterosomes, Robertsonian rearrangements cannot explain the changes that occur simultaneously in autosomes and heterosomes.
An alternative scenario are non-reciprocal translocations affecting the rDNA clusters which could lead to changes in position (translocation) or numbers of rDNA sites (transposition retaining a copy at the origin). These changes can be facilitated by the presence of transposable elements, as e.g. in the Type I and II ribosomal gene insertions in Drosophila melanogaster, Bombyx mori [41], Apis mellifera and other Hymenoptera [42]. Translocations between chromosomes bearing rDNA loci and those lacking them have been invoked to explain the rearrangements observed in Paeonia [43] and Allium [44], and also may occur in the ground beetle genus Zabrus which presents the highest variability in the number of rDNA sites (2–12) found so far in insects [45]. However, these rearrangements occur among autosomes only, not affecting the sex chromosomes. The peculiar situation in C. marginata, where translocation of rDNA copies between chromosomes and even within a chromosome may be responsible for the unusual number of rDNA loci, might be a good model for elucidating the cytogenetic basis for the changes in number of X chromosomes and gene content.
Whatever the mechanisms that produce the change of number and position of rDNA clusters, they seem to be affected by various constraints to the rearrangements of karyotypes. Careful inspection of the chromosome preparations suggested that the rDNA loci were not always in the same pair of autosomes, and may be localized on different homeologous pairs, even in closely related species. Therefore, numerous additional chromosomal rearrangements between autosomes and heterosomes may be present which are not detected here when scoring chromosome numbers and rDNA autosomal-heterosomal localization only. Yet, the overall gestalt of the karyotype was maintained despite these apparent changes in gene content. Nine pairs of autosomes were found in all species, showing very similar sizes and phenotypes. This degree of conservation appears to be specific to the multiple-X karyotype, as such constraints on chromosome morphology are not seen in the ancestral simple-X system of distantly related cicindelids which differ greatly in chromosome number and overall morphology [25]. Similarly, there are apparent constraints to the number and gene content of heterosomes. For example, the X1X2Y and X1X2 X3 X4Y karyotypes represented derived states which were confined to a single species (or perhaps small clades, had denser taxon sampling been available), indicating that deviations from the modal X1X2X3Y system are evolutionarily short-lived and unstable. Finally, the chromosomal positions of rDNA clusters also appeared to be constrained in the multiple-X system. The autosomal number of clusters was two (one pair of allelic copies), but never exceeded this number, and when more than two rDNA copies were found in Cicindela they always appeared on the heterosomes, most commonly on the X only, and in some species a further rDNA copy on the Y chromosome. This is in contrast to the single-X chromosome systems in the basal groups of Cicindelidae which exhibit between four and eight (two to four pairs) autosomal rDNA clusters [25]. The nature of these constraints on the karyotype remains unknown but they may be linked to the evolutionary stability of this multiple-X system [25].
Despite the morphological conservation of the multiple-X system, the frequent movements of genes between autosomal to heterosomal positions expose the affected loci to greatly altered dynamics of gene evolution and recombination. As there is no cross-over in the male heterosomes in cicindelids, rates of homologous recombination in the sex chromosomes are reduced by half for the X chromosomes (recombination only in females) and to virtually zero for the Y (resulting in their inevitable degradation; [13]). The hemizygous nature of the X will greatly increase selection on recessive mutations, altering the rate and kind of mutational changes. This would cause the rearranged genes to diverge quickly, even if rearranged gene regions are duplicated. In the case of the rDNA clusters, this could reduce the rate of homogenization of copies in different parts of the genome. For example, an analysis of sequence variation in the ITS1 region of the rDNA cluster in C. dorsalis, a species shown here to exhibit rDNA copies on a pair of autosomes plus a single copy on the X, exhibited greatly divergent ITS types, possibly corresponding to heterosomal and autosomal copies [46]. This supports the idea that the translocation to the sex chromosomes results in changes of evolutionary dynamics.
These kinds of chromosomal rearrangements might also have an effect on speciation. Translocations of genes between autosomes and sex chromosomes will greatly change the possibilities for gene flow, and changes in the number of X chromosomes may alter the production of balanced gametes. Incompatibility of gametes with different numbers of X chromosomes, or indeed changed position of rDNA clusters, could lead to incorrect separation of chromosomes during anaphase I in a hybrid, and produce a number of unbalanced gametes resulting in reproductive disadvantage. In the case of changes in X chromosome numbers this effect could be exacerbated by altering the sex determination control and the gene regulation associated with changes in heterosome number. As pointed out in the recent literature [1,6,7,47], it is not likely that these 'underdominant' variants become established in a population. This has provided a strong argument against the stasipatric model of speciation [5] which suggests that chromosomal rearrangements cause reproductive isolation due to hybrid dysfunction. However, when involving sex chromosomal unidirectional rearrangements as those in Cicindela, this model may still be valid. Depending on the precise genotypes participating in a mating, the combination of certain gametes could lead to a significant proportion of inviable (e.g., no rDNA clusters, unbalanced number of X chromosomes) zygotes, selecting against heterozygotes and maintenance of polymorphisms in a population. These effects may be exacerbated by the reduction of gene flow from suppressed recombination and extending the effect of linked isolation genes, a mechanism proposed as the main driver for the fixation of novel karyotypes under more recent models [8,48]. As gene flow is more restricted between sex chromosomes than autosomes, sex linked genes are particularly efficient to produce such postzygotic barriers [49], and hence rearrangements involving sex chromosomal portions of the genome may be a particularly effective isolating mechanism.
Therefore, the high level of apparent chromosome rearrangements and the deposition of a substantial portion of the genome in the low-recombining sex may promote speciation in Cicindela. With some 1,000 species world-wide, this is one of the largest genera of insects. In particular, the observation of evolutionarily short-lived X1X2Y and X1X2X3X4Y lineages suggests that these chromosomal changes could initiate reproductive isolation. If these rearrangements are frequent relative to other (ecological or geographical) processes influencing speciation rates, the cytogenetic parameters could drive speciation and possibly be responsible for the great species richness in Cicindela. In addition, the population structure of Cicindela is also favoring the fixation of chromosomal mutations locally, as most species are early succession specialists frequently occurring in isolated habitat patches where a dynamic system of colonization and extinction may enhance the separation of local genetic entities. In support of this possibility, we found two Iberian species, C. flexuosa and C. littoralis, where in each case a single population was fixed for an rDNA localization different from all other populations [29], indicating that local genetic races arise frequently.
The evolutionary significance of elaborate multiple-X systems compared with simpler sex chromosomes is still poorly understood. Phylogenetic approaches can greatly increase the power of comparative cytogenetic analyses, and have revealed the great fluidity of the cicindelid multiple-X chromosomes, while also establishing evolutionarily conserved features. Even more variable than the X-chromosome numbers are translocations of the rDNA clusters. This provides the background for future investigations to understand the evolutionary forces operating on the sex chromosomes. Whereas the current study uses a macroevolutionary approach, establishing the framework of character variation over greater evolutionary distances, this can be combined with the specific effects of rearrangements on the population level.
Sex chromosomes are of specific interest to speciation, not least because they have been shown to accumulate genes determining species specific traits such as host plant use and pheromone response in butterflies [50]. The observed cytogenetic phenomena should also be studied because of their functional consequences, with regard to the control of sex determination, chromosome size and morphology, and the mechanisms of gene repositioning. Further investigation will require a targeted approach to the comparative genomics of tiger beetles and the construction of chromosomal homology maps, using reciprocal chromosome painting with sex chromosome specific probes obtained by microdissection. Molecular cytogenetics studies, assisted by comparative genomics and phylogenetics, may help to investigate the evolutionary dynamics of gene content of chromosomes and may reveal karyotypic changes that remain unnoticed in conventional cytogenetics analysis. These studies are the basis for tests of how variation in sex chromosomes can drive population differentiation and speciation rates.
Samples for study and chromosome preparations
Taxon sampling for this study was limited to Cicindela from North America. There are some 147 described species recorded for the North American continent, grouped in 11 subgenera [51]. A recent phylogenetic analysis based on 1897 base pairs of mtDNA and taxon sampling which is 75% complete at the species level, revealed that most species can be ascribed to four monophyletic groups representing radiations endemic to North and Central America [30]. A representative sample of all major clades in the mtDNA tree was selected for chromosome analysis, with good representation of all four endemic radiations. Character variation in X chromosome number and rDNA localization was assessed on this tree, using parsimony optimization as implemented in MacClade [52]. Adult beetles were obtained in the field, and chromosome preparations were obtained from male gonads. Mitotic and meiotic chromosomes can be obtained at different stages of development in the tubular testes, and can be observed as described previously [28].
Silver staining and in situ hybridization
Active NORs (i.e., regions of active rRNA transcription) were detected with silver according to the [53] technique, with slight modifications [29]. In situ hybridization was performed as previously reported in Proença and Galián (2003) using a 555 bp rDNA probe obtained after PCR amplification of the conserved 18S rRNA gene region from C. campestris total genomic DNA using the universal primers NSI and NSII [54], labelled with biotin-11dUTP by a second PCR reaction.
Authors' contributions
JG carried out the design of the study, the collecting of most samples, the molecular cytogenetic analysis, and drafted the manuscript. SJRP participated in the cytogenetic studies. APV conducted the character analysis, and participated in the design of the study and in the writing of the manuscript. All authors read and approved the final manuscript.
We are grateful to D. Maddison, D. Pearson, A. Cardoso and T. Barraclough for providing specimens, to J. Serrano for advice and comments, and E. Martínez-Navarro for technical assistance. We are grateful for useful comments from three anonymous reviewers. This work was supported by Acciones Integradas Hispano Britanicas/The British Council No. 193A, the Project No BOS2002-02870 of the DGI of the Spanish Ministry of Science and Technology to JG, the Large Scale Facility Program to visit the NHM to JG, the Museum Research Fund of the Natural History Museum, and NERC (NER/A/S/2000/00489) to APV and T. Barraclough.
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Journey to Everest Cafe and Curry HouseJune 9, 2014Indian • Semaphore, Australia
Best curry in SA.
There is a pub attached for those just looking to have a cold beer. Good local wine list as well.
Pretty pissed off that I come to this winery for the sea and vines festival and they sell me wine from NSW, WTF! Oh yes no other reds available.
The porterhouse kilpatrick is not a steak as you know it. It's a schnitzel with a 4mm thin steak in the middle. Not to happy. The staff said "oh this happens all the time"! WTF!
The coffee is very good - I had a double shot flat white. The staff is also friendly.
Have a platter on the lawn or bring your own picnic.
Good service, good food and nice coffee.
Beer at 1973 prices, gold!
The double shot flat white was good and the new owners are friendly.
Not only is the beer good, the breakfast is good.