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Related Concept Videos

Trihybrid Crosses02:27

Trihybrid Crosses

Trihybrid Crosses
Some of Mendel’s crosses examined three pairs of contrasting characteristics. Such a cross is called a trihybrid cross. A trihybrid cross is a combination of three individual monohybrid crosses. For example, plant height (tall vs. short), seed shape (round vs. wrinkled), and seed color (yellow vs. green).
The F1 generation plants of a trihybrid cross are heterozygous for all three traits and produce eight gametes. Upon self-fertilization, these gametes have an equal chance to...

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Related Experiment Video

Updated: May 8, 2026

An Array-based Comparative Genomic Hybridization Platform for Efficient Detection of Copy Number Variations in Fast Neutron-induced Medicago truncatula Mutants
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An Array-based Comparative Genomic Hybridization Platform for Efficient Detection of Copy Number Variations in Fast Neutron-induced Medicago truncatula Mutants

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Reverse genetics in Medicago truncatula using a TILLING mutant collection.

Maria Carelli1, Ornella Calderini, Francesco Panara

  • 1Consiglio per la Ricerca e la sperimentazione in Agricoltura (C.R.A.) - Centro di ricerca per le Produzioni Foraggere e Lattiero-casearie, Lodi, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|September 3, 2013
PubMed
Summary

This study details a TILLING mutant collection in Medicago truncatula, a model legume. This resource aids in identifying gene functions and studying metabolic pathways, specifically triterpenic saponins.

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Published on: April 13, 2012

Area of Science:

  • Plant Molecular Genetics
  • Legume Genomics
  • Reverse Genetics

Background:

  • Medicago truncatula is a key model organism for legume molecular genetics research.
  • Development of mutant populations is crucial for identifying and characterizing gene functions.
  • TILLING (Targeting Induced Local Lesions IN Genomes) is an efficient reverse-genetic approach.

Purpose of the Study:

  • To describe the creation and characterization of a TILLING mutant collection in Medicago truncatula.
  • To demonstrate the utility of this resource for gene discovery and functional analysis.
  • To validate the application of the mutant collection in studying secondary metabolite biosynthesis.

Main Methods:

  • Creation of a mutant population of approximately 2,300 M2 individuals.
  • Genomic DNA extraction and M3 seed preservation.
  • Implementation of a two-dimensional DNA pooling strategy for efficient screening.
  • Application of PCR-based screening for identifying point mutations.
  • Genotypic and phenotypic analysis of M3 progeny.

Main Results:

  • A validated TILLING mutant collection of Medicago truncatula was established.
  • The two-dimensional pooling strategy effectively reduced screening complexity.
  • The mutant collection proved valuable for studying the triterpenic saponin biosynthetic pathway.
  • Successful identification of mutations and subsequent gene function studies.

Conclusions:

  • The developed Medicago truncatula TILLING resource is effective for forward and reverse genetic screens.
  • This mutant collection facilitates the functional characterization of genes in legumes.
  • The study highlights the importance of such resources for investigating complex metabolic pathways.