Dataset Information

High resolution microscopy to evaluate the efficiency of surface sterilization of Zea Mays seeds.

ABSTRACT: Surface sterilization of seeds is a key step in providing microorganisms-free seeds for numerous applications like understanding the role of seed-borne microorganisms in plant development, studying microbial cells-plant interactions by inoculating model microorganisms in a simplified system or selective cultivation of seed endobionts. However applying efficient treatment for surface sterilization of seeds without affecting the plant growth is not an easy task. In this study we aimed to provide an efficient surface sterilization treatment for maize seeds using i) hydrogen peroxide (HP), ii) sodium hypochlorite (SH) and iii) ethanol-sodium hypochlorite (EtOH-SH) under stirring (st) and vacuum-stirring (va-st) conditions. We used fluorescence microscopy and ultra-high resolution Helium Ion Microscopy (HIM) as powerful imaging approaches in combination with macroscopic techniques to visualize, quantify and evaluate the efficiency of seed sterilization, quality of root germination, seedlings and root hair development as well as the presence or absence of microorganisms on the root surface. Our results showed a strong reduction in microbial cell numbers of 4 orders of magnitude after the EtOH-SH treatments. Moreover, seeds exposed to EtOH-SH treatments displayed the lowest percentage of microbial growth (50%) and the highest percentage of germinated seeds (100%) compared to other sterilization treatments. HIM imaging proved the absence of microbial cells on the roots grown from seeds exposed to EtOH-SH treatments. Moreover, root hair development seemed not to be affected by any of the sterilization treatments. Our findings demonstrated that EtOH-SH treatments are significantly reducing the abundance of microbial cells from the surface of maize seeds and can be used with high confidence in future studies.

SUBMITTER: Davoudpour Y

PROVIDER: S-EPMC7703986 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

High resolution microscopy to evaluate the efficiency of surface sterilization of Zea Mays seeds.

Davoudpour Yalda Y Schmidt Matthias M Calabrese Federica F Richnow Hans Hermann HH Musat Niculina N

PloS one 20201130 11

Surface sterilization of seeds is a key step in providing microorganisms-free seeds for numerous applications like understanding the role of seed-borne microorganisms in plant development, studying microbial cells-plant interactions by inoculating model microorganisms in a simplified system or selective cultivation of seed endobionts. However applying efficient treatment for surface sterilization of seeds without affecting the plant growth is not an easy task. In this study we aimed to provide a ...[more]

PMID: 33253171

Similar Datasets

Project description:Root hydraulic properties play a central role in the global water cycle, in agricultural systems productivity, and in ecosystem survival as they impact the canopy water supply. However, the existing experimental methods to quantify root hydraulic conductivities, such as the root pressure probing, are particularly challenging, and their applicability to thin roots and small root segments is limited. Therefore, there is a gap in methods enabling easy estimations of root hydraulic conductivities in diverse root types. Here, we present a new pipeline to quickly estimate root hydraulic conductivities across different root types, at high resolution along root axes. Shortly, free-hand root cross-sections were used to extract a selected number of key anatomical traits. We used these traits to parametrize the Generator of Root Anatomy in R (GRANAR) model to simulate root anatomical networks. Finally, we used these generated anatomical networks within the Model of Explicit Cross-section Hydraulic Anatomy (MECHA) to compute an estimation of the root axial and radial hydraulic conductivities (k x and k r , respectively). Using this combination of anatomical data and computational models, we were able to create a root hydraulic conductivity atlas at the root system level, for 14-day-old pot-grown Zea mays (maize) plants of the var. B73. The altas highlights the significant functional variations along and between different root types. For instance, predicted variations of radial conductivity along the root axis were strongly dependent on the maturation stage of hydrophobic barriers. The same was also true for the maturation rates of the metaxylem vessels. Differences in anatomical traits along and across root types generated substantial variations in radial and axial conductivities estimated with our novel approach. Our methodological pipeline combines anatomical data and computational models to turn root cross-section images into a detailed hydraulic atlas. It is an inexpensive, fast, and easily applicable investigation tool for root hydraulics that complements existing complex experimental methods. It opens the way to high-throughput studies on the functional importance of root types in plant hydraulics, especially if combined with novel phenotyping techniques such as laser ablation tomography.

Project description:Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology. Moreover, our striped-illumination approach is able to improve the resolution of any laser-scanning-microscope, including confocal microscopes, by simply choosing an appropriate detector.

Project description:BackgroundLaser applications in agriculture have recently gained much interest due to improved plant characteristics following laser treatment before the sowing of seeds. In this study, maize seeds were exposed to different levels of laser treatment prior to sowing to improve their field performance. The aim of this study is to evaluate the impact of pre-sowing laser photobiomodulation on the field emergence and growth of treated maize seeds.MethodsThe maize seeds were first photobiomodulated with two lasers: 1) a helium-neon (He-Ne) red laser (632.8 nm), and 2) a neodymium-doped yttrium aluminum garnet (Nd:YAG) green laser (532 nm). Following three replications of randomized complete block design (RCBD), four irradiation treatments were applied (45 s, 65 s, 85 s, and 105 s) at two power intensities (2 mW/cm2 and 4 mW/cm2).ResultsBased on the results, maize seeds pretreated with a green laser and 2 mW/cm2 power intensity for 105 s exhibited the highest rate of seed emergence (96%) compared to the untreated control seeds with a lower seed emergence rate (62.5%). Furthermore, maize seeds treated with a red laser for 45 s showed an increased vigor index compared to the other treatment options and the control (P < 0.01). The treatment groups also showed statistically significant differences in seedling growth characteristics compared to the control group p < 0.01. The green laser produced a significant enhancement of about 24.20 cm in seedling length, 8.2 leaves/plant, and 3.4 cm in stem diameter compared to the untreated seeds. Moreover, the green laser treatment showed 57.4 days to anthesis, which was earlier than the untreated seeds (61.4 days). The results showed that the protein, oil, and starch contents of the seeds irradiated with the green laser were 17.54%, 6.18%, and 73.32%, respectively, compared to the seeds irradiated by the red laser with 16.51%, 6.33%, and 71.05%, respectively.ConclusionsThe photo biomodulation of maize seeds using a green laser light can improve the field emergence, seedling growth, and seed quality of the treated seed compared to the red laser treatment.

Dataset Information

High resolution microscopy to evaluate the efficiency of surface sterilization of Zea Mays seeds.

Publications

High resolution microscopy to evaluate the efficiency of surface sterilization of Zea Mays seeds.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets