scholarly journals Transcriptome analysis of the differential effect of the NADPH oxidase gene RbohB in Phaseolus vulgaris roots following Rhizobium tropici and Rhizophagus irregularis inoculation

BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Citlali Fonseca-García ◽  
Alejandra E. Zayas ◽  
Jesús Montiel ◽  
Noreide Nava ◽  
Federico Sánchez ◽  
...  
Keyword(s):  
Cell Wall ◽  
Phaseolus Vulgaris ◽  
Genetic Interaction ◽  
Am Fungi ◽  
Rhizophagus Irregularis ◽  
Ros Scavenging ◽  
Rhizobium Tropici ◽  
Oxidase Gene ◽  
Transgenic Roots ◽  
Early Stages

Abstract Background Reactive oxygen species (ROS) are generated by NADPH oxidases known as respiratory burst oxidase homologs (RBOHs) in plants. ROS regulate various cellular processes, including the mutualistic interactions between legumes and nitrogen-fixing bacteria or arbuscular mycorrhizal (AM) fungi. Rboh is a multigene family comprising nine members (RbohA–I) in common bean (Phaseolus vulgaris). The RNA interference-mediated silencing of RbohB (PvRbohB-RNAi) in this species diminished its ROS production and greatly impaired nodulation. By contrast, the PvRbohB-RNAi transgenic roots showed early hyphal root colonization with enlarged fungal hypopodia; therefore, we proposed that PvRbohB positively regulates rhizobial infection (Rhizobium tropici) and inhibits AM colonization by Rhizophagus irregularis in P. vulgaris. Results To corroborate this hypothesis, an RNA-Seq transcriptomic analysis was performed to identify the differentially expressed genes in the PvRbohB-RNAi roots inoculated with Rhizobium tropici or Rhizophagus irregularis. We found that, in the early stages, root nodule symbioses generated larger changes of the transcriptome than did AM symbioses in P. vulgaris. Genes related to ROS homeostasis and cell wall flexibility were markedly upregulated in the early stages of rhizobial colonization, but not during AM colonization. Compared with AM colonization, the rhizobia induced the expression of a greater number of genes encoding enzymes involved in the metabolism of auxins, cytokinins, and ethylene, which were typically repressed in the PvRbohB-RNAi roots. Conclusions Our research provides substantial insights into the genetic interaction networks in the early stages of rhizobia and AM symbioses with P. vulgaris, as well as the differential roles that RbohB plays in processes related to ROS scavenging, cell wall remodeling, and phytohormone homeostasis during nodulation and mycorrhization in this legume.

scholarly journals PvRACK1 Loss-of-Function Impairs Cell Expansion and Morphogenesis in Phaseolus vulgaris L. Root Nodules

2011 ◽  
Vol 24 (7) ◽  
pp. 819-826 ◽  
Author(s):  
Tania Islas-Flores ◽  
Gabriel Guillén ◽  
Xóchitl Alvarado-Affantranger ◽  
Miguel Lara-Flores ◽  
Federico Sánchez ◽  
...  
Keyword(s):  
Phaseolus Vulgaris ◽  
Cell Expansion ◽  
Root Nodules ◽  
Microscopic Analysis ◽  
Nodule Development ◽  
Loss Of Function ◽  
Rhizobium Tropici ◽  
Transcript Accumulation ◽  
Transgenic Roots ◽  
Plant Signal Transduction

Receptor for activated C kinase (RACK1) is a highly conserved, eukaryotic protein of the WD-40 repeat family. Its peculiar β-propeller structure allows its interaction with multiple proteins in various plant signal-transduction pathways, including those arising from hormone responses, development, and environmental stress. During Phaseolus vulgaris root development, RACK1 (PvRACK1) mRNA expression was induced by auxins, abscissic acid, cytokinin, and gibberellic acid. In addition, during P. vulgaris nodule development, PvRACK1 mRNA was highly accumulated at 12 to 15 days postinoculation, suggesting an important role after nodule meristem initiation and Rhizobium nodule infection. PvRACK1 transcript accumulation was downregulated by a specific RNA interference construct which was expressed in transgenic roots of composite plants of P. vulgaris inoculated with Rhizobium tropici. PvRACK1 downregulated transcript levels were monitored by quantitative reverse-transcription polymerase chain reaction analysis in individual transgenic roots and nodules. We observed a clear phenotype in PvRACK1-knockdown nodules, in which nodule number and nodule cell expansion were impaired, resulting in altered nodule size. Microscopic analysis indicated that, in PvRACK1-knockdown nodules, infected and uninfected cells were considerably smaller (80 and 60%, respectively) than in control nodules. In addition, noninfected cells and symbiosomes in silenced nodules showed significant defects in membrane structure under electron microscopy analysis. These findings indicate that PvRACK1 has a pivotal role in cell expansion and in symbiosome and bacteroid integrity during nodule development.

scholarly journals An NADPH oxidase regulates carbon metabolism and the cell cycle during root nodule symbiosis in common bean (Phaseolus vulgaris)

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Citlali Fonseca-García ◽  
Noreide Nava ◽  
Miguel Lara ◽  
Carmen Quinto
Keyword(s):  
Cell Cycle ◽  
Phaseolus Vulgaris ◽  
Common Bean ◽  
Carbon Metabolism ◽  
Root Nodule ◽  
Transgenic Roots ◽  
Early Stages ◽  
Root Nodule Symbiosis ◽  
Legume Symbiosis ◽  
Nodule Symbiosis

Abstract Background Rhizobium–legume symbiosis is a specific, coordinated interaction that results in the formation of a root nodule, where biological nitrogen fixation occurs. NADPH oxidases, or Respiratory Burst Oxidase Homologs (RBOHs) in plants, are enzymes that generate superoxide (O2•−). Superoxide produces other reactive oxygen species (ROS); these ROS regulate different stages of mutualistic interactions. For example, changes in ROS levels are thought to induce ROS scavenging, cell wall remodeling, and changes in phytohormone homeostasis during symbiotic interactions. In common bean (Phaseolus vulgaris), PvRbohB plays a key role in the early stages of nodulation. Results In this study, to explore the role of PvRbohB in root nodule symbiosis, we analyzed transcriptomic data from the roots of common bean under control conditions (transgenic roots without construction) and roots with downregulated expression of PvRbohB (by RNA interference) non-inoculated and inoculated with R. tropici. Our results suggest that ROS produced by PvRBOHB play a central role in infection thread formation and nodule organogenesis through crosstalk with flavonoids, carbon metabolism, cell cycle regulation, and the plant hormones auxin and cytokinin during the early stages of this process. Conclusions Our findings provide important insight into the multiple roles of ROS in regulating rhizobia–legume symbiosis.

scholarly journals Genetic Interaction Studies Reveal Superior Performance of Rhizobium tropici CIAT899 on a Range of Diverse East African Common Bean (Phaseolus vulgaris L.) Genotypes

2019 ◽  
Vol 85 (24) ◽  
Author(s):  
A. H. Gunnabo ◽  
R. Geurts ◽  
E. Wolde-meskel ◽  
T. Degefu ◽  
K. E. Giller ◽  
...  
Keyword(s):  
Phaseolus Vulgaris ◽  
Common Bean ◽  
Genetic Interaction ◽  
A Priori ◽  
Superior Performance ◽  
Rhizobium Tropici ◽  
East African ◽  
Positive Interaction ◽  
Content Type ◽  
Strain Interaction

ABSTRACT We studied symbiotic performance of factorial combinations of diverse rhizobial genotypes (GR) and East African common bean varieties (GL) that comprise Andean and Mesoamerican genetic groups. An initial wide screening in modified Leonard jars (LJ) was followed by evaluation of a subset of strains and genotypes in pots (contained the same, sterile medium) in which fixed nitrogen was also quantified. An additive main effect and multiplicative interaction (AMMI) model was used to identify the contribution of individual strains and plant genotypes to the GL × GR interaction. Strong and highly significant GL × GR interaction was found in the LJ experiment but with little evidence of a relation to genetic background or growth habits. The interaction was much weaker in the pot experiment, with all bean genotypes and Rhizobium strains having relatively stable performance. We found that R. etli strain CFN42 and R. tropici strains CIAT899 and NAK91 were effective across bean genotypes but with the latter showing evidence of positive interaction with two specific bean genotypes. This suggests that selection of bean varieties based on their response to inoculation is possible. On the other hand, we show that symbiotic performance is not predicted by any a priori grouping, limiting the scope for more general recommendations. The fact that the strength and pattern of GL × GR depended on growing conditions provides an important cautionary message for future studies. IMPORTANCE The existence of genotype-by-strain (GL × GR) interaction has implications for the expected stability of performance of legume inoculants and could represent both challenges and opportunities for improvement of nitrogen fixation. We find that significant genotype-by-strain interaction exists in common bean (Phaseolus vulgaris L.) but that the strength and direction of this interaction depends on the growing environment used to evaluate biomass. Strong genotype and strain main effects, combined with a lack of predictable patterns in GL × GR, suggests that at best individual bean genotypes and strains can be selected for superior additive performance. The observation that the screening environment may affect experimental outcome of GL × GR means that identified patterns should be corroborated under more realistic conditions.

scholarly journals Rhizobium tropici Genes Involved in Free-Living Salt Tolerance are Required for the Establishment of Efficient Nitrogen-Fixing Symbiosis with Phaseolus vulgaris

2002 ◽  
Vol 15 (3) ◽  
pp. 225-232 ◽  
Author(s):  
Joaquina Nogales ◽  
Rosario Campos ◽  
Hanaa BenAbdelkhalek ◽  
José Olivares ◽  
Carmen Lluch ◽  
...  
Keyword(s):  
Salt Tolerance ◽  
Phaseolus Vulgaris ◽  
Environmental Changes ◽  
Nitrogenase Activity ◽  
Regulation Of Gene Expression ◽  
Elongation Factor ◽  
Regulatory System ◽  
Trna Synthetase ◽  
Nitrogen Fixing ◽  
Rhizobium Tropici

Characterization of nine transposon-induced mutants of Rhizobium tropici with decreased salt tolerance (DST) allowed the identification of eight gene loci required for adaptation to high external NaCl. Most of the genes also were involved in adaptation to hyperosmotic media and were required to overcome the toxicity of LiCl. According to their possible functions, genes identified could be classified into three groups. The first group included two genes involved in regulation of gene expression, such as ntrY, the sensor element of the bacterial ntrY/ntrX two-component regulatory system involved in regulation of nitrogen metabolism, and greA, which encodes a transcription elongation factor. The second group included genes related to synthesis, assembly, or maturation of proteins, such as alaS coding for alanine-tRNA synthetase, dnaJ, which encodes a molecular chaperone, and a nifS homolog probably encoding a cysteine desulfurase involved in the maturation of Fe-S proteins. Genes related with cellular build-up and maintenance were in the third group, such as a noeJ-homolog, encoding a mannose-1-phosphate guanylyltransferase likely involved in lipopolysaccharide biosynthesis, and kup, specifying an inner-membrane protein involved in potassium uptake. Another gene was identified that had no homology to known genes but that could be conserved in other rhizobia. When inoculated on Phaseolus vulgaris growing under nonsaline conditions, all DST mutants displayed severe symbiotic defects: ntrY and noeJ mutants were impaired in nodulation, and the remaining mutants formed symbiosis with very reduced nitrogenase activity. The results suggest that bacterial ability to adapt to hyper-osmotic and salt stress is important for the bacteroid nitrogen-fixing function inside the legume nodule and provide genetic evidence supporting the suggestion that rhizobia face severe environmental changes after their release into plant cells.

Microtubular configurations during endosperm development in Phaseolus vulgaris

1994 ◽  
Vol 72 (10) ◽  
pp. 1489-1495 ◽  
Author(s):  
X. XuHan ◽  
A. A. M. Van Lammeren
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Phaseolus Vulgaris ◽  
Key Words ◽  
Cell Walls ◽  
Endosperm Development ◽  
Initial Cell ◽  
Related Cell ◽  
Cellular Endosperm ◽  
Cell Shaping

Microtubular cytoskeletons in nuclear, alveolar, and cellular endosperm of bean (Phaseolus vulgaris) were analyzed immunocytochemically and by electron microscopy to reveal their function during cellularization. Nuclear endosperm showed a fine network of microtubules between the wide-spaced nuclei observed towards the chalazal pole. Near the embryo, where nuclei were densely packed, bundles of microtubules radiated from nuclei. They were formed just before alveolus formation and functioned in spacing nuclei and in forming internuclear, phragmoplast-like structures that gave rise to nonmitosis-related cell plates. During alveolus formation cell plates extended and fused with other newly formed walls, thus forming the walls of alveoli. Growing wall edges of cell plates exhibited arrays of microtubules perpendicular to the plane of the wall, initially. When two growing walls were about to fuse, microtubules of both walls interacted, and because of the interaction of microtubules, the cell walls changed their position. When a growing wall was about to fuse with an already existing wall, such interactions between microtubules were not observed. It is therefore concluded that interactions of microtubules of fusing walls influence shape and position of walls. Thus microtubules control the dynamics of cell wall positioning and initial cell shaping. Key words: cell wall, cellularization, endosperm, microtubule, Phaseolus vulgaris.

scholarly journals Aminoethoxyvinylglycine Inhibits Fruit Abscission Induced by Naphthaleneacetic Acid and Associated Relationships with Expression of Genes for Ethylene Biosynthesis, Perception, and Cell Wall Degradation in ‘Delicious’ Apples

2008 ◽  
Vol 133 (6) ◽  
pp. 727-734 ◽  
Author(s):  
Hong Zhu ◽  
Eric P. Beers ◽  
Rongcai Yuan
Keyword(s):  
Cell Wall ◽  
Ethylene Production ◽  
Acc Oxidase ◽  
Ethylene Biosynthesis ◽  
Naphthaleneacetic Acid ◽  
Cell Wall Degradation ◽  
Oxidase Gene ◽  
Young Fruit ◽  
Fruit Abscission ◽  
Expression Of Genes

Effects of naphthaleneacetic acid (NAA) and aminoethoxyvinylglycine (AVG) on young fruit abscission, leaf and fruit ethylene production, and expression of genes related to ethylene biosynthesis and cell wall degradation were examined in ‘Delicious’ apples (Malus ×domestica Borkh.). NAA at 15 mg·L−1 increased fruit abscission and ethylene production of leaves and fruit when applied at the 11-mm stage of fruit development, whereas AVG, an inhibitor of ethylene biosynthesis, at 250 mg·L−1 reduced NAA-induced fruit abscission and ethylene production of leaves and fruit. NAA also increased expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase genes (MdACS5A and MdACS5B), ACC oxidase gene (MdACO1), and ethylene receptor genes (MdETR1a, MdETR1b, MdETR2, MdERS1, and MdERS2) in fruit cortex and fruit abscission zones. However, AVG reduced NAA-induced expression of these genes except for MdERS2 in fruit abscission zones. NAA increased expression of the polygalacturonase gene MdPG2 in fruit abscission zones but not in fruit cortex, whereas AVG reduced NAA-enhanced expression of MdPG2 in fruit abscission zones. The expression of β-1,4-glucanase gene MdCel1 in fruit abscission zones was decreased by NAA but was unaffected by AVG. Our results suggest that ethylene biosynthesis, ethylene perception, and the MdPG2 gene are involved in young fruit abscission caused by NAA.

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