Comparison of Sample Preparation Techniques for Inspection of Leaf Epidermises Using Light Microscopy and Scanning Electronic Microscopy

Yuan, Jinhong; Wang, Xiaoduan; Zhou, Huihui; Li, Yulin; Zhang, Jing; Yu, Shuxin; Wang, Mengni; Hao, Menghan; Zhao, Qian; Liu, Le; Li, Mingjun; Li, Junhua

doi:10.3389/fpls.2020.00133

Cited by 27 publications

(22 citation statements)

References 47 publications

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“…In ltration, nally resin embedding and polymerization, the tissue block was taken out and stored in a desiccator. The sample was cut into ultra-thin sections of 50-70 nm, double-stained with uranyl acetate-lead citrate, observed and photographed by transmission electron microscope (H-7650; Hitachi, Tokyo, Japan) [51,52] [53]. Nitro blue tetrazole (NBT) can react with oxygen-free radical O 2 ¯ to produce dark blue insoluble formazan, which can be used to detect the O 2 ¯ content in plant tissues [54].…”

Section: Plant Materials and Low Fe Tress Treatmentmentioning

confidence: 99%

Changes in the Transcriptomic Profiles of Allohexaploid Wheat in Response to Low Fe Stress

Wang

Hua

Zhou

et al. 2022

Preprint

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Background: Around the world, especially in aerobic soils, because the solubility of iron (Fe) is very low, plants are often stressed by low Fe tress, and the growth, yield, and quality of crops will be inhibited in the case of severe Fe deficiency. The metabolism of Fe in plants is controlled by a series of complex transport, storage, and regulatory mechanisms to maintain the homeostasis of Fe in cells. Allohexaploid wheat (Triticum aestivum L.) is an important food crop and is more sensitive to low Fe tress. Although some studies have been conducted on the low Fe tress response of different plant species, these mechanisms are still unclear in wheat. Results: According to the results of transmission electron microscope, and paraffin section, under low Fe stress, leaf chlorosis was due to damage to the chloroplast structure, and the effect on root structure was mediated by reducing the rate of cell division in meristems and reducing cell elongation in the elongation zone. ICP-MS showed that low Fe stress significantly limited the absorption of essential elements, including N, Pi, K, Ca, Mg, Fe, Mn, Cu, Zn, and B nutrients. RNA sequencing revealed the transcriptomic changes of wheat under low Fe stress. The results showed that 378 and 2,619 differentially expressed genes (DEGs) were identified in the shoots and roots, respectively. These DEGs were mainly involved in the synthesis of Fe chelating agents, ion transport, photosynthesis, amino acid metabolism, protein synthesis, and other processes. Next, to find the core genes responding to low Fe stress, the gene co-expression network diagram was constructed. The results indicated that TaIRT1b-4A, TaNAS2-6D, TaNAS1a-6A, TaNAS1-6B, and TaNAAT1b-1D might play a key role in dealing with low-Fe stress. Conclusions: These research results might help to fully understand the morphological and molecular responses of plants to low Fe stress, and provide excellent genetic resources for the genetic modification of Fe deficiency crops.

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Section: Plant Materials and Low Fe Tress Treatmentmentioning

confidence: 99%

Changes in the Transcriptomic Profiles of Allohexaploid Wheat in Response to Low Fe Stress

Wang

Hua

Zhou

et al. 2022

Preprint

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“…An electron beam has a much shorter wavelength than that of visible light of an optical microscope. Thus, compared with LMs, both the resolution and magnification of SEMs are greatly improved (Yuan et al, 2020). Caldwell and Iyer‐Pascuzzi (2019) used SEM to study restriction of Fusarium oxysporum f. sp.…”

Section: Introductionmentioning

confidence: 99%

Effect of necrotrophic fungus and PGPR on the comparative histochemistry of Vigna radiata by using multiple microscopic techniques

Javed

Javaid

Hanif

et al. 2021

Microscopy Res & Technique

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Rapid advances in the field of pathogen detection have opened new opportunities and better understanding for their management approaches. Aim of this study was to elucidate histopathological observations of different tissues affected by Macrophomina phaseolina and to observe the defense responses of plant growth promoting rhizobacteria (PGPR) in mungbean plants. Sections of the stem and root were prepared and stained with ferric chloride, Lugol's iodine and Wiesner's reagent and were then observed under multiple microscopic techniques. Results revealed that both pathogen and PGPR produce responses on the plant that include colonization of xylem vessels by hyphae and sclerotia, hypertrophy and hyperplasia of the cells, destruction of xylem fibers and amyloplasts in parenchymatous cells; and production of gels by the plant were observed. There was a significant increase in lignin and phenolic compounds deposition in stem and root sections of PGPR treated and non‐treated mungbean plants. Whereas the soil amended with PGPR showed very less to no starch production. Moreover, production of gels and gums were also observed in both stem and root sections. Compared to light microscopy, scanning electron microscope provided greater depth of focus and resolution of the pathogen attack on plant tissues, associated bacteria. As a whole, the data demonstrated that inoculation of PGPR can be an effective strategy to stimulate plant growth and they could significantly activate disease resistance against M. phaseolina.

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“…The perishable nature of fresh leaf samples demands a preservation technique while dealing with many samples. Several studies mentioned that using a preservation technique allowed cross-sections to be further used with orchids (Orchidaceae) [23] and Arabidopsis leaves [24]. The concentration of clearing agents is a factor for determining the time to become transparent leaves, which varies between a few hours and several days based on plant species [25].…”

Section: Introductionmentioning

confidence: 99%

Optimizing the Experimental Method for Stomata-Profiling Automation of Soybean Leaves Based on Deep Learning

Sultana

Park

Choi

et al. 2021

Plants

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Stomatal observation and automatic stomatal detection are useful analyses of stomata for taxonomic, biological, physiological, and eco-physiological studies. We present a new clearing method for improved microscopic imaging of stomata in soybean followed by automated stomatal detection by deep learning. We tested eight clearing agent formulations based upon different ethanol and sodium hypochlorite (NaOCl) concentrations in order to improve the transparency in leaves. An optimal formulation—a 1:1 (v/v) mixture of 95% ethanol and NaOCl (6–14%)—produced better quality images of soybean stomata. Additionally, we evaluated fixatives and dehydrating agents and selected absolute ethanol for both fixation and dehydration. This is a good substitute for formaldehyde, which is more toxic to handle. Using imaging data from this clearing method, we developed an automatic stomatal detector using deep learning and improved a deep-learning algorithm that automatically analyzes stomata through an object detection model using YOLO. The YOLO deep-learning model successfully recognized stomata with high mAP (~0.99). A web-based interface is provided to apply the model of stomatal detection for any soybean data that makes use of the new clearing protocol.

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Comparison of Sample Preparation Techniques for Inspection of Leaf Epidermises Using Light Microscopy and Scanning Electronic Microscopy

Cited by 27 publications

References 47 publications

Changes in the Transcriptomic Profiles of Allohexaploid Wheat in Response to Low Fe Stress

Changes in the Transcriptomic Profiles of Allohexaploid Wheat in Response to Low Fe Stress

Effect of necrotrophic fungus and PGPR on the comparative histochemistry of Vigna radiata by using multiple microscopic techniques

Optimizing the Experimental Method for Stomata-Profiling Automation of Soybean Leaves Based on Deep Learning

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