Rapid Analysis of Legume Root Nodule Development Using Confocal Microscopy
Date: 2004
Creator: Haynes, Janine G.; Czymmek, Kirk J.; Carlson, Carol A.; Veereshlingam, Harita; Dickstein, Rebecca & Sherrier, D. Janine
Description: This article discusses the rapid analysis of legume root nodule development using confocal microscopy. A rapid method for detailed analysis of nodule formation has been developed. Inoculated root tissues were stained with SYTO 13, a cell-permeant fluorescent nucleic acid-binding dye, and visualized using confocal laser scanning microscopy (CLSM). Structures with high concentrations of DNA and RNA, such as plant cell nuclei and bacteria, labeled strongly. The autofluorescent properties of cell walls made it possible to use CLSM to visualize both plant and rhizobial structures and generate a three-dimensional reconstruction of the root and invading bacteria. This method allowed clear observation of stages and structures important in nodule formation, such as rhizobial attachment to root hairs, hair deformation, infection thread ramification, nodule primordium development and nodule cell invasion. Bacteroid structures were easily were easily assessed without the need for fixation that might alter cellular integrity. Plant nodulation mutants with phenotypic differences in thread growth, cellular invasion and plant defense response were also documented. Multiple samples can be assessed using detailed microscopy without the need for extensive preparative work, labor-intensive analysis, or the generation of genetically modified samples.
Contributing Partner: UNT College of Arts and Sciences
Permallink:digital.library.unt.edu/ark:/67531/metadc40415/
Functional Assessment of the Medicago truncatula NIP/LATD Protein Demonstrates That It Is a High-Affinity Nitrate Transporter
Date: October 2012
Creator: Bagchi, Rammyani; Salehin, Mohammad; Adeyemo, O. Sarah; Salazar, Carolina; Shulaev, Vladimir; Sherrier, D. Janine et al
Description: This article discusses a functional assessment of the Medicago truncatula NIP/LATD. Abstract: The Medicago truncatula NIP/LATD (for Numerous Infections and Polyphenolics/Lateral root-organ Defective) gene encodes a protein found in a clade of nitrate transporters within the large NRT1(PTR) family that also encodes transporters of dipeptides and tripeptides, dicarboxylates, auxin, and abscisic acid. Of the NRT1(PTR) members known to transport nitrate, most are low-affinity transporters. Here, the authors show that M. truncatula nip/latd mutants are more defective in their lateral root responses to nitrate provided at low (250 μм) concentrations than at higher (5mм) concentrations; however, nitrate uptake experiments showed no discernible differences in uptake in the mutants. Heterologous expression experiments showed that MtNIP/LATD encodes a nitrate transporter: expression in Xenopus laevis oocytes conferred upon the oocytes the ability to take up nitrate from the medium, but oocytes expressing the less severe Mtnip-3 allele were proficient in nitrate transport. M. truncatula nip/latd mutants have pleiotropic defects in nodulation and root architecture defects but not for nodulation defects. This suggests that the spectrum of activities inherent in AtNRT1.1 is different from that possessed by MtNIP/LATD, but it could also reflect stability differences of each protein in M. truncatula. Collectively, the data show ...
Contributing Partner: UNT College of Arts and Sciences
Permallink:digital.library.unt.edu/ark:/67531/metadc130185/
A Putative Transporter is Essential for Integrating Nutrient and Hormone Signaling with Lateral Root Growth and Nodule Development in Medicago truncatula
Date: 2010
Creator: Yendrek, Craig R.; Lee, Yi-Ching; Morris, Viktoriya; Liang, Yan; Pislariu, Catalina I.; Burkart, Graham et al
Description: This article discusses a putative transporter for integrating nutrient and hormone signaling with lateral root growth and nodule development in Medicago truncatula. Legume root architecture involves not only elaboration of the root system by the formation of lateral roots but also the formation of symbiotic root nodules in association with nitrogen-fixing soil rhizobia. The Medicago truncatula LATD/NIP gene plays an essential role in the development of both primary and lateral roots as well as nodule development. We have cloned the LATD/NIP gene and show that it encodes a member of the NRT1(PTR) transporter family. LATD/NIP is expressed throughout the plant. pLATD/NIP-GFP promoter-reporter fusions in transgenic roots establish the spatial expression of LATD/NIP in primary root, lateral root and nodule meristems and the surrounding cells. Expression of LATD/NIP is regulated by hormones, in particular by abscisic acid which has been previously shown to rescue the primary and lateral root meristem arrest of latd mutants. latd mutants respond normally to ammonium but have defects in responses of the root architecture to nitrate. Taken together, these results suggest that LATD/NIP may encode a nitrate transporter or transporter of another compound.
Contributing Partner: UNT College of Arts and Sciences
Permallink:digital.library.unt.edu/ark:/67531/metadc40412/