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since 4/20/2004

Last updated 10/01/09

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Mutant List


Mutants for DMI3: 9
Mutant Symbol Allele or Genotype Mutant Gene Phenotype Detail Symbiotic Phenotype Brief Mutagenesis Method Mutant Class WT Strain or Line
TRV25 dmi3-1 dmi3 Nod-; defective in calcium spiking; With induced touch response, the mutant showed root hair tip swelling (Has) but blocked for branching (Hab-), lack cortical cell division (Ccd-), no root curling (Hac-) and no infection threads (Inf-) in response to live bacteria; blocked in the induction of ENOD11 and ENOD40; impaired for ENOD12 expression; dramatically reduced induction of RIP1; Myc- phenotype; Recessive. A more recent paper by Esseling et al., 2004 showed that when care is taken not to induce touch response, root hairs reorientate upon NF spot application. In Morandi et al. 2004 paper, TRV25 belongs to the mycorrhizal phenotype 2, presenting only lens-shaped appressoria with no further development of AM colonisation, regardless of the time after inoculation or the mycorrhizal fungus involved. Calcium flux+, Calcium spiking+, Has+, Hab-(touch response), Enod gene expression-, Ccd-, Hac-(touch response), Hac+(no touch response), Inf-, Nod-, Myc- gamma rays Nod- Jemalong J5 
dmi3-1 35S-DMI3 1-311 dmi3-1 35S-DMI3 1-311 dmi3 Roots transformed with DMI3 1-311 showed strong ENOD11-GUS expression in epidermal cells, primarily of lateral roots, in the absence of Nod factor treatment 46% of root systems. The ENOD11-GUS expression induced by the deletion construct was observed in a similar zone of epidermal cells to that of the ENOD11 induction by Nod factor. No bacterially infected nodules were found in complementation tests with dmi3-1. Nod+Fix- deletion Nod+Fix- Jemalong J5 
dmi3-1 35S-DMI3 1-326 dmi3-1 35S-DMI3 1-326 dmi3 Roots transformed with DMI3 1-326 showed strong ENOD11-GUS expression in epidermal cells, primarily of lateral roots, in the absence of Nod factor treatment 53% of root systems. The ENOD11-GUS expression induced by the deletion construct was observed in a similar zone of epidermal cells to that of the ENOD11 induction by Nod factor. No bacterially infected nodules were found in complementation tests with dmi3-1. Nod+Fix- deletion Nod+Fix- Jemalong J5 
dmi3-1 35S-DMI3 T271A dmi3-1 35S-DMI3 T271A dmi3 DMI3 T271A activated ENOD11-GUS in the absence of Nod factor indicating gain-of-function activities. Unlike the deletion constructs, DMI3 T271A was able to complement dmi3-1 and formed bacterial infected nodules, indicating that the gain-of-function mutant was also fully functional proteins. Authors predicted that DMI3 T271A and DMI3 T271D would also lead to spontaneous nodulation if expressed from the native promoter. Nod+ Site-directed Mutagenesis Nod+Fix+ Jemalong J5 
dmi3-1 35S-DMI3 T271D dmi3-1 35S-DMI3 T271D dmi3 DMI3 T271D activated ENOD11-GUS in the absence of Nod factor indicating gain-of-function activities. Unlike the deletion constructs, DMI3 T271D was able to complement dmi3-1 and formed bacterial infected nodules, indicating that the gain-of-function mutant was also fully functional proteins. Authors predicted that DMI3 T271A and DMI3 T271D would also lead to spontaneous nodulation if expressed from the native promoter. Nod+ Site-directed Mutagenesis Nod+Fix+ Jemalong J5 
dmi3-1 35S-DMI3 delta(328-355) dmi3-1 35S-DMI3 delta(328-355) dmi3 Roots transformed with DMI3 delta(328-355) showed strong ENOD11-GUS expression in epidermal cells, primarily of lateral roots, in the absence of Nod factor treatment 36% of root systems. The ENOD11-GUS expression induced by the deletion construct was observed in a similar zone of epidermal cells to that of the ENOD11 induction by Nod factor. No bacterially infected nodules were found in complementation tests with dmi3-1. Nod+Fix- deletion Nod+Fix- Jemalong J5 
dmi3-1 DMI3p-DMI3 1-311 dmi3-1 DMI3p-DMI3 1-311 dmi3 43% of dmi3-1 plants (20 of 47) transformed with DMI3p-DMI3 1-311 showed spontaneous nodules, with an average of 5.4 nodules per plant. Spontaneous nodules induced by DMI3p-DMI3 1-311 showed appropriate ENOD11-GUS induction. When S. meliloti was inoculated onto dmi3-1 roots transformed with DMI3p-DMI3 1-311, no bacteria was detected inside the nodules and little indication of root hair responses and no development of infection threads was observed. Hence, even though DMI3p-DMI3 1-311 leads to the formation of nodules, it is not sufficient to fully complement the dmi3-1 mutation to allow bacterial entry. Nod+Fix- deletion Nod+Fix- Jemalong J5 
dmi3-1 DMI3p-DMI3 1-326 dmi3-1 DMI3p-DMI3 1-326 dmi3 53% of dmi3-1 plants (16 of 30) transformed with DMI3p-DMI3 1-326 showed spontaneous nodules, with an average of 4.4 nodules per plant. When S. meliloti was inoculated onto dmi3-1 roots transformed with DMI3p-DMI3 1-326, no bacteria was detected inside the nodules and little indication of root hair responses and no development of infection threads was observed. Hence, even though DMI3p-DMI3 1-326 leads to the formation of nodules, it is not sufficient to fully complement the dmi3-1 mutation to allow bacterial entry. Nod+Fix- deletion Nod+Fix- Jemalong J5 
TRV25 pRNAiIPD3 dmi3-1 ipd3-RNAi dmi3ipd3 Nodules were never observed on control nontransgenic roots of dmi3 plants nor on dmi3 plants transformed with pRNAiIPD3. Nod- RNAi Nod- Jemalong A17 

References:
Authors Year Title Locator
Ane JM, Levy J, Thoquet P, Kulikova O, de Billy F, Penmetsa V, Kim DJ, Debelle F, Rosenberg C, Cook DR, Bisseling T, Huguet T and Denarie J 2002 Genetic and cytogenetic mapping of DMI1, DMI2, and DMI3 genes of Medicago truncatula involved in Nod factor transduction, nodulation, and mycorrhization Mol Plant Microbe Interact. 15(11):1108-18 
Catoira R, Galera C, de Billy F, Penmetsa RV, Journet EP, Maillet F, Rosenberg C, Cook D, Gough C and Denarie J 2000 Four genes of Medicago truncatula controlling components of a nod factor transduction pathway Plant Cell. 12(9):1647-66 
Esseling JJ, Lhuissier FG and Emons AM 2004 A nonsymbiotic root hair tip growth phenotype in NORK-mutated legumes: implications for nodulation factor-induced signaling and formation of a multifaceted root hair pocket for bacteria Plant Cell. 16(4):933-44 
Gleason C, Chaudhuri S, Yang T, Munoz A, Poovaiah BW and Oldroyd GE 2006 Nodulation independent of rhizobia induced by a calcium-activated kinase lacking autoinhibition Nature. 441(7097):1149-52 
Levy J, Bres C, Geurts R, Chalhoub B, Kulikova O, Duc G, Journet EP, Ane JM, Lauber E, Bisseling T, Denarie J, Rosenberg C and Debelle F 2004 A putative Ca2+ and calmodulin-dependent protein kinase required for bacterial and fungal symbioses Science. 303(5662):1361-4 
Messinese E, Mun JH, Yeun LH, Jayaraman D, Rouge P, Barre A, Lougnon G, Schornack S, Bono JJ, Cook DR and Ane JM 2007 A novel nuclear protein interacts with the symbiotic DMI3 calcium- and calmodulin-dependent protein kinase of Medicago truncatula Mol Plant Microbe Interact. 20(8):912-21 
Mitra RM, Gleason CA, Edwards A, Hadfield J, Downie JA, Oldroyd GE and Long SR 2004 A Ca2+/calmodulin-dependent protein kinase required for symbiotic nodule development: Gene identification by transcript-based cloning Proc Natl Acad Sci U S A. 101(13):4701-5 
Mitra RM, Shaw SL and Long SR 2004 Six nonnodulating plant mutants defective for Nod factor-induced transcriptional changes associated with the legume-rhizobia symbiosis Proc Natl Acad Sci U S A. 101(27):10217-22 
Morandi D, Prado E, Sagan M, and Duc G 2004 Characterisation of new symbiotic Medicago truncatula (Gaertn.) mutants, and phenotypic or genotypic complementary information on previously described mutants. Mycorrhiza 
Sagan M, de Larembergue H and Morandi D 1998 Genetic analysis of symbiosis mutants in Medicago truncatula Elmerich C, Kondorosi A, Newton WE (eds) Biological nitrogen fixation for the 21st century. Kluwer, Dordrecht, pp 317?318 
Shaw SL and Long SR 2003 Nod factor elicits two separable calcium responses in Medicago truncatula root hair cells Plant Physiol. 131(3):976-84 
Wais RJ, Galera C, Oldroyd G, Catoira R, Penmetsa RV, Cook D, Gough C, Denarie J and Long SR 2000 Genetic analysis of calcium spiking responses in nodulation mutants of Medicago truncatula Proc Natl Acad Sci U S A. 97(24):13407-12