scholarly journals Label-Free Comparative Proteomic Analysis Combined with Laser-Capture Microdissection Suggests Important Roles of Stress Responses in the Black Layer of Maize Kernels

2020 ◽  
Vol 21 (4) ◽  
pp. 1369
Author(s):  
Quanquan Chen ◽  
Ran Huang ◽  
Zhenxiang Xu ◽  
Yaxin Zhang ◽  
Li Li ◽  
...  
Keyword(s):  
Laser Capture Microdissection ◽  
Stress Responses ◽  
Label Free ◽  
Kernel Development ◽  
Flight Mass Spectrometry ◽  
Specific Proteins ◽  
Response To Stress ◽  
Black Layer ◽  
Maize Kernels ◽  
Laser Capture

The black layer (BL) is traditionally used as an indicator for kernel harvesting in maize, as it turns visibly dark when the kernel reaches physiological maturity. However, the molecular roles of BL in kernel development have not been fully elucidated. In this work, microscopy images showed that BL began to appear at a growth stage earlier than 10 days after pollination (DAP), and its color gradually deepened to become dark as the development period progressed. Scanning electron microscopy observations revealed that BL is a tissue structure composed of several layers of cells that are gradually squeezed and compressed during kernel development. Laser-capture microdissection (LCM) was used to sample BL and its neighboring inner tissue, basal endosperm transfer layer (BETL), and outer tissue, inner epidermis (IEP), from 20 DAP of kernels. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry profiling (MALDI-TOF MS profiling) detected 41, 104, and 120 proteins from LCM-sampled BL, BETL, and IEP, respectively. Gene ontology (GO) analysis indicated that the 41 BL proteins were primarily involved in the response to stress and stimuli. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that the BL proteins were enriched in several defense pathways, such as the ascorbate and aldarate metabolic pathways. Among the 41 BL proteins, six were BL-specific proteins that were only detected from BL. Annotations of five BL-specific proteins were related to stress responses. During kernel development, transcriptional expression of most BL proteins showed an increase, followed by a decrease, and reached a maximum zero to 20 DAP. These results suggest a role for BL in stress responses for protecting filial tissue against threats from maternal sides, which helps to elucidate the biological functions of BL.

scholarly journals Laser Capture Microdissection-Assisted Protein Biomarker Discovery from Coccidioides-Infected Lung Tissue

2020 ◽  
Vol 6 (4) ◽  
pp. 365
Author(s):  
Natalie M. Mitchell ◽  
Surendra Dasari ◽  
Thomas E. Grys ◽  
Douglas F. Lake
Keyword(s):  
Laser Capture Microdissection ◽  
Biomarker Discovery ◽  
Pathogenic Fungus ◽  
Dependent Manner ◽  
Label Free ◽  
Human Lungs ◽  
Human Proteins ◽  
Laser Capture

Laser capture microdissection (LCM) coupled to label-free quantitative mass spectrometry is a viable strategy to identify biomarkers from infected tissues. In this study, LCM was employed to take a “snapshot” of proteins produced in vivo during Coccidiodies spp. infection in human lungs. Proteomic analysis of LCM lung sections revealed hundreds of hosts and Coccidioidal proteins. Twenty-seven highly abundant Coccidioides spp. proteins were identified which do not share significant sequence orthology with human proteins. Three of the 27 Coccidioidal proteins are also potential Coccidoides-specific biomarkers, as they also do not share sequence homology to any other pathogenic fungus or microbe. Gene ontology analysis of the 27 biomarker candidate proteins revealed enriched hydrolase activity and increased purine and carbohydrate metabolism functions. Finally, we provide proteomic evidence that all 27 biomarker candidates are produced by the fungus when grown in vitro in a media- and growth-phase dependent manner.

scholarly journals A proteomic analysis of grain yield-related traits in wheat

AoB Plants ◽  
2020 ◽  
Vol 12 (5) ◽  
Author(s):  
Sintayehu D Daba ◽  
Xiaoqin Liu ◽  
Uma Aryal ◽  
Mohsen Mohammadi
Keyword(s):  
Grain Yield ◽  
Quantitative Proteomics ◽  
Label Free ◽  
Leaf Sample ◽  
Kernel Development ◽  
Argonaute Proteins ◽  
Organ Specific ◽  
Specific Proteins ◽  
Important Trait ◽  
Spike Initiation

Abstract Grain yield, which is mainly contributed by tillering capacity as well as kernel number and weight, is the most important trait to plant breeders and agronomists. Label-free quantitative proteomics was used to analyse yield-contributing organs in wheat. These were leaf sample, tiller initiation, spike initiation, ovary and three successive kernel development stages at 5, 10 and 15 days after anthesis (DAA). We identified 3182 proteins across all samples. The largest number was obtained for spike initiation (1673), while the smallest was kernel sample at 15 DAA (709). Of the 3182 proteins, 296 of them were common to all seven organs. Organ-specific proteins ranged from 148 in ovary to 561 in spike initiation. When relative protein abundances were compared to that of leaf sample, 347 and 519 proteins were identified as differentially abundant in tiller initiation and spike initiation, respectively. When compared with ovary, 81, 35 and 96 proteins were identified as differentially abundant in kernels sampled at 5, 10 and 15 DAA, respectively. Our study indicated that two Argonaute proteins were solely expressed in spike initiation. Of the four expansin proteins detected, three of them were mainly expressed during the first 10 days of kernel development after anthesis. We also detected cell wall invertases and sucrose and starch synthases mainly during the kernel development period. The manipulation of these proteins could lead to increases in tillers, kernels per spike or final grain weight, and is worth exploring in future studies.

Identification of Vascular Breast Tumor Markers by Laser Capture Microdissection and Label-Free LC−MS

2011 ◽  
Vol 10 (5) ◽  
pp. 2479-2493 ◽  
Author(s):  
Jennifer J. Hill ◽  
Tammy-Lynn Tremblay ◽  
Ally Pen ◽  
Jie Li ◽  
Anna C. Robotham ◽  
...  
Keyword(s):  
Tumor Markers ◽  
Laser Capture Microdissection ◽  
Breast Tumor ◽  
Label Free ◽  
Laser Capture

Adaptation to stress in yeast: to translate or not?

2012 ◽  
Vol 40 (4) ◽  
pp. 794-799 ◽  
Author(s):  
Clare E. Simpson ◽  
Mark P. Ashe
Keyword(s):  
Protein Synthesis ◽  
Translation Initiation ◽  
Stress Responses ◽  
Translational Regulation ◽  
Protein Production ◽  
Yeast Cells ◽  
Specific Proteins ◽  
Response To Stress ◽  
Eukaryotic Organisms ◽  
The Impact

For most eukaryotic organisms, including Saccharomyces cerevisiae, the rapid inhibition of protein synthesis forms part of a response to stress. In order to balance the changing conditions, precise stress-specific alterations to the cell's proteome are required. Therefore, in the background of a global down-regulation in protein synthesis, specific proteins are induced. Given the level of plasticity required to enable stress-specific alterations of this kind, it is surprising that the mechanisms of translational regulation are not more diverse. In the present review, we summarize the impact of stress on translation initiation, highlighting both the similarities and distinctions between various stress responses. Finally, we speculate as to how yeast cells generate stress-responsive programmes of protein production when regulation is focused on the same steps in the translation pathway.

scholarly journals A quantitative proteomic workflow for characterization of frozen clinical biopsies: Laser capture microdissection coupled with label-free mass spectrometry

2012 ◽  
Vol 77 ◽  
pp. 433-440 ◽  
Author(s):  
John P. Shapiro ◽  
Sabyasachi Biswas ◽  
Anand S. Merchant ◽  
Anjali Satoskar ◽  
Cenny Taslim ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Capture Microdissection ◽  
Label Free ◽  
Laser Capture

Isolating Human Notochord Using Laser Capture Microdissection

2016 ◽  
Vol 77 (S 02) ◽  
Author(s):  
Youssef Yakkioui ◽  
Remco Santegoeds ◽  
Koo van Overbeeke ◽  
Andreas Herrler ◽  
Yasin Temel
Keyword(s):  
Laser Capture Microdissection ◽  
Laser Capture
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