Rapid analysis of seed size in Arabidopsis for mutant and QTL discovery.

Department of Biochemistry, University of Otago, PO Box 56, Dunedin 9054, New Zealand. .
Plant Methods (Impact Factor: 2.67). 02/2011; 7(1):3. DOI: 10.1186/1746-4811-7-3
Source: PubMed

ABSTRACT Arabidopsis thaliana is a useful model organism for deciphering the genetic determinants of seed size; however the small size of its seeds makes measurements difficult. Bulk seed weights are often used as an indicator of average seed size, but details of individual seed is obscured. Analysis of seed images is possible but issues arise from variations in seed pigmentation and shadowing making analysis laborious. We therefore investigated the use of a consumer level scanner to facilitate seed size measurements in conjunction with open source image-processing software.
By using the transmitted light from the slide scanning function of a flatbed scanner and particle analysis of the resulting images, we have developed a method for the rapid and high throughput analysis of seed size and seed size distribution. The technical variation due to the approach was negligible enabling us to identify aspects of maternal plant growth that contribute to biological variation in seed size. By controlling for these factors, differences in seed size caused by altered parental genome dosage and mutation were easily detected. The method has high reproducibility and sensitivity, such that a mutant with a 10% reduction in seed size was identified in a screen of endosperm-expressed genes. Our study also generated average seed size data for 91 Arabidopsis accessions and identified a number of quantitative trait loci from two recombinant inbred line populations, generated from Cape Verde Islands and Burren accessions crossed with Columbia.
This study describes a sensitive, high-throughput approach for measuring seed size and seed size distribution. The method provides a low cost and robust solution that can be easily implemented into the workflow of studies relating to various aspects of seed development.

  • [Show abstract] [Hide abstract]
    ABSTRACT: A variety of imaging methodologies are being used to collect data for quantitative studies of plant growth and development from living plants. Multi-level data, from macroscopic to molecular, and from weeks to seconds, can be acquired. Furthermore, advances in parallelized and automated image acquisition enable the throughput to capture images from large populations of plants under specific growth conditions. Image-processing capabilities allow for 3D or 4D reconstruction of image data and automated quantification of biological features. These advances facilitate the integration of imaging data with genome-wide molecular data to enable systems-level modeling.
    Trends in Plant Science 01/2014; · 11.81 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Plant phenotyping technology has become more advanced with the capacity to measure many morphological and physiological traits on a given individual. With increasing automation, getting access to various traits on a high number of genotypes over time raises the need to develop systems for data storage and analyses, all congregating into plant phenotyping pipelines. In this review, we highlight several studies that illustrate the latest advances in plant multi-trait phenotyping and discuss future needs to ensure the best use of all these quantitative data. We assert that the next challenge is to disentangle how plant traits are embedded in networks of dependencies (and independencies) by modelling the relationships between them and how these are affected by genetics and environment.
    Current Opinion in Plant Biology 03/2014; 18:96-102. · 8.46 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Stimulus-specific calcium (Ca(2+) ) signals crucially function in developmental processes in many organisms and are deciphered by various Ca(2+) binding proteins. In Arabidopsis thaliana, a signaling network consisting of Calcineurin B-like protein (CBL) calcium sensors and CBL-interacting protein kinases (CIPKs) has been shown to fulfill pivotal functions at the plasma membrane in regulating ion fluxes and abiotic stress responses. However, the role of tonoplast localized CBL proteins and especially their function in regulating developmental programs has remained largely unknown. In this study we analyzed single and double mutants of the closely-related tonoplast localized calcium sensors CBL2 and CBL3 with either reduction of function (rf) or complete loss of function (lf). While single cbl2 or cbl3 mutants did not display discernable phenotypes, cbl2/cbl3 mutants exhibited defects in vegetative growth and were severely impaired in seed development and morphology. Seeds of the cbl2/3rf mutant were smaller in size and exhibited reduced weight and fatty acid content compared to wild type while accumulation of sucrose was not altered. Moreover, accumulation of inositol hexakisphosphate (InsP6 ), the major storage form of phosphorus in seeds, was significantly reduced in mutant seeds. In addition, complete loss of CBL2 and CBL3 function in cbl2/3lf resulted in a high frequency of severe defects in embryonic development. Together our findings uncover a crucial function of Ca(2+) controlled processes at the vacuolar membrane as determinants of seed yield and size and reveal the importance of vacuolar CBL calcium sensors for plant embryogenesis. This article is protected by copyright. All rights reserved.
    The Plant Journal 01/2014; · 6.58 Impact Factor

Full-text (2 Sources)

Available from
Jun 1, 2014

Similar Publications