Use of Hairy Root System to Study Signaling Pathways During Nodule Formation

Hairy Roots ◽  
2018 ◽  
pp. 243-274
Author(s):  
Swarup Roy Choudhury ◽  
Sona Pandey
Keyword(s):  
Signaling Pathways ◽  
Root System ◽  
Hairy Root ◽  
Nodule Formation

Lateral root formation and nutrients: nitrogen in the spotlight

2021 ◽  
Author(s):  
Pierre-Mathieu Pélissier ◽  
Hans Motte ◽  
Tom Beeckman
Keyword(s):  
Signaling Pathways ◽  
Root System ◽  
Root Development ◽  
Lateral Root ◽  
Molecular Mechanisms ◽  
Root Formation ◽  
Lateral Roots ◽  
Nutrient Use Efficiency ◽  
Lateral Root Formation ◽  
Lateral Root Development

Abstract Lateral roots are important to forage for nutrients due to their ability to increase the uptake area of a root system. Hence, it comes as no surprise that lateral root formation is affected by nutrients or nutrient starvation, and as such contributes to the root system plasticity. Understanding the molecular mechanisms regulating root adaptation dynamics towards nutrient availability is useful to optimize plant nutrient use efficiency. There is at present a profound, though still evolving, knowledge on lateral root pathways. Here, we aimed to review the intersection with nutrient signaling pathways to give an update on the regulation of lateral root development by nutrients, with a particular focus on nitrogen. Remarkably, it is for most nutrients not clear how lateral root formation is controlled. Only for nitrogen, one of the most dominant nutrients in the control of lateral root formation, the crosstalk with multiple key signals determining lateral root development is clearly shown. In this update, we first present a general overview of the current knowledge of how nutrients affect lateral root formation, followed by a deeper discussion on how nitrogen signaling pathways act on different lateral root-mediating mechanisms for which multiple recent studies yield insights.

scholarly journals Accumulation and Distribution of Fertilizer Nitrogen and Nodule-Fixed Nitrogen in Soybeans with Dual Root Systems

Agronomy ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 397 ◽  
Author(s):  
Rui Zhang ◽  
Cong Wang ◽  
Wenzhi Teng ◽  
Jing Wang ◽  
Xiaochen Lyu ◽  
...  
Keyword(s):  
Root System ◽  
Root Nodules ◽  
Culture Conditions ◽  
Sand Culture ◽  
Nodule Formation ◽  
N Fixation ◽  
Fertilizer N ◽  
N Accumulation ◽  
Biological N Fixation ◽  
Nodule Growth

The soybean (Glycine max L. Merr.) is a crop with a high demand for nitrogen (N). The root nodules that form in soybeans can fix atmospheric N effectively, yet the goal of achieving high yields cannot be met by relying solely on nodule-fixed N. Nonetheless, the application of N fertilizer may inhibit nodule formation and biological N fixation (BNF), but the underpinning mechanisms are still unclear. In this study, we grafted the roots of non-nodulated soybeans onto nodulated soybeans to generate plants with dual root system. The experiment included three treatments conducted under sand culture conditions with NO 3 − and NH 4 + as N sources. Treatment I: The non-nodulated roots on one side received 50 mg·L−1 15 NO 3 − or 15NH4+, and the nodulated roots on the other side were not treated. Treatment II: The non-nodulated roots received 50 mg·L−1 15 NO 3 − or 15 NH 4 + , and the nodulated roots received 50 mg·L−1 14 NO 3 − or 14 NH 4 + . Treatment III: Both non-nodulated and nodulated roots received 50 mg·L−1 15 NO 3 − or 15 NH 4 + . The results showed the following: (1) Up to 81.5%–87.1% of the N absorbed by the soybean roots and fixed by the root nodules was allocated to shoot growth, leaving 12.9%–18.5% for root and nodule growth. Soybeans preferentially used fertilizer N in the presence of a NO 3 − or NH 4 + supply. After the absorbed fertilizer N and nodule-fixed N was transported to the shoots, a portion of it was redistributed to the roots and nodules. The N required for root growth was primarily derived from the NO 3 − or NH 4 + assimilated by the roots and the N fixed by the nodules, with a small portion translocated from the shoots. The N required for nodule growth was primarily contributed by nodule-fixed N with a small portion translocated from the shoots, whereas the NO 3 − or NH 4 + that was assimilated by the roots was not directly supplied to the nodules. (2) Based on observations of the shoots and one side of the roots and nodules in the dual root system as an N translocation system, we proposed a method for calculating the N translocation from soybean shoots to roots and nodules during the R1–R5 stages based on the difference in the 15N abundance. Our calculations showed that when adding N at a concentration of 50 mg·L−1, the N translocated from the shoots during the R1–R5 stages accounts for 29.6%–52.3% of the N accumulation in nodulated roots (Rootn) and 9.4%–16.6% of the N accumulation in Nodulen of soybeans. Through the study of this experiment, the absorption, distribution and redistribution characteristics of fertilizer N and root nodule N fixation in soybean can be clarified, providing a theoretical reference for analyzing the mechanisms of the interaction between fertilizer N and nodule-fixed N.

scholarly journals Highly efficient Agrobacterium rhizogenes-mediated hairy root transformation for gene functional and gene editing analysis in soybean

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Yuanyuan Cheng ◽  
Xiaoli Wang ◽  
Li Cao ◽  
Jing Ji ◽  
Tengfei Liu ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Subcellular Localization ◽  
Root System ◽  
Hairy Root ◽  
Protein Interactions ◽  
Gene Editing ◽  
Transformation Frequency ◽  
Highly Efficient ◽  
Root Transformation ◽  
Gene Functional Analysis

Abstract Background Agrobacterium-mediated genetic transformation is a widely used and efficient technique for gene functional research in crop breeding and plant biology. While in some plant species, including soybean, genetic transformation is still recalcitrant and time-consuming, hampering the high-throughput functional analysis of soybean genes. Thus we pursue to develop a rapid, simple, and highly efficient hairy root system induced by Agrobacterium rhizogenes (A. rhizogenes) to analyze soybean gene function. Results In this report, a rapid, simple, and highly efficient hairy root transformation system for soybean was described. Only sixteen days were required for the whole workflow and the system was suitable for various soybean genotypes, with an average transformation frequency of 58–64%. Higher transformation frequency was observed when wounded cotyledons from 1-day-germination seeds were inoculated and co-cultivated with A. rhizogenes in 1/2 B5 (Gamborg’ B-5) medium. The addition of herbicide selection to root production medium increased the transformation frequency to 69%. To test the applicability of the hairy root system for gene functional analysis, we evaluated the protein expression and subcellular localization in transformed hairy roots. Transgenic hairy roots exhibited significantly increased GFP fluorescence and appropriate protein subcellular localization. Protein–protein interactions by BiFC (Bimolecular Fluorescent Complimentary) were also explored using the hairy root system. Fluorescence observations showed that protein interactions could be observed in the root cells. Additionally, hairy root transformation allowed soybean target sgRNA screening for CRISPR/Cas9 gene editing. Therefore, the protocol here enables high-throughput functional characterization of candidate genes in soybean. Conclusion A rapid, simple, and highly efficient A. rhizogenes-mediated hairy root transformation system was established for soybean gene functional analysis, including protein expression, subcellular localization, protein–protein interactions and gene editing system evaluation.

scholarly journals Effects of soil temperature and moisture on biological nitrogen fixation in soybean crop

2019 ◽  
pp. 1327-1334
Author(s):  
Evandro Ademir Deak ◽  
Thomas Newton Martin ◽  
Glauber Monçon Fipke ◽  
Jessica Deolinda Leivas Stecca ◽  
Luciane Almeri Tabaldi ◽  
...  
Keyword(s):  
Soil Temperature ◽  
Root System ◽  
Azospirillum Brasilense ◽  
Water Retention ◽  
Nodule Formation ◽  
Water Retention Capacity ◽  
Soil Water Retention ◽  
Retention Capacity ◽  
Soybean Crop ◽  
Temperature And Humidity

Soil temperature and humidity are the chief determinants for good nodule formation at the time of sowing and emergence. The aim of this study was to estimate the soil temperature and humidity range, at which the Bradyrhizobium spp., and Azospirillum brasilense may have the highest effect on root enhancement and development of nodulation in soybean. Two experiments were conducted, the first of which was done in the seed laboratory performing the treatments listed as non-inoculated; inoculation with Bradyrhizobium; root enhancers; inoculation with Bradyrhizobium + root enhancers; co-inoculation with Bradyrhizobium + Azospirillum brasilense; co-inoculation with Bradyrhizobium + Azospirillum brasilense + root enhancers, and assessed at temperatures of 15, 20, 25, 30 and 35ºC. The experimental design was completely randomized with distribution in a factorial. The second experiment was executed in a greenhouse, employing the same treatments affected in the seed laboratory experiment, but with the addition of non-inoculated control with mineral nitrogen. Tests were done at the soil moisture levels of 25, 50, 75 and 100% water retention capacity, forming a factorial 7 x 4 (only second experiment). All co-inoculated treatments induced the soybean root system to improve, in terms of length, volume, surface area and root diameter, exhibiting superiority to the uninoculated control in the 20 to 30ºC temperature range. The co-inoculation raised nodulation in the soybean crop, when soil water retention capacity was at the range of 56 - 96%, achieving higher means compared to the standard inoculation. The root planter added no improvement to either the root system or nodulation in soybean.

scholarly journals The application of CRISPR/Cas9 in hairy roots to explore the functions of AhNFR1 and AhNFR5 genes during peanut nodulation

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Hongmei Shu ◽  
Ziliang Luo ◽  
Ze Peng ◽  
Jianping Wang
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Root Nodules ◽  
Crop Improvement ◽  
Functional Study ◽  
Nodule Formation ◽  
Transformation System ◽  
Knock Out ◽  
Root Transformation ◽  
Peanut Hairy Root

Abstract Background Peanut is an important legume crop growing worldwide. With the published allotetraploid genomes, further functional studies of the genes in peanut are very critical for crop improvement. CRISPR/Cas9 system is emerging as a robust tool for gene functional study and crop improvement, which haven’t been extensively utilized in peanut yet. Peanut plant forms root nodules to fix nitrogen through a symbiotic relationship with rhizobia. In model legumes, the response of plants to rhizobia is initiated by Nod factor receptors (NFRs). However, information about the function of NFRs in peanut is still limited. In this study, we applied the CRISPR/Cas9 tool in peanut hairy root transformation system to explore the function of NFR genes. Results We firstly identified four AhNFR1 genes and two AhNFR5 genes in cultivated peanut (Tifrunner). The gene expression analysis showed that the two AhNFR1 and two AhNFR5 genes had high expression levels in nodulating (Nod+) line E5 compared with non-nodulating (Nod-) line E4 during the process of nodule formation, suggesting their roles in peanut nodulation. To further explore their functions in peanut nodulation, we applied CRISPR technology to create knock-out mutants of AhNFR1 and AhNFR5 genes using hairy root transformation system. The sequencing of these genes in transgenic hairy roots showed that the selected AhNFR1 and AhNFR5 genes were successfully edited by the CRISPR system, demonstrating its efficacy for targeted mutation in allotetraploid peanut. The mutants with editing in the two AhNFR5 genes showed Nod- phenotype, whereas mutants with editing in the two selected AhNFR1 genes could still form nodules after rhizobia inoculation. Conclusions This study showed that CRISPR-Cas9 could be used in peanut hairy root transformation system for peanut functional genomic studies, specifically on the gene function in roots. By using CRISPR-Cas9 targeting peanut AhNFR genes in hairy root transformation system, we validated the function of AhNFR5 genes in nodule formation in peanut.

scholarly journals Hairy Root Induction in Linum mucronatum ssp. mucronatum, an Anti-Tumor Lignans Producing Plant

2012 ◽  
Vol 40 (1) ◽  
pp. 125 ◽  
Author(s):  
Afsaneh SAMADI ◽  
Jirair CARAPETIAN ◽  
Reza HEIDARI ◽  
Morad JAFARI ◽  
Abdollah HASSANZADEH GORTTAPEH
Keyword(s):  
Root System ◽  
Hairy Root ◽  
Hairy Roots ◽  
Culture Media ◽  
Basal Medium ◽  
Cotyledonary Node ◽  
Root Induction ◽  
Hairy Root Induction ◽  
Cancer Agent ◽  
Promising Source

Transgenic hairy root system is a promising source of secondary metabolites in medicinal plants with high pharmaceutical value.For the first time, hairy roots were established in different explants of Linum mucronatum, an anti-cancer agent producing plant, via amikimopine type strain of Agrobacterium rhizogenes, ‘A13’. The percentage of hairy root induction varied from 0 to 60% depended onthe explants and hypocotyl (including cotyledonary node) explants were found to be highly susceptible to A. rhizogenes infection withthe highest (60%) rate of hairy root induction. four different Murashige and Skoog (MS)-based liquid culture media were used for wellestablishment of hairy roots. Hairy root growth medium D (HRGM-D) containing hormone-free MS basal medium with an extra oneday pre-incubation period at 35°C was found to be more efficient for profuse growth (fresh weight; 8500 mg per 25 ml culture medium)of hairy roots. Hairy root system presented in this study may offer a suitable platform for optimization and production of satisfactorylevel of aryltetralin lignans like podophyllotoxin and its derivatives from L. mucronatum.

Establishment of a transgenic hairy root system in wild and domesticated watermelon (Citrullus lanatus) for studying root vigor under drought

2010 ◽  
Vol 29 (7) ◽  
pp. 771-778 ◽  
Author(s):  
Masataka Kajikawa ◽  
Kaoru Morikawa ◽  
Yosuke Abe ◽  
Akiho Yokota ◽  
Kinya Akashi
Keyword(s):  
Root System ◽  
Hairy Root ◽  
Citrullus Lanatus ◽  
Transgenic Hairy Root ◽  
Root Vigor
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