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Science Behind “Anthesis”
…
and …
2015 Grower Results
Matthew DeBacco
Anthesis
Cucurbit Maxus
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
http://news.bbcimg.co.uk/media/images/44014000/jpg/_44014297_gloc.jpg
Cucumber Mosaic Virus
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
This led to two years of research into the scientific literature.
• Mostly, it led to dead ends.
http://sellingsimplified.com/main/wp-content/uploads/Dead-End-Good-Ways-To-Make-Money1.jpg
http://superscholar.org/wp-content/uploads/2011/04/dead-end-jobs.jpg
https://aidanmkeith.files.wordpress.com/2012/11/piles.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.
• 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://revivechurch.org/revivechurchorg/wp-content/uploads/2013/10/what-i-learned-11.png
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)
Fruit Size is determined by…
• Number of cells and
• Size of individual cells
http://img.webmd.com/dtmcms/live/webmd/consumer_assets/site_images/articles/health_tools/fat_fighting_foods_slideshow/webmd_rf_photo_of_grapes_and_raisins.jpg http://images.calorieking.com.au/branding/dc/runtime/portionsense/140.jpg
https://m1.behance.net/rendition/modules/45655451/disp/bbaa72c9b1867efdfd1e3504eb55f657.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
http://openi.nlm.nih.gov/detailedresult.php?img=2685393_1471-2229-9-49-6&req=4 Figure 6: Pericarp growth following anthesis.
Final Pericarp Mass
• The duration of cell division had the largest influence on final cell number and final pericarp mass.
http://upload.wikimedia.org/wikipedia/commons/thumb/1/11/Drupe_fruit_diagram-en.svg/2000px-Drupe_fruit_diagram-en.svg.png http://www.fruitsinfo.com/images/fruit3.gif
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://rbcweb.shopfactory.com/contents/media/limited_time.gif http://3.bp.blogspot.com/-DxHXoZ70QWw/TfaPndvpj5I/AAAAAAAAAOQ/GcFuJjwxTtw/s1600/16DaysLogo-1.png
http://static.flickr.com/50/159047357_4cba94d98f.jpg
Summary
• * Cell division is of great importance in the variation of final fruit size.
http://www.leadingmenonly.com/wp-content/uploads/2013/09/What-Matters-Most-Important.jpg
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
So….. Now what?
• How can I apply this knowledge?
http://cdn2.hubspot.net/hub/282640/file-1146954527-jpg/thumbnail.jpg https://cdn.empowernetwork.com/user_images/post/2013/02/01/c/7d/d038/540_293_resize_20130201_c7dd03893e28d07fb03906a4421b74d1_jpg.jpg
http://i.kinja-img.com/gawker-media/image/upload/s--HyceRyTr--/c_fit,fl_progressive,q_80,w_636/18uyj7qd47anujpg.jpg
Plant Hormones Influence Cell Signaling
http://www.mhhe.com/biosci/genbio/enger/student/olc/art_quizzes/genbiomedia/0486.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-dimensions-2D.png/160px-Ethylene-CRC-MW-dimensions-2D.png
http://i.ytimg.com/vi/A2qoQgrkE_U/hqdefault.jpg
Why not Gibberellic acid (GA)?
http://www.extension.org/sites/default/files/w/e/e1/GA_cluster.jpg
http://5e.plantphys.net/images/ch20/wt2002b_s.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://img.pandawhale.com/post-8936-Woman-at-beach-Perfect-timing-N0Ig.jpeg http://www.macforensicslab.com/ProductsAndServices/images/icon_Timing.jpg
Ethylene = Ripening*
• *When applied later in development
http://cen.acs.org/content/cen/articles/92/web/2014/11/Engineered-Bacteria-Ripen-Fruit-Belching/_jcr_content/articlebody/subpar/articlemedia_0.img.jpg/1416932919785.jpg
http://ripening-fruit.com/yahoo_site_admin/assets/images/tomato_ripening_cherries_big.121132938_std.jpg
http://www.scielo.br/img/revistas/bjpp/v17n3/a02fig01.jpg
http://www.californiaavocado.com/blog/wp-content/uploads/2014/06/ripen-avocado-in-bag.png
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
*Anthesis*
• Anthesis = the period of blooming in flowers
http://www.bigpumpkins.com/DiaryImages/2014/Lrg/219464.jpg
https://media.licdn.com/media/p/6/005/00f/151/07d5b5d.png
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://dictionary.reference.com/browse/anthesis http://www.merriam-webster.com/dictionary/anthesis
http://www.scielo.br/img/revistas/aabc/2011nahead/aop4411fig02.jpg
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://www.press.uchicago.edu/dms/ucp/books/jacket/978/02/26/31/9780226316567.jpeg
http://images.sciencedaily.com/2008/06/080628065632-large.jpg
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://www.bigpumpkins.com/Diary/DiaryViewOne.asp?eid=221669 http://tvbythenumbers.zap2it.com/wp-content/uploads/2010/06/48-Hours_mystery.png
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://bonnieplants.com/wp-content/uploads/bonnie-original-tomato.jpg http://i.bnet.com/blogs/srqpix.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
https://s-media-cache-ak0.pinimg.com/236x/6d/7d/c8/6d7dc84c3366eeb29fd59e283d7ab0e2.jpg
Terms Defined
• ACC = 1-Aminocyclopropane-1-carboxylic acid • 1-Aminocyclopropane-1-carboxylic acid (ACC) is a disubstituted cyclic alpha-
amino 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-1-
carboxylate 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
Ethylene and Cytokinins
• Ethylene production was stimulated by submicromolar concentrations of cytokinins in seedlings
http://www.barnaor.com/images/NewsImages/strips.jpg
http://bcs.whfreeman.com/thelifewire8e/content/cat_010/f37018.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
https://media.licdn.com/mpr/mpr/p/7/005/092/256/3a950e9.jpg
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
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
http://upload.wikimedia.org/wikipedia/commons/2/21/Mosaicr_seagull.jpg
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*
http://www.keepcalmandposters.com/posters/2413221.png 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=0-0-1629-1224&r=x404&c=534x401/local/-/media/GreatFalls/2014/08/11/leadphotogroupchat.jpg http://earlylearningcentral.ca/wp-content/uploads/2012/10/pumpkin-measuring.jpg
R² = 0.9965
R² = 0.9902
R² = 0.9983
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Anthesis and Untreated (Control) Pumpkins in Andy Box's 2015 Patch
Anthesis Treated
Untreated 1
Untreated 2
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 Anthesis-treated pumpkin continued to grow while the other fruits decreased in growth rate.
6. Trend lines use the Logarithmic Regression with R2 values provided.
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Growth Curves for Anthesis and Non-Treated Pumpkins Grown in 150sq.ft.
Anthesis Treated '15
Untreated '14
Untreated '13
Graph 2 Summary
1. 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.
2. 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.
3. 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.
4. Day 60 and beyond, there is a continual upward trend of growth with no indication of a plateau.
5. 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.
6. 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 8
Total Number of Top 2015 pumpkins
using Anthesis
(not counting personal bests)
6
… …
Pumpkins over 2,000 pounds 2
Pumpkins 1,900-2,000 pounds 2
… …
Tomatoes grown over 5 pounds 4
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
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://www.polyvore.com/cgi/img-thing?.out=jpg&size=l&tid=57680386
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Cost of Cucurbit
• $3.00 per application
•*Do not allow mixed solution to stand overnight.
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http://thumbs.dreamstime.com/x/three-dollars-13693976.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://www.cleanfax.com/wp-content/uploads/2015/07/Tech-tip-frozen-gauge-website.jpg
http://facultyfiles.deanza.edu/images/gawrychjeff/Frozencar.jpg
http://i.ebayimg.com/images/g/~H4AAOxy6MBSN1uG/s-l300.jpg
https://s-media-cache-ak0.pinimg.com/736x/c7/1c/60/c71c60967e20d353144af1748355f549.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
http://img0135.psstatic.com/186784127_eppendorf-022363514-polypropylene-flex-tubes-.jpg https://upload.wikimedia.org/wikipedia/commons/thumb/6/69/US-Series-2006G-$5-obverse.jpg/1024px-US-Series-2006G-$5-obverse.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
The only ingredient not included… Distilled Water
• Only need ~29 mL per application
https://s3.amazonaws.com/mws-assets-production/app/public/spree/products/5142/large/distilled-water-case-of-6-one-gallon-bottles.jpg?1332783919 https://www.aquagrade.com/wp-content/uploads/2013/12/Arrowhead-Distilled-Water.jpg
http://canigivemybaby.com/wp-content/uploads/2011/09/can-i-give-my-baby-distilled-water.jpg
http://www.distilleddeionizedwater.com/wp-content/uploads/2012/09/Distilled-Water1.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
http://cultivateprayer.com/wp-content/uploads/2011/12/Basic-Instructions.jpg
• 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!
Anthesis
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