Miguel Meza Lara - Academia.edu (original) (raw)

Papers by Miguel Meza Lara

Genes, 2019

Receptor-like kinases (RLKs) are conserved upstream signaling molecules that regulate several bio... more Receptor-like kinases (RLKs) are conserved upstream signaling molecules that regulate several biological processes, including plant development and stress adaptation. Cysteine (C)-rich receptor-like kinases (CRKs) are an important class of RLK that play vital roles in disease resistance and cell death in plants. Genome-wide analyses of CRK genes have been carried out in Arabidopsis and rice, while functional characterization of some CRKs has been carried out in wheat and tomato in addition to Arabidopsis. A comprehensive analysis of the CRK gene family in leguminous crops has not yet been conducted, and our understanding of their roles in symbiosis is rather limited. Here, we report the comprehensive analysis of the Phaseolus CRK gene family, including identification, sequence similarity, phylogeny, chromosomal localization, gene structures, transcript expression profiles, and in silico promoter analysis. Forty-six CRK homologs were identified and phylogenetically clustered into fiv...

Genes, Jan 3, 2018

BYPASS1 (), which is a well-conserved gene in plants, is required for normal root and shoot devel... more BYPASS1 (), which is a well-conserved gene in plants, is required for normal root and shoot development. In the absence of gene function, overproduces a mobile signalling compound (the signal) in roots, and this transmissible signal arrests shoot growth and causes abnormal root development. In addition to the shoot and root meristem activities, the legumes also possess transient meristematic activity in root cortical cells during symbiosis. We explored the role of during nodule primordium development using an RNA-interference (RNAi) silencing approach. Our results show that upon infection, the transgenic roots failed to induce cortical cell divisions without affecting the rhizobia-induced root hair curling and infection thread formation. The transcript accumulation of early nodulin genes, cell cyclins, and cyclin-dependent kinase genes was affected in RNAi lines. Interestingly, the root nodule phenotype was partially rescued by exogenous application of fluridone, a carotenoid biosyn...

Journal of visualized experiments : JoVE, Dec 23, 2017

The upstream sequences of gene coding sequences are termed as promoter sequences. Studying the ex... more The upstream sequences of gene coding sequences are termed as promoter sequences. Studying the expression patterns of promoters are very significant in understanding the gene regulation and spatiotemporal expression patterns of target genes. On the other hand, it is also critical to establish promoter evaluation tools and genetic transformation techniques that are fast, efficient, and reproducible. In this study, we investigated the spatiotemporal expression pattern of the rhizobial symbiosis-specific nodule inception (NIN) promoter of Phaseolus vulgaris in the transgenic hairy roots. Using plant genome databases and analysis tools we identified, isolated, and cloned the P. vulgaris NIN promoter in a transcriptional fusion to the chimeric reporter β-glucuronidase (GUS) GUS-enhanced::GFP. Further, this protocol describes a rapid and versatile system of genetic transformation in the P. vulgaris using Agrobacterium rhizogenes induced hairy roots. This system generates ≥2 cm hairy roots...

Symbiosis, 2017

Expansins are non-enzymatic cell wall proteins that mediate plant growth by catalyzing loosening ... more Expansins are non-enzymatic cell wall proteins that mediate plant growth by catalyzing loosening of cell walls without lysing the wall polymers. Advances in the field of bioinformatics have facilitated the prediction of the members of expansin gene family across several model plants. Expansins constitutes into four sub-families; α-expansin, βexpansin, expansin-like A and expansin-like B. Biological functions of expansin gene family include diverse aspects of plant growth and development, shoot and root elongation, leaf morphogenesis, flower and fruit development, embryogenesis, pollen tube growth, stress tolerance, etc. Recent studies have demonstrated the role of expansins in plantsymbiotic interactions. The present review reveals the factors that govern plant-arbuscular mycorrhizal fungi (AMF) and legume-rhizobia symbioses; and the genes that participate in these diverse symbiont interactions. Further, we focus on the expression profiles and the functions of expansins during plant-AMF and legume-rhizobia interactions. The key roles of expansin proteins during AMF invasion, arbuscule formation, rhizobial infection and nodule organogenesis were uncovered during symbioses. This review summarizes discoveries that support the key and versatile roles of various expansin members in the plant-mycorrhizal and legumerhizobial symbioses.

PLOS ONE, 2017

S5 Fig. DEGs of defense-responsive genes during root symbioses. Expression profile of unique defe... more S5 Fig. DEGs of defense-responsive genes during root symbioses. Expression profile of unique defense-responsive genes in P. vulgaris roots colonized by (A) AMF and (B) rhizobia. Expression profile showing upregulated and downregulated DEGs obtained form GO analysis. Statistically significant DEGs were identified using an unpaired t-test (p<0.05) in symbiont treatment over controls (S3 Table). Fold-change values (over control) were used to plot heat maps. Color bar scale shows the fold-change range with red and green representing downregulation and upregulation, respectively.

Plant Physiology, 2016

The target of rapamycin (TOR) protein kinase regulates metabolism, growth, and life span in yeast... more The target of rapamycin (TOR) protein kinase regulates metabolism, growth, and life span in yeast, animals, and plants in coordination with nutrient status and environmental conditions. The nutrient-dependent nature of TOR functionality makes this kinase a putative regulator of symbiotic associations involving nutrient acquisition. However, TOR's role in these processes remains to be understood. Here, we uncovered the role of TOR during the bean (Phaseolus vulgaris)-Rhizobium tropici (Rhizobium) symbiotic interaction. TOR was expressed in all tested bean tissues, with higher transcript levels in the root meristems and senesced nodules. We showed TOR promoter expression along the progressing infection thread and in the infected cells of mature nodules. Posttranscriptional gene silencing of TOR using RNA interference (RNAi) showed that this gene is involved in lateral root elongation and root cell organization and also alters the density, size, and number of root hairs. The suppression of TOR transcripts also affected infection thread progression and associated cortical cell divisions, resulting in a drastic reduction of nodule numbers. TOR-RNAi resulted in reduced reactive oxygen species accumulation and altered CyclinD1 and CyclinD3 expression, which are crucial factors for infection thread progression and nodule organogenesis. Enhanced expression of TORregulated ATG genes in TOR-RNAi roots suggested that TOR plays a role in the recognition of Rhizobium as a symbiont. Together, these data suggest that TOR plays a vital role in the establishment of root nodule symbiosis in the common bean.

BMC Biotechnology, 2016

Background: Phaseolus vulgaris is one of the most extensively studied model legumes in the world.... more Background: Phaseolus vulgaris is one of the most extensively studied model legumes in the world. The P. vulgaris genome sequence is available; therefore, the need for an efficient and rapid transformation system is more imperative than ever. The functional characterization of P. vulgaris genes is impeded chiefly due to the non-amenable nature of Phaseolus sp. to stable genetic transformation. Transient transformation systems are convenient and versatile alternatives for rapid gene functional characterization studies. Hence, the present work focuses on standardizing methodologies for protoplast isolation from multiple tissues and transient transformation protocols for rapid gene expression analysis in the recalcitrant grain legume P. vulgaris. Results: Herein, we provide methodologies for the high-throughput isolation of leaf mesophyll-, flower petal-, hypocotyl-, root-and nodule-derived protoplasts from P. vulgaris. The highly efficient polyethylene glycol-mannitol magnesium (PEG-MMG)-mediated transformation of leaf mesophyll protoplasts was optimized using a GUS reporter gene. We used the P. vulgaris SNF1-related protein kinase 1 (PvSnRK1) gene as proof of concept to demonstrate rapid gene functional analysis. An RT-qPCR analysis of protoplasts that had been transformed with PvSnRK1-RNAi and PvSnRK1-OE vectors showed the significant downregulation and ectopic constitutive expression (overexpression), respectively, of the PvSnRK1 transcript. We also demonstrated an improved transient transformation approach, sonication-assisted Agrobacterium-mediated transformation (SAAT), for the leaf disc infiltration of P. vulgaris. Interestingly, this method resulted in a 90 % transformation efficiency and transformed 60-85 % of the cells in a given area of the leaf surface. The constitutive expression of YFP further confirmed the amenability of the system to gene functional characterization studies.

Limnology and Oceanography: Fluids and Environments, 2013

In low-nutrient, macrophyte-dominated coastal zones, benthic ammonium (NH 4 +) uptake may be infl... more In low-nutrient, macrophyte-dominated coastal zones, benthic ammonium (NH 4 +) uptake may be influenced by the structural properties of plant canopies via their effect on near-bed hydrodynamics. Using a dual-tracer (uranine and 15 NH 4 +) method that does not require enclosures, we examined how this process affects nutrient uptake rates within a tidally dominated, patchy Caulerpa prolifera-Cymodocea nodosa landscape. NH 4 + uptake was determined by calculating tissue 15 N excesses and correcting for 15 N enrichment as derived from uranine concentration. Vertical hydrodynamic profiles were measured in the downstream flow direction from outside to inside of the C. nodosa bed by using an array of acoustic Doppler velocimeters. The transition from a C. prolifera to a C. nodosa bed included a change in both benthic canopy properties (short and dense to tall and sparse) and sediment topography (0.2-m increase in water column depth) that resulted in an increase in longitudinal advection and turbulent diffusivity within the C. nodosa canopy between 0.5 and 1.5 m from the leading edge. Vertical differences in canopy water exchange appeared to explain variations in uptake between biotic functional groups; however, no clear differences in longitudinal uptake were found. Using in situ labeling, this study demonstrated for the first time the role of hydrodynamics in structuring NH 4 + uptake within an undisturbed, patchy macrophyte landscape.

Scientia Horticulturae, 2014

The paper presents a versatile, high throughput, and a reliable in planta transformation protocol... more The paper presents a versatile, high throughput, and a reliable in planta transformation protocol. This method is advantageous over the conventional regeneration-based transformation protocols as it is genotype-independent and expandable to a variety of crop species. The methodology involves in planta inoculation of the differentiating cells of apical meristem with vir gene induced Agrobacterium tumefaciens. The study deals with the feasibility of the strategy in bell pepper by introducing a non-expresser of pathogenesis-related gene (NPR1) for resistance to powdery mildew pathogen (Leveillula taurica (Lev.) Arn.,). With the development of the transformation strategy presented in the paper, we could not only transform recalcitrant crop species with ease but also generate large number of independent transgenic events with improved transformation efficiencies. The technology can be used as a versatile tool in the development of transgenic plants in economically important crop species. Furthermore, this method is rapid as it takes 3-5 months from Agrobacterium transformation to characterization of transgenics.

Este documento representa una herramienta de diagnóstico simplificada del estado de salud de las ... more Este documento representa una herramienta de diagnóstico simplificada del estado de salud de las praderas de fanerógamas marinas, con el estudio de una serie de indicadores biológicos que permitan dicha estimación. Por tanto, este documento sirve de guía para decidir qué ...

Plants, 2021

Macroautophagy/autophagy is a fundamental catabolic pathway that maintains cellular homeostasis i... more Macroautophagy/autophagy is a fundamental catabolic pathway that maintains cellular homeostasis in eukaryotic cells by forming double-membrane-bound vesicles named autophagosomes. The autophagy family genes remain largely unexplored except in some model organisms. Legumes are a large family of economically important crops, and knowledge of their important cellular processes is essential. Here, to first address the knowledge gaps, we identified 17 ATG families in Phaseolus vulgaris, Medicago truncatula and Glycine max based on Arabidopsis sequences and elucidated their phylogenetic relationships. Second, we dissected ATG18 in subfamilies from early plant lineages, chlorophytes to higher plants, legumes, which included a total of 27 photosynthetic organisms. Third, we focused on the ATG18 family in P. vulgaris to understand the protein structure and developed a 3D model for PvATG18b. Our results identified ATG homologs in the chosen legumes and differential expression data revealed th...