Marker Assisted Breeding and Selection for Disease Resistance in Tulips

Paul J.J. Bijman, Arwa Shahin, Paul Arens and Jaap M. van Tuyl

Introduction: Yearly 10.000 hectares of tulip bulbs are being grown. During tulip bulb cultivation excessive amounts of fungicides are being used to reduce loses of Fusarium oxysporum and Botrytis tulipae, and insecticides are heavily being applied to reduce the transmission of the Tulip Breaking Virus (TBV) by aphids. Because of the long juvenile phase (5 years), there is a need to select for the resistance traits at an early phase and to introgress these disease resistance traits in a cost effective manner. In this study, a population segregating for these three diseases is used to locate QTLs associated with these traits. With the use of new sequencing technology a genetic map will be developed.
Plant material: In 1989 a cross between cultivars T.
gesneriana Kees Nelis (Fig 1a) and T. fosteriana Cantata (Fig 1b) was made. The obtained population consists of 125 individuals (Fig 1c). This population is segregating for the three main diseases. In Cantata resistance against TBV and B. tulipae can be found. In Kees Nelis resistance against F. oxysporum can be found. a

b c Fig 3: a. Wound inoculation four wounds first leaf two wound second leaf b. Cantata after 4 days showing hardly any infection c. Kees Nelis heavily infected after 4 days.

Tulip Breaking Virus: TBV is being transmitted in a

non-persistent way by aphids. Resistance against TBV seems to be based on a single gene. The trait segregates in a 1:1 ratio. The population has already been tested for TBV resistance by growing them in a plot with high amounts of TBV infected tulips without aphid control. Virus a b c Fig 1: a. T. gesneriana Kees Nelis b. T. fosteriana Cantata c. siblings of the cross infection has been determined in the population visually between Kees Nelis crossed with Cantata. and by ELISA assay.

Disease tests
Fusarium oxysporum: Two different experiments will be
performed to determine the resistance level against F. oxysporum (Fig 2a). The first barrier of defence against this pathogen is the bulb skin. Therefore, skin quality will be scored in the population (Fig 2b, c). A correlation between Fusarium resistance and skin quality will be evaluated.

a b c Fig 4: a. Symptoms of TBV infection in the flower b. the leaves c. the flower stigma. (Pictures: PPO-Bloembollen)

Molecular markers: SNP markers will be developed

from sequencing data of five tulip cultivars. These markers will be used to construct a genetic map. The population, different species within the section Tulipa and a collection of popular cultivars will be screened for SNPs with the use of the Golden Gate Assay. QTL mapping will be performed with the results of the disease experiments in order to identify genomic regions associated with resistance. Based on these results identified markers linked to the traits will be used in marker assisted selection.

a b c Fig 2: a. Result after the Fusarium experiment b. good skin quality c. poor skin quality.

Botrytis tulipae: A new disease test has been developed to

determine B. tulipae resistance. After forcing early flowering in the greenhouse, detached leaves are wound-inoculated with B. tulipae. Lesion diameter is measured after 4 days (Fig 3 a). Complete resistance can be found in Cantata (Fig 3 b), whereas Kees Nelis is susceptible (Fig 3 c).
For correspondence: Paul Bijman Plant Research International, Plant Breeding Wageningen UR P.O. Box 16, 6700 AA Wageningen, The Netherlands Tel: +31 317 486081 Fax: +31 317 483457 Email: paul2.bijman@wur.nl