Abstract

The main objective of this research project is to use and further develop molecular tools as screening methods to determine the genetic identity of polyploid and introgressed populations.

The use of different complementary approaches on the same carefully sampled material will enable accurate appraisal of the usefulness of the different techniques in:

• Genotyping polyploid individuals.
• Revealing hybrid identity.
• Estimating the extent of introgression in natural populations.

All of these are very important issues in conservation genetics. Willow species and hybrids at a European scale will be used as a case-study. The deliverables are at both the level of the molecular tools and their performance and at the landscape restoration level. Willows dominate the landscape in many riparian ecosystems, valleys, meadows and act as a green screen in agricultural landscapes. The evaluation of these techniques in hybridisation-related research will allow different end-users to apply this knowledge.

Objectives

The objective is to use and further develop molecular tools as screening techniques to determine the genetic identity of introgressed populations of polyploid plants. Complementary approaches on the same material will enable accurate appraisal of the utility of each techniques in:

• genotyping polyploid individuals,
• revealing hybrid identity, and
• estimating the extent of introgression in natural populations.

These objectives are highly important issues for the progression of genetics in polyploids. In this project, Salix (willow) species and their hybrids will be used as the model plants. Recommendations for appropriate use of techniques will be invaluable in the selection of material for stock establishment, breeding lines and germplasm collections and in the development of management strategies for the utilisation of germplasm at the landscape level. The project will evaluate the molecular techniques in answering questions relating to polyploid heterozygosities and introgressive hybridisation.

Description of the work

The molecular tools that will be included as screening methods in this study are:

• A selection of AFLPs (Amplified Fragment Length Polymorphisms) and RAPDs (Random Amplified Polymorphic DNAs) (partner 01).
• Enzyme consensus primers of conserved sequences (partner 01).
• Nuclear SSRs (Simple Sequence Repeats) (partner 02).
• cpDNA (chloroplast) and mtDNA (mitochondrial) sequence analysis (partner 03).
• cpSSRs (chloroplast Simple Sequence Repeats) (partner 04).

The total time span of the project is 36 months, including 30 months for the fulfilment of the experimental work packages and the remaining 6 months for accomplishing the data treatment, reports and working out guidelines.

Deliverables

Species-specific markers will be selected. F1 hybrids and introgression will be genotyped. The quality, complementarity, compatibility and comparability of the molecular tools will be evaluated for RAPD and manual AFLP profiles, consensus primers of particular enzymes (ADH), nuclear willow SSRs, polymorphic cpSSR markers and cp- and mtDNA-related data using consensus primers. The geographical pattern of these landscape-dominating trees and the dynamic nature of hybridisation will be assessed from discriminative molecular information. Dissemination of the results through the BBP of the EU will inform a broad range of end-users.

Contacts

Coordinator

• Ludwig TRIEST / Vrije Universiteit Brussel Group of General Botany and Nature Management / BELGIUM