Acetocarmine squash method for observing sugar beet chromosomes

The protein content and its type are principal factors affecting wheat (Triticum aestivum) end product quality. Among the wheat proteins, glutenin proteins, especially, high molecular weight glutenin subunits (HMW-GS) are major determinants of processing quality. Wheat and its primary gene pool have limited variation in terms of HMW-GS alleles. Wild relatives of wheat are an important source of genetic variation. For improvement of wheat processing quality its wild relative Thinopyrum elongatum with significant potential was utilized. An attempt was made to replace Th. elongatum chromosome long arm (1EL) carrying HMW-GS genes related to high dough strength with chromosome 1AL of wheat with least or negative effect on dough strength while retaining the chromosomes 1DL and 1BL with a positive effect on bread making quality. To create chromosome specific translocation line [1EL(1AS)], double monosomic of chromosomes 1E and 1A were created and further crossed with different cultivars and homoeologous pairing suppressor mutant line PhI. The primary selection was based upon glutenin and gliadin protein profiles, followed by sequential genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH). These steps significantly reduced time, efforts, and economic cost in the generation of translocation line. In order to assess the effect of translocation on wheat quality, background recovery was carried out by backcrossing with recurrent parent for several generations and then selfing while selecting in each generation. Good recovery of parent background indicated the development of almost near isogenic line (NIL). Morphologically also translocation line was similar to recipient cultivar N61 that was further confirmed by seed storage protein profiles, RP-HPLC and scanning electron microscopy. The processing quality characteristics of translocation line (BC4F6) indicated significant improvement in the gluten performance index (GPI), dough mixing properties, dough strength, and extensibility. Our work aims to address the challenge of limited genetic diversity especially at chromosome 1A HMW-GS locus. We report successful development of chromosome 1A specific translocation line of Th. elongatum in wheat with improved dough strength.

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“…The Acetocarmine squash method (Tsuchiya and Nakamura, 1979 ) was utilized for the preparation of mitotic chromosome spreads. Th.…”

Section: Methodsmentioning

confidence: 99%

Rapid Development and Characterization of Chromosome Specific Translocation Line of Thinopyrum elongatum with Improved Dough Strength

et al. 2017

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“…Cytological analysis was made using root-tips and anthers fixed with acetic alcohol (1:3) and stained with alcohol-HCl-carmine (SNOW 1963). Preparations were made according to a squash method (TSUCHIYA and NAKAMURA 1979). Pollen fertility was determined in 100 pollen grains stained with 0.2 % acetocarmine.…”

Section: Methodsmentioning

confidence: 99%

Nematode Resistance and Meiotic Abnormality in Diploid Sugar Beet Selected from Interspecific Beta vulgaris×B. procumbens Hybrids*

Nakamura

Tsuchiya

1988

Plant Breeding

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T"w' O nematode-resistant trisomic lines which were derived from interspecific Beta vuigans X ii. procumbens hybrids were intercrossed or backcrossed with susceptible diploid sugar beet and their progenies were screened for nematode resistance. The transmission rate of resistance varied from 1.5 % to 47.6 % with an average of 20.4 % in the progenies of individual trisomics derived from the two trisomic hnes. Eleven resistant diploids were selected with a frequency of 0.2 %. These resistant diploids were classified mto two groups, i.e., one group showed relatively high transmission rates of resistance with an average of 25.4 % and the other extremely low with an average of 1.2 % in their backcrossed and selfed progenies. Meiotic chromosome behavior in a resistant diploid group with high transmission rates was considerably normal as compared to that in a resistant diploid group with low transmission rates. Chromatid bridges and acentric fragments were detected in 93 % of resistant diploids and in 46 % of susceptible diploids. Two different sized fragments occurred in resistant diploids, while only a smaller fragment was present in susceptible diploids. A frequency of sporocytes with bridges-fragments was 17.4 % at anap'hase I and 13.9 % at anaphase II in resistant diploids, while in susceptible diploids a frequency was 2.9 % and 5.3 % at the respective stages. These results suggest that at least two paracentric inversions are present in resistant diploids, one of which is linked to nematode resistance and may be responsible for the low transmission rate of resistance.

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“…Cytological analyses were performed on root tips of the plants used for flow cytometry. Chromosome counts were done on slides made according to the acetocarmine squash protocol as described by Tsuchiya and Nakamura [32] with some modifications. Root tips, approximately 1.5 cm in length, were cut and immediately pretreated with 8-hydroxyquinoline (Sigma, USA) for 4 h at room temperature and then fixed in Farmer's solution (3:1 absolute ethanol:glacial acetic acid) and stored at 4°C for at least 24 h. For mitotic analysis, root tips were stained with 2% acetocarmine and kept at least 3-4 days at 4°C.…”

Section: Chromosome Countsmentioning

confidence: 99%

In vitro propagation of Silene bolanthoides Quézel, Contandr. & Pamukç. and assessment of genetic stability by flow cytometry

Çördük¹,

Yücel²,

Akıncı³

et al. 2018

ARCH BIOL SCI BELGRA

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Silene bolanthoides Quézel, Contandr. & Pamukç. is an endemic species from Kazdagi (Mt. Ida), Canakkale-Balikesir, Turkey. In order to develop an efficient shoot regeneration protocol, the leaf, nodal and internodal explants of S. bolanthoides were cultured on Murashige and Skoog (MS) medium containing benzyladenine (BA) alone or in combination with α-naphthaleneacetic acid (NAA). The highest number of regenerated shoots (5.75±0.1) was obtained from nodal explants that were cultured on MS medium with 8.8 µM BA+0.54 µM NAA. Regenerated shoots were rooted on MS medium without plant growth regulators (PGRs). Rooted plants (2-3 cm) were separately transferred to pots containing a mixture of peat and perlite (3:1 v/v) and acclimatized successfully in a growth chamber. Genetic stability of the propagated plants was assessed by flow cytometry and cytological analysis. Flow cytometry analysis demonstrated that regenerated plants had 2.61±0.01 pg nuclear DNA (2C) and seed-derived plants had on average 2.58±0.02 pg/2C. Cytological analysis showed that the regenerated plants had the same chromosome number as seed-derived plants of S. bolanthoides (2n=24). It was determined that regenerated plants were uniform in chromosome number and had a similar DNA content to the seed-derived ones, indicating that the described efficient shoot regeneration protocol can be applied for ex situ conservation of this species.

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Acetocarmine squash method for observing sugar beet chromosomes

Cited by 15 publications

References 16 publications

Rapid Development and Characterization of Chromosome Specific Translocation Line of Thinopyrum elongatum with Improved Dough Strength

Rapid Development and Characterization of Chromosome Specific Translocation Line of Thinopyrum elongatum with Improved Dough Strength

Nematode Resistance and Meiotic Abnormality in Diploid Sugar Beet Selected from Interspecific Beta vulgaris×B. procumbens Hybrids*

In vitro propagation of Silene bolanthoides Quézel, Contandr. & Pamukç. and assessment of genetic stability by flow cytometry

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