A rapid and universal method for isolating starch granules in plant tissues

2019 ◽  
Vol 42 (12) ◽  
pp. 3355-3371
Author(s):  
Liangjie Niu ◽  
Huiying Ding ◽  
Ruiqi Hao ◽  
Hui Liu ◽  
Xiaolin Wu ◽  
...  
Keyword(s):  
Plant Tissues ◽  
Universal Method ◽  
Starch Granules

scholarly journals A universal method for high-quality RNA extraction from plant tissues rich in starch, proteins and fiber

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Amaranatha R. Vennapusa ◽  
Impa M. Somayanda ◽  
Colleen J. Doherty ◽  
S. V. Krishna Jagadish
Keyword(s):  
Rna Extraction ◽  
Poor Quality ◽  
High Yield ◽  
Plant Tissues ◽  
Universal Method ◽  
High Quality ◽  
Yield And Quality ◽  
Extraction Buffer ◽  
Co Precipitation

Abstract Using existing protocols, RNA extracted from seeds rich in starch often results in poor quality RNA, making it inappropriate for downstream applications. Though some methods are proposed for extracting RNA from plant tissue rich in starch and other polysaccharides, they invariably yield less and poor quality RNA. In order to obtain high yield and quality RNA from seeds and other plant tissues including roots a modified SDS-LiCl method was compared with existing methods, including TRIZOL kit (Invitrogen), Plant RNeasy mini kit (Qiagen), Furtado (2014) method, and CTAB-LiCl method. Modifications in the extraction buffer and solutions used for RNA precipitation resulted in a robust method for extracting RNA in seeds and roots, where extracting quality RNA is challenging. The modified SDS-LiCl method revealed intense RNA bands through gel electrophoresis and a nanodrop spectrophotometer detected ratios of ≥ 2 and 1.8 for A260/A230 and A260/A280, respectively. The absence of starch co-precipitation during RNA extraction resulted in enhanced yield and quality of RNA with RIN values of 7–9, quantified using a bioanalyzer. The high-quality RNA obtained was demonstrated to be suitable for downstream applications, such as cDNA synthesis, gene amplification, and RT-qPCR. The method was also effective in extracting RNA from seeds of other cereals including field-grown sorghum and corn. The modified SDS-LiCl method is a robust and highly reproducible RNA extraction method for plant tissues rich in starch and other secondary metabolites. The modified SDS-LiCl method successfully extracted high yield and quality RNA from mature, developing, and germinated seeds, leaves, and roots exposed to different abiotic stresses.

A universal method for direct PCR amplification of plant tissues

2017 ◽  
Vol 9 (11) ◽  
pp. 1800-1805 ◽  
Author(s):  
Yuping Li ◽  
Huanhuan Zhao ◽  
Xuefen Yan ◽  
Meng Li ◽  
Peng Chen ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Dna Purification ◽  
Plant Tissues ◽  
Universal Method ◽  
Direct Pcr ◽  
Modern Biology

PCR is a vital tool in modern biology; however, it can be costly owing to the price of commercial DNA purification kits.

scholarly journals A study of starch gelatinisation behaviour in hydrothermally-processed plant food tissues and implications for in vitro digestibility

2015 ◽  
Vol 6 (12) ◽  
pp. 3634-3641 ◽  
Author(s):  
Cathrina H. Edwards ◽  
Frederick J. Warren ◽  
Grant M. Campbell ◽  
Simon Gaisford ◽  
Paul G. Royall ◽  
...  
Keyword(s):  
In Vitro Digestibility ◽  
Plant Tissues ◽  
Particle Sizes ◽  
Starch Granules ◽  
Plant Food ◽  
Starch Gelatinisation

Within plant tissues of different particle sizes, the extent of gelatinisation revealed by DSC was related to thein vitrodigestion of encapsulated starch granules.

Microscopy studies of powdery mildew on summer squash

1991 ◽  
Vol 49 ◽  
pp. 520-521
Author(s):  
John S. Gardner ◽  
W. M. Hess
Keyword(s):  
Powdery Mildew ◽  
Tip Growth ◽  
Hyphal Growth ◽  
Plant Tissues ◽  
Vegetative Hypha ◽  
Powdery Mildews ◽  
Summer Squash ◽  
Conidial Formation ◽  
Polar Tip Growth ◽  
Short Conidiophore

Powdery mildews are characterized by the appearance of spots or patches of a white to grayish, powdery, mildewy growth on plant tissues, entire leaves or other organs. Ervsiphe cichoracearum, the powdery mildew of cucurbits is among the most serious parasites, and the most common. The conidia are formed similar to the process described for Ervsiphe graminis by Cole and Samson. Theconidial chains mature basipetally from a short, conidiophore mother-cell at the base of the fertile hypha which arises holoblastically from the conidiophore. During early development it probably elongates by polar-tip growth like a vegetative hypha. A septum forms just above the conidiophore apex. Additional septa develop in acropetal succession. However, the conidia of E. cichoracearum are more doliform than condia from E. graminis. The purpose of these investigations was to use scanning electron microscopy (SEM) to demonstrate the nature of hyphal growth and conidial formation of E. cichoracearum on field-grown squash leaves.

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.

Representation of information in SEM by equal signal lines

1978 ◽  
Vol 36 (1) ◽  
pp. 144-145
Author(s):  
E. Rau ◽  
N. Karelin ◽  
V. Dukov ◽  
M. Kolomeytsev ◽  
S. Gavrikov ◽  
...  
Keyword(s):  
Electronic System ◽  
Space Distribution ◽  
Specimen Surface ◽  
Universal Method ◽  
Base Level ◽  
Signal Value ◽  
Information Signal ◽  
Fixed Base ◽  
Scanning Line ◽  
Amplitude Analyzer

There are different methods and devices for the increase of the videosignal information in SEM. For example, with the help of special pure electronic [1] and opto-electronic [2] systems equipotential areas on the specimen surface in SEM were obtained. This report generalizes quantitative universal method for space distribution representation of research specimen parameter by contour equal signal lines. The method is based on principle of comparison of information signal value with the fixed levels.Transformation image system for obtaining equal signal lines maps was developed in two versions:1)In pure electronic system [3] it is necessary to compare signal U (see Fig.1-a), which gives potential distribution on specimen surface along each scanning line with fixed base level signals εifor obtaining quantitative equipotential information on solid state surface. The amplitude analyzer-comparator gives flare sport videopulses at any fixed coordinate and any instant time when initial signal U is equal to one of the base level signals ε.

Cryosectioning plant roots prepared by high-pressure freezing

1987 ◽  
Vol 45 ◽  
pp. 662-663
Author(s):  
R.E. Crang ◽  
M. Mueller ◽  
K. Zierold
Keyword(s):  
High Pressure ◽  
Cell Walls ◽  
Plant Tissues ◽  
Ice Crystal ◽  
High Pressure Freezing ◽  
Vitreous State ◽  
Radial Depth ◽  
Irregular Topography ◽  
Considerable Depth ◽  
Conventional Freezing

Obtaining frozen-hydrated sections of plant tissues for electron microscopy and microanalysis has been considered difficult, if not impossible, due primarily to the considerable depth of effective freezing in the tissues which would be required. The greatest depth of vitreous freezing is generally considered to be only 15-20 μm in animal specimens. Plant cells are often much larger in diameter and, if several cells are required to be intact, ice crystal damage can be expected to be so severe as to prevent successful cryoultramicrotomy. The very nature of cell walls, intercellular air spaces, irregular topography, and large vacuoles often make it impractical to use immersion, metal-mirror, or jet freezing techniques for botanical material.However, it has been proposed that high-pressure freezing (HPF) may offer an alternative to the more conventional freezing techniques, inasmuch as non-cryoprotected specimens may be frozen in a vitreous, or near-vitreous state, to a radial depth of at least 0.5 mm.

The role of silicon in forages: A quantitative X-Ray study

1987 ◽  
Vol 45 ◽  
pp. 416-417
Author(s):  
Janet H. Woodward ◽  
D. E. Akin
Keyword(s):  
Sodium Acetate ◽  
Dry Matter ◽  
Acetate Buffer ◽  
Plant Tissues ◽  
Sodium Acetate Buffer ◽  
X Ray ◽  
Dry Matter Digestibility ◽  
Percent Composition

Silicon (Si) is distributed throughout plant tissues, but its role in forages has not been clarified. Although Si has been suggested as an antiquality factor which limits the digestibility of structural carbohydrates, other research indicates that its presence in plants does not affect digestibility. We employed x-ray microanalysis to evaluate Si as an antiquality factor at specific sites of two cultivars of bermuda grass (Cynodon dactvlon (L.) Pers.). “Coastal” and “Tifton-78” were chosen for this study because previous work in our lab has shown that, although these two grasses are similar ultrastructurally, they differ in in vitro dry matter digestibility and in percent composition of Si.Two millimeter leaf sections of Tifton-7 8 (Tift-7 8) and Coastal (CBG) were incubated for 72 hr in 2.5% (w/v) cellulase in 0.05 M sodium acetate buffer, pH 5.0. For controls, sections were incubated in the sodium acetate buffer or were not treated.

An Electron Diffraction Study on Litnerized Potato Starch

1990 ◽  
Vol 48 (1) ◽  
pp. 580-581
Author(s):  
Jean-Claude Jésior ◽  
Roger Vuong ◽  
Henri Chanzy
Keyword(s):  
Electron Microscope ◽  
Electron Diffraction ◽  
Signal To Noise Ratio ◽  
Potato Starch ◽  
Image Intensifier ◽  
Electron Diffraction Study ◽  
Starch Granules ◽  
X Ray ◽  
Individual Grains ◽  
Diamond Knife

Starch is arranged in a crystalline manner within its storage granules and should thus give sharp X-ray diagrams. Unfortunately most of the common starch granules have sizes between 1 and 100μm, making them too small for an X-ray study on individual grains. There is only one instance where an oriented X-ray diagram could be obtained on one sector of an individual giant starch granule. Despite their small size, starch granules are still too thick to be studied by electron diffraction with a transmission electron microscope. The only reported study on starch ultrastructure using electron diffraction on frozen hydrated material was made on small fragments. The present study has been realized on thin sectioned granules previously litnerized to improve the signal to noise ratio.Potato starch was hydrolyzed for 10 days in 2.2N HCl at 35°C, dialyzed against water until neutrality and embedded in Nanoplast. Sectioning was achieved with a commercially available low-angle “35°” diamond knife (Diatome) after a very carefull trimming and a pre-sectioning with a classical “45°” diamond knife. Sections obtained at a final sectioning angle of 42.2° (compared with the usual 55-60°) and at a nominal thickness of 900Å were collected on a Formvar-carbon coated grid. The exact location of the starch granules in their sections was recorded by optical microscopy on a Zeiss Universal polarizing microscope (Fig. 1a). After rehydration at a relative humidity of 95% for 24 hours they were mounted on a Philips cryoholder and quench frozen in liquid nitrogen before being inserted under frozen conditions in a Philips EM 400T electron microscope equipped with a Gatan anticontaminator and a Lhesa image intensifier.

Sign in / Sign up

Export Citation Format

Share Document