Cranberry Domestication and Challenges of the Changing

'Climate' Nick Vorsa, Rutgers University Dept. Plant Biology & Pathology

‘Climate’

Physiological Cranberry adaptation – chilling hrs?; heat stress

Disease & Insect Threats Evolving insects pressure Evolving disease pressure

Business/sociological Juice vs. sweetend-dried Sugar Farming efficiency/sustainability

Acid soils pH 3 – 5.5

Highbush

blueberry

V. corymbosum

Tetraploid

2n = 4x = 48

American cranberry

V. macrocarpon

Diploid

2n = 2x = 24

Asexual - stolons

Sexual - seeds

Dispersal

buoyancy

Blueberry

animal

+/-

Cranberry

Water

+

Brix (soluble solids)

Glucose+Fructose

12 – 15%

7 – 10%

7 – 10%`

2 – 7%

main volatiles Linalool

Blueberry flavor

α – terpineol

antiseptic

Acidity

Benzoic Acid

Storage life

Low TA

(0.5 CAE)

0 %

2 - 3 wks

High TA

(2.5 CAE)

~0.1%

3 - 6 months

Proanthocyanidins +/- ++++

Fruit chemistry - Cranberry seed dispersal based on animal or water?

2n = 2x = 24

diakinesis

anaphase I

Cranberry Meiosis in cv. Wilcox

genome 570 Mbp

protandry

S6 Pilgrim line

S8 Ben Lear line

cv. Early Black – 1852 Harwich, MA

Industry still cultivates native cultivars

Early Black 1852 MA Howes 1843 MA Searles 1893 WI McFarlin 1874 MA Lemunyon 1870’s NJ Ben Lear 1907 WI

Flavonoid biosynthetic

pathway

• Anthocyanins

• Flavonols

• Flavan-3-ols

• Proanthocyanidins

Shikimic acid pathway

mg

/g (

FW

)

0

2

4

6

8

10

12

14

16

flavo

nols

proan

thocy

anid

ins

anth

ocyan

ins

Trite

rpen

oids

resv

erat

rol

Constituents of cranberry fruit

germplasm

variation

RBC, P type E.coli

- Cranberry extract

RBC, P type E.coli

+ Cranberry extract

Hahn et al. 2002 J. Mol. Biol. 323:845

Urinary tract disease

E. coli

E. Coli tyes type-1 cystitis P-type pyelonephritis

A-type proanthocyanidins

1st breeding and selection cycle – initiated in 1929

USDA with New Jersey and Massachusetts Agricultural Experiment Stations

> 10,000 seedlings from 46 crosses

6 Cultivars named

Stevens, Pilgrim, Bergman, Beckwith, Wilcox, Franklin

40 selections

Year

1880 1900 1920 1940 1960 1980 2000

Acre

ag

e

0

5000

10000

15000

20000

25000

New Jersey

Wisconsin

False Blossom disease

Fruit rot

2000-present cranberry breeding objectives

Desirable horticultural traits Fruit Rot Resistance (warmer climate) Yield Season of ripening Juice vs. sweeten-dried cranberry (SDC) Processing requirements Tacy (fruit anthocyanidins) Round fruit shape Large fruit size Fruit firmness Early/late season ripening

Cranberry Germplasm

V. macrocarpon

Germplasm – genetic resources

Gene pool

genetic resources,

or more precisely the DNA of an organism

and collections of that material

Domesticated germplasm – NJ, MA and WI Native germplasm - DE, NJ, NY, MA, ME, WV, PA, MI, WI, MN > 600 accessions

Canada - NS, NB, PEI, NF

Berry Wt Total Phenolics

Yield TotalP PACs

TA Tacy

Citric Acid Cyan3Gal

Malic Acid Cyan3Glu

Quinic Acid Cyan3Arab

Brix Peon3Gal

Glucose Peon3Glu

Fructose PeonArab

Sucrose

Fruit of germplasm evaluated in 1998 for:

0.0 5.0 10.0 15.0 20.0 25.0 30.0 -500

1,000

2,000

3,000

Stevens mAU

min

C-3

-gal

C-3

-glc

C

-3-a

rab

P-3

-gal

P-3

-glc

P

-3-a

rab

RT (min)

Absorb

ance

HPLC @ OSC, Inc.

Breeding cranberry for fruit rot

resistance – The impossible

dream? Nicholi Vorsa

Jennifer Johnson-Cicalese

Resistance to

Fungi causing cranberry fruit rot disease complex

• Coleophoma empetri

• Colletotrichum accutatum

• Colletotrichum cf. gloeosporioides

• Fusicoccum putrefaciens

• Monilinia oxycocci

• Phomopsis vaccinii

• Phyllosticta vaccinii

• Physalospora vaccinii

Oudemans et al. Plant Disease 82:1176-84, 1998

Germplasm Evaluation

0

100

200

300

400

500

1 2 3 4 5

Rot ratings (1=no rot; 5=severe rot)

# o

f p

lots

2003

2004

When fungicides were withheld from our germplasm collection, most accessions were highly susceptible to fruit rot (rating of 5), but a few showed resistance both years.

34 simple sequence repeats (SSR)

NJ

MA

NS

?

Resistant Susceptible

Bog 4 germplasm

Budd’s Blues

0

10

20

30

40

50

60

BB xBB

BB x89-3

BB x86-45

BB x86-46

BB xCum.

Cum. xBB

BB xHoll.

Cum. x89-3

Cum. xHoll.

Pro

gen

y w

ith

eac

h r

atin

g (%

)

Resistant x resistant

1 2 3 4 5Rot ratings, 1=no rot, 5=all rot

0

10

20

30

40

50

60

BB xCQ

BB xDE

BB xMQ

BB x98-76

98-62 xBB

89-3 xMQ

MQ x89-3

DE xCum.

Cum. xDE

Pro

gen

y w

ith

eac

h r

atin

g (%

)

Resistant x susceptible

0

10

20

30

40

50

60

70

80

MQ x 98-62

98-62 xMQ

ST x 98-62

98-62 x ST

Pro

gen

y w

ith

eac

h r

atin

g (%

)

Susceptible x susceptible

Res. Clones BB – Budds Blues Holl. – Holliston 89-3

Populations segregating for fruit rot

20 - ½ acre beds 2 – 5 acre beds

ST

ST

ST ST

35

CQ

MQ

B B

P B

B

E

P

EB

B

E

B B

DE

EB - W

2009 12 ac of progeny plots, Marucci Center

3-5 years to establish maturity

1

1.5

2

2.5

3

3.5

4

4.5

5

1 1.5 2 2.5 3 3.5 4 4.5 5

Mea

n r

ot

rati

ng

of

pro

gen

y

Mean rot rating of parents

A

R2 = 0.79 h2 = 0.81 SE = 0.21

46 crosses 20

30

40

50

60

70

80

90

100

20 30 40 50 60 70 80 90 100

Mea

n r

ott

ed f

ruit

of

pro

gen

y (%

)

Mean rotted fruit of parents (%)

B

R² = 0.64 h2 = 0.90 SE = 6.8

25 crosses

Heritability based on mid-parent - offspring

2012 Results - 1 ft2 fruit samples from resistant progeny and susceptible controls, Oct. 2.

Mullica Queen

Sound fruit Partial Rotten

CNJ06-30-31*

CNJ06-27-11 Stevens

Sound Partial Rotten fruit

Added sugar Sugar/acid ratio

acids or sugars

Cranberry TA – 2.5 CE 8-10 mg/g Citric Acid 6-10 mg/g Malic Acid

Citric acid mg/g

0 2 4 6 8 10 12

TA

(C

E)

1.6

1.8

2.0

2.2

2.4

2.6

2.8

1% CA

Acidity in a breeding population

Current cultivars

Scleroracus vaccinii

Blunt-nosed leafhopper

New subgroup 16SrIII-Yphytoplasmas associated with false-blossom diseased cranberry (Vaccinium macrocarpon) plants and with known and potential insect vectors in New Jersey Lee et al. 2014 Eur. J. Plant Pathol.

Figure 2. Phytoplasma-like particles (red arrows) in a leaf midrib phloem cell of the diseased Rhododendron plant (bar = 500 nm).

et al. 2014

Toad bug – first found in NJ in 2013

Pre-bloom Post-bloom Bloom

Eggs (September-October)

Nymphs (end of June-August)

Adults (August-October)

Harvest

Cranberry Toad-bug

• Phylloscelis atra (Dictyopharidae).

Described 1914

• Feeds only on cranberries.

• Single generation a year.

• Overwinters as eggs.

Global warming

Chilling hrs. Union of Concerned Scientists?

Heat Stress

days over 90 °F expected to increase

fruit rot pressure

scald

Days of treatment

0 5 10 15

Pn

(u

mo

l C

O2 m

-2s

-1)

0

2

4

6

8

10

12

14

16

Demoranville 25 oC

Stevens 25 oC

Demoranville 35 oC

Stevens 35 oC

.

Demoranville® (selected in the 1990’s)

Stevens (selected in the 1940’s)

ST

ST

Dem

Dem

Ph

oto

syn

thes

is

Cranberry cultivar tolerance to heat stress

25 C

35 C

Unpublished data Huang and Xu

City Point, WI

Planted

1st rep – 1999

2nd rep – 2000

Oct 9, 2003

Bandon, OR planted in

March 2009

August 5, 2009

June 15, 2011

June 15, 2011

scf45d0.0scf191a0.9vccj1c1.4scf55c5.5scf34s8.6scf32j13.1scf306f17.1

scf210s33.9scf210w34.2scf21n35.8Ig1296a40.4scf6i40.7

scf275d52.9

scf13a61.1

scf35k66.3NA61968.3

scf43976.8scf200179.4vccj981.6

NA104088.5

scf1655c102.8

ctg259107.9

200

9R

R

Vm1

scf32980.0scf105g4.42ms4d10b4.9

5ms2b1217.1

scf117222.7

scf217730.7

CA855F37.4

scf6955c49.5scf144d49.6SCAR565751.4

SCAR585969.2

NA171377.1

scf24k82.8

ctg60089.1

scf43g107.2scf222a108.1scf26r111.2

Vm2

scf203h0.0

Ig51a9.3

scf303c23.5scf5k24.2NA179229.63ms2g0933.2

scf36l60.9

scf31h69.7

scf137c77.1

6ms4e4b84.3

scf1594100.1scf2s100.6

Vm3

Ig6523b0.02ms2g090.8

CA1413b14.6

CA1413a27.2

scf486031.5

NA800F38.3

scf22m57.9

scf19e68.3

scf351478.5scf2505a80.6

Ig729b86.8

scf1p96.3

Vm4

scf239d0.0

2ms2a0210.4

scf283b28.4scf207d32.0Ig13662a33.4

scf27L48.4

vccj555.8NA82458.1scf112c61.7

ctg65287.1

Ig9279a92.0

scf17d97.1

ctg428102.1scf44a105.0

MF

W1

Vm5

scf37h0.0

scf15b22.7

scf79c27.4

scf621336.6

scf24844.1

CA39R48.8

Vm6

NA1720.0NA800c5.1ctg4806.1

scf1068822.7scf142e23.9scf530428.9SCAR212230.5

scf902555.1

scf2253d67.7

200

7R

R

ctg1300.0scf5111.2scf20g2.8

Vm7

scf28l0.0

scf41c12.0CA32513.8

scf2000b27.4

scf1291636.5

scf300f48.4scf25m50.2

scf14j55.5

CA187F69.2ctg70470.7scf23c71.6vccb376.4Ig15420a82.1

MF

W2

201

0S

FY

Vm8

scf262a0.0

scf11l6.7

scf72c13.8

scf8l21.8

scf258d36.4

Vm9

CA9330.0

vccj38.6

scf9e18.7Ig21768b19.8

CA794F41.5CA42142.4

Vm10

scf15903c0.0CA1413d3.8

scf94a8.2SCAR505111.8

vccj1a0.0vccj1b0.1

scf6341b6.3

scf3072b20.6

scf171f44.0

scf46g50.5scf108b52.2scf30g57.0scf12i57.4

MR

R

Vm11

scf4b0.0scf28823.5scf63555.7Ig28559a7.6vccj1d9.4scf39e10.0

Vm12

genome size of about 570 Mbp SSRs SCARs ESTs

SOLiD mate-paired Illumina Blueberry ESTs

scf203h0.01TC4503091.1SCF105142.4SCF1556373.6SCF468247.6Ig51a9.4SCF15089810.5SCF91515.1SCF16779317.9419834_K6318.3SCF18797919.3scf5k24.6SCF11016828.0SCF19310329.6ct9534530.9SCF12830731.1NA179233.0SCF13801437.7311291_K7037.8SCF430537.91TC33778038.1SCF20888341.3SCF987242.8ct14521743.0ct14786444.3SCF13749444.5SCF34010SCF72379

45.4

409618_K6346.576326_K70SCF34513

46.9

1TC21715852.2SCF9423780734_K70ct145906

55.7

SCF11114560.8SCF11254061.3SCF18487361.7SCF11686463.4SCF8594665.1SCF2522167.8scf31h69.5SCF8924771.8scf137c73.4SCF715573.8SCF2068176.1SCF3674578.96ms4e4b79.61TC43562082.6SCF2594483.9SCF248386.2SCF5671790.7scf2s95.3scf159498.8ct13955399.8

1

scf203h 0.0

Ig51a 9.3

scf303c 23.5scf5k 24.2

NA1792 29.6

3ms2g09 33.2

scf36l 60.9

scf31h 69.7

scf137c 77.2

6ms4e4b 84.2

scf2s 99.3scf1594 100.1

1

418294_K630.0SCF565611.8SCF266972.01TC1763032.1ct1195233.12ms4d10b3.6ct945044.5SCF830368.55ms2b1212.2ct83024SCF162175

12.6

SCF915718.2scf117223.7416328_K6329.4ct11959030.2SCF7222930.6SCF11338931.5CA855F33.5SCF14062833.9SCF4673935.5ct5268236.9SCF653040.6SCF12407543.4scf6955c44.1ct5220444.2SCF2810046.6305731_K6347.9SCF15454149.5SCF3855353.1SCF1584555.6SCF19455259.4SCF9653969.7NA171373.4SCF1635973.7SCF10082074.9SCF7901475.2SCF20497976.0scf24k76.1SCF10106480.2contig60081.8SCF4959883.7SCF4691288.3SCF112888.7SCF10829488.8SCF1108489.0SCF5328295.0SCF14573995.4SCF1836395.7scf26r97.2SCF2956097.3SCF5181098.1SCF7489599.2281884_K7099.7SCF14119100.1

TY

2

scf3298 0.0

scf105g 4.42ms4d10b 4.9

5ms2b12 17.1

scf1172 22.7

scf2177 30.7

CA855F 37.4

scf6955c 49.5scf144d 49.6

SCAR5657 51.4

SCAR5859 69.2

NA1713 77.1

scf24k 82.8

contig600 89.1

scf43g 107.2scf222a 108.1

scf26r 111.2

2

Crimson Queen x Mullica Queen

FUNDING

NJAES USDA-NIFA-SCRI USDA-NIFA-AFRI NIH Ocean Spray Cranberries, Inc. NJ Blueberry & Cranberry Research Council

Dr. G. Daverdin Yifei Wang R. DeStefano Stephanie Fong K. Destefano Jennifer Johnson-Cicalese Dr. L. Georgi Jim White Dr. J. Polashock Mariusz Tadych S. Vancho Dr. J. Zalapa

Acknowledgements

V. darrowii BNJ01-8 V. macrocarpon