scholarly journals A Root Tip-Specific Expressing Anthocyanin Marker for Direct Identification of Transgenic Tissues by the Naked Eye in Symbiotic Studies

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 605
Author(s):  
Yiting Ruan ◽  
Ke Chen ◽  
Yangyang Su ◽  
Suyu Jiang ◽  
Ping Xu ◽  
...  
Keyword(s):  
Root Cap ◽  
Root Tip ◽  
Root Tips ◽  
Reporter System ◽  
Specific Expression ◽  
Transgenic Roots ◽  
Fluorescent Reporters ◽  
Naked Eye ◽  
Root Transformation ◽  
Transgenic Hairy Roots

The Agrobacterium rhizogenes hairy root transformation system is widely used in symbiotic studies of model legumes. It typically relies on fluorescent reporters, such as DsRed, for identification of transgenic roots. The MtLAP1 transcription factor has been utilized as a reporter system in Medicago truncatula based on production of anthocyanin pigment. Here, we describe a version of this reporter driven by a root-cap specific promoter for direct observation of anthocyanin accumulation in root tips, which allows the identification of transgenic hairy roots by the naked eye. Results from our analysis suggest that the reporter had no significant effects on nodulation of M. truncatula. This approach, by virtue of its strong and specific expression in root cap cells, greatly reduces false positives and false negatives, and its use of an easily scored visible pigment should allow greater versatility and efficiency in root biology studies.

scholarly journals The relationship of root-cap slimes to proteins

1974 ◽  
Vol 139 (3) ◽  
pp. 525-534 ◽  
Author(s):  
Kenneth Wright ◽  
D. H. Northcote
Keyword(s):  
Chemical Characteristic ◽  
Wheat Root ◽  
Root Cap ◽  
Water Soluble ◽  
Maize Root ◽  
Root Tip ◽  
Root Tips ◽  
Maize Roots ◽  
Neutral Sugars ◽  
Relationship Of

1. The patterns of incorporation of radioactivity from d-[U-14C]glucose into the pectic components of sections of sycamore roots changed so that sections nearer the tip incorporated relatively more label into arabinose and galactose compared with uronic acid. 2. Radioactive maize root-cap slime was prepared and found to contain three water-soluble component polymers which were electrophoretically (i) neutral, (ii) weakly acidic and (iii) strongly acidic at pH6.5. The neutral component was a glucan. The other components, which could be degraded by trans-elimination, consisted of an acidic backbone chain composed of galacturonic acid and glucose, attached to which were different proportions of neutral sugars. Arabinose, galactose and fucose, the main neutral sugars of the weakly and strongly acidic materials, were absent from the neutral fraction. 3. Fucose was a major sugar in maize-root slime and in a slime of similar composition synthesized by a maize callus of shoot origin. Only trace amounts were found in sycamore, pea and wheat root tips, and in pectin prepared from maize roots and coleoptiles. A high proportion of fucose is therefore a chemical characteristic of maize slime, and slime synthesis indicated a state of differentiation of the tissue. 4. The similarity between the slime and pectin is discussed; slime is a form of pectin modified in such a way as to provide a hydrated protective coating around the root tip.

Localization of acid phosphatases in root cap of rice plant

1991 ◽  
Vol 49 ◽  
pp. 224-225
Author(s):  
Y. R. Chen ◽  
Y. F. Huang ◽  
W. S. Chen
Keyword(s):  
Rice Plant ◽  
Developmental Stages ◽  
Uranyl Acetate ◽  
Root Cap ◽  
Plant Tissues ◽  
Acid Phosphatases ◽  
Root Tips ◽  
Buffer Solution ◽  
Cacodylate Buffer ◽  
Ethanol Series

Acid phosphatases are widely distributed in different tisssues of various plants. Studies on subcellular localization of acid phosphatases show they might be present in cell wall, plasma lemma, mitochondria, plastid, vacuole and nucleus. However, their localization in rice cell varies with developmental stages of cells and plant tissues. In present study, acid phosphatases occurring in root cap are examined.Sliced root tips of ten-day-old rice(Oryza sativa) seedlings were fixed in 0.1M cacodylate buffer containing 2.5% glutaraldehyde for 2h, washed overnight in same buffer solution, incubated in Gomori's solution at 37° C for 90min, post-fixed in OsO4, dehydrated in ethanol series and finally embeded in Spurr's resin. Sections were doubly stained with uranyl acetate and lead citrate, and observed under Hitachi H-600 at 75 KV.

Histochemical Examination of Invertase Activities in the Growing Tip of the Plant Root

2017 ◽  
Vol 10 (1) ◽  
pp. 35-45
Author(s):  
N.F. Lunkova ◽  
N.A. Burmistrova ◽  
M.S. Krasavina
Keyword(s):  
Plant Root ◽  
Invertase Activity ◽  
Root Tip ◽  
Elongation Zone ◽  
Root Tips ◽  
Phloem Unloading ◽  
Meristem Cell ◽  
Transition To Elongation ◽  
Different Tissues

Background:A growing part of the root is one of the most active sinks for sucrose coming from source leaves through the phloem. In the root, sucrose is unloaded from conducting bundles and is distributed among the surrounding cells. To be involved in the metabolism, sucrose should disintegrate into hexoses by means of degrading enzymes.Aims:The aim of this research was to explore the possibility of the involvement of one such enzymes, invertase, in phloem unloading as well as distribution of its activity in the functionally different tissues of the plant root tips.Method:To estimate the enzyme activities in root tissues, we applied two techniques: the histochemical method using nitro blue tetrazolium. The localization of phloem unloading was studied with carboxyfluorescein, a fluorescent marker for symplastic transport.Results:Invertase activity was not detected in the apical part of the meristem. It appeared only between the basal part of this zone and the beginning of the elongation zone. There is the root phloem unloading in that area. Invertase activity increased with increasing the distance from the root tip and reached the highest values in the region of cell transition to elongation and in the elongation zone. The activities of the enzyme varied in different tissues of the same zone and sometimes in the neighboring cells of the same tissue. Biochemical determination of invertase activity was made in the maize root segments coincident to the zones of meristem, cell elongation and differentiation. The results of both methods of determination of invertase activity were in agreement.Conclusion:It was concluded that phloem unloading correlated with invertase activity, possibly because of the activation of invertase by unloaded sucrose. Invertase is one of the factors involved in the processes preparing the cells for their transition to elongation because the concentration of osmotically active hexoses increases after cleavage of sucrose, that stimulates water entry into the cells, which is necessary for elongation growth.

scholarly journals Effect of earthworms on mycorrhization, root morphology and biomass of silver fir seedlings inoculated with black summer truffle (Tuber aestivum Vittad.)

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tina Unuk Nahberger ◽  
Gian Maria Niccolò Benucci ◽  
Hojka Kraigher ◽  
Tine Grebenc
Keyword(s):  
Root System ◽  
Root Biomass ◽  
Fine Root ◽  
Abies Alba ◽  
Fine Root Biomass ◽  
Root Tip ◽  
Silver Fir ◽  
Tuber Aestivum ◽  
Root Tips ◽  
Root Parameters

AbstractSpecies of the genus Tuber have gained a lot of attention in recent decades due to their aromatic hypogenous fruitbodies, which can bring high prices on the market. The tendency in truffle production is to infect oak, hazel, beech, etc. in greenhouse conditions. We aimed to show whether silver fir (Abies alba Mill.) can be an appropriate host partner for commercial mycorrhization with truffles, and how earthworms in the inoculation substrate would affect the mycorrhization dynamics. Silver fir seedlings inoculated with Tuber. aestivum were analyzed for root system parameters and mycorrhization, how earthworms affect the bare root system, and if mycorrhization parameters change when earthworms are added to the inoculation substrate. Seedlings were analyzed 6 and 12 months after spore inoculation. Mycorrhization with or without earthworms revealed contrasting effects on fine root biomass and morphology of silver fir seedlings. Only a few of the assessed fine root parameters showed statistically significant response, namely higher fine root biomass and fine root tip density in inoculated seedlings without earthworms 6 months after inoculation, lower fine root tip density when earthworms were added, the specific root tip density increased in inoculated seedlings without earthworms 12 months after inoculation, and general negative effect of earthworm on branching density. Silver fir was confirmed as a suitable host partner for commercial mycorrhization with truffles, with 6% and 35% mycorrhization 6 months after inoculation and between 36% and 55% mycorrhization 12 months after inoculation. The effect of earthworms on mycorrhization of silver fir with Tuber aestivum was positive only after 6 months of mycorrhization, while this effect disappeared and turned insignificantly negative after 12 months due to the secondary effect of grazing on ectomycorrhizal root tips.

scholarly journals Molecular Identification of Ectomycorrhizal Mycelium in Soil Horizons

2003 ◽  
Vol 69 (1) ◽  
pp. 327-333 ◽  
Author(s):  
Renske Landeweert ◽  
Paula Leeflang ◽  
Thom W. Kuyper ◽  
Ellis Hoffland ◽  
Anna Rosling ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Sequence Similarity ◽  
Mineral Soil ◽  
Molecular Techniques ◽  
Root Tip ◽  
Root Tips ◽  
Soil Horizons ◽  
Novel Approach ◽  
Total Dna ◽  
Identification Techniques

ABSTRACT Molecular identification techniques based on total DNA extraction provide a unique tool for identification of mycelium in soil. Using molecular identification techniques, the ectomycorrhizal (EM) fungal community under coniferous vegetation was analyzed. Soil samples were taken at different depths from four horizons of a podzol profile. A basidiomycete-specific primer pair (ITS1F-ITS4B) was used to amplify fungal internal transcribed spacer (ITS) sequences from total DNA extracts of the soil horizons. Amplified basidiomycete DNA was cloned and sequenced, and a selection of the obtained clones was analyzed phylogenetically. Based on sequence similarity, the fungal clone sequences were sorted into 25 different fungal groups, or operational taxonomic units (OTUs). Out of 25 basidiomycete OTUs, 7 OTUs showed high nucleotide homology (≥99%) with known EM fungal sequences and 16 were found exclusively in the mineral soil. The taxonomic positions of six OTUs remained unclear. OTU sequences were compared to sequences from morphotyped EM root tips collected from the same sites. Of the 25 OTUs, 10 OTUs had ≥98% sequence similarity with these EM root tip sequences. The present study demonstrates the use of molecular techniques to identify EM hyphae in various soil types. This approach differs from the conventional method of EM root tip identification and provides a novel approach to examine EM fungal communities in soil.

Proteomic Analyses of Soybean Root Tips During Germination

2014 ◽  
Vol 21 (12) ◽  
pp. 1308-1319
Author(s):  
Setsuko Komatsu ◽  
Myeong W. Oh ◽  
Hee Y. Jang ◽  
Soo J. Kwon ◽  
Hye R. Kim ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Proteomic Analysis ◽  
Plant Root ◽  
Root Tip ◽  
Root Tips ◽  
Form Complex ◽  
Molecular Weights ◽  
Soybean Seedlings ◽  
2D Gel ◽  
Proteomic Techniques

Plant root systems form complex networks with the surrounding soil environment and are controlled by both internal and external factors. To better understand the function of root tips of soybean during germination, three proteomic techniques were used to analyze the protein profiles of root tip cells. Proteins were extracted from the root tips of 4-dayold soybean seedlings and analyzed using two-dimensional (2D) gel electrophoresis-based proteomics, SDS-gel based proteomics, and gel-free proteomics techniques. A total of 121, 862, and 341 proteins were identified in root tips using the 2D gel-based, SDS gel-based, and gel-free proteomic techniques, respectively. The proteins identified by 2D gel-based proteomic analysis were predominantly localized in the cytoplasm, whereas nuclear-localized proteins were most commonly identified by the SDS gel-based and gel-free proteomics techniques. Of the 862 proteins identified in the SDS gelbased proteomic analysis, 190 were protein synthesis-related proteins. Furthermore, 24 proteins identified using the 2Dgel based proteomic technique shifted between acidic and basic isoelectric points, and 2 proteins, heat shock protein 70.2 and AAA-type ATPase, displayed two different molecular weights at the same isoelectric point. Taken together, these results suggest that a number of proteins related to protein synthesis and modification are activated in the root tips of soybean seedlings during germination.

scholarly journals The Jacalin-Related Lectin HvHorcH Is Involved in the Physiological Response of Barley Roots to Salt Stress

2021 ◽  
Vol 22 (19) ◽  
pp. 10248
Author(s):  
Katja Witzel ◽  
Andrea Matros ◽  
Uwe Bertsch ◽  
Tariq Aftab ◽  
Twan Rutten ◽  
...  
Keyword(s):  
Salt Stress ◽  
Salt Tolerance ◽  
Extracellular Fluid ◽  
Root Tip ◽  
Salinity Treatment ◽  
Root Tips ◽  
Salt Stress Response ◽  
Cdna Sequences ◽  
Barley Cultivars ◽  
Protein Levels

Salt stress tolerance of crop plants is a trait with increasing value for future food production. In an attempt to identify proteins that participate in the salt stress response of barley, we have used a cDNA library from salt-stressed seedling roots of the relatively salt-stress-tolerant cv. Morex for the transfection of a salt-stress-sensitive yeast strain (Saccharomyces cerevisiae YSH818 Δhog1 mutant). From the retrieved cDNA sequences conferring salt tolerance to the yeast mutant, eleven contained the coding sequence of a jacalin-related lectin (JRL) that shows homology to the previously identified JRL horcolin from barley coleoptiles that we therefore named the gene HvHorcH. The detection of HvHorcH protein in root extracellular fluid suggests a secretion under stress conditions. Furthermore, HvHorcH exhibited specificity towards mannose. Protein abundance of HvHorcH in roots of salt-sensitive or salt-tolerant barley cultivars were not trait-specific to salinity treatment, but protein levels increased in response to the treatment, particularly in the root tip. Expression of HvHorcH in Arabidopsis thaliana root tips increased salt tolerance. Hence, we conclude that this protein is involved in the adaptation of plants to salinity.

scholarly journals THE CYTOCHEMICAL LOCALIZATION OF ASCORBIC ACID IN ROOT TIP CELLS

1956 ◽  
Vol 2 (1) ◽  
pp. 87-92 ◽  
Author(s):  
William A. Jensen ◽  
Leroy G. Kavaljian
Keyword(s):  
Ascorbic Acid ◽  
Cell Surface ◽  
Silver Nitrate ◽  
Ribonucleic Acid ◽  
Root Tip ◽  
Root Tips ◽  
Cytochemical Localization ◽  
Meristematic Cells ◽  
Tip Cells ◽  
Root Tip Cells

The intracellular distribution of ascorbic acid was studied in frozen-dried root tips of Allium cepa and Vicia faba by the silver nitrate procedure. The sites of the ascorbic acid as indicated by the deposited silver appear as spherical (0.2 to 0.6 µ in diameter) cytoplasmic particles. The site appears to have small amounts of lipides and to be rich in ribonucleic acid. These particles are concluded to be submicroscopic in size and associated, in the elongating cell, with the cell surface. In the meristematic cells they appear fewer in number and are distributed throughout the cytoplasm.

scholarly journals Combined Transcriptome Profiling Reveals a Novel Family of Arbuscular Mycorrhizal-Specific Medicago truncatula Lectin Genes

2005 ◽  
Vol 18 (8) ◽  
pp. 771-782 ◽  
Author(s):  
André Frenzel ◽  
Katja Manthey ◽  
Andreas M. Perlick ◽  
Folker Meyer ◽  
Alfred Pühler ◽  
...  
Keyword(s):  
Cdna Library ◽  
Medicago Truncatula ◽  
Arbuscular Mycorrhizal ◽  
Am Fungi ◽  
Host Cells ◽  
Genomic Research ◽  
Cdna Libraries ◽  
Specific Expression ◽  
Transgenic Roots ◽  
Lectin Genes

The large majority of plants are capable of undergoing a tight symbiosis with arbuscular mycorrhizal (AM) fungi. During this symbiosis, highly specialized new structures called arbuscules are formed within the host cells, indicating that, during interaction with AM fungi, plants express AM-specific genetic programs. Despite increasing efforts, the number of genes known to be induced in the AM symbiosis is still low. In order to identify novel AM-induced genes which have not been listed before, 5,646 expressed sequence tags (ESTs) were generated from two Medicago truncatula cDNA libraries: a random cDNA library (MtAmp) and a suppression subtractive hybridization (SSH) library (MtGim), the latter being designed to enhance the cloning of mycorrhiza-upregulated genes. In silico expression analysis was applied to identify those tentative consensus sequences (TCs) of The Institute for Genomic Research M. truncatula gene index (MtGI) that are composed exclusively of ESTs deriving from the MtGim or MtAmp library, but not from any other cDNA library of the MtGI. This search revealed 115 MtAmp- or MTGim-specific TCs. For the majority of these TCs with sequence similarities to plant genes, the AM-specific expression was verified by quantitative reverse-transcription polymerase chain reaction. Annotation of the novel genes induced in mycorrhizal roots suggested their involvement in different transport as well as signaling processes and revealed a novel family of AM-specific lectin genes. The expression of reporter gene fusions in transgenic roots revealed an arbuscule-related expression of two members of the lectin gene family, indicating a role for AM-specific lectins during arbuscule formation or functioning.

scholarly journals Histological Observation of Primary and Secondary Aerenchyma Formation in Adventitious Roots of Syzygium kunstleri (King) Bahadur and R.C.Gaur Grown in Hypoxic Medium

Forests ◽  
2019 ◽  
Vol 10 (2) ◽  
pp. 137 ◽  
Author(s):  
Hong-Duck Sou ◽  
Masaya Masumori ◽  
Hiroyuki Kurokochi ◽  
Takeshi Tange
Keyword(s):  
Formation Process ◽  
Rainy Season ◽  
Adventitious Roots ◽  
Root Tip ◽  
Histological Observation ◽  
Root Tips ◽  
Aerenchyma Formation

Trees growing in wetlands develop adventitious roots from the trunk during the rainy season and adapt to the flooded environment by forming primary (schizogenous or lysigenous) and secondary aerenchyma in the roots. Therefore, it is necessary to clarify the formation process of each type of aerenchyma in these adventitious roots. In this study, saplings of Syzygium kunstleri (King) Bahadur and R.C.Gaur were grown under four different treatments, and a total of 12 adventitious roots generated from trunks were used to clarify the distribution of each aerenchyma type in the roots using light or epi-florescence microscopy. Schizogenous aerenchyma was observed in the root tips where the root color was white or light brown, whereas lysigenous aerenchyma was found at some distance from the root tip where the root color gradually changed from light to dark brown. The secondary aerenchyma and periderm were observed in dark brown parts near the root base. None or only one layer of phellem cells was detected in the white roots near the root tip, but dark brown roots near the root base had at least three layers of phellem cells. Considering these results, oxygen transportation may occur between primary and secondary aerenchyma at the point where two or more layers of phellem cells are formed.

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