Using CWR and pre-breeding potato

Paulina Smyda-Dajmund, Jadwiga Śliwka, Ewa Zimnoch-Guzowska

Plant Breeding and Acclimatization Institute – NationalResearch Institute (IHAR-PIB), Młochów Research Center,

Poland

Potato parental line breeding in Poland

• Diploid level

• Tetraploid level –parents of 72 Polishvarieties

IHAR-PIB Młochów ResearchCenter 2

The scheme of diploid potato breeding at IHAR-PIB

3Zimnoch-Guzowska and Flis 2021 Potato Research

4

Main and supporting programmes of diploid breeding at IHAR-PIB

Zimnoch-Guzowska and Flis 2021 Potato Research

TraitChromosome, location of the R gene or QTL

I II III IV V VI VII VIII IX X XI XII

Resistance to viruses NsNy-Smira

Ny-1Gm

PLRV1Ny-2Rm

PLRV4

Ryf-sto

Resistance toS. endobioticum

Sen2

Resistance to pectinolytic bacteria 2000 (QTL)

Resistance to pectinolytic bacteria 2021 (QTL)

Resistance toP. infestans (QTL)

Resistance to P. infestans R2-like Rpi-mch1 Rpi-phu1 Rpi-rzc1 Rpi-Smira1

Potato resistance genes and QTL mapped atIHAR-PIB

Marczewski et al. 2001, 2002, 2004, 2006; Flis et al. 2005; Szajko et al. 2019; Plich et al. 2018; Zimnoch-Guzowska et al. 2000; Lebecka et al. 2021; Śliwka et al. 2007, 2006, 2012a, 2012b

5

TraitChromosome, location of the major genes or QTL

I II III IV V VI VII VIII IX X XI XII

Flower colour F

Maturity

Tuber morphology and dormancy

Tuber morphology

Tuber blackspot bruiseand enzymaticdiscoloration

Crisp colour

Starch-corrected crispcolour

Leaf sucrose content

Tuber starch content

Tuber greening

QTL for other traits mapped at IHAR-PIB

Śliwka et al. 2008, 2012, 2016; Sołtys-Kalina et al. 2020; Hara-Skrzypiec et al. 2018; Plich et al. 2020 6

Resistance to P. infestans

7

Genes for late blight resistance on potato genetic map

Śliwka and Zimnoch-Guzowska 2013 updated

Rpi-mcq1

Rpi-edn2 R9

Rpi-dlc1

R8/Rpi-Smira2

I

VIIIVII

VIVIVIIIII

XIIXIXIX

R1

R3a R3b R5

R6 R7 R10

R11 Rpi-sto2

Rpi-pcs1

Rpi-Smira1

Rpi-vnt1.1

Rpi-vnt1.2

Rpi-vnt1.3

Rpi-nrs1

Rpi-phu1

R2, R2-like

R2-likeBzura

Rpi-blb3

Rpi-abpt

Rpi-bst1

Rpi-mcd1

Rpi-amr3i

Rpi-blb2

Rber,

Rpi-ber1

Rpi-ber2

Rpi-blb1/RB

Rpi-sto1

Rpi-pta1

Rpi-plt1

Rpi-ver1-8

Rpi1

Rpi-rzc1

Rpi-mch1

Rpi-avl1

Rpi-cap1

Rpi-qum1

Rpi-edn3

Rpi-pur1

Rpi-vnt2

8

Virulence of P. infestans isolates from Poland (2017)

9

• 1970 S. stenotomum x S. phureja comes from CIP, Lima, Peru

crosses with S. tuberosum 2x

• 1992 first resistant tetraploid (unreduced gametes)

• 2006 mapping the gene (Śliwka et al. 2006, TAG)

• 2009 sequencing, virulent isolate P. infestans EC1; MP1162; = Rpi-vnt1.1

(Foster et al. 2009, MPMI)

• 2010 MAS (Śliwka et al. 2010, JAG)

• 2012 pyramiding with Sárpo Mira

• 2014 GMO field tests in The Sainsbury Laboratory [Jones et al.]

• 2018 Cultivar Gardena by Zamarte Potato Breeding Ltd Group IHAR

Rpi-phu1

10Solanum phureja Solanum stenotomumhttp://www.cgn.wageningen-ur.nl/pgr/collections/crops/potato/species.htm

• Originates from:Bolivia, Peru

• Syn. S. sparsipilum, re-identified as • S. brevicaule

• Selected clones:- resistant to P. infestans

– low tendency to enzymatic blackeningand after-cooking darkening

Solanum ruiz-ceballosii Cárd. 2x (2EBN) VIR8664 (VIR7370)

Ochoa 199011

rzc flower colourdH Balbina x 99-10/36 rzc

Śliwka et al. 2012 BMC Genet 13:1112

Two linkage maps:

Brylińska et al. 2015

Flower colour locus F

13Śliwka et al. 2012

Chromosom X

• Originates from:Mexico, state: Michoácan,

• Natural hybrid:S. bulbocastanum x S. pinnatisectum

(Hawkes 1990)

• Selected clones:

- resistant to P. infestans (whole leaf test)- suitable for cold chipping

Solanum × michoacanum (Bitter.) Rydb. 2x (1EBN) VIR5763

14Resistant genotypes with mean scores ≥ 6

Tuber sizes

Tetraploid Diploid S. × michoacanum

15

Transfer of resistance to cultivated potato

S. × michoacanum

somatichybridization with

S. tuberosum

mappingR genecloning

MAS backcrossing

cisgenesispotato cultivar

intraspecificcross

16

Smyda et al. 2012

Smyda-Dajmund et al. 2017

Śliwka et al. 2012 TAG

Rpi-mch1 gene was mapped to the chromosome VII of the potato genome

17

dHBardcv. Rywal

DG 81-68

99-12/899-12/39

0

5

10

15

20

25

30

35

40

45

50

1.9 2.9 3.9 4.9 5.9 6.9 7.9 8.9 9.0

Resistant genotypes with mean scores ≥ 6

autofused 4x mch

somatic hybrids

Out of 97 S. × michoacanum (+) S. tuberosum somatichybrids 2 were resistant to P. infestans

18

Nu

mb

er

of

fusi

on

pro

du

cts

Classes of resistance(Smyda-Dajmund et al. 2016)

Composition of 12 chromosomes of somatic hybrids

0

50

100

150

200

250

I II III IV V VI VII VIII IX X XI XII

99-12/39 (+) DG 81-68

mch del

tbr

mch

tbr del

19

No.

of

mark

ers

0

200

400

600

800

1000

1200

1400

1600

1800

2000

MS

43

MS

31

MS

8

MS

15

MS

10

MS

16

MS

35

MS

38

MS

32

MS

28

MS

42

MS

18

MS

37

MS

23

MS

30

MS

20

MS

24

MS

25

MS

45

MS

21

No.

of

mark

ers

Nuclear genome composition of somatic hybrids

Nuclear composition of S. × michoacanum (+) S. tuberosumsomatic hybrids based on DArT markers

Smyda-Dajmund et. al. 2016

Epigenetic analysis

20

• MSAP and HPLC methods indicated an increase in DNA methylation in the somatic hybrids in comparison to theirparents

• The somatic hybridization changed the level of cytosinemethylation in the studied potato somatic hybrids

Smyda-Dajmund et al. 2021 Agronomy 11,845

BC1 progeny from S. × michoacanum (+) S. tuberosum somatic hybrids and cv. Flaming

21Smyda-Dajmund et al. 2017

IHAR-PIB, Młochów Research CenterDepartment of Potato Genetics and Parental

LinesLaboratory of Potato Pathogens

Jadwiga Śliwka

Marta Janiszewska (Brylińska)

Zhimin Yin

Sylwester Sobkowiak

Paulina Chmiel

Mirella Ludwiczewska

Laboratory of Parental Lines and Breeding Methods

Bogdan Flis

Jarosław Plich

Beata Tatarowska

Dorota Milczarek

Laboratory of Phytopathology

Renata Lebecka

Anna Grupa-Urbańska

Krystyna Michalak

Laboratory of Biotechnology

Waldemar Marczewski

Dorota Sołtys-Kalina

Katarzyna Szajko

Paulina Smyda-Dajmund

Iwona Wasilewicz-Flis

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Thank you for your attention