Expression Analysis of PIN Genes in Root Tips and Nodules of Lotus japonicus

Sańko-Sawczenko, Izabela; Dmitruk, Dominika; Łotocka, Barbara; Różańska, Elżbieta; Czarnocka, Weronika

doi:10.3390/ijms20020235

Cited by 12 publications

(11 citation statements)

References 63 publications

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“…It was shown that auxin transport is altered upon early stages of the symbiosis (Mathesius et al, 1998; Boot et al, 1999; Pacios-Bras et al, 2003), and that expression patterns of some PIN-encoding genes are distinct in nodule primordia (Huo et al, 2006; Sanko-Sawczenko et al, 2019). Application of gravitropic stimulation alters auxin response localization and has commonly been used in studies of lateral root emergence.…”

Section: Discussionmentioning

confidence: 99%

A Toolkit for High Resolution Imaging of Cell Division and Phytohormone Signaling in Legume Roots and Root Nodules

2019

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Legume plants benefit from a nitrogen-fixing symbiosis in association with rhizobia hosted in specialized root nodules. Formation of root nodules is initiated by de novo organogenesis and coordinated infection of these developing lateral root organs by rhizobia. Both bacterial infection and nodule organogenesis involve cell cycle activation and regulation by auxin and cytokinin is tightly integrated in the process. To characterize the hormone dynamics and cell division patterns with cellular resolution during nodulation, sensitive and specific sensors suited for imaging of multicellular tissues are required. Here we report a modular toolkit, optimized in the model legume Lotus japonicus , for use in legume roots and root nodules. This toolkit includes synthetic transcriptional reporters for auxin and cytokinin, auxin accumulation sensors and cell cycle progression markers optimized for fluorescent and bright field microscopy. The developed vectors allow for efficient one-step assembly of multiple units using the GoldenGate cloning system. Applied together with a fluorescence-compatible clearing approach, these reporters improve imaging depth and facilitate fluorescence examination in legume roots. We additionally evaluate the utility of the dynamic gravitropic root response in altering the timing and location of auxin accumulation and nodule emergence. We show that alteration of auxin distribution in roots allows for preferential nodule emergence at the outer side of the bend corresponding to a region of high auxin signaling capacity. The presented tools and procedures open new possibilities for comparative mutant studies and for developing a more comprehensive understanding of legume-rhizobia interactions.

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Section: Discussionmentioning

confidence: 99%

A Toolkit for High Resolution Imaging of Cell Division and Phytohormone Signaling in Legume Roots and Root Nodules

2019

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“…RNA was obtained from two biological replicates of L. japonicus and M. truncatula non-inoculated roots and root nodules either subjected to drought for two (T2) or four (T4) days or well-watered control nodules (T0). Total RNA extraction was performed as described previously [ 104 ]. In the case of samples derived from root nodules, an additional rRNA subunit was observed after gel electrophoresis, which proved that, along with plant RNA, bacterial RNA had been extracted ( Figure S5 ).…”

Section: Methodsmentioning

confidence: 99%

“…cDNA synthesis was performed as described previously [ 104 ]. Real-time qPCR was performed using the Connect CFX Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with Power SYBR Green Master Mix (Thermo Fisher Scientific, Waltham, MA, USA) or SsoFast EvaGreen Supermix (Bio-Rad, Hercules, CA, USA) for plant or bacterial genes, respectively.…”

Section: Methodsmentioning

confidence: 99%

Transcriptomic Changes in Medicago truncatula and Lotus japonicus Root Nodules during Drought Stress

Sańko-Sawczenko

Łotocka

Mielecki

et al. 2019

IJMS

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Drought is one of the major environmental factors limiting biomass and seed yield production in agriculture. In this research, we focused on plants from the Fabaceae family, which has a unique ability for the establishment of symbiosis with nitrogen-fixing bacteria, and are relatively susceptible to water limitation. We have presented the changes in nitrogenase activity and global gene expression occurring in Medicago truncatula and Lotus japonicus root nodules during water deficit. Our results proved a decrease in the efficiency of nitrogen fixation, as well as extensive changes in plant and bacterial transcriptomes, shortly after watering cessation. We showed for the first time that not only symbiotic plant components but also Sinorhizobium meliloti and Mesorhizobium loti bacteria residing in the root nodules of M. truncatula and L. japonicus, respectively, adjust their gene expression in response to water shortage. Although our results demonstrated that both M. truncatula and L. japonicus root nodules were susceptible to water deprivation, they indicated significant differences in plant and bacterial response to drought between the tested species, which might be related to the various types of root nodules formed by these species.

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“…Auxin also plays a critical role in controlling the development of medicago indeterminate nodules as well as lotus and soybean determinate nodules [43,60,61]. Sanko-Sawczenko et al recently described the central role of polar auxin transport and auxin homeostasis in controlling determinate nodule development and differentiation [62].…”

Section: Introductionmentioning

confidence: 99%

Plant Hormones Differentially Control the Sub-Cellular Localization of Plasma Membrane Microdomains during the Early Stage of Soybean Nodulation

Qiao

Zogli

Libault

2019

Genes

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Phytohormones regulate the mutualistic symbiotic interaction between legumes and rhizobia, nitrogen-fixing soil bacteria, notably by controlling the formation of the infection thread in the root hair (RH). At the cellular level, the formation of the infection thread is promoted by the translocation of plasma membrane microdomains at the tip of the RH. We hypothesize that phytohormones regulate the translocation of plasma membrane microdomains to regulate infection thread formation. Accordingly, we treated with hormone and hormone inhibitors transgenic soybean roots expressing fusions between the Green Fluorescent Protein (GFP) and GmFWL1 or GmFLOT2/4, two microdomain-associated proteins translocated at the tip of the soybean RH in response to rhizobia. Auxin and cytokinin treatments are sufficient to trigger or inhibit the translocation of GmFWL1 and GmFLOT2/4 to the RH tip independently of the presence of rhizobia, respectively. Unexpectedly, the application of salicylic acid, a phytohormone regulating the plant defense system, also promotes the translocation of GmFWL1 and GmFLOT2/4 to the RH tip regardless of the presence of rhizobia. These results suggest that phytohormones are playing a central role in controlling the early stages of rhizobia infection by regulating the translocation of plasma membrane microdomains. They also support the concept of crosstalk of phytohormones to control nodulation.

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Expression Analysis of PIN Genes in Root Tips and Nodules of Lotus japonicus

Cited by 12 publications

References 63 publications

A Toolkit for High Resolution Imaging of Cell Division and Phytohormone Signaling in Legume Roots and Root Nodules

A Toolkit for High Resolution Imaging of Cell Division and Phytohormone Signaling in Legume Roots and Root Nodules

Transcriptomic Changes in Medicago truncatula and Lotus japonicus Root Nodules during Drought Stress

Plant Hormones Differentially Control the Sub-Cellular Localization of Plasma Membrane Microdomains during the Early Stage of Soybean Nodulation

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