Science Behind “Anthesis” and 2015 Grower - Team

Anthesis
Cucurbit Maxus
Science Behind
“Anthesis”
…
and
…
2015 Grower
Results
Matthew DeBacco
About “Mad Scientist,”
Matt D.
• Master’s in Agronomy from University of Connecticut
• Thesis on Organic Suppression of Powdery Mildew on Cucurbit & Assessing
Decomposition of Fiber Pots
• Worked for UConn Extension Service with their Nutrient
Management Program and was their in-field diagnostician
for vegetable growers
http://assets.sbnation.com/assets/2459443/UConnHusky-Logo.jpg
About Mad Scientist, Matt D.
• Growing pumpkins for last 16 years
• Has grown pumpkins continuously
for 2,367 days
• Two-time CT State Record Holder
– Personal Best: 1,766.5 pounds
Making the best of a ‘bad’ situation
• 2012 my patch was infected with Mosaic Virus
Cucumber Mosaic Virus
http://news.bbcimg.co.uk/media/images/44014000/jpg/_44014297_gloc.jpg
Zucchini Yellow
Mosaic Virus
Matt DeBacco’s patch, 2012
http://fatalii.net/chile/kuvat/kirvasika3532532.jpg
What the Virus in my patch taught me
• Pumpkin cells, after 20 days post pollination,
are no longer actively dividing.
• All younger pumpkins (under 20 days) developed viral mosaic
patterns, but the one older pumpkin in the patch never
showed visual symptoms even though the entire plant was
severely infected.
http://schoolworkhelper.net/wp-content/uploads/2010/07/Viral-replication.gif
http://www.biology.iupui.edu/biocourses/N100/images/8mitosiscropped.jpg
http://superscholar.org/wp-content/uploads/2011/04/dead-end-jobs.jpg
This led to two years of research into
the scientific literature.
• Mostly, it led to dead ends.
https://aidanmkeith.files.wordpress.com/2012/11/piles.jpg
http://sellingsimplified.com/main/wp-content/uploads/Dead-End-Good-Ways-To-Make-Money1.jpg
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Putting the pieces together
was a journey
http://www.jonathanrosenbaum.net/wp-content/uploads/2012/06/ijandthelostcrusade.jpg
Two Key Articles
Tomato Fruit Growth
…
Integrating Cell Division,
Cell Growth and
Endoreduplication by
Experimentation and
Modeling
• 145 pg Doctoral Thesis
by Julienne Fanwoua
• Early Application of
Ethrel Extends Tomato
Fruit Cell Division and
Increases Fruit Size and
Yield with Ripening
Delay
Atta-Aly, M.A., G.S. Riad, Z.S. El-Lacheene, and
A.S. El-Beltagy. 1999. Early application of ethrel
extends tomato fruit cell division and increases
fruit size and yield with ripening delay. J. Plant
Growth Regulat.18:15–24.
http://revivechurch.org/revivechurchorg/wp-content/uploads/2013/10/what-i-learned-11.png
• Cell division, cell growth, and cell
endoreduplication are three fundamental
cellular processes responsible for fruit growth.
http://www.biology.iupui.edu/biocourses/N100/images/8mitosiscropped.jpg
http://www.biw.kuleuven.be/biosyst/mebios/OT2012_MAP/WP1
http://plantsinaction.science.uq.edu.au/book/export/html/70
Cell cycle
http://www2.le.ac.uk/departments/genetics/vgec/diagrams/22-Cell-cycle.gif
Mitosis
http://www2.le.ac.uk/departments/genetics/vgec/diagrams/38%20mitotis%20phases.jpg
Mitosis
http://media1.shmoop.com/images/biology/biobook_dna_graphik_26.png
Cell Growth = Cell Expansion
• Same number of cells, but each cell is
increasing in size (swelling).
http://openi.nlm.nih.gov/imgs/512/151/2530862/2530862_gb-2008-9-7-226-1.png
Cell Expansion
• Contributes by far the most to fruit growth
http://postharvest.tfrec.wsu.edu/pages/N4I1B
Cell Expansion (alone)
• Can increase cell size up to 11,000 fold (of initial volume)
https://m1.behance.net/rendition/modules/45655451/disp/bbaa72c9b1867efdfd1e3504eb55f657.jpg
Fruit Size is determined by…
• Number of cells and
• Size of individual cells
http://images.calorieking.com.au/branding/dc/runtime/portionsense/140.jpg
http://img.webmd.com/dtmcms/live/webmd/consumer_assets/site_images/articles/health_tools/fat_fig
hting_foods_slideshow/webmd_rf_photo_of_grapes_and_raisins.jpg
Cell Division
Mitosis
• *Genetic variation in
final fruit size (large vs. small
fruited melons) was mainly
accounted for by
variation in final pericarp
cell number caused by
differences in cell
division duration
Figure 6: Pericarp growth following anthesis.
http://openi.nlm.nih.gov/detailedresult.php?img=2685393_1471-2229-9-49-6&req=4
Final Pericarp Mass
• The duration of cell division had the largest
influence on final cell number and final
pericarp mass.
http://www.fruitsinfo.com/images/fruit3.gif
http://upload.wikimedia.org/wikipedia/commons/thumb/1/11/Drupe_fruit_diagram-en.svg/2000pxDrupe_fruit_diagram-en.svg.png
http://static.flickr.com/50/159047357_4cba94d98f.jpg
Limited Time for Cell Division
• Duration of cell division after anthesis ranges
from 1-2 weeks in cucurbits and tomatoes
– Age at which cells cannot divide = 16 days
• Under normal conditions
http://3.bp.blogspot.com/-DxHXoZ70QWw/TfaPndvpj5I/AAAAAAAAAOQ/GcFuJjwxTtw/s1600/16DaysLogo-1.png
http://rbcweb.shopfactory.com/contents/media/limited_time.gif
Summary
• * Cell division is of great importance in the
variation of final fruit size.
http://1.bp.blogspot.com/-V-hSF9LKAVE/TzYhENKokKI/AAAAAAAAAGk/8YGbf4xjdHc/s1600/Elodea.jpg
http://www.mbg.com.my/MBG/images/stories/promotion/mango-size01.jpg
http://www.leadingmenonly.com/wp-content/uploads/2013/09/What-Matters-Most-Important.jpg
http://i.kinja-img.com/gawker-media/image/upload/s--HyceRyTr--/c_fit,fl_progressive,q_80,w_636/18uyj7qd47anujpg.jpg
So….. Now what?
• How can I apply this knowledge?
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_c7dd03893e28d07fb03906a4421b74d1_jpg.jpg
Plant Hormones Influence Cell Signaling
http://www.mhhe.com/biosci/genbio/enger/student/olc/art_quizzes/genbiomedia/0486.jpg
http://i.ytimg.com/vi/A2qoQgrkE_U/hqdefault.jpg
Ethylene
• Often thought of as just a ripening hormone…
http://labs.bio.unc.edu/Kieber/Figures/Ethylene.jpg
http://upload.wikimedia.org/wikipedia/commons/thumb/3/3b/Ethylene-CRC-MW-dimensions2D.png/160px-Ethylene-CRC-MW-dimensions-2D.png
http://5e.plantphys.net/images/ch20/wt2002b_s.jpg
Why not Gibberellic acid (GA)?
http://www.extension.org/sites/default/files/w/e/e1/GA_cluster.jpg
http://www.sciencephoto.com/image/442065/350wm/C0118720-Effect_of_Gibberellic_Acid_on_Cabbage-SPL.jpg
Why Ethylene and not Gibberellin?
-Direction of cell growth is
influenced by effects on
cellulose microfibril
orientation.
-Giberrellins lead to growth
mainly to the longitudinal
direction (elongation)
-Ethylene promotes a more
random orientation and
growth in all three
dimensions (swelling)
(Based on Raven et al. 1992)
http://plantsinaction.science.uq.edu.au/edition1/?q=figure_view/290
Timing is Very Important
• Applying the same
product at different
stages of plant
development results in
very different outcomes
http://www.macforensicslab.com/ProductsAndServices/images/icon_Timing.jpg
http://img.pandawhale.com/post-8936-Woman-at-beach-Perfect-timing-N0Ig.jpeg
http://ripening-fruit.com/yahoo_site_admin/assets/images/tomato_ripening_cherries_big.121132938_std.jpg
Ethylene = Ripening*
• *When applied later in
development
http://www.californiaavocado.com/blog/wp-content/uploads/2014/06/ripen-avocado-in-bag.png
http://cen.acs.org/content/cen/articles/92/web/2014/11/Engineered-Bacteria-Ripen-FruitBelching/_jcr_content/articlebody/subpar/articlemedia_0.img.jpg/1416932919785.jpg
http://www.scielo.br/img/revistas/bjpp/v17n3/a02fig01.jpg
Ethylene = Causes plants to only produce
female flowers* (when applied at the 1-5 leaf stage)
• *For cucumbers, apply 250 to 500 ppm
when plants have 1,3 and 5 leaves
• Induces gynoecious female line for
hybrid seed production
http://www.harvesttotable.com/wp-content/uploads/2009/05/Cucumber-flower1.jpg
https://media.licdn.com/media/p/6/005/00f/151/07d5b5d.png
*Anthesis*
• Anthesis = the period of blooming in flowers
http://www.bigpumpkins.com/DiaryImages/2014/Lrg/219464.jpg
http://www.scielo.br/img/revistas/aabc/2011nahead/aop4411fig02.jpg
Anthesis = [an-thee-sis]
• The period or act of expansion in flowers,
especially the maturing of the stamens
• The action or period of opening of a flower
• Origin of Anthesis
• New Latin, from Greek anthēsis bloom, from anthein to flower, from anthos
– First Known Use: circa 1823
http://www.merriam-webster.com/dictionary/anthesis
http://dictionary.reference.com/browse/anthesis
Early Ethrel Application Extends Tomato Fruit Cell Division
and Increases Fruit Size and Yield with Ripening Delay
• This document provides specific materials and methods
• Ethrel (2, chloroethyl phosphonic acid) is an ethylene-releasing
substance
http://images.sciencedaily.com/2008/06/080628065632-large.jpg
http://www.press.uchicago.edu/dms/ucp/books/jacket/978/02/26/31/9780226316567.jpeg
http://www.candocareersolutions.ca/marketing-plan.jpg
Ethrel® Application
Extends Tomato Fruit
Cell Division
• Flowers were sprayed in the
early morning two days after
anthesis with 100ppm Ethrel®
– Tomato fruits produce their
highest ethylene level at the stage
of fruit set (48 hrs. after anthesis)
http://tvbythenumbers.zap2it.com/wp-content/uploads/2010/06/48-Hours_mystery.png
http://www.bigpumpkins.com/Diary/DiaryViewOne.asp?eid=221669
Ethrel® Application Extends Tomato
Fruit Cell Division
• 100ppm applied two days after anthesis
• Cell Division was extended up to 16-18 days
– 10 -12 days = just water (control)
http://wid.wisc.edu/content/uploads/2012/09/cells-web-550x367.jpg
Ethrel® Application Increases Fruit
Size and Yield with Ripening Delay
• Weight was significantly
increased by 8% to 34%
– and ripened 10 days later
than the (water) control.
http://i.bnet.com/blogs/srqpix.jpg
http://bonnieplants.com/wp-content/uploads/bonnie-original-tomato.jpg
https://s-media-cache-ak0.pinimg.com/236x/6d/7d/c8/6d7dc84c3366eeb29fd59e283d7ab0e2.jpg
Trial #2
• When same treatment was applied
to the entire plant one week after
anthesis of the 1st flower cluster,
yield was increased by 15%
http://grandpacliff.com/Plants/Img-Plants/plant-whole-pt.jpg
Terms Defined
• ACC = 1-Aminocyclopropane-1-carboxylic acid
• 1-Aminocyclopropane-1-carboxylic acid (ACC) is a disubstituted cyclic alphaamino acid in which a three-membered cyclopropane ring is fused to the
C(alpha)-atom of the amino acid.
• ACC plays an important role in the biosynthesis of the plant hormone
ethylene. It is synthesized by the enzyme ACC synthase from methionine and
converted to ethylene by ACC oxidase
• AOA = Aminooxyacetic acid
• Aminooxyacetic acid (AOA) can also inhibit 1-aminocyclopropane-1carboxylate synthase preventing ethylene synthesis, which can increase the
vase life of cut flowers.
http://defined.com/themes/site_themes/defined/default_site/img/defined-logo.png
What about Cytokinins?
• Increase fruit size (in apples) by increasing
number of cell layers
– not cell expansion
• Play an important role in determining
the eventual cell number of the fruit
http://education-portal.com/cimages/multimages/16/Key_Concept.png
http://www.efoodsdirect.com/media/catalog/product/cache/1/image/9df78eab33525d08d6e5fb8d27136e95/a/p/apples2.jpg
http://bcs.whfreeman.com/thelifewire8e/content/cat_010/f37018.jpg
Ethylene and Cytokinins
• Ethylene production was stimulated
by submicromolar concentrations
of cytokinins in seedlings
http://www.barnaor.com/images/NewsImages/strips.jpg
https://media.licdn.com/mpr/mpr/p/7/005/092/256/3a950e9.jpg
Site of application is important
• Localized applications of cytokinins to tomatoes
is known to increase…
• Sink strength
• Fruit size and
• Fruit dry matter at harvest
– How? By enabling them to attract photoassimilates
and nutrients from foliage
Source : Sink
Relationship
http://www.frontiersin.org/files/Articles/73624/fpls-05-00011-HTML/image_m/fpls-05-00011-g001.jpg
http://cnx.org/resources/cd8839949717d6b04f4fade2e6b0f7cf/Figure_30_05_07.jpg
Cytokinins are partially miscible in water
-Slightly soluble in hot water
-Soluble in dilute acid
• Ethrel is an acid that could aid in the incorporation
of cytokinis to water for even application to fruit
http://www.chem1.com/acad/webtext/solut/solut-images/miscibility.png
http://www.uiowa.edu/~c004131a/f8_18.gif
http://upload.wikimedia.org/wikipedia/commons/2/21/Mosaicr_seagull.jpg
So what does this all mean?
http://image.slidesharecdn.com/tg-intea14complexity-dung-beetles-talerevd-140227093950-phpapp01/95/the-dungbeetles-tale-systemsthinking-complexity-and-the-realworld-18-638.jpg?cb=1393964982
Research Conclusions
• Ethylene, when applied 48 hrs. post pollination, can
extend cellular division about 6 more days
• End of season weight can increase by 15% (range of 8-34%)
• The pumpkin should ripen 10 days later
• Cytokinins can increase fruit sink strength, size and
dry matter
http://image.slidesharecdn.com/researchconclusions5-111020040315-phpapp02/95/research-conclusions-1-728.jpg?cb=1319083428
The Directions are Simple
(Supporting science is complex)
• Apply Ethylene + Cytokinins 48 hours after pollination
*directly to the fruit*
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http://biznology.com/wp-content/uploads/2014/05/artworks-000058593031-9xetyb-original.jpg
Anthesis Specific Observations
• Pumpkins should be smaller than untreated
controls up to about day 35
– This indicates a potential delay in cell expansion due to the
increased time of cell division
• Anthesis-treated pumpkins should have
improved late season growth rates
– The theory is there are more cells present in the pumpkin
and also a ripening delay
https://7373-presscdn-0-43-pagely.netdna-ssl.com/wp-content/uploads/2013/06/binoculars1.jpg
2015 Grower Reported Data
“I will say the Anthesis-applied pumpkin was still growing,
where the other fruits slowed way down.”
-Andy Box
“The Anthesis-treated pumpkins that I had going grew better
than anything I ever had”
-Allen Bradly
http://www.gannett-cdn.com/-mm-/382e7a583110dfb611e22bc73f3f8339b9df2203/c=0-0-16291224&r=x404&c=534x401/local/-/media/GreatFalls/2014/08/11/leadphotogroupchat.jpg
http://earlylearningcentral.ca/wp-content/uploads/2012/10/pumpkin-measuring.jpg
1600
Anthesis and Untreated (Control) Pumpkins in
Andy Box's 2015 Patch
Estimated Weight (pounds) Using the
Team-Pumpkin Chart
1400
R² = 0.9965
R² = 0.9902
1200
R² = 0.9983
1000
800
Anthesis Treated
600
Untreated 1
400
Untreated 2
200
20
30
40
50
Days Old
60
70
80
Graph 1 Summary
1. Figure 1: It is clearly evident that the Anthesis-treated pumpkin was
of comparable size up to the critical day 35.
2. After day 35, the Anthesis-treated pumpkin had an increased growth
rate that was sustained for the remainder of the season, resulting in
his top producing pumpkin for the year.
3. Both of the untreated control pumpkins have a very similar growth
curve for the season and ended up within 4.9% of each other
4. The Anthesis-treated is noticeably greater past day 40 and finished
13.0% greater than the average of the untreated pumpkins.
5. The graph supports the grower’s comments that the Anthesistreated pumpkin continued to grow while the other fruits decreased
in growth rate.
6. Trend lines use the Logarithmic Regression with R2 values provided.
700
650
Estimated Weight (pounds) using the
Team-Pumpkin Chart
600
Growth Curves for Anthesis and Non-Treated Pumpkins
Grown in 150sq.ft.
550
500
450
400
350
300
Anthesis Treated '15
250
Untreated '14
200
Untreated '13
150
100
50
0
10
20
30
40
50
Days Old
60
70
80
Graph 2 Summary
1.
2.
3.
4.
5.
6.
Growth curves for Anthesis-treated and untreated (control) pumpkins
grown in the same 150 sq.ft. location by Bart Toftness during three
different years with almost daily measurements taken.
The growth curve shows that the Anthesis-treated pumpkin (diamond
shape) was smaller up to day 37 than the untreated ones, suggesting a
delay in cell expansion.
After day 37, the growth rate is accelerated and continues for an extended
period of time, leading to an improvement in late season growth that
increases final size of the pumpkin.
Day 60 and beyond, there is a continual upward trend of growth with no
indication of a plateau.
Even though the Anthesis-treated pumpkin is the oldest, the delay in
harvest was grower’s choice, as it was still showing positive gains and was
allowed to remain on the vine until a later weigh-off.
The Anthesis-applied pumpkin grew the biggest pumpkin out of this space
even though, up to day 30, it was the smallest.
Anthesis 2015 Harvest Data Summary
Top 10 Average for Anthesis-Treated Pumpkins 1819.85 lbs.
Top 20 Average for Anthesis-Treated Pumpkins 1643.25 lbs.
…
…
Total Number of Personal Best
for growers using Anthesis
Total Number of Top 2015 pumpkins
using Anthesis
(not counting personal bests)
8
…
…
Pumpkins over 2,000 pounds
Pumpkins 1,900-2,000 pounds
2
2
…
…
Tomatoes grown over 5 pounds
4
6
Improvements for 2016
• Ethrel liquid will be replaced with a powder
– Due to the small quantity of Ethrel needed, some growers
reported evaporated samples.
• Small tubes will be of improved quality
– Small blue tubes last year had inconsistent quality
• Only pre-orders will be taken (Jan. 1st-March 15th)
– Makes it easier to know how much I need to prepare
• Orders will be mailed out in mid-April
– This will ensure growers have fresh product that will fit
the 4 month self-life.
http://www.ddubcustomsportswear.com/wp-content/uploads/2014/08/improve.jpg
Anthesis Specifics
• The directions are not hard, but must be
followed for effectiveness.
http://images3.cliqueclack.com/p/files/2014/07/INSTRUCTIONS-e1404879204784.jpg
http://www.polyvore.com/cgi/img-thing?.out=jpg&size=l&tid=57680386
Cucurbit (Part A)
• This is a fine grain powder
– (very small quantity)
• Will easily dissolve in distilled water
• *Store in the refrigerator
• *Keep away from water until ready
to use
• Does come in a sealed container
• Shelf-life of about 2 years @
refrigeration temps.
http://blogs.ext.vt.edu/eatsmart-movemore/files/2013/11/95102395.jpg
Cost of Cucurbit
• $3.00 per application
•*Do not allow mixed solution to stand overnight.
http://thumbs.dreamstime.com/x/three-dollars-13693976.jpg
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http://lghttp.41526.nexcesscdn.net/8015C4D/magento/media/catalog/product/cache/1/image/1400x/9d
f78eab33525d08d6e5fb8d27136e95/G/S/GS-111700-2.jpg
http://www.cleanfax.com/wp-content/uploads/2015/07/Tech-tip-frozen-gauge-website.jpg
Maxus (Part B)
• This is a clear liquid suspension
• *Store in freezer*
• Comes in a brown light block container to
reduce breakdown
• Frozen has a 5 month shelf life
– (In refrigerator it has only a 3 week shelf-life)
• Requires cold shipping from supplier
http://i.ebayimg.com/images/g/~H4AAOxy6MBSN1uG/s-l300.jpg
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http://facultyfiles.deanza.edu/images/gawrychjeff/Frozencar.jpg
Cost of Maxus
• $5.00 per application
• Must be placed immediately in
the freezer upon arrival
• Take out of the freezer and
place in the refrigerator the
day of pollination to provide
two days to defrost
https://upload.wikimedia.org/wikipedia/commons/thumb/6/69/US-Series-2006G-$5obverse.jpg/1024px-US-Series-2006G-$5-obverse.jpg
http://img0135.psstatic.com/186784127_eppendorf-022363514-polypropylene-flex-tubes-.jpg
Buyer Beware
of other hormone suppliers
• There are slightly different types of hormones
that have different modes of action.
(Example: there are at least three different types of cytokinins)
• Hormones have a shelf life, and other suppliers
do not provide shelf life information or state
proper storage conditions, which will reduce the
effectiveness.
http://jacobsinjurylaw.com/wp-content/uploads/It%E2%80%99s-Buyer-Beware.png
Spray bottle + Mixing container
$5.00
http://files.hgsitebuilder.com/hostgator884163/image/1550mlnesttubes.jpg
http://www.distilleddeionizedwater.com/wp-content/uploads/2012/09/Distilled-Water1.jpg
The only ingredient not included…
Distilled Water
• Only need ~29 mL per application
http://canigivemybaby.com/wp-content/uploads/2011/09/can-i-give-my-baby-distilled-water.jpg
https://www.aquagrade.com/wp-content/uploads/2013/12/Arrowhead-Distilled-Water.jpg
https://s3.amazonaws.com/mws-assets-production/app/public/spree/products/5142/large/distilledwater-case-of-6-one-gallon-bottles.jpg?1332783919
http://cultivateprayer.com/wp-content/uploads/2011/12/Basic-Instructions.jpg
Basic Instructions
• Keep brown vial in freezer
• Keep blue-capped vial in refrigerator
• Mix both with distilled water and
immediately apply directly to the pumpkin
48 hrs. after pollination
• Mist entire pumpkin, do not apply to petals
or inner flower parts (stigmas/lobes)
• Discard what you do not use the same day
Anthesis
• Contains plant hormones known to
increase cell division
•
Duration of cell division has the greatest influence on final mass.
• Must be applied 48 hours post pollination
• Requires only one application
• Get yours today!