A Medicago truncatula Tobacco Retrotransposon Insertion Mutant Collection with Defects in Nodule Development and Symbiotic Nitrogen Fixation

A Tnt1-insertion mutant population of Medicago truncatula ecotype R108 was screened for defects in nodulation and symbiotic nitrogen fixation. Primary screening of 9,300 mutant lines yielded 317 lines with putative defects in nodule development and/or nitrogen fixation. Of these, 230 lines were resc...

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Published in:Plant physiology (Bethesda) 2012-08, Vol.159 (4), p.1686-1699
Main Authors: Pislariu, Catalina I., Murray, Jeremy D., Wen, JiangQi, Cosson, Viviane, Muni, RajaSekhara Reddy Duvvuru, Wang, Mingyi, Benedito, Vagner A., Andriankaja, Andry, Cheng, Xiaofei, Jerez, Ivone Torres, Mondy, Samuel, Zhang, Shulan, Taylor, Mark E., Tadege, Million, Ratet, Pascal, Mysore, Kirankumar S., Chen, Rujin, Udvardi, Michael K.
Format: Article
Language:English
Subjects:
Agronomy. Soil science and plant productions
Biological and medical sciences
Economic plant physiology
Fundamental and applied biological sciences. Psychology
Genes, Plant - genetics
Infections
Legumes
Medicago truncatula - genetics
Medicago truncatula - microbiology
Medicago truncatula - physiology
Morphogenesis - genetics
Mutagenesis, Insertional - genetics
Mutation - genetics
Mycorrhizae - physiology
Nicotiana - genetics
Nitrogen fixation
Nitrogen Fixation - genetics
Nodulation
Nodules
Parasitism and symbiosis
Phenotype
Phenotypes
Plant cells
Plant physiology and development
Plant Root Nodulation - genetics
Plants
PLANTS INTERACTING WITH OTHER ORGANISMS
Retroelements - genetics
Retrotransposons
Root Nodules, Plant - growth & development
Symbiosis
Symbiosis (nodules, symbiotic nitrogen fixation, mycorrhiza...)
Symbiosis - genetics
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Summary:A Tnt1-insertion mutant population of Medicago truncatula ecotype R108 was screened for defects in nodulation and symbiotic nitrogen fixation. Primary screening of 9,300 mutant lines yielded 317 lines with putative defects in nodule development and/or nitrogen fixation. Of these, 230 lines were rescreened, and 156 lines were confirmed with defective symbiotic nitrogen fixation. Mutants were sorted into six distinct phenotypic categories: 72 nonnodulating mutants (Nod-), 51 mutants with totally ineffective nodules (Nod+ Fix-), 17 mutants with partially ineffective nodules (Nod+ Fix+/-), 27 mutants defective in nodule emergence, elongation, and nitrogen fixation (Nod+/- Fix-), one mutant with delayed and reduced nodulation but effective in nitrogen fixation (dNod+/- Fix+), and 11 supernodulating mutants (Nod++Fix+/-). A total of 2,801 flanking sequence tags were generated from the 156 symbiotic mutant lines. Analysis of flanking sequence tags revealed 14 insertion alleles of the following known symbiotic genes: NODULE INCEPTION (NIN), DOESN'T MAKE INFECTIONS3 (DMI3/CCaMK), ERF REQUIRED FOR NODULATION, and SUPERNUMERARY NODULES (SUNN). In parallel, a polymerase chain reaction-based strategy was used to identify Tnt1 insertions in known symbiotic genes, which revealed 25 additional insertion alleles in the following genes: DMI1, DMI2, DMI3, NIN, NODULATION SIGNALING PATHWAY1 (NSP1), NSP2, SUNN, and SICKLE. Thirty-nine Nod- lines were also screened for arbuscular mycorrhizal symbiosis phenotypes, and 30 mutants exhibited defects in arbuscular mycorrhizal symbiosis. Morphological and developmental features of several new symbiotic mutants are reported. The collection of mutants described here is a source of novel alleles of known symbiotic genes and a resource for cloning novel symbiotic genes via Tnt1 tagging.
ISSN:0032-0889
1532-2548
1532-2548
DOI:10.1104/pp.112.197061