diversity in the breadfruit complex (artocarpus, moraceae...

ORIGINAL PAPER

Diversity in the breadfruit complex (Artocarpus, Moraceae):genetic characterization of critical germplasm

Nyree Zerega & Tyr Wiesner-Hanks & Diane Ragone &Brian Irish & Brian Scheffler & Sheron Simpson & Francis Zee

Received: 29 August 2014 /Revised: 7 October 2014 /Accepted: 2 December 2014# Springer-Verlag Berlin Heidelberg 2015

Abstract Breadfruit (Artocarpus altilis, Moraceae) is a tradi-tional staple crop in Oceania and has been introduced through-out the tropics. This study examines important germplasmcollections of breadfruit and its closest wild relatives and aimsto (1) characterize genetic diversity, including identification ofunknown and duplicate accessions, (2) evaluate genetic struc-ture and hybridization within the breadfruit complex, and (3)compare utility of microsatellite markers to previously report-ed amplified fragment length polymorphism (AFLP) and iso-zyme markers in differentiating among cultivars. Data for 19microsatellite loci were collected for 349 individuals

(representing 255 accessions) including breadfruit (A. altilis),two wild relatives (Artocarpus camansi and Artocarpusmariannensis) , and putat ive hybrids (A. alt i l is ×A. mariannensis). Accessions were of mixed ploidy and re-gional origin, but predominantly from Oceania. Microsatelliteloci collectively had a polymorphic information content (PIC)of 0.627 and distinguished 197 unique genotypes sorted into129 different lineages, but a single genotype accounts for49 % of all triploid breadfruit examined. Triploid hybridsand diploid A. altilis exhibited the highest levels of diversityas measured by allele number and gene diversity. Most acces-sions (75 %) of unknown origin matched either a knowngenotype or lineage group in the collection. Putative hybridsall had genetic contributions from A. mariannensis but rangedin the level of genetic contribution from A. altilis. Microsat-ellite markers were found to be more informative than iso-zyme markers and slightly less informative, with regard toaccession discrimination, than AFLP markers. This set ofmicrosatellite markers and the dataset presented here will bevaluable for breadfruit germplasm management andconservation.

Keywords Breadfruit .Artocarpus . Germplasmconservationmanagement . Microsatellites . Plant geneticresources . Underutilized crops

Introduction

Breadfruit (Artocarpus altilis (Parkinson) Fosberg, Moraceae)is a traditional staple in Oceania and has many uses rangingfrom construction, medicine, and animal feed to insect repel-lent (Ragone 1997; Jones et al. 2011). However, it is princi-pally grown as a starch food crop and is an important compo-nent of agroforestry systems. Breadfruit has been recognizedas a crop with great potential for increasing food security and

Communicated by W. Ratnam

Electronic supplementary material The online version of this article(doi:10.1007/s11295-014-0824-z) contains supplementary material,which is available to authorized users.

N. Zerega (*) : T. Wiesner-HanksPlant Biology and Conservation, Northwestern University, Evanston,IL 60208, USAe-mail: [email protected]

N. ZeregaDepartment of Plant Science, Chicago Botanic Garden, Glencoe,IL 60022, USA

D. RagoneBreadfruit Institute, National Tropical Botanical Garden, Kalaheo,HI 96741, USA

B. IrishTropical Agriculture Research Station, USDA-ARS,Mayagüez 00680, Puerto Rico

B. Scheffler : S. SimpsonGenomics and Bioinformatics Research Unit, USDA-ARS,Stoneville, MS 38776-0350, USA

F. ZeePacific Basin Agricultural Research Center (PBARC), USDA-ARS,Hilo, HI 96720, USA

Tree Genetics & Genomes (2015) 11:4 DOI 10.1007/s11295-014-0824-z

http://dx.doi.org/10.1007/s11295-014-0824-z

alleviating malnutrition in Oceania, the Caribbean, tropicalAfrica, and beyond (Wootton and Tumaalii 1984; Morton1990; Adebowale et al. 2005; Omubuwajo 2007; Roberts-Nkrumah 2007; Taylor et al. 2009; Jones et al. 2011). Itproduces large starchy compound fruits with high yields andhigh levels of minerals and provitamin A carotenoids ascompared to other important staples like wheat, maize, andrice (Jones et al. 2011, 2013). Although most cultivars pro-duce fruit from August to January, growing a set of cultivarswith diverse fruit-bearing seasons could allow for year-roundharvest of nutrient-rich fruit (Fownes and Raynor 1991; Joneset al. 2010, 2013). Since Europeans first encountered bread-fruit in the Pacific nearly 400 years ago (Markham 1904), asmall number of cultivars have been introduced to tropicalregions throughout the tropics, including the Caribbean (Pow-ell 1977; Leakey 1977; Roberts-Nkrumah 2007), Africa(Omubuwajo 2007), and India (Ragone 1997). The geneticdiversity and importance of breadfruit, however, remaingreatest in Oceania, where breadfruit was domesticated(Zerega et al. 2004, 2005, 2006).

Breadfruit’s wild relatives have been identified asArtocarpus camansi Blanco and Artocarpus mariannensisTrécul, and hybrids also exist (Fosberg 1960; Zerega et al.2004, 2005). Over millennia, Pacific Islanders have select-ed and named hundreds of traditional cultivars based onfruiting season, fruit shape, color and texture of the fleshand skin, presence or absence of seeds, flavor, cooking andstorage qualities, leaf shape, and horticultural needs (Wil-der 1928; Ragone 1997). Cultivars include vegetativelypropagated seedless triploids, vegetatively or seed-propagated fertile diploids, and diploid and triploid hybrids(Ragone 2001, 2007; Zerega et al. 2004). Over 2000 nameshave been collected for breadfruit cultivars in Oceania,where breadfruit was domesticated (Ragone 1991). Be-cause names are typically based on morphological traitsof the tree and fruit, which may be environmentally influ-enced across islands (Ragone and Wiseman 2007), andbecause the same name may be used for different typesor different names may be used for the same type, geneticcharacterization of cultivars is important.

To help conserve, study, and improve breadfruit, manygermplasm collections have been assembled over the lastseveral decades throughout the tropics (Ragone 1997).Most cultivars are seedless, and even when seeds arepresent, they are recalcitrant and cannot be dried, frozen,or stored, so collections must be maintained as livingtrees in field genebanks. This is a time-consuming andexpensive task, and for this reason, many collections areno longer being maintained (Ragone 1997). Understand-ing the genetic diversity of collections is critical forinformed germplasm management and prioritizing con-servation efforts and is an important step in the increasedutilization of this crop.

The largest and most extensive collection of breadfruit ishoused at the Breadfruit Institute at the National TropicalBotanical Garden (NTBG). It contains breadfruit, hybrids,and wild relatives from throughout Oceania and beyond (Ta-ble 1, see also Online Resource 1 and www.http://ntbg.org/breadfruit). USDA-ARS National Plant Germplasm System(NPGS) repositories (Pacific Basin Agricultural ResearchCenter (PBARC) in Hilo, HI, and Tropical Agriculture Re-search Station (TARS) in Mayagüez, Puerto Rico) includeduplicates from NTBG as well as some additional uniqueaccessions (http://www.ars-grin.gov/npgs/acc/acc_queries.html) (Table 1). This study analyzed the accessions from theseimportant breadfruit collections. Previous genetic diversitystudies conducted on a subset of these collections used iso-zymes (Ragone 1991) and amplified fragment length poly-morphism (AFLPs) (Zerega et al. 2004, 2005, 2006). Thepresent study characterized 349 breadfruit individuals andwild congeners from the NTBG and NPGS collections using19 microsatellite loci (Witherup et al. 2013) in order to (1)characterize genetic diversity, including identification of un-known and replicate accessions, (2) evaluate genetic structureand hybridization within the breadfruit complex, and (3) com-pare microsatellite utility to AFLP and isozyme markers inassessing breadfruit diversity and differentiating amongcultivars.

Methods and materials

Plant materials

Leaf tissue samples from 349 individuals (representing 255accessions) were collected on silica for 229 A. altilis, 70A. altilis×A. mariannensis hybrids, 36 A. camansi, and 14A. mariannensis specimens. Samples came from the followingsites: NTBG (Kahanu Garden, Maui, and the McBryde,Allerton, and Limahuli Gardens in Kauai, HI), USDA-ARS,TARS, and USDA-ARS, PBARC (Table 1). Provenance lo-calities were predominantly in Oceania (Fig. 1, Table 1).Taxon name, cultivar name, and additional provenance infor-mation are also publicly available for most of the accessionsthrough NTBG (http://ntbg.org/breadfruit/database) and theARS’s Germplasm Resources Information Network (GRIN)databases (http://www.ars-grin.gov/npgs/acc/acc_queries.html). An accession number was frequently represented byonly one tree; however, in some cases, multiple individualswere represented from the same base accession number (thesix-digit number preceding the decimal point). If individualsshared the same base accession number, it indicated thatthey were either vegetatively propagated from the sameparent tree or that they were grown from seed from thesame mother tree.

4 Page 2 of 26 Tree Genetics & Genomes (2015) 11:4

http://www.http//ntbg.org/breadfruithttp://www.http//ntbg.org/breadfruithttp://www.ars-grin.gov/npgs/acc/acc_queries.htmlhttp://www.ars-grin.gov/npgs/acc/acc_queries.htmlhttp://ntbg.org/breadfruit/databasehttp://www.ars-grin.gov/npgs/acc/acc_queries.htmlhttp://www.ars-grin.gov/npgs/acc/acc_queries.html

Tab

le1

Accession

dataforspecim

ensused

instudy

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

TARS6732

11

3nTA

RS

Mayaguez

Plot2

2Unk

Caribbean

Barbados

H4

Aa

890472.001

11

3nNTBG

Kahanu

W4

Enua

EPoly

CookIslands

H4

Aa

890472.002

11

3nNTBG

Kahanu

U4

Enua

EPoly

CookIslands

H4

Aa

900256.001

11

3nNTBG

Kahanu

T3

Enua

EPoly

CookIslands

H4

Aa

HART56

11

3nHART

Hilo

Kahaluu

EPoly

Haw

aii

H4

Aa

070101.001

11

3nNTBG

Kahanu

O10

Ulu

EPo

lyHaw

aii

H4

Aa

070882.002

11

3nNTBG

McB

ryde

AU2

Ulu

EPoly

Haw

aii

H4

Aa

090739.001

11

3nNTBG

Lim

ahuli

LM1

Ulu

EPo

lyHaw

aii

H4

Aa

100346.001

11

3nNTBG

Allerton

AG4

Ulu

EPo

lyHaw

aii

H4

Aa

100347.001

11

3nNTBG

Allerton

AG1

Ulu

EPo

lyHaw

aii

H4

Aa

100348.001

11

3nNTBG

Allerton

AG2

Ulu

EPo

lyHaw

aii

H4

Aa

100349.001

11

3nNTBG

Allerton

AG3

Ulu

EPo

lyHaw

aii

H4

Aa

970274.001

11

3nNTBG

McB

ryde

CG1

Ulu

EPoly

Haw

aii

H4

Aa

900240.001

11

3nNTBG

Kahanu

M8

Meikauhiva

EPoly

Marquesas

H4

Aa

900238.001

11

3nNTBG

Kahanu

F5

Meikiiahi

EPoly

Marquesas

H4

Aa

890462.001

11

3nNTBG

Kahanu

U2

Meipuau

EPo

lyMarquesas

H4

Aa

HART23

(900237)

11

3nHART

Hilo

Meipuou

EPoly

Marquesas

H4

Aa

780332.001

11

3nNTBG

Kahanu

33Aarue

EPoly

Society

Islands

H4

Aa

040051.001

11

3nNTBG

McB

ryde

McB

8Afara

EPo

lySo

cietyIslands

H4

Aa

780325.001

11

3nNTBG

Kahanu

32Afara

EPoly

Society

Islands

H4

Aa

910267.001

11

3nNTBG

Kahanu

V8

Afara

EPoly

Society

Islands

H4

Aa

780333.001

11

3nNTBG

Kahanu

30Ahani

EPoly

Society

Islands

H4

Aa

900249.002

11

3nNTBG

Kahanu

I8Anahonaho

EPoly

Society

Islands

H4

Aa

890157.001

11

3nNTBG

Kahanu

42Apu

EPoly

Society

Islands

H4

Aa

HART29

(890157)

11

3nHART

Hilo

Apu

EPoly

Society

Islands

H4

Aa

900232.001

11

3nNTBG

Kahanu

A7

Atu

EPoly

Society

Islands

H4

Aa

890147.001

11

3nNTBG

Kahanu

21Aue

EPoly

Society

Islands

H4

Aa

890147.002

11

3nNTBG

Kahanu

L7

Aue

EPoly

Society

Islands

H4

Aa

780335.001

11

3nNTBG

Kahanu

23Aum

eeEPoly

Society

Islands

H4

Aa

780330.001

11

3nNTBG

Kahanu

56Fafai

EPoly

Society

Islands

H4

Aa

890154.001

11

3nNTBG

Kahanu

Y1

Ham

oaEPoly

Society

Islands

H4

Aa

HART32

(890154)

11

3nHART

Hilo

Ham

oaEPoly

Society

Islands

H4

Aa

780291.001

11

3nNTBG

Kahanu

47Havanapataitai

EPo

lySo

cietyIslands

H4

Aa

900245.001

11

3nNTBG

Kahanu

G8

Huero

EPoly

Society

Islands

H4

Aa

890150.001

11

3nNTBG

Kahanu

Y8

Ioio

EPo

lySo

cietyIslands

H4

Aa

HART42

(890150)

11

3nHART

Hilo

IoIo

EPo

lySo

cietyIslands

H4

Aa

890149.001

11

3nNTBG

Kahanu

46Mam

aha

EPo

lySo

cietyIslands

H4

Tree Genetics & Genomes (2015) 11:4 Page 3 of 26 4

Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

890464.001

11

3nNTBG

Kahanu

P7

Ouo

EPoly

Society

Islands

H4

Aa

780328.001

11

3nNTBG

Kahanu

14Puaa

EPo

lySo

cietyIslands

H4

Aa

890460.001

11

3nNTBG

Kahanu

T6

Puaa

EPo

lySo

cietyIslands

H4

Aa

900244.001

11

3nNTBG

Kahanu

S8

Puaa

EPo

lySo

cietyIslands

H4

Aa

HART18

(900244)

11

3nHART

Hilo

Puaa

EPo

lySo

cietyIslands

H4

Aa

HART16

11

3nHART

Hilo

Puero

EPoly

Society

Islands

H4

Aa

890152.001

11

3nNTBG

Kahanu

W1

Puurea

EPoly

Society

Islands

H4

Aa

890152.002

11

3nNTBG

Kahanu

S7

Puurea

EPoly

Society

Islands

H4

Aa

780329.001

11

3nNTBG

Kahanu

29Rare

EPo

lySo

cietyIslands

H4

Aa

040518.001

11

3nNTBG

McB

ryde

McB

14Rareautia

EPoly

Society

Islands

H4

Aa

780345.001

11

3nNTBG

McB

ryde

BB1

Raumae

EPoly

Society

Islands

H4

Aa

780338.001

11

3nNTBG

Kahanu

51Tapehaa

EPoly

Society

Islands

H4

Aa

100370.001

11

3nNTBG

Lim

ahuli

LM3

Tuutou

EPoly

Society

Islands

H4

Aa

900246.001

11

3nNTBG

Kahanu

H7

Tuutouauena

EPo

lySo

cietyIslands

H4

Aa

900247.001

11

3nNTBG

Kahanu

I6Tuutouooa

EPoly

Society

Islands

H4

Aa

890186.001

11

3nNTBG

Kahanu

Z2

Tuutoutaatoe

EPoly

Society

Islands

H4

Aa

890186.002

11

3nNTBG

Kahanu

B7

Tuutoutaatoe

EPoly

Society

Islands

H4

Aa

890165.001

11

3nNTBG

Kahanu

Z3

Meichon

Micro

Chuuk,F

SMH

4

Aa

HART51

(890165)

11

3nHART

Hilo

Meichon

Micro

Chuuk,F

SMH

4

Aa

890162.001

11

3nNTBG

Kahanu

X4

Lem

aeMicro

Mariana

Islands

H4

Aa

890159.001

11

3nNTBG

Kahanu

X5

Meriaur

Micro

Palau

H4

Aa

HART35

(890159)

11

3nHART

Hilo

Meriaur

Micro

Palau

H4

Aa

890478.001

11

3nNTBG

Kahanu

R5

Meikalak

Micro

Pohnpei,FS

MH

4

Aa

890478.002

11

3nNTBG

Kahanu

O4

Meikalak

Micro

Pohnpei,FS

MH

4

Aa

790497.002

11

3nNTBG

Kahanu

49Meinuwe

Micro

Pohnpei,FS

MH

4

Aa

890167.001

11

3nNTBG

Kahanu

41Meisaip

Micro

Pohnpei,F

SMH

4

Aa

890167.002

11

3nNTBG

Kahanu

R6

Meisaip

Micro

Pohnpei,F

SMH

4

Aa

890479.001

11

3nNTBG

Kahanu

Q7

Meisei

Micro

Pohnpei,FS

MH

4

Aa

890479.002

11

3nNTBG

Kahanu

O7

Meisei

Micro

Pohnpei,FS

MH

4

Aa

030033.001

11

3nNTBG

McB

ryde

McB

16Meitehid

Micro

Pohnpei,FS

MH

4

Aa

910273.002

11

3nNTBG

Kahanu

G9

Meitehid

Micro

Pohnpei,FS

MH

4

Aa

910271.001

11

3nNTBG

Kahanu

D9

Meiuhpw

Micro

Pohnpei,F

SMH

4

Aa(A

a×Am)

890480.003

11

3nNTBG

Kahanu

R8

Lipet2

Micro

Pohnpei,FS

MH

4

Aa

810290.002

11

3nNTBG

Kahanu

45White

Seychelles

Seychelles

H4

Aa

Unknown

11

3nTA

RS

Mayaguez

Plot1

8Unk

Unk

(Epoly

orMicro)

Unknown

H4

Aa

910290.001

11

3nNTBG

Kahanu

L10

Unk

04Unk

(Epoly

orMicro)

Unknown

H4

Aa

910287.001

11

3nNTBG

Kahanu

R9

Unk

09Unk

(Epoly

orMicro)

Unknown

H4


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

910286.001

11

3nNTBG

Kahanu

T8

Unk

11Unk

(Epoly

orMicro)

Unknown

H4

Aa

800269.001

13

3nNTBG

Kahanu

36Mahani

EPo

lySo

cietyIslands

H4

Aa

TARS17990

13

3nTA

RS

Mayaguez

Plot5

Samoa

WPo

lySamoa

H4

Aa

900242.001

15

3nNTBG

Kahanu

B8

Meikopumoko

EPoly

Marquesas

H4

Aa

HART28

(900242)

15

3nHART

Hilo

Meikopumoku

EPoly

Marquesas

H4

Aa

900249.001

15

3nNTBG

Kahanu

I7Anahonaho

EPoly

Society

Islands

H4

Aa

030028.001

15

3nNTBG

McB

ryde

McB

13White

Seychelles

Seychelles

H4

Aa

810290.001

15

3nNTBG

Kahanu

43White

Seychelles

Seychelles

H4

Aa

810290.003

15

3nNTBG

Kahanu

48White

Seychelles

Seychelles

H4

Aa

810290.004

15

3nNTBG

McB

ryde

BB3

White

Seychelles

Seychelles

H4

Aa

810289.001

15

3nNTBG

McB

ryde

BB2

Yellow

Seychelles

Seychelles

H4

Aa

810289.002

15

3nNTBG

Kahanu

38Yellow

Seychelles

Seychelles

H4

Aa

890158.001

16

3nNTBG

Kahanu

Z6

Apuapua

EPoly

Society

Islands

H4

Aa

890158.002

16

3nNTBG

Kahanu

H8

Apuapua

EPoly

Society

Islands

H4

Aa

HART34

(890158)

16

3nHART

Hilo

Apuapua

EPoly

Society

Islands

H4

Aa

900243.001

17

3nNTBG

Kahanu

D7

Araarahaari

EPo

lySo

cietyIslands

H4

Aa

780330.002

19

3nNTBG

Kahanu

40Fafai

EPoly

Society

Islands

H4

Aa

100369.001

19

3nNTBG

Lim

ahuli

LM2

Fafai(unk)

EPoly

Society

Islands

H4

Aa

890162.002

114

3nNTBG

Kahanu

S4

Lem

aeMicro

Mariana

Islands

H4

Aa

HART38

(890162)

114

3nHART

Hilo

Lem

aeMicro

Mariana

Islands

H4

Aa

890459.001

115

3nNTBG

Kahanu

R7

Maire

EPoly

Society

Islands

H4

Aa

HART44

(890459)

115

3nHART

Hilo

Maire

EPoly

Society

Islands

H4

Aa

HART54

(890459)

115

3nHART

Hilo

Maire

EPoly

Society

Islands

H4

Aa

900267.001

115

3nNTBG

Kahanu

N8

Maire

(unk

05)

EPoly(unk)

SocietyIslands(unk)

H4

Aa

890459.002

116

3nNTBG

Kahanu

J7Maire

EPoly

Society

Islands

H4

Aa

900262.001

117

3nNTBG

Kahanu

M6

Manua

WPo

lySam

oaH

4

Aa

900241.001

118

3nNTBG

Kahanu

F6

Meiaueka

EPo

lyMarquesas

H4

Aa

900239.001

119

3nNTBG

Kahanu

T5

Meimaoi

EPoly

Marquesas

H4

Aa

HART11

(900239)

119

3nHART

Hilo

Meimaoi

EPoly

Marquesas

H4

Aa

900237.001

120

3nNTBG

Kahanu

B6

Meipuou

EPoly

Marquesas

H4

Aa

080439.001

123

3nNTBG

Kahanu

Heiau1

Ulu

EPoly

Haw

aii

H4

Aa

080858.001

123

3nNTBG

Kahanu

CG3

Ulu

EPo

lyHaw

aii

H4

Aa

080863.001

123

3nNTBG

Kahanu

CG8

Ulu

EPo

lyHaw

aii

H4

Aa

080881.001

123

3nNTBG

Kahanu

CG11

Ulu

EPoly

Haw

aii

H4

Aa

070132.001

123

3nNTBG

Kahanu

CG1

Ulu

EPo

lyH

4

Aa

080440.001

123

3nNTBG

Kahanu

Heiau2

Ulu

EPoly

H4

Aa

890159.002

123

3nNTBG

Kahanu

V4

Meriaur

Micro

Palau

H4


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

000534.001

123

3nNTBG

Kahanu

GG4

Meikalaken

meikuet

Micro

Pohnpei,FS

MH

4

Aa

790493.001

125

3nNTBG

Kahanu

4Meitehid

Micro

Pohnpei,FS

MH

4

Aa

HART57

127

3nHART

Hilo

PoahomoPrickly

EPoly

H4

Aa

030035.001

132

3nNTBG

McB

ryde

McB

7Otea

EPoly

Society

Islands

H4

Aa

780327.001

132

3nNTBG

Kahanu

39Otea

EPo

lySo

cietyIslands

H4

Aa

910266.001

136

3nNTBG

Kahanu

H9

Piip

iiaEPoly

Society

Islands

H3

Aa

30037.001

137

3nNTBG

McB

ryde

McB

10Porohiti

EPoly

Society

Islands

H4

Aa

790486.001

138

3nNTBG

Kahanu

20Roihaa

EPo

lySo

cietyIslands

H4

Aa

890465.001

139

3nNTBG

Kahanu

V7

Teahim

atoa

EPoly

Society

Islands

H4

Aa

HART40

(890465)

139

3nHART

Hilo

Teahim

atoa

EPoly

Society

Islands

H4

Aa

070882.001

142

3nNTBG

McB

ryde

AU1

Ulu

EPoly

Haw

aii

H4

Aa

080859.001

143

3nNTBG

Kahanu

CG4

Ulu

EPo

lyHaw

aii

H4

Aa

080864.001

144

3nNTBG

Kahanu

CG9

Ulu

EPo

lyHaw

aii

H4

Aa

080864.002

144

3nNTBG

Kahanu

CG10

Ulu

EPoly

Haw

aii

H4

Aa

900268.001

146

3nNTBG

Kahanu

E4

Unk

01Unk

Unknown

H4

Aa

900225.001

148

3nNTBG

Kahanu

S5

Unk

10Unk

Unknown

H4

Aa

890148.001

150

3nNTBG

Kahanu

Y6

Meipuou

(unk

14)

EPoly

Marquesas

(Society)

H4

Aa

070883.001

152

3nNTBG

McB

ryde

Pump6

Unk

EPoly

Unknown

H4

Aa

910265.001

169

3nNTBG

Kahanu

V9

Rotum

aEPo

lySo

cietyIslands

H4

Aa

Unknown

22

3nTA

RS

Mayaguez

Plot11

Unk

Epoly

(unk)

SocietyIslands(unk)

G3

Aa

890151.001

210

3nNTBG

Kahanu

X7

Fafai

EPoly

Society

Islands

G3

Aa

880690.001

213

3nNTBG

Kahanu

P8

Kea

WPo

lyTo

nga

G3

Aa

890153.002

233

3nNTBG

Kahanu

2Paea

EPoly

CookIslands

G3

Aa

890463.001

234

3nNTBG

Kahanu

V3

Patara

EPo

lySo

cietyIslands

G3

Aa

890463.002

234

3nNTBG

Kahanu

G6

Patara

EPo

lySo

cietyIslands

G3

Aa

HART33

(890463)

234

3nHART

Hilo

Patara

EPo

lySo

cietyIslands

G3

Aa

790485.001

234

3nNTBG

Kahanu

16Puupuu

EPoly

Society

Islands

G3

Aa

790491.001

234

3nNTBG

Kahanu

13Tuutou

EPoly

Society

Islands

G3

Aa

900236.001

34

3nNTBG

Kahanu

D6

Abareba

Mela

Solomon

Islands

C3

Aa

HART45

(890186)

48

3nHART

Hilo

Tuutou

EPoly

Society

Islands

F4/1

Aa

HART30

(900245)

612

2nHART

Hilo

Huero

EPoly

Society

Islands

E3

Aa

900266.001

721

3nNTBG

Kahanu

M5

Meiarephe

Micro

Pohnpei,F

SMD

4

Aa

900266.002

721

3nNTBG

Kahanu

E5

Meiarephe

Micro

Pohnpei,F

SMD

4

Aa

000531.001

824

3nNTBG

Kahanu

58Meipw

etMicro

Pohnpei,F

SMD

4

Aa

030042.001

1140

3nNTBG

McB

ryde

McB

11To

neno

EPoly

Society

Islands

F4

Aa

790488.001

1141

3nNTBG

Kahanu

12To

neno

EPoly

Society

Islands

F4

Aa

30044.001

1245

3nNTBG

Offsite

Offsite

Ulu

tala

WPo

lySam

oaE

3


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

770524.001

1245

3nNTBG

Kahanu

54Ulu

tala

WPo

lySamoa

E3

Aa

910288.001

1347

3nNTBG

Kahanu

Q8

Uto

vula(unk

08)

Mela(unk)

Fiji(unk)

F2

Aa

890476.002

1351

3nNTBG

Kahanu

S6Uto

vula

Mela

Fiji

F2

Aa

020354.001

2576

2nNTBG

Kahanu

KM1

Aveloloa

WPo

lySamoa

G3

Aa

910278.001

2677

2nNTBG

Kahanu

M9

Forari

Mela

Vanuatu

E3

Aa

910279.001

26103

2nNTBG

Kahanu

E9

Siviri2

Mela

Vanuatu

E3

Aa

910279.002

26103

2nNTBG

Kahanu

C8

Siviri2

Mela

Vanuatu

E3

Aa

890470.002

2778

2nNTBG

Kahanu

N5

Furau

Mela

Rotum

aG

2

Aa

040063.001

2778

2nNTBG

McB

ryde

McB

15Ulu

fiti

Mela

Rotum

aG

2

Aa

890258.001

2778

2nNTBG

Kahanu

35Ulu

fiti

Mela

Rotum

aG

2

Aa

890458.001

2778

2nNTBG

Kahanu

T4

Ulu

fiti

Mela

Rotum

aG

2

Aa

890458.002

2778

2nNTBG

Kahanu

Q4

Ulu

fiti

Mela

Rotum

aG

2

Aa

900368.001

2778

2nNTBG

Kahanu

C4

Ulu

fiti

Mela

Rotum

aG

2

Aa

970236.001

2778

2nNTBG

Kahanu

AA4

Ulu

fiti

Mela

Rotum

aG

2

Aa

890469.002

2778

2nNTBG

Kahanu

S3Kukum

utasi

Mela

Solom

onIslands

G2

Aa

020353.001

2778

2nNTBG

Kahanu

KM5

Unk

18W

Poly

Sam

oaG

2

Aa

020353.002

2778

2nNTBG

Kahanu

KM4

Unk

18W

Poly

Sam

oaG

2

Aa

900260.001

27110

2nNTBG

Kahanu

K7

Ulu

fiti,

Sam

oan

Mela

Rotum

aG

2

Aa

900260.002

27110

2nNTBG

Kahanu

K5

Ulu

fiti,

Sam

oan

Mela

Rotum

aG

2

Aa

890470.001

2879

2nNTBG

Kahanu

V6

Furau

Mela

Rotum

aG

2

Aa

890469.001

2879

2nNTBG

Kahanu

V1

Kukum

utasi

Mela

Solom

onIslands

G2

Aa

HART26

2980

2nHART

Hilo

G.W

ilder-Brash

EPoly

E3

Aa

900248.001

3081

2nNTBG

Kahanu

G5

Huero

ninamu

EPo

lySo

cietyIslands

E3

Aa

890461.001

3089

2nNTBG

Kahanu

W2

Meikakano

EPo

lyMarquesas

E3

Aa

890461.002

3089

2nNTBG

Kahanu

C7

Meikakano

EPo

lyMarquesas

E3

Aa

HART21

(890461)

3089

2nHART

Hilo

Meikakano

EPoly

Marquesas

E3

Aa

890457.001

3182

2nNTBG

Kahanu

W5

Karaw

aMela

Rotum

aG

2

Aa

020356.001

3182

2nNTBG

Kahanu

KM7

Ulu

fiti

Mela

Rotum

aG

2

Aa

900265.001

3283

2nNTBG

Kahanu

C5

Karaw

aMela

Fiji

F2

Aa

900224.001

3283

2nNTBG

Kahanu

U7

Karaw

a(unk

12)

Mela(unk)

Fiji(unk)

F2

Aa

900234.001

32102

2nNTBG

Kahanu

N6

Samoan1

Mela

Fiji

F2

Aa

070659.021

3384

2nNTBG

McB

ryde

CHC1

Ma’afala

WPo

lySam

oaE

3

Aa

070659.022

3384

2nNTBG

McB

ryde

HQ1

Ma’afala

WPo

lySam

oaE

3

Aa

770517.001

3384

2nNTBG

Kahanu

55Ma’afala

WPo

lySam

oaE

3

Aa

HART1

3385

2nHART

Hilo

Ma’afala

E3

Aa

890454.001

3393

2nNTBG

Kahanu

W6

Niue

EPoly

CookIslands

E3

Aa

900231.001

3393

2nNTBG

Kahanu

L6

Niue

EPoly

CookIslands

E3


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

890153.001

3393

2nNTBG

Kahanu

25Niue(Paea)

EPoly

CookIslands

E3

Aa

900259.001

3486

2nNTBG

Kahanu

C6

Malphang

Mela

Vanuatu

G3

Aa

890473.001

3587

2nNTBG

Kahanu

U5

Manang

Mela

Vanuatu

E3

Aa

900263.001

3688

2nNTBG

Kahanu

J8Masee

WPo

lySamoa

E3

Aa

890156.001

3790

2nNTBG

Kahanu

Y4

Tahitian

EPoly

CookIslands

E3

Aa

890156.002

3790

2nNTBG

Kahanu

W7

Tahitian

EPoly

CookIslands

E3

Aa

890477.001

3790

2nNTBG

Kahanu

R4

Uto

samoa

Mela

Fiji

E3

Aa

910275.001

3790

2nNTBG

Kahanu

K8

Puou

Mela

Vanuatu

E3

Aa

960575.001

3790

2nNTBG

McB

ryde

LF1

Mos

enSamoa

Micro

Kosrae

E3

Aa

070246.002

3790

2nNTBG

Kahanu

61Meiuhpw

enSamoa

Micro

Pohnpei,FS

ME

3

Aa

030039.001

3790

2nNTBG

McB

ryde

McB

17Pu

ouW

Poly

Sam

oaE

3

Aa

770520.001

3790

2nNTBG

Kahanu

53Pu

ouW

Poly

Sam

oaE

3

Aa

890474.001

3798

2nNTBG

Kahanu

U6

Puou

WPo

lySam

oaE

3

Aa

910289.001

3798

2nNTBG

Kahanu

O9

Puou

(unk

06)

Wpoly

(unk)

Samoa

(unk)

E3

Aa

880691.001

3799

2nNTBG

Kahanu

O8

Puou

WPo

lyTo

nga

E3

Aa

770519.001

3891

2nNTBG

Kahanu

57Mom

olega

WPo

lySam

oaE

3

Aa

HART50

(890469)

3992

2nHART

Hilo

Kukum

uTasi

Mela

Solom

onIslands

C3

Aa

910266.002

4094

2nNTBG

Kahanu

F8Piipiia

EPoly

Society

Islands

E4

Aa

790492.001

4195

2nNTBG

Kahanu

9Porohiti

EPoly

Society

Islands

C1

Aa

900261.001

4296

2nNTBG

Kahanu

F7Samoan2

Mela

Fiji

G2

Aa

900261.002

4296

2nNTBG

Kahanu

P9Samoan2

Mela

Fiji

G2

Aa

890471.001

4296

2nNTBG

Kahanu

W3

Uto

dina

Mela

Fiji

G2

Aa

890471.002

4296

2nNTBG

McB

ryde

MV4

Uto

dina

Mela

Fiji

G2

Aa

900233.002

4296

2nNTBG

Kahanu

8Pulupulu

Mela

Rotum

aG

2

Aa

900233.001

4297

2nNTBG

Kahanu

J6Pu

lupulu

Mela

Rotum

aG

2

Aa

900226.001

42114

2nNTBG

Kahanu

P5

Unk

07Mela(unk)

Fiji(unk)

G2

Aa

020352.001

42117

2nNTBG

Kahanu

KM3

Unk

17W

Poly

Samoa

G2

Aa

900264.001

42121

2nNTBG

Kahanu

A8

Uto

niviti

Mela

Fiji

G2

Aa

900257.001

43100

2nNTBG

Kahanu

A6

Rauulu

Mela

Rotum

aF

2

Aa

890475.002

44101

2nNTBG

Kahanu

S9Sagosago

WPo

lySamoa

E3

Aa

910276.001

45104

2nNTBG

Kahanu

K9

Siviri3

Mela

Vanuatu

E3

Aa

910277.001

46105

2nNTBG

Kahanu

E8

Tedailir

Mela

Vanuatu

G3

Aa

900281.001

47106

2nNTBG

Kahanu

P4Tehelewa

Mela

Solomon

Islands

C3

Aa

900281.002

47107

2nNTBG

Kahanu

D4

Tehelewa

Mela

Solomon

Islands

C3

Aa

890456.001

48108

2nNTBG

Kahanu

T7

Toro

Mela

Solomon

Islands

C3

Aa

770521.001

49109

2nNTBG

Kahanu

52Ulu

eaW

Poly

Tokelau

E3

Aa

890155.001

50111

2nNTBG

Kahanu

17Ulu

sina

WPo

lySam

oaE

3


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa

890155.002

50111

2nNTBG

Kahanu

K6

Ulu

sina

WPo

lySam

oaE

3

Aa

900235.001

52113

2nNTBG

Kahanu

L5

Unk

03Mela

Solomon

Islands

E3

Aa

020347.001

53115

2nNTBG

Kahanu

KM2

Unk

15W

Poly

Samoa

C3

Aa

020348.001

54116

2nNTBG

Kahanu

KM8

Unk

16W

Poly

Samoa

C3

Aa

020355.001

55118

2nNTBG

Kahanu

KM6

Unk

19W

Poly

Samoa

E3

Aa

020498.001

56119

2nNTBG

Kahanu

59Unk

21Mela

Solomon

Islands

F3

Aa

020500.001

57120

2nNTBG

Kahanu

60Unk

22Mela

Solom

onIslands

F3

Aa×

Am

HART37

(890160)

48

3nHART

Hilo

Ebechab

Micro

Palau

F4/1

Aa×

Am

890183.001

426

3nNTBG

Kahanu

Y3

Midolab

Micro

Palau

0.735(0.579–0.863)

F4/1

Aa×

Am

890183.003

426

3nNTBG

McB

ryde

MV3

Midolab

Micro

Palau

0.752(0.586–0.883)

F4/1

Aa×

Am

(Aa)

HART46

(890183)

426

3nHART

Hilo

Midolab

Micro

Palau

0.845(0.693–0.953)

F4/1

Aa×

Am

890183.002

467

3nNTBG

Kahanu

L8

Midolab

Micro

Palau

0.775(0.615–0.9)

F4/1

Aa×

Am

(Aa)

790487.001

511

3nNTBG

Kahanu

27Unk

(Huehue)

Micro

(EPo

ly)

Pohnpei,F

SM(Society)

0.722(0.566–0.853)

D5

Aa×

Am

HART52

(890480)

529

3nHART

Hilo

Lipet

Micro

Pohnpei,F

SM0.76

(0.59–0.894)

D4

Aa×

Am

(Aa)

790489.001

535

3nNTBG

Kahanu

1Unk

(Piip

iia)

Micro

(EPoly)

Pohnpei,FS

M(Society)

0.768(0.617–0.887)

D4

Aa×

Am

030034.001

554

3nNTBG

McB

ryde

McB

9Meinpadahk

Micro

Pohnpei,F

SM0.768(0.617–0.887)

D4

Aa×

Am

030034.002

554

3nNTBG

McB

ryde

McB

18Meinpadahk

Micro

Pohnpei,F

SM0.251(0.115–0.433)

D4

Aa×

Am

790494.001

554

3nNTBG

Kahanu

Z9

Meinpadahk

Micro

Pohnpei,F

SM0.752(0.586–0.883)

D4

Aa×

Am

030041.001

555

3nNTBG

McB

ryde

McB

20Rotum

aEPo

lySo

cietyIslands

0.773(0.625–0.89)

D5/4

Aa×

Am

890163.002

561

3nNTBG

Kahanu

U9

Lem

aeMicro

Mariana

Islands

0.802(0.652–0.915)

D5/4

Aa×

Am

890163.001

562

3nNTBG

Kahanu

X2

Lem

aeMicro

Mariana

Islands

0.802(0.652–0.915)

D4

Aa×

Am

890480.001

563

3nNTBG

Kahanu

O6

Lipet

Micro

Pohnpei,F

SM0.694(0.5–0.856)

D4

Aa×

Am

890480.002

563

3nNTBG

Kahanu

N7

Lipet

Micro

Pohnpei,F

SM0.812(0.668–0.92)

D4

Aa×

Am

910270.001

564

3nNTBG

Kahanu

D8

Lipet

Micro

Pohnpei,F

SM0.27

(0.131–0.451)

D4

Aa×

Am

890467.001

566

3nNTBG

Kahanu

W9

Meinpwuht

Micro

Pohnpei

0.845(0.693–0.953)

D4

Aa×

Am

790490.001

568

3nNTBG

Kahanu

15Rotum

aEPo

lySo

cietyIslands

0.775(0.615–0.9)

D5/4

Aa×

Am

910272.001

928

3nNTBG

Kahanu

F9

Meinpohnsakar

Micro

Pohnpei,F

SM0.734(0.573–0.867)

D5

Aa×

Am

910272.002

928

3nNTBG

Kahanu

E6

Meinpohnsakar

Micro

Pohnpei,F

SM0.719(0.562–0.85)

D5

Aa×

Am

HART49

(910272)

928

3nHART

Hilo

Meinpohnsakar

Micro

Pohnpei,F

SM0.815(0.667–0.924)

D5

Aa×

Am

HART53

(910274)

1030

3nHART

Hilo

Nahnm

wal

Micro

Pohnpei,FS

M0.768(0.601–0.898)

D4

Aa×

Am

910274.001

1031

3nNTBG

Kahanu

Q10

Nahnm

wal

Micro

Pohnpei,FS

M0.804(0.647–0.922)

D4

Aa×

Am

980210.001

1553

3nNTBG

Kahanu

EE3

Ebechab

Micro

Palau

0.768(0.617–0.887)

F4

Aa×

Am

890160.001

1558

3nNTBG

Kahanu

X8

Ebechab

Micro

Palau

0.736(0.575–0.866)

F4

Aa×

Am

910652.001

1559

3nNTBG

Kahanu

U8

Errud

Micro

Palau

0.792(0.644–0.905)

F4

Aa×

Am

910268.001

1656

3nNTBG

Kahanu

J9Meion

Micro

Chuuk,F

SM0.85

(0.701–0.955)

D4

Aa×

Am

990781.001

1656

3nNTBG

McB

ryde

LF2

Meion

(unk)

Micro

Pohnpei,FS

M0.798(0.637–0.919)

D4

Aa×

Am

030045.001

1757

3nNTBG

McB

ryde

McB

19Yap

Micro

Palau

0.804(0.647–0.922)

F4


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa×

Am

900250.001

1774

3nNTBG

Kahanu

A5

Yap

Micro

palau

0.845(0.693–0.953)

F4

Aa×

Am

910269.001

1860

3nNTBG

Kahanu

A9

Faine

Micro

Chuuk,F

SM0.768(0.617–0.887)

D4

Aa×

Am

900255.001

1965

3nNTBG

Kahanu

B5

Meinpwahr

Micro

Pohnpei,FS

M0.773(0.625–0.89)

D5

Aa×

Am

890164.001

2070

3nNTBG

Kahanu

22Sewan

Micro

Chuuk,F

SM0.717(0.553–0.853)

D4

Aa×

Am

890164.002

2070

3nNTBG

Kahanu

T9

Sewan

Micro

Chuuk,F

SM0.491(0.319–0.667)

D4

Aa×

Am

890468.002

2171

3nNTBG

Kahanu

L9

Tebukiraro

Micro

Kiribati

0.555(0.378–0.726)

D4

Aa×

Am

890453.001

2272

3nNTBG

Kahanu

Q5

Ulu

afa

WPo

lyTo

kelau

0.356(0.2–0.539)

D5

Aa×

Am

890185.001

2373

3nNTBG

Kahanu

28Ulu

elise

WPo

lyTo

kelau

0.428(0.264–0.607)

D5

Aa×

Am

890161.001

2475

3nNTBG

Kahanu

19Yuley

Micro

Yap

0.783(0.629–0.902)

F4/5

Aa×

Am

890452.001

58122

2nNTBG

Kahanu

U3

Temai

Micro

Kiribati

0.926(0.791–0.997)

D5

Aa×

Am

890184.001

59123

2nNTBG

Kahanu

10Luthar

Micro

Yap

0.263(0.127–0.44)

C5

Aa×

Am

900253.002

60124

2nNTBG

Kahanu

J5Meichocho

Micro

Chuuk,F

SM0.254(0.122–0.427)

D5

Aa×

Am

900253.001

61125

2nNTBG

Kahanu

Q6

Meichocho

Micro

Chuuk,F

SM0.204(0.085–0.378)

D5

Aa×

Am

890166.001

62126

2nNTBG

Kahanu

Y7

Meikoeng

Micro

Chuuk,F

SM0.768(0.617–0.887)

D5

Aa×

Am

890466.002

63127

2nNTBG

Kahanu

Q9

Meikoeng

Micro

Chuuk,F

SM0.752(0.586–0.883)

D5

Aa×

Am

890177.003

64128

2nNTBG

Kahanu

3Ulu

afa

WPo

lyTo

kelau

0.491(0.319–0.667)

D5

Aa×

Am

890178.001

65129

2nNTBG

Kahanu

26Ulu

afa

WPo

lyTo

kelau

0.366(0.212–0.543)

D5

Aa×

Am

890177.001

66130

2nNTBG

Kahanu

31Ulu

afa

WPo

lyTo

kelau

0.649(0.469–0.807)

D5

Aa×

Am

890172.001

67131

2nNTBG

Kahanu

37Ulu

afa

WPo

lyTo

kelau

0.432(0.266–0.611)

D5

Aa×

Am

890179.001

68132

2nNTBG

Kahanu

44Ulu

afa

WPo

lyTo

kelau

0.462(0.293–0.64)

D5

Aa×

Am

890257.001

69133

2nNTBG

Kahanu

X3

Ulu

afa

WPo

lyTo

kelau

0.213(0.095–0.376)

D5

Aa×

Am

890171.001

70134

2nNTBG

Kahanu

X6

Ulu

afa

WPo

lyTo

kelau

0.628(0.465–0.776)

D5

Aa×

Am

890176.001

71135

2nNTBG

Kahanu

Z8

Ulu

afa

WPo

lyTo

kelau

0.527(0.351–0.702)

D5

Aa×

Am

890174.001

72136

2nNTBG

Kahanu

18Ulu

afa

WPo

lyTo

kelau

0.274(0.14–0.446)

D5

Aa×

Am

890172.002

73137

2nNTBG

Kahanu

ZZ5

Ulu

afa

WPo

lyTo

kelau

0.429(0.265–0.608)

D5

Aa×

Am

890176.002

74138

2nNTBG

Kahanu

ZZ8

Ulu

afa

WPo

lyTo

kelau

0.588(0.409–0.757)

D5

Aa×

Am

890168.002

75139

2nNTBG

Kahanu

ZZ9

Ulu

afa

WPo

lyTo

kelau

0.814(0.644–0.934)

D5

Aa×

Am

890177.002

76140

2nNTBG

Kahanu

5Ulu

afa1

WPo

lyTo

kelau

0.622(0.442–0.787)

D5

Aa×

Am

890171.002

77141

2nNTBG

Kahanu

7Ulu

afa2

WPo

lyTo

kelau

0.556(0.379–0.728)

C5/3

Aa×

Am

890181.002

78142

2nNTBG

Kahanu

B4

Ulu

afa3

WPo

lyTo

kelau

0.554(0.377–0.726)

D5

Aa×

Am

890181.001

79143

2nNTBG

Kahanu

Z4

Ulu

afa4

WPo

lyTo

kelau

0.491(0.32–0.668)

D5

Aa×

Am

890173.001

80144

2nNTBG

Kahanu

11Ulu

afaelise

WPo

lyTo

kelau

0.685(0.505–0.839)

D5

Aa×

Am

890173.002

81145

2nNTBG

Kahanu

ZZ7

Ulu

afaelise

WPo

lyTo

kelau

0.428(0.264–0.607)

D5

Aa×

Am

890175.001

81147

2nNTBG

Kahanu

34Ulu

afahamoa

WPo

lyTo

kelau

0.429(0.264–0.607)

D5

Aa×

Am

890182.002

81149

2nNTBG

Kahanu

ZZ6

Ulu

elise2

WPo

lyTo

kelau

0.554(0.377–0.725)

D5

Aa×

Am

900230.001

82146

2nNTBG

Kahanu

6Ulu

afahamoa

WPo

lyTo

kelau

0.511(0.359–0.664)

C5

Aa×

Am

890182.001

83148

2nNTBG

Kahanu

X9

Ulu

elise1

WPo

lyTo

kelau

0.651(0.471–0.811)

D5


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Aa×

Am

890180.001

84150

2nNTBG

Kahanu

24Ulu

hamoa

WPo

lyTo

kelau

0.926(0.791–0.997)

D5

Aa×

Am

890170.002

85151

2nNTBG

Kahanu

ZZ3

Ulu

hamoa

WPo

lyTo

kelau

0.872(0.728–0.966)

D5

Aa×

Am

(Am)

000528.001

129

197

2nNTBG

Kahanu

AA5

Meikole

Micro

Pohnpei,FS

M0.064(0.01–0.188)

D5

Ac

TARS18009

86152

2nTA

RS

Mayaguez

Plot1

4Unk

Unk

A1

Ac

960576.001

86153

2nNTBG

McB

ryde

MV2

Breadnut

C.A

m.

Honduras

A1

Ac

770444.001

86153

2nNTBG

Kahanu

50Cam

ansi

Epoly

SocietyIslands

A1

Ac

980212.001

87154

2nNTBG

Kahanu

EE5

Cam

ansi

Micro

Palau

A1

Ac

910281.001

88155

2nNTBG

Kahanu

M10

Kam

ansi

Phil.

A1

Ac

000390.002

89156

2nNTBG

McB

ryde

McB

4Kapiak

Mela

Papua

New

Guinea

A1

Ac

000394.002

90157

2nNTBG

McB

ryde

McB

2Kapiak

Mela

Papua

New

Guinea

A1

Ac

000395.002

91158

2nNTBG

McB

ryde

McB

21Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000398.005

92159

2nNTBG

McB

ryde

McB

25Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000499.003

93160

2nNTBG

McB

ryde

McB

3Kapiak

Mela

Papua

New

Guinea

A1

Ac

000500.002

94161

2nNTBG

McB

ryde

McB

23Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000501.004

95162

2nNTBG

McB

ryde

McB

5Kapiak

Mela

Papua

New

Guinea

A1

Ac

000501.005

96163

2nNTBG

McB

ryde

McB

1Kapiak

Mela

Papua

New

Guinea

A1

Ac

000502.002

97164

2nNTBG

McB

ryde

McB

22Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000503.004

98165

2nNTBG

McB

ryde

McB

24Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000398.001

99166

2nNTBG

Kahanu

F10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000501.001

100

167

2nNTBG

Kahanu

F11

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000398.003

101

168

2nNTBG

Kahanu

H10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000399.001

102

169

2nNTBG

Kahanu

I10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000398.002

103

170

2nNTBG

Kahanu

K10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000502.001

104

171

2nNTBG

Kahanu

D10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000501.001

105

172

2nNTBG

Kahanu

EE4

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000499.001

106

173

2nNTBG

Kahanu

EE7

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000503.001

107

174

2nNTBG

Kahanu

E10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000499.002

108

175

2nNTBG

Kahanu

E11

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000395.001

109

176

2nNTBG

Kahanu

FF6

Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000390.001

110

177

2nNTBG

Kahanu

FF7

Kapiak

Mela

PapuaNew

Guinea

A1

Ac

000503.002

111

178

2nNTBG

Kahanu

G10

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000389.001

112

179

2nNTBG

Kahanu

GG3

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000398.004

113

180

2nNTBG

Kahanu

HH3

Kapiak

Mela

Papua

New

Guinea

A1

Ac

000500.001

114

181

2nNTBG

Kahanu

HH4

Kapiak

Mela

Papua

New

Guinea

A1

Ac

910280.001

115

182

2nNTBG

Kahanu

B9

Meikole

Micro

Pohnpei,FS

MA

1

Ac

HART63

116

183

2nHART

Hilo

N90-148

Malaysia

A1

Ac

910283.001

117

184

2nNTBG

Kahanu

R10

Tim

bul

Indonesia

A1


Tab

le1

(contin

ued)

Taxon

Accession

number

Lin

Gn

PlGC

GS

Grid

Cultiv

arRegion

Island

HybridIndex

UP

ST

Ac(A

a)890455.001

1449

2nNTBG

Kahanu

V5

Ulu

fatu

WPo

lySamoa

A1

Ac(A

a)900228.001

51112

2nNTBG

Kahanu

E7

Ulu

fatu

(unk

02)

WPo

lySam

oaA

1

Am

000523.002

118

185

2nNTBG

McB

ryde

McB

6Dugdug

Micro

Mariana

Islands

B5

Am

900252.001

119

186

2nNTBG

Kahanu

H5

Dugdug

Micro

Mariana

Islands

B5

Am

900252.002

120

187

2nNTBG

Kahanu

A4

Dugdug

Micro

Mariana

Islands

B5

Am

900252.003

120

187

2nNTBG

Kahanu

N9

Dugdug

Micro

Mariana

Islands

B5

Am

000522.002

121

188

2nNTBG

Kahanu

BB6

Dugdug

Micro

Mariana

Islands

B5

Am

000523.001

122

189

2nNTBG

Kahanu

CC4

Dugdug

Micro

Mariana

Islands

B5

Am

HART67

122

198

2nHART

Hilo

N05-8

Micro

B5

Am

000521.002

123

190

2nNTBG

Kahanu

CC5

Dugdug

Micro

Mariana

Islands

B5

Am

000521.001

124

191

2nNTBG

Kahanu

CC7

Dugdug

Micro

Mariana

Islands

B5

Am

000522.001

125

192

2nNTBG

Kahanu

DD5

Dugdug

Micro

Mariana

Islands

B5

Am

000521.003

126

193

2nNTBG

Kahanu

DD6

Dugdug

Micro

Mariana

Islands

B5

Am

940010.002

126

194

2nNTBG

McB

ryde

MV1

Dugdug

Micro

Mariana

Islands

B5

Am

970306.001

127

195

2nNTBG

Kahanu

CC3

Kurukiniti

Micro

Pohnpei,F

SMB

5

Am

970306.002

128

196

2nNTBG

Kahanu

DD4

Kurukiniti

Micro

Pohnpei,FS

MB

5

Taxon(A

a=A.altilis,A

c=A.cam

ansi,A

m=A.m

ariannensis,andAa×

Am=A.altilis×A.m

ariannensis),accession

number(N

TBGno.isgiveninparenthesesifthecollectionishoused

somew

hereelse

butisaduplicatefrom

NTBG),lin

eage

(lin),genotype

(gn),p

loidy(pl),g

ermplasm

code

(GC),germ

plasm

site(G

S),g

ridlocationwith

ingerm

plasm

(Grid),cultiv

arname(U

nk=unknow

n),regionof

provenance

(MelaMelanesia,M

icro

Micronesia,EPolyEastern

Polynesia,W

Poly,Western

Polynesia,P

hil.Philip

pines,C.A

m.C

entralAmerica,Unk

unknow

n),islandgroupof

provenance,hybridindex

forp

utativehybrids(num

berscloserto1indicategreatergeneticcontributio

nfrom

A.altilis,and

numberscloserto0indicategreatergeneticcontributio

nfrom

A.m

ariannensis),U

PGMA(U

P),and

STRU

CTURE(ST)clustersareprovided.For

anindividualtobe

assigned

toaST

RUCTUREcluster,itmusthavegreaterthan60

%lik

elihoodof

beingamem

bero

fthe

cluster.Ifan

individualisassigned

totwo

clusters,thatindividualhadless

than

60%

likelihoodof

beingamem

berof

asinglecluster.Ifaspeciesor

cultivaridentificationwas

changedas

aresultof

thestudy,theprevious

misidentificationis

indicatedin

parentheses.Fo

raccessions

thathadunknow

nprovenance

orname,thelik

elyidentificationisindicatedbasedon

lineage

and/or

genotype

match,and

“unk”isin

parentheses


DNA extraction and SSR marker analysis

DNA extraction, PCR amplification of 19 microsatellite loci,electrophoresis, and scoring of the samples were performed asdescribed in Witherup et al. (2013). Ploidy levels of someindividuals were known from chromosome squashes (Ragone2001). To assess ploidy in the other individuals, any individ-ual with three alleles at multiple microsatellite regions wasdesignated as triploid. Genotype-based ploidy assignment wasconsidered to be accurate, as the vast majority of knowntriploid individuals displayed three alleles for at least threeloci (deviations are discussed in “Results”). In addition, 174 ofthe individuals were known to be either seeded (indicatingdiploidy) or seedless (frequently indicating triploidy), andthese data matched ploidy assignments in virtually allinstances (deviations discussed in “Results”).

Data analyses

Resolving allele dosage in partial heterozygotes is challengingin triploid samples. The presence of two alleles at a givenlocus could signify either a genotype with allele x representedtwice or allele y represented twice (i.e., xxy vs. xyy). Variousmethods exist for estimating allele copy number, but they aremeant for samples with uniform, even-numbered ploidy and aknown selfing rate (Clark and Jasieniuk 2011; De Silva et al.2005), assume that either allele has an equal chance of beingpresent in two copies (Hardy and Vekemans 2002; Tomiuket al. 2009), or require data that can be normalized to completeheterozygotes (Esselink et al. 2004). Therefore, we did notimpute a third allele in the case of partial heterozygotes(McGregor et al. 2000; Mengoni et al. 2000; Creste et al.2004).

GenoDive (Meirmans and Van Tienderen 2004) was usedto calculate genetic diversity measures and genetic distance.Genetic distances were calculated using the Bruvo method, adistance measure suited to codominant marker data in popu-lations of mixed ploidy (Bruvo et al. 2004). UPGMA andneighbor-joining trees were then constructed using the neigh-bor program from PHYLIP package 3.69 (Felsenstein 2005)and visualized using FigTree 1.4.0. Analyses were run with all349 samples and also excluding hybrids. Principal componentanalysis was performed in GenoDive (Meirmans and VanTienderen 2004) and visualized using the ggplot2 visualiza-tion package for R (Wickham 2009).

The Bayesian clustering analysis software STRUCTUREv2.3.4 (Pritchard et al. 2000; Falush et al. 2007) was used tovisualize genetic structure and subdivisions (number of genet-ic clusters, K) among samples. In order to analyze the alleledata from both diploid and triploid samples simultaneously,the recessive alleles option in STRUCTURE was set to one toallow for ambiguity in genotypes, and diploid individualswere scored as missing data for the third allele. We carried

out 20 independent runs per K using a burn-in period of 100,000 and collected data for 100,000 iterations forK=1–15. Theminimum value of K that can explain the data was assessedusing the rate of change in the log likelihood probability ofdata between corresponding K values (DK) as detailed inEvanno et al. (2005).

The hybrid index, a quantitative estimate of the geneticcontribution of two parental species, was calculated for hybridcollections using GenoDive (Meirmans and Van Tienderen2004), extending the maximum likelihood approach ofBuerkle (2005) to include polyploid individuals.

GenoDive (Meirmans and Van Tienderen 2004) was usedto assign lineage groups and genotypes. Samples wereassigned the same genotype only if they were identical at allloci. Since allele dosage in triploids with two alleles could notbe reliably distinguished, any instances of such partial hetero-zygotes were considered identical, leading to a possible un-derestimation of unique genotypes. Samples were assignedthe same lineage group if the genotype of one could betransformed to the genotype of the other within a thresholdnumber of mutation steps. The threshold distance was setindependently for each ploidy level, since the triploids weremuch less diverse. Pairwise genetic distances clustered asexpected for both ploidy levels, with a distribution boundedat zero from those relationships that are putatively clonal and anormal distribution consisting of the distances between sib-lings. Thresholds were set to include all contiguous distancesgreater than 0 with frequency greater than 1. The lineagethreshold for triploids was set at 16, while the diploid thresh-old was set at 8.

The polymorphic information content (PIC) for each locuswas calculated as follows:

1−Xn

i¼1p2i −

Xn−1

i¼1

Xn

j¼1þ12p2i p

2j

where pi and pj are the frequency of two given alleles i and j(Botstein et al. 1980). In order to evaluate data from bothploidy levels concurrently, the allele frequency for a givenallele i was calculated as pi ¼ c12þci3N , where cin is the numberof occurrences of allele i among individuals with ploidy n andN is the number of all non-null alleles observed.

Results

Diversity characterization

When considering each taxon separately, the average numberof alleles across all 19 loci ranged from 1.74 in


A. mariannensis to 7.37 in diploid breadfruit (Table 2). Allelesper locus ranged from 1 (in MAA3 for all taxa exceptA. camansi) to 16 (in MAA201 for A. camansi), and acrossall taxa, the average number of alleles was 11.63, with a rangeof 2 (in MAA3) to 24 (in MAA201) (Table 2). Gene diversitywas greatest in triploid hybrids (He=0.62), followed by dip-loid A. altilis (He=0.61), triploid A. altilis (He=0.52),A. camansi (He=0.41), diploid hybrids (He=0.38), andA. mariannensis (He=0.18). Averaged across all taxa, genediversity was 0.44 (Table 2).

Lineage and genotype groups

The 349 individuals analyzed represented 197 distinct geno-types sorted into 129 lineages (Table 1). All lineage groupsand genotypes were comprised of individuals of the samespecies, with the exception of one lineage group (lineage 4)that included A. altilis as well as hybrids. Among the 189triploid individuals (representing 140 accessions of A. altilisand hybrids) there were 74 genotypes, 50 of which weresingletons present in only a single individual (Table 1).Among the 150 A. altilis triploid individuals (representing112 accessions), there were 42 genotypes sorted into 8 lineagegroups. However, there was a single dominant genotype

(genotype 1) that accounted for 75 individuals (representing62 accessions) and was part of lineage group 1, whichaccounted for 130 individuals (representing 111 accessions).The members of this lineage group have provenance fromEastern Polynesia and Micronesia, with a few from Barbados,the Seychelles, or of unknown provenance. Among the 39A. altilis×A. mariannensis triploid individuals (representing27 accessions), there were 30 genotypes sorted into 15 lineagegroups.

Compared to triploids, the diploid accessions were morereadily distinguishable from one another based on microsat-ellite profiles (Table 1). Among the 160 diploid individuals(representing 113 accessions across all four taxa), there were115 genotypes, 111 of which were represented by a singleindividual in the collection. The genotypes were sorted into106 lineages, 89 of which contained only one accession. Therewere no genotypes or lineages shared across different taxaamong the diploids. Among the 36 A. camansi individuals(representing 21 accessions), there were 35 genotypes sortedinto 34 lineage groups. Among the 14 A. mariannensis indi-viduals (representing 7 accessions), there were 13 genotypessorted into 11 lineage groups. Among the 79 A. altilis diploidindividuals (representing 63 accessions), there were 47 geno-types sorted into 33 lineage groups. Among the 31 diploid

Fig. 1 Map of collections used in the study. Inset in lower left showscollections outside of Oceania with sample size in parentheses. The areahighlighted in gray on the inset map is the focus of the larger Oceania

panel, which has island groups indicated and the following regionsoutlined: East Polynesia. West Polynesia, Micronesia, and Melanesia


Tab

le2

Geneticdiversity

inbreadfruitandrelativ

esbasedon

19microsatelliteloci

Primer

A.altilis

N=231

A.altilis×A.m

ariannensisN=69

Diploid

N=79

TriploidN=152

Diploid

N=31

TriploidN=38

NA

ASR

He

NA

ASR

He

PIC(%

MD)

NA

ASR

He

NA

ASR

He

PIC

(%MD)

MAA3

1216

01

216

00(0)

1216

01

216

00(0)

MAA9

4153–173

0.1

5153–171

0.103

0.176(2.9)

4163–171

0.361

6161–171

0.602

0.375(3.2)

MAA40

8170–188

0.792

6180–192

0.52

0.579(0.3)

2182–186

0.032

3180–190

0.417

0.478(0)

MAA54a

9167–195

0.774

10173–195

0.583

0.657(0)

2173–187

0.228

4173–187

0.654

0.482(0.6)

MAA54b

3205–215

0.311

2207–215

0.5

0.309(0.8)

2205–207

0.032

3205–207

0.368

0.214(0)

MAA71

10154–184

0.808

8152–178

0.561

0.646(0.5)

4152–182

0.238

5152–182

0.694

0.516(1.3)

MAA85

7154–178

0.773

5154–164

0.54

0.573(2.1)

3154–164

0.448

4158–164

0.54

0.625(0.6)

MAA96

8176–214

0.785

6204–214

0.581

0.638(2.2)

4204–220

0.548

6204–220

0.751

0.717(1.3)

MAA122

9241–293

0.699

7277–295

0.71

0.750(0.2)

3285–291

0.404

6277–291

0.711

0.596(0)

MAA135

11258–300

0.804

10268–322

0.722

0.770(0.2)

7270–320

0.5

14268–328

0.846

0.770(0.6)

MAA145

9256–304

0.662

9262–328

0.538

0.595(0.9)

6282–328

0.698

8268–304

0.76

0.740(3.9)

MAA156

10273–307

0.523

6273–307

0.547

0.600(2.4)

4279–307

0.384

6277–309

0.692

0.730(1.3)

MAA178a

9207 –235

0.788

7209–229

0.549

0.582(10.7)

5209–229

0.582

8209–229

0.779

0.798(2.5)

MAA178b

9241–259

0.729

8241–259

0.558

0.563(1.7)

5241–253

0.706

6241–253

0.793

0.690(1.3)

MAA182

6182–214

0.526

7182–212

0.541

0.619(1.7)

5200–210

0.72

6182–210

0.72

0.719(1.9)

MAA201

9238–288

0.491

11262–294

0.704

0.773(1.1)

4262–278

0.585

9262–296

0.798

0.767(0.6)

MAA219

6247–277

0.708

5259–277

0.518

0.577(0.5)

4259–271

0.292

4259–277

0.413

0.595(0.6)

MAA287

9179–223

0.686

7179–215

0.546

0.501(1.1)

3183–211

0.153

5179–199

0.695

0.519(0)

MAA293

3158–166

0.591

5154–174

0.519

0.428(0.9)

3160–166

0.371

3160–166

0.646

0.485(2.5)

Average

7.368

0.608

6.579

0.518

0.544(1.6)

3.737

0.383

5.632

0.625

0.569(1.2)

A.cam

ansiN=36

A.m

ariannensisN=14

Overall

Primer

NA

ASR

He

PIC(%

MD)

NA

ASR

He

PIC(%

MD)

NA

PIC

(%MD)

He

MAA3

2216–218

0.08

0.077(0)

1216

00(0)

20.007(0)

0.02

MAA9

2161–171

0.131

0.077(0)

2164–168

0.423

0.238(0)

100.430(2.6)

0.305

MAA40

2180–182

0.106

0.054(0)

1182

00(0)

100.664(0.2)

0.271

MAA54a

8173–185

0.797

0.627(0)

1173

00(0)

120.712(0.1)

0.528

MAA54b

1207

00(0)

1207

00(0)

40.278(0.6)

0.186

MAA71

5154–160

0.322

0.245(0)

1152

00(0)

140.736(0.6)

0.41

MAA85

2156

0.081

0(0)

1162

00(0)

80.694(1.6)

0.366

MAA96

4208–218

0.666

0.534(0)

1204

00(0)

100.713(1.8)

0.513

MAA122

3279

0.055

0(0)

2289–291

0.349

0.218(0)

110.780(0.1)

0.448

MAA135

9278–302

0.806

0.603(0)

4280–326

0.64

0.221(0)

220.814(0.2)

0.75

MAA145

7268–320

0.522

0.424(0)

3282–304

0.071

0.166(0)

200.704(1.3)

0.518

MAA156

6257–279

0.724

0.579(0)

2281–307

0.254

0.178(0)

140.718(1.9)

0.549


hybrid individuals (representing 22 accessions), there were 31genotypes sorted into 29 lineage groups.

Only a few lineages or genotypes were shared acrossregions. When considering only the breadfruit (A. altilis) andhybrids (A. altilis×A. mariannensis), one lineage was sharedacross all regions in Oceania (lineage 37); another lineage wasshared acrossWestern and Eastern Polynesia, Micronesia, andareas outside of Oceania (lineage 1); two lineages were sharedbetween Western and Eastern Polynesia (lineages 2 and 33),two lineages were shared across Eastern Polynesia and Mi-cronesia (lineages 4 and 5), and two lineages were sharedacross Western Polynesia and Melanesia (Lineages 27 and42) (Fig. 2a). When considering genotypes, one genotype(lineage 37/genotype 90) was shared across all regions inOceania; one genotype was shared across Micronesia andEastern Polynesia (lineage 1/genotype 23); one genotypewas shared across Eastern Polynesia, Micronesia, and areasoutside of Oceania (lineage 1/genotype 1); one genotype wasshared across Melanesia and Western Polynesia (lineage 27/genotype 78); one genotype was shared across Western andEastern Polynesia (lineage 1/genotype 3); and one genotypewas shared across Eastern Polynesia and areas outside ofOceania (lineage 1/genotype 5) (Fig. 2b).

Identification of unknowns

Twenty-eight accessions had unknown cultivar names, and 13of them were also of unknown provenance. Among the 28unknowns, 12 (43 %) shared identical genotypes to an acces-sion(s) with known provenance and cultivar name(s)(Table 1). Nine (32 %) of the unknown accessions shared alineage group but not a genotype with other known acces-sions, and seven (25 %) unknowns shared neither lineagenor genotypes with other accessions, suggesting they aregenetically unique within the collection.

Cultivar names

There were 136 different cultivar names recorded for thecultivated breadfruit and hybrids analyzed in this study. Halfof these names (68) were singletons, only represented by asingle individual. Among the other 68 names with multipleindividuals, 40 names were represented each by 2 individuals,16 names were represented each by 3 individuals, 5 nameswere represented each by 4 individuals, and 3 names wererepresented each by 5 individuals. There were four differentnames that were represented by 6, 7, 13, and 18 individuals.Only one name occurred in more than one island group withina region: Puou (Tonga and Samoa in Western Polynesia).Puou also occurred in Vanuatu (Melanesia), but this is knownto be an introduction from a Samoan variety. Of the 68 namesrepresented by more than 1 individual, only 33 names consis-tently shared the same genotypes with other individualsTa

ble2

(contin

ued)

MAA178a

5211–245

0.16

0.104(0)

2223–245

0.304

0.207(7.1)

140.741(8.1)

0.477

MAA178b

6241–257

0.621

0.519(3.0)

3251–255

0.588

0.367(7.1)

100.648(1.9)

0.661

MAA182

6182–210

0.785

0.712(0)

2202–204

0.304

0.201(0)

100.662(1.6)

0.604

MAA201

16268–312

0.925

0.808(0)

3266–276

0.405

0.274(0)

240.838(0.9)

0.695

MAA219

4256–277

0.607

0.461(0)

1260

00(0)

90.654(0.4)

0.405

MAA287

2179

0.028

0(0)

1183

00(0)

110.632(0.8)

0.319

MAA293

3162–166

0.319

0.214(0)

1160

00(0)

60.492(1.1)

0.363

Average

4.895

0.407

0.318(0.2)

1.737

0.176

0.109(1.4)

11.632

0.627(1.4)

0.441

Na,Num

berof

alleles;ASR

,allelesize

range(bp),H

e=expected

heterozygosity,P

IC(%

MD)=

polymorphicinform

ationcontent(%

missing

data.)


bearing the same name, and 24 of those 33 were members ofthe same base accession number. Eight of those 33 names allshared the most ubiquitous lineage group and genotype (1/1).The remaining nine names, represented by more than oneindividual and consistently sharing the same genotype, share

their respective genotypes across different base accessionnumbers: Afara, Enua, Mei chon, Meion, Meinpadahk, Niue,Otea, Puaa, and Ulu tala. Among the 75 individuals with theubiquitous genotype (lineage 1/genotype 1), 44 differentnames were represented.

250

500

1000

1500km

Genotype Distribution

B

250

500

1000

1500km

Lineage Distribution

A

East Polynesia (119)

East Polynesia (119)

West Polynesia (53)

West Polynesia (53)

Micronesia (66)

Micronesia (66)

Melanesia (41)

Melanesia (41)

Non-Oceania (Barbados-1/Seychelles-7)

Non-Oceania (Barbados-1/Seychelles-7)

Equator

Equator

Fig. 2 Distribution across Oceania of lineages and genotypes in A. altilisand A. altilis×A. mariannensis hybrids based on microsatellite data from19 loci. Pie charts show regional distributions of lineages (a) andgenotypes (b). Within a pie chart, solid gray shades indicate uniquegenotypes that are not shared across regions. Patterned wedges that are

pulled out indicate lineages or genotypes that are shared across regions.Arrows indicate shared lineages or genotypes across regions (thinarrows=one shared group and thick arrows indicate two sharedgroups). Patterened circles on the arrows indicate the wedge(s) sharedacross regions. Sample sizes for each region are indicated in parentheses


Genetic distance

The results of the UPGMA and neighbor-joining analyseswere nearly identical and only the UPGMA tree is discussedhere. Analyses without hybrids resulted in each taxon makingup its own cluster with the A. camansi and A. altilis clusterstogether forming a larger cluster with the A. mariannensiscluster next to it (not shown). This is consistent with previousstudies based on AFLP (Zerega et al. 2005). In the UPGMAtree with all samples included, each taxon largely clusteredwith other members of the same taxon, with hybrids largelyclustered between A. altilis and A. mariannensis (Fig. 3). Allsamples identified as A. camansi clustered together with twospecimens (890455.001 of unknown provenance and900228.001 from Samoa) that had previously been classifiedas A. altilis. Upon closer examination of the two specimens,we determined that they share many characteristics withA. camansi and their identifications have been changed toA. camansi. The A. camansi cluster was split into twosmaller clusters, one cluster made up of samples fromPapua New Guinea and the other cluster made up of

samples from elsewhere. With the exception of one sam-ple (000528.001 from Pohnpei), all specimens previouslyidentified as A. mariannensis clustered together. Theexception was nested in a cluster containing mostly hy-brids as well as triploid and diploid A. altilis fromislands throughout Oceania. Upon close examination ofthe specimen, we determined that his accession shareshybrid characteristics with greater contribution fromA. mariannensis.

Principal component analysis

Plotting of the first two principal components of the microsat-ellite profiles shows segregation by species, with hybridsclustering to some degree between parental species (Fig. 4).When regional samples are analyzed separately, most Micro-nesian and Western Polynesian hybrids cluster between theparental species or cluster more closely to A. mariannensis(Fig. 5), and in Eastern Polynesia, hybrids cluster much moreclosely to A. altilis (Fig. 5).

Fig. 3 UPGMA tree of A. altilis, A. altilis×A. mariannensis, A. camansi,and A. mariannensis based on data from 19 microsatellite loci. Clustermembers are indicated in the legend. The area within cluster G that is inthe shaded gray circle is the 75 individuals belonging to the ubiquitous

lineage 1/genotype 1. The filled circle on the node in cluster A denotessamples from outside of Papua New Guinea and the open circle denotessamples from Papua New Guinea. Cluster designations are also includedin Table 1. Bar of Bruvo distance 0.03 shown for scale


Hybrid index

Examining the hybrid index (h) across the 69 individualsidentified as hybrids based on morphological characters(Table 1) revealed a similar pattern evident from princi-pal component analysis (PCA). Individuals showinggreater contributions from A. mariannensis (h0.7) were predominantly from Micronesiaand Eastern Polynesia.

Structure

In the STRUCTURE analysis, the modal value of the distri-bution of the true K identified a peak at K=5, which wassupported by large shifts in L(K) and Ln’(K) from K=5 toK=6 associated with true value of K, as described in Evannoet al. (2005). All A. mariannensis individuals group togetheras do all the A. camansi individuals (Fig. 6). A. altilis speci-mens were subdivided into three groups that can generally becharacterized as Melanesian breadfruit (predominantly dip-loid), breadfruit from throughout Oceania (diploid and

triploid), and triploid breadfruit from Eastern Polynesia andMicronesia. Diploid Micronesian hybrids almost exclusivelyshared the same group as A. mariannensis, while triploidMicronesian hybrids were admixtures of A. mariannensisand the triploid Micronesian/Eastern Polynesian breadfruitgroups. Western Polynesian hybrids either had dominant con-tributions from A. mariannensis or were admixtures betweenA. mariannensis and the diploid/triploid trans-Oceania bread-fruit group. Some of the diploid hybrids from Western Poly-nesia and Micronesia only shared the A. mariannensis groupwith no apparent admixture with A. altilis.

Utility of microsatellite markers

The PIC of the microsatellite loci across all taxa ranged from0.007 to 0.838, with an average value of 0.627 (Table 2). Theextent to which each locus was informative varied by taxon,with loci generally being the most informative for A. altilisand hybrids. A codominant marker is generally classified ashighly informative if its PIC >0.5 (Botstein et al. 1980;Ghislain et al. 2004). Using this criterion, 15 markers werehighly informative within A. altilis, 13 within hybrids, 7within A. camansi, and none within A. mariannensis. If onlythe genotypes from the 10 most informative loci were ana-lyzed, 111 out of 123 unique diploid genotypes (90 %) and

Fig. 4 PCA results based on datafrom 19 microsatellite loci ofbreadfruit, hybrids, and wildrelatives from throughoutOceania. Taxon coding isindicated in the legend, and 95 %bivariate confidence intervalellipses are shown for taxonclustering


100 % of 75 triploid genotypes could still be distinguished,and all taxa could be fully distinguished (i.e., no two acces-sions of different species shared the samemicrosatellite profilebased on these 10 markers). Only primer MAA3 had zerodistinguishing utility.

A previous study examined AFLP profiles of breadfruitand its relatives using three primer pairs across 313 individ-uals (Zerega et al. 2004), 181 of which were included in thepresent study. Among this 181 subset of individuals, 156distinct AFLP and 110 distinct microsatellite genotypes weredistinguished. Given the nature of the PIC equation,

comparing PICs between microsatellites and AFLPs is notinformative. Because AFLP fragments are not mutually ex-clusive, the information of an AFLP primer pair can changedrastically while the relative frequencies of fragments remainthe same. On the basis of number of markers, the AFLP assayswere generally more informative (Table 3).

Zymotypes established from the isozyme profiles of sixenzymes were available for 140 of the individuals character-ized here (Ragone 1991; Zerega et al. 2005). Among these140 individuals, there were 54 distinct zymotypes and 88distinct microsatellite genotypes. While the average enzyme

Fig. 5 PCA results based on data from 19 microsatellite loci ofbreadfruit, hybrids, and wild relatives from throughout Oceania.Samples were analyzed by region with A. mariannensis and A. camansi

included in each regional analysis. Taxon coding is indicated in thelegend, and 95 % bivariate confidence interval ellipses are shown fortaxon clustering


was somewhat informative (Table 3), 51 of the 140 individ-uals shared the same zymotype profile. Zymotype profiles foroverlapping individuals were less informative thanmicrosatellites, as measured by both PIC and number ofpolymorphisms (Table 3).

Comparison of lineage and genotype groups across the sameaccession

Not all individuals of the same base accession number sharedthe same lineage or genotype group. This was especially trueamong the seed-propagated diploids, which never shared thesame genotype, and frequently represented different lineagegroups. This was expected given that the diploid accessionswere largely seed

diversity in the breadfruit complex (artocarpus, moraceae...

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