NUCLEAR DNA CONTENT IN RICE (Oryza sativa L.) REGENERANTS DERIVED FROM ANTHER CULTURE in vitro

2018 ◽  
Vol 53 (3) ◽  
Author(s):  
M.V. Ilyushko ◽  
Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 17-25 ◽  
Author(s):  
Nicola J. Winston ◽  
Martin H. Johnson ◽  
Peter R. Braude

SummaryThe nuclear DNA content of intact, live or fixed, human and mouse oocytes and blastomeres has been measured rapidly and reliably. Chromosomal DNA has been stained with DAPI, the fluorescent emission from which has been measured photocytometrically.In vitrofertilised mouse oocytes and embryos at various stages of development were assessed for their DNA content. The mean values of 1C, 2C and 4C DNA content were clearly different, and it was possible to assign correctly individual values for DNA content to each class with 92%, 61% and 81% confidence respectively. Maintaining the cells as whole mounts allowed other morphological and structural features to be examined. When formation of multiple micronuclei was induced in mouse oocytes by their insemination in the presence of nocodazole, the additive signal from all the micronuclei in one zygote was equivalent to the expected DNA content. Application to early human blastomeres of this photocytometric technique for measurement of the total cellular DNA content revealed that multinucleated blastomeres contained 2C to 4C DNA levels, consistent with a diploid DNA content.


Author(s):  
M. V. Ilyushko ◽  
M. V. Romashova

Morphological variability of haploid plants and doubled haploids of rice obtained on one callus line in anther culture in vitro was studied. The work was carried out on rice plants Oryza sativa L. subspecies japonica Kato varietу Cascade. Regenerant plants of one callus line obtained from one rice anther (four in total) were divided into two or three groups of 20-30 plants, depending on the sample size in order of their differentiation on callus and transplantation on the rooting medium. Two callus lines (15.1 and 18.1) formed half of the haploids, half of the doubled haploids, and two other callus lines (5.1 and 7.2) numerous haploids. On callus lines with numerous haploids (5.1 and 7.2), as the regenant number increases, the size of plants decreases (plant height, number of flowers on the main panicle, number of panicles). On the lines 15.1 and 18.1 between groups of haploids and between groups of doubled haploids statistically significant differences absent. In breeding purposes for the induced doubling of the number of chromosomes in haploid regenerants with antitubulin substances such as colchicine, it is advisable to use plants that form on callus among the first. Between haploids of four callus lines and doubled haploids of two callus lines, statistically significant differences (at p=0.001) were revealed using the Hotelling's T2-criterion, calculated for the whole complex of biometric features. Haploids of different lines differed in three or four of them, doubled haploids on three of the five signs (length of panicle, productive bushiness and plant height). Varieties of interest to breeders may be improved by anther culture in vitro.


2019 ◽  
Vol 6 (1) ◽  
pp. 1-7
Author(s):  
Diego Pandeló José ◽  
José Marcello Salabert De Campos ◽  
Lyderson Facio Viccini ◽  
Emilly Ruas Alkimim ◽  
Marcelo De Oliveira Santos

Lippia lacunosa is a Brazilian savanna plant that belongs to the Verbenaceae family. It has been used in folk medicine as a treatment for different diseases. This species represents an endangered Brazilian medicinal plant, and this is the first report documenting a reliable protocol for the in vitro propagation and regeneration of L. lacunosa. Axenic explants were cultivated in MS medium containing different concentrations of naphthalene acetic acid (NAA) to induce root growth. The mean shoot length and the number of roots were highest with 0.06 mg·L-1 NAA. The highest number of buds in shoot regeneration was induced with 2 mg·L-1 6-benzylaminopurine (BA). To obtain a long-term culture, the dwarf shoots were elongated on MS media containing 0.5 mg·L-1 BA alternated with MS containing 2 mg·L-1 BA every 40 days. In the present protocol, the long-term shoots retained the ability to root even after long periods of BA treatment. In addition, we evaluated the nuclear DNA content and ploidy levels, including the occurrence of endopolyploidy, in long-term micropropagated plant leaves using flow cytometry analysis. The plants propagated in vitro over several years possessed nuclear DNA contents ranging from 2.940 to 3.095 pg, and no differences in DNA content were found among in vitro plants or between these plants and the control (L. lacunosa from a greenhouse with a DNA content of 3.08 pg). The flow cytometry analysis also demonstrated that there was no polyploidization. The present study will be useful for biotechnological approaches and provides the first estimate of the nuclear DNA content of this species using flow cytometry.


1991 ◽  
Vol 69 (2) ◽  
pp. 239-244 ◽  
Author(s):  
Narender S. Nehra ◽  
Kutty K. Kartha ◽  
Cecil Stushnoff

Callus cultures of strawberry cv. Redcoat (2n = 8x = 56) initiated from greenhouse and in vitro leaf explants were examined at various culture periods for morphogenic response, changes in nuclear DNA content, and isozyme banding patterns of four enzymes. The flow cytometric analysis of nuclear DNA content revealed the occurrence of polyploid and aneuploid changes as a function of ageing of callus cultures. The calli initiated from in vitro leaf explants were more prone to such changes than those initiated from greenhouse leaf explants. The in vitro morphogenic ability of callus cultures was affected by the ploidy changes, but the latter were not the only cause for loss in regeneration potential of long-term callus cultures. The isozyme phenotypes of esterase, phosphoglucomutase, phosphoglucoisomerase, and leucine aminopeptidase did not change with the chromosomal variation in callus cultures. Key words: strawberry, Fragaria × ananassa, callus culture, flow cytometry, nuclear DNA content, isozyme.


1994 ◽  
Vol 34 (7) ◽  
pp. 911 ◽  
Author(s):  
AM Callegarin ◽  
K Perfanov ◽  
G Dorotea ◽  
G Baldi

Increasing market demand has led to efforts to select for non-sticky, long-grain rice varieties suited to the Italian environment. The variability of recombinant populations derived from crosses between Italian and USA genotypes was exploited. Together with the conventional breeding methods, in vitro anther culture was used to obtain homozygous lines from crosses more quickly. Moreover, hybrid lines were obtained from crosses otherwise impossible to exploit owing to sterility in the F, generation (namely those resulting from crosses involving semi-dwarf USA varieties). Plant regeneration was obtained from 6 F, hybrids whose parents were chosen for their grain quality or their suitability to the Italian environment. About 5500 plantlets were regenerated. Of these, 1000 were albino, and about 50% of the green ones died after transplanting into soil; 65% of the surviving plants were sterile, so about 1000 progeny were obtained (30-445 for each cross). Progeny with desirable characteristics were evaluated for agronomic traits. On the basis of these results, limits and possibilities of anther-culture breeding in rice are discussed.


2008 ◽  
Vol 94 (1) ◽  
pp. 65-71 ◽  
Author(s):  
Joanna Makowczyńska ◽  
Emilia Andrzejewska-Golec ◽  
Elwira Sliwinska

2016 ◽  
Vol 14 (2) ◽  
pp. 178-180
Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
D. V. Shpak ◽  
A. O. Dobrova ◽  
S. O. Ignatova

Aims. To study the effect of chemically modified starch D–5aM in the culture medium on the efficiency of androgenesis in vitro in anther culture of rice. Methods. Obtaining of rice double haploid lines by anther culture in vitro. The statistical methods. Results The influence different variants of gellatyne source in culture medium on the processes of induction and regeneration in anther culture of rice were studied. The 119 green plants-regenerants were received. Conclusions. The negative effect on the formation of green regenerants using a gel-forming components of the chemically modified starch D–5aM was shown. Keywords: rice, anther culture in vitro, callus, regeneration, chemically modified starch.


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