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Drone samples wanted for research into NZ honeybee diversity


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I work with Associate Professor Peter Dearden at the University of Otago. We have funding through a sustainable farming fund grant to screen the genetic diversity of honeybees in New Zealand. For this study we are doing a survey of sex alleles present in New Zealand's bee population.

 

To be able to successfully carry out this study we need the help of New Zealand beekeepers. We need honeybee samples from different regions of New Zealand and from different beekeepers. We are particularly interested in samples from commercial beekeepers and queen breeders, as these beekeepers supply a very high percentage of queens within New Zealand.

 

What will the samples be used for?

DNA will be extracted from the drones collected and we will use this to determine the sex allele carried by that drone. This information will be used to map the sex allele diversity throughout New Zealand (and this information will be publicly available at the conclusion of the study). These samples will only be used for the currently funded study and will be destroyed at the conclusion of this study.

 

The sampling is very straight forward.

 

Sampling method:

1) Collect some drones from one apiary into a queen cage (multiple hives if possible).

2) Label the queen cage with location details (the more accurate the better).

3) Place the queen cage in the freezer overnight.

4) Send the queen cage/s with dead drones to us.

 

If you are willing to help and send us samples please get in touch with me through email at:

otto.hyink@otago.ac.nz

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  • 3 months later...

We are still looking for drone samples. Still especially interested in drones from hives being bred from (breeder queens) and from individual hives or groups of hives.

 

If you are willing to send some please collect as described above and get in touch with me, either through the forum or by email at the above address.

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Hi Otto, if we send in samples; do we get to know what the results were, not that it would mean much to us

Absolutely. It won't mean much by itself no but a little further down the track we'll produce a map of what we've found where. If we get enough samples from beekeepers around New Zealand it should be a useful database for those wanting to add some new genetics to their breeding operation.

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Otto If it was me I would wait until Janice's hazels are out before going round.

 

You might be lucky and get a free sample.:D

Don't need Janice's hazels as I have an apiary in a block of hazels elsewhere and I'm allowed to help myself.

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Otto, come around any time and sample whatever you like (from the hives, that is).

Hi Janice. If the offer is still open, any chance you're home Thu or Fri for me to come and grab a few drones?

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Hi Otto, if we send in samples; do we get to know what the results were, not that it would mean much to us

 

No offence Otto .... this sounds like a really good way of getting screeds of data for many permutations of spreadsheet, but I'm struggling to imagine what the end benefit might be and how such a project is a worthy use of sustainable farming fund dosh.

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but I'm struggling to imagine what the end benefit might be and how such a project is a worthy use of sustainable farming fund dosh.

 

end benefit = enormous. one of the most worthwhile SFF projects I've seen.

 

Right now we have no real idea about the genetic variability or viability of our populations. As we move more towards selected breeding programmes to try and combat varroa and such we need to know that what we have out there and that we can maintain enough genetic variability that we don't breed ourselves into a corner of overly homogenous bees.

Or for that matter, that we know how much genetic material we have explored to find solutions before we go reaching for other solutions (eg, imports).

 

Otto, I'm small commercial = 75 hives currently. Only do queens for a proportion of my own hives, but happy to get you a sample next weekend if you're interested.

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end benefit = enormous. one of the most worthwhile SFF projects I've seen.

 

Right now we have no real idea about the genetic variability or viability of our populations. As we move more towards selected breeding programmes to try and combat varroa and such we need to know that what we have out there and that we can maintain enough genetic variability that we don't breed ourselves into a corner of overly homogenous bees.

Or for that matter, that we know how much genetic material we have explored to find solutions before we go reaching for other solutions (eg, imports).

 

Otto, I'm small commercial = 75 hives currently. Only do queens for a proportion of my own hives, but happy to get you a sample next weekend if you're interested.

Yes please Dee. Don't have any from Hawke's Bay as yet. Are you happy to collect a random sample or two (2-3 drones from around 10 hives in an apiary)? If you have any breeder queens a sample of 8 or so drones from this hive would be great too.

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Weeell alright then, as long as no drones are harmed ......

It depends on what you mean by 'harmed'. If you mean anything but getting euthanased and blended into bee soup before purifying DNA . . .then no, the drones won't be harmed.

 

Otto has developed a great assay for this work - no screes of excel data but easy to observe 'is this bee the same of different' by following a DNA trace (in colour even for no extra cost!). If two traces the same = bees the same. If trace different . . .you get the picture. Talking of which - how about posting one Otto??

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It depends on what you mean by 'harmed'. If you mean anything but getting euthanased and blended into bee soup before purifying DNA . . .then no, the drones won't be harmed.

 

Otto has developed a great assay for this work - no screes of excel data but easy to observe 'is this bee the same of different' by following a DNA trace (in colour even for no extra cost!). If two traces the same = bees the same. If trace different . . .you get the picture. Talking of which - how about posting one Otto??

Good idea. Will have to be Wednesday as I'm away from work tomorrow. I'll use the results from the drones you provided.

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Yes please Dee. Don't have any from Hawke's Bay as yet. Are you happy to collect a random sample or two (2-3 drones from around 10 hives in an apiary)? If you have any breeder queens a sample of 8 or so drones from this hive would be great too.

 

 

:) Will do. Probably have a bit of running around to do, so I can possibly get you a couple of samples from different origins

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Hi Janice. If the offer is still open, any chance you're home Thu or Fri for me to come and grab a few drones?

Yes, I will be home both mornings. After 10am works best for me because I work evenings.

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Hi Otto, if we send in samples; do we get to know what the results were, not that it would mean much to us
Without being asked to, Otto gave me great feedback on the genetics of my drones, very interesting from my perspective.

 

I am very much looking forward to seeing the results of the whole study.

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  • 2 weeks later...

Some more information for anyone who's interested.

We are looking at the genetic variation (ie DNA differences) in the gene responsible for sex determination. Many of you will know that fertilised eggs result in a female bee while unfertilised eggs result in drone bees. The way this works is through the action of a gene known as csd (for Complementary Sex Determiner). This is a gene for which over 100 different sequences have been identified in Apis mellifera so far (each allele is a different version of the csd gene). When a bee develops with two different alleles of this gene it becomes a female (queen or worker) while a single copy results in a drone.

There are two main objectives for this initial part of our research.

1) Identifying the alleles of csd that are present in queen being used for queen breeding in New Zealand. This tells us the number of different sequences actively being propagated by beekeepers (the more sequences, the better gene pool depth).

2) Identifying the different alleles present in New Zealand bees as a whole to give us a measure of the genetic variation present in New Zealand bees (that can potentially be compared to the variation present in other parts of the world).

The assay we are using to run this test uses DNA purified from drones. The most variable part of the csd gene is amplified out using a PCR test. This is simply a way to get many millions of copies of the specific part of the csd gene we are interested in so that we can analyse the sequence of it and see how it varies from other alleles.

There are several ways of analysing the sequence of a given allele. We can sequence it which gives us exact information for that allele. This is ideal but is prohibitively expensive to do on a larger scale. Instead we analyse the sequence with a technique called High Resolution Melting (HRM). Different sequences will melt differently and give a different fluorescent trace. The same method was used to differentiate bad Psa-V in kiwifruit from a benign Psa species also detected.

The pictures below show some DNA fluorescent traces from some of the assays I have done on some New Zealand drone samples. The first one are the melt curves from 9 drones samples from one of Alastairs breeder queens. A queen has two alleles and her progeny drones will have either one of these two. In this test 5 drones had one distinct melt curve while the other 4 had a different one. The second picture show the results from the drones collected from the hive containing Alastairs second breeder queen. Here the results show 4 drones with one melt curve, 3 drones with a second curve and 1 each with different melt curves. This suggests that 7 drones were the progeny from the queen in this hive while the other 2 were drones that originated in different hives. The last picture show the curves obtained from 12 drones collected as a 'random' sample from an apiary. Two drones were collected from 6 different hives in one apiary. Here there are seven different curves (meaning seven different alleles).

All up, the more samples we receive from beekeepers around the country the more useful the information from this study will be for the beekeeping industry. So keep the samples coming please!

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Slide1.jpg.9ef75b050c230c362ff784490e3fe1f9.jpg

Slide2.jpg.92c0cb74f6f2e459aafd6ee46e2d334f.jpg

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RFU stands for relative fluorescent units.

DNA is made up of two strands bound to each other through four bases (A's bind to T's and C's bind to G's). The binding of the two strands comes apart as DNA is heated. The way a piece of DNA melts is dependent on it's sequence, the combination of A's, C's, G's and T's that make it up, and their order. With this method we take our amplified DNA and very slowly heat it up and as we do so measure the amount of unmelted DNA left using a fluorescent dye. The higher this number the more double-stranded DNA there is.

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I would send you some, but I don't have a queen cage, and my bees are all self-bred (all my queens are at least 2-3 generations away from a bred queen), so I don't know if thats what you want anyway.

Do the drones need to be alive when you get them?

Could I just stick them in a small tin and post them?

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Otto, the latest NZ Journal of Forestry (which I no longer have), has an illuminating paper which has some relevance to your work. The subject matter is the presentation of information for the masses.........

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I would send you some, but I don't have a queen cage, and my bees are all self-bred (all my queens are at least 2-3 generations away from a bred queen), so I don't know if thats what you want anyway.

Do the drones need to be alive when you get them?

Could I just stick them in a small tin and post them?

Any container is fine. I only suggest a queen cage because it is something many beekeepers have lying around. Thanks:)

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Clear as mud Otto thankyou :)

 

That just reinforces my decision to become a beekeeper not a scientist I thought RFU stood for Rugby Football Union :)

Take home message from the graphs is that if two curves have a different shape they are genetically different.

 

If RFU was a performance measure for my local rugby football union team I think they'd be flat-lining at zero.

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