Growing a Bigger Pumpkin by

Increasing Cell Division

Matthew DeBacco
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 About “Mad Scientist”
Matt D.
• Masters 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 as 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 15 years

• Growing pumpkins continuously for

the last 2,100+ days

• Two time and current CT State

Record Holder @ 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 and the one older pumpkin in the patch never
showed visual symptoms even though the entire plant was
severely infected

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This lead to two years of research into

the scientific literature
• Mostly it lead to dead ends

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 Then there was a break through

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 Two Key Articles

Tomato Fruit Growth • Early Application of

… Ethrel Extends Tomato
Integrating Cell Division, Fruit Cell Division and
Cell Growth and Increases Fruit Size and
Endoreduplication by Yield with Ripening
Experimentation and Delay
Modelling
• 145 pg Doctor Thesis by
• Julienne Fanwoua 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.
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• 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

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 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)
 Endoreduplication
• The replication of DNA during the S phase of the
cell cycle without the subsequent completion of
mitosis and/or cytokinesis.

http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/E/Endoreplication.html
Endoreduplication compared to Mitosis
 Endoreduplication- Does not need
reorganization and patterning of the tissue.

http://www.rv-orchidworks.com/orchidtalk/content/172-scientific-behind-how-orchid-flowers-keep-growing-even-after-buds-open.html
Endoreduplication
 Grand Gala apples, at right, undergo
endoreduplication, which causes cells to grow
larger than in the regular Gala apples, at left.
(Purdue University photo/Peter Hirst)

*Grand Gala
Regular
apples undergo
Gala
endoreduplication
apples

http://www.purdue.edu/newsroom/research/2010/100630HirstGala.html
 Endoreduplication Benefits

1. Allows an organ to enlarge in size even when it has lost

the capacity for multiplication.

2. Does not need reorganization and patterning of the tissue.

3. Is much more energy efficient.

– It involves much less protein synthesis and energy expenditure
on the part of the cells.

4. The resulting cells are polyploidy

– (Study on chickpeas does not show a decrease in seed yield)
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Fruit Size is determined by…

• Number of cells and
• Size of individual cells

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 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://upload.wikimedia.org/wikipedia/commons/thumb/1/11/Drupe_fruit_diagram-en.svg/2000px-
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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

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 Summary
• *There is great importance of cell division in
the variation of final fruit size.

http://1.bp.blogspot.com/-V-hSF9LKAVE/TzYhENKokKI/AAAAAAAAAGk/8YGbf4xjdHc/s1600/Elodea.jpg

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So….. Now what

• How can I apply this knowledge?

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Plant Hormones Influence Cell Signaling

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Why not Gibberellic acid (GA)?

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Ethylene
• Often thought of as just a ripening hormone…

http://upload.wikimedia.org/wikipedia/commons/thumb/3/3b/Ethylene-CRC-MW-dimensions-
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 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 result in
very different outcomes

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Ethylene = Ripening*

• *When applied later in

development

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 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
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*Anthesis*

• Anthesis = the period of blooming in flowers

http://www.bigpumpkins.com/DiaryImages/2014/Lrg/219464.jpg
 Early Ethrel Application Extends Tomato Fruit Cell Division
and Increase Fruit Size and Yield with Ripening Delay

• This document provides specific materials and methods

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 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 (2 days after anthesis)

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 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 Increase Fruit Size
and Yield with Ripening Delay
• Weight was significantly
increased by 8% to 34%
– and ripened 10 days later
than the (water) control.

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Trial #2
• When same treatment was applied
to the entire plant one week after
flowers anthesis of the 1st 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 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.

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So what does this all mean?

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 The Directions are Simple
(supporting science complex)
• Apply 100ppm of Ethrel® 2 days after pollination
*directly to the fruit*

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 Warning very small tolerance of error
• To much = negative effects (deformities, no seeds…)
• To little = not effective

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 Ethrel® (Ethephon)
• Ethrel (2, chloroethyl phosphonic acid) is an ethylene releasing substance

• Aqueous solution of ethrel is stable below pH 3.5

• Above pH 3.5 the hydrolysis of ethrel begins with the

release of free ethylene along with chloride and phosphate ions

• Ethrel, can enter into the fruits cells

http://www.cropscience.bayer.ca/~/media/Bayer%20CropScience/Country-Canada-Internet/Products/Ethrel/Ethrel.ashx
 What should happen after
Ethrel® application
• Cell Division should be extended up to 16-18 days
• End of season weight can increase by 15% (range of 8-34%)
• The pumpkin should ripen 10 days later
Pumpkin Weight (lbs) 15% increase
100 115
500 575
1000 1150
1500 1725
1650 1897.5
1900 2185
2009 2310
2323.7 2672
 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

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Ethylene and Cytokinins

• Ethylene production was stimulated
by submicromolar concentrations
of cytokinins in seedlings

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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

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Tank mix of Ethrel® + Cytokinin

• One study looking at apples
and thinning related to
applying both of these
• Note applied at different times
in this study

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 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

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 Now the cost…

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 Estimated cost of Ethrel®
• About $1.00 per application
• Easy procedure
– (It is like someone circled Waldo)

•*Mix only the amount of spray you expect to use each day.

•*Do not allow mixed solution to stand overnight.

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Cytokinins are more difficult

• Often need to be stored at very low temperatures
and require special “Cold shipping”
• About $5.00 per application
– *So, when you see them online the odds are you are
not getting a properly stored product

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 Conclusion
• Ethrel has the potential to be very effective

• Cytokinins may be well worth their special requirements

– *There are different cytokinins so it is important to select the “best” match

• Cost to benefit ratio is the greatest that I have seen

• If you are growing giant tomatoes this is worth a look

 • Contains plant hormones known to
increase cell division
• Duration of cell division has the greatest influence on final mass.

• Must be applied 2 days post pollination

• Requires only one application

• Get yours today!

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