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Isolation of genes in peas involved in the transition to flowering - AR0125

Description
This proposal is relevant to ROAME A Pea Breeding R&D policy to improve the economic competitiveness and output of legumes to meet the likely increase in demand for vegetable-based proteins in the future. The objectives, to improve crop yield potential and harvestability through the development of lines with a shorter, defined, pod-forming period, will be met by isolating genes involved in the transition to flowering. Characterisation of the role of each gene in the floral transition will lead to an improved understanding of the synchrony of flowering in pea and this knowledge will be exploitable for the development of more competitive varieties. It is anticipated that the genes characterised will be present in other crop species, thus the resulting understanding will be widely applicable.

Objective
Objective 01 Isolation of genes essential to the transition to flowering.
The UNI gene will be cloned into a yeast expression vector (Objective 01/01/01) and cDNA synthesised from shoot apex mRNA will be cloned into a different vector (Objective 01/01/02). Both vectors will be used in a yeast two hybrid system [8] that will assay for interaction between UNI and a cDNA product (Objective 01/01/03). Interacting cDNA products will be mapped to ascertain their position relative to known flowering transition genes eg. Veg2, Det (Objective 01/02/01) and any that correspond will be characterised further (see 02 below). Bona fide gene products will be assayed for interactions with each other using the same yeast two hybrid assay (Objective S01/03/01).

Objective 02 Characterisation of genes essential to the transition to flowering.
Clones will be sequenced to enable a function search of DNA databases to be made (Objective 02/01/01). Full-length cDNAs will be obtained (and sequenced) if the interacting cDNA products isolated were truncated versions (Objective 02/02/01). Cosegregation of a cDNA with a floral transition mutation will be tested in an independent, large segregating population (Objective S02/03/01). Sequence analysis of mutant and wild-type alleles will be carried out to characterise the genetic lesion corresponding to the mutation (Objective S02/04/01). RNA in situ hybridisation analysis will be carried out to visualise the timing and spatial distribution of response gene activity in wild-type shoot apices before and during flower development, under long and short day light conditions (Objective 02/05/01). In situ analysis of response gene expression in different mutant shoot apices will determine further gene interactions in the floral transition cascade (Objective 02/05/02).
Project Documents
• Final Report : Isolation of genes in peas involved in the transition to flowering   (758k)
Time-Scale and Cost
From: 1998

To: 2002

Cost: £229,691
Contractor / Funded Organisations
John Innes Centre (BBSRC)
Keywords
Arable Farming              
Biotechnology              
Crop Improvement              
Crops              
Farming              
Fields of Study
Arable Crops