A Sorghum bicolor × S. macrospermum hybrid recovered by embryo rescue and culture

2005 ◽  
Vol 53 (6) ◽  
pp. 579 ◽  
Author(s):  
H. James Price ◽  
George L. Hodnett ◽  
Byron L. Burson ◽  
Sally L. Dillon ◽  
William L. Rooney
Keyword(s):  
Nuclear Dna ◽  
Embryo Rescue ◽  
Nuclear Dna Content ◽  
Female Parent ◽  
Male Sterile ◽  
Leaf Pubescence ◽  
Culture Techniques ◽  
Close Phylogenetic Relationship ◽  
Improvement Programs

Although exotic germplasm is extensively used in sorghum improvement programs, Sorghum species classified in sections other than Eu-sorghum have not been utilised as germplasm because of strong reproductive barriers involving pollen–pistil incompatibilities. S. macrospermum is of particular interest to sorghum breeders because of its close phylogenetic relationship and cytogenetic similarities to S. bicolor and its resistance to important sorghum pests and pathogens, such as sorghum midge and sorghum downy mildew. A vegetatively vigorous interspecific hybrid was obtained from a cross between a cytoplasmic male-sterile S. bicolor plant and S. macrospermum by using embryo rescue and in vitro culture techniques. The hybrid was morphologically intermediate to S. bicolor and S. macrospermum in leaf width, leaf pubescence, plant height, inflorescence morphology, chromosome number and nuclear DNA content. It was male-sterile like its ATx623 parent. The hybrid produced no offspring when used as the female parent in a backcross with S. bicolor. This is the first confirmed hybrid between S. bicolor and S. macrospermum, and to our knowledge, it is the first reported hybrid between S. bicolor and any Sorghum species outside the Eu-sorghum section.

scholarly journals Sex-Linked Molecular Markers Identify Female Lines in Endosperm-Derived Kiwifruit Callus and in Regenerants

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 526
Author(s):  
Iwona Chłosta ◽  
Dagmara Kwolek ◽  
Elwira Sliwinska ◽  
Grzegorz Góralski ◽  
Marzena Popielarska-Konieczna
Keyword(s):  
Molecular Markers ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Dioecious Species ◽  
Culture Techniques ◽  
Flow Cytometric ◽  
Regenerated Plants ◽  
Tissue Culture Techniques ◽  
Selection Of

This is the first report of molecular markers application for the analysis of endosperm-derived callus and nonaploid kiwifruit (Actinidia chinensis var. deliciosa, formerly: Actinidia deliciosa) plants. As a source of explants, fruits of ‘Hayward’, the most popular cultivar, were used. Additionally, analyses of the nuclear DNA content and sex were conducted on the regenerated plants. Hexaploid seedlings were used as control for the flow cytometric analyses. Most of the plants (about 90%) regenerated via endosperm-derived callus possessed 2C = 9Cx DNA, which confirmed their endosperm origin and nonaploidy. Because Actinidia is a dioecious species, and female plants bearing fruits are desired by breeders, it is crucial to identify the sex of an individual at early stages of development. Analyses were conducted with ex vitro and in vitro samples. Results revealed that specific markers for a Y-chromosome applied at the callus stage allowed us to reliably predict the sex of plants regenerated from it. This is a novel application of sex-linked markers for early selection of female and male callus lines when the sex of the initial explants is still unknown, such as fresh isolated embryos and endosperm. It may have significant importance for breeding kiwifruit programs, which involve tissue culture techniques.

Analysis of nuclear DNA content and ploidy levels in 3x × 4x Lilium (Lilium × elegans L.) progenies by flow cytometry

2019 ◽  
Vol 66 (3-4) ◽  
pp. 127-135 ◽  
Author(s):  
Mast Ram Dhiman ◽  
Siddharth Moudgil ◽  
Chander Parkash ◽  
Raj Kumar ◽  
Sandeep Kumar ◽  
...  
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Female Parent ◽  
Morphological Characterization ◽  
Male Sterile ◽  
Aesthetic Value ◽  
Ploidy Levels ◽  
Interploidy Crosses ◽  
Lilium Lancifolium

Interploidy crosses between Lilium lancifolium (3x) and Asiatic lily cultivar ‘Brunello’ (4x) were attempted for creating genetic variability and to analyse the progenies for different ploidy levels. Experimental results revealed that most of the crosses attempted were developed into fruits, confirming that male-sterile triploid lilies can be used as the female parent for crossing with a suitable male parent. Wide variation in chromosome numbers (28 to 38) was obtained in different plant progenies, indicating that aneuploidy is generated by 3x × 4x crosses. The nuclear DNA content analysis of 13 plant progenies showed that the 2C nuclear DNA content has increased (range = 32.60 pg to 41.32 pg) as compared to Lilium lancifolium, while it was found lower than the cultivar ‘Brunello. Further, morphological characterization of different plant progenies revealed significant differences among themselves, which confirmed the dependence of these traits on cultivars ploidy level. Therefore, present findings will be instrumental for development of new Lilium cultivars with high aesthetic value and utility.

scholarly journals Embryo rescue from interspecific crosses in apple rootstocks

2006 ◽  
Vol 41 (6) ◽  
pp. 969-973 ◽  
Author(s):  
Adriana Cibele de Mesquita Dantas ◽  
José Itamar Boneti ◽  
Rubens Onofre Nodari ◽  
Miguel Pedro Guerra
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Embryo Rescue ◽  
Casein Hydrolysate ◽  
Female Parent ◽  
Interspecific Crosses ◽  
Open Pollination ◽  
Apple Rootstocks ◽  
Hand Pollination

The objetive of this work was to rescue immature embryos of apple rootstocks Malus prunifolia (Marubakaido) and Malus pumila (M9) after 40-60 days of pollination and to put them into MS culture media supplemented with agar (6 g L-1) and casein hydrolysate (500 mg L-1). Embryos originated from interspecific crosses and open pollination showed differences in the in vitro responses, depending on the female parent, the developmental stage of the embryo, and the culture medium composition. Embryos of the M. pumila rootstock, rescued within 40 days after pollination and put in culture medium supplemented with indolacetic acid (IAA), gibberellic acid (GA3), kinetin and maltose, resulted in a normal development of plantlets. However, embryos originating from hand-pollination, cultivated in medium supplemented with 14 µM IAA, 5 µM kinetin and 1.5 µM Ga3 (MS1), mainly those of M. prunifolia x M. pumila, showed a high percentage of rusted embryos (96.2%). Embryos from open pollination of M. prunifolia and M. pumila formed calluses. It was possible to identify the influence of the female parent by the enhanced development of M. pumila shoots derived from open or hand-pollination. The crossing of responsive species and the use of the technique of embryo culture provided a rapid and uniform germination and, consequently, the development of fully normal seedlings.

Factors influencing gene introgression into the allotetraploid Coffea arabica L. from its diploid relatives

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1053-1060 ◽  
Author(s):  
Juan C Herrera ◽  
Marie C Combes ◽  
Hernando Cortina ◽  
Philippe Lashermes
Keyword(s):  
Coffea Arabica ◽  
Nuclear Dna ◽  
Flow Cytometric Analysis ◽  
Diploid Species ◽  
Nuclear Dna Content ◽  
Female Parent ◽  
Coffea Canephora ◽  
Triploid Hybrid ◽  
Gene Introgression ◽  
Coffea Arabica L

Factors controlling gene introgression into cultivated arabica coffee (Coffea arabica L.) were investigated. Interspecific triploid hybrid plants between the tetraploid species C. arabica (2n = 44) and a diploid species (2n = 22), either Coffea canephora or Coffea eugenioides, were backcrossed to C. arabica (male parent). Flow cytometric analysis of the nuclear DNA content revealed that most of the BC1 individuals derived from triploid hybrids involving C. eugenioides were tetraploid or nearly tetraploid. Among the gametes produced by the interspecific triploid hybrids, those possessing approximately 22 chromosomes appeared strongly favored. The amount of introgression in BC1 individuals (21 and 43 for the BC1 progenies involving C. canephora and C. eugenioides, respectively) was estimated using species-specific microsatellite markers. A large number of introgressed markers was observed in all BC1 individuals. Nevertheless, while the frequency of introgressed markers seemed as expected, assuming random chromosome segregation and diploid gamete formation, in the BC1 derived from triploid hybrids involving C. canephora, this frequency appeared significantly lower in the BC1 derived from triploid hybrids involving C. eugenioides. Furthermore, the comparison of reciprocal progenies between C. arabica and triploid interspecific hybrids (C. arabica × C. canephora) used as male or female parent revealed a very strong effect of the backcross direction.Key words: irregular meiosis, coffee, reciprocal crosses, molecular marker, triploid hybrids.

Assessment of the cellular DNA content of whole mounted mouse and human oocytes and of blastomeres containing single or multiple nuclei

Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 17-25 ◽  
Author(s):  
Nicola J. Winston ◽  
Martin H. Johnson ◽  
Peter R. Braude
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Structural Features ◽  
Chromosomal Dna ◽  
Individual Values ◽  
Mean Values ◽  
Mouse Oocytes ◽  
Cellular Dna

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.

scholarly journals MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT

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
Keyword(s):  
Flow Cytometry ◽  
Medicinal Plant ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Naphthalene Acetic Acid ◽  
Flow Cytometry Analysis ◽  
Ploidy Levels

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.

Nuclear DNA content and isozyme variation in relation to morphogenic potential of strawberry (Fragaria × ananassa) callus cultures

1991 ◽  
Vol 69 (2) ◽  
pp. 239-244 ◽  
Author(s):  
Narender S. Nehra ◽  
Kutty K. Kartha ◽  
Cecil Stushnoff
Keyword(s):  
Dna Content ◽  
Nuclear Dna ◽  
Flow Cytometric Analysis ◽  
Nuclear Dna Content ◽  
Leaf Explants ◽  
Callus Cultures ◽  
Fragaria Ananassa ◽  
Flow Cytometric ◽  
Ploidy Changes

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.

scholarly journals Creation of Large Hybrid Populations Using Male-sterile Germplasm as the Female Parent in Jujube

HortScience ◽  
2018 ◽  
Vol 53 (5) ◽  
pp. 609-612 ◽  
Author(s):  
Fenfen Yan ◽  
Zhiguo Liu ◽  
Mengjun Liu ◽  
Xingjuan Zheng ◽  
Zhi Luo ◽  
...  
Keyword(s):  
Molecular Mapping ◽  
Embryo Rescue ◽  
Genetic Research ◽  
Female Parent ◽  
Hybrid Population ◽  
Chinese Jujube ◽  
Male Sterile ◽  
Cross Compatibility ◽  
Bona Fide ◽  
Ziziphus Jujuba

The establishment of large hybrid populations is the key basis to breeding new cultivars and genetic research, such as molecular mapping. However, it is extremely difficult to generate hybrid progeny using conventional artificial hybridization in Chinese jujube (Chinese date) (Ziziphus jujuba Mill.) because of its low fruit set of ≈0.01%. The objective of this study was to create large hybrid populations in Chinese jujube using bona fide male-sterile germplasm as the female parent. Two male-sterile varieties that lacked viable pollen (JMS1 and JMS2) previously discovered by our research group and three male-fertile genotypes (‘Xing16’, ‘Jiao5’, and ‘Wuhefeng’) were used as female and male parents, respectively. Bee-aided controlled hybridization of five combinations was conducted from 2014 to 2016. The results indicated that both JMS1 and JMS2 were stable male-sterile germplasm across the years examined. JMS2 showed more effective compatibility with the male parents tested than JMS1. A total of 7681 fruits were obtained from the five cross combinations, and 3120 of them contained seeds. The mean rate of the fruit with seed varied from 32.62% to 64.21%, and the highest rate (73.38%) was obtained from the cross of JMS2 × ‘Xing16’ in 2016. A total of 831 seedlings were obtained and consisted of 602 from JMS2 × ‘Xing16’, 221 from JMS2 × ‘Jiao5’, two from JMS1 × ‘Xing16’, five from JMS1 × ‘Wuhefeng’, and one from JMS2 × ‘Wuhefeng’. The 118 randomly selected progeny seedlings, including 96 from JMS2 × ‘Xing16’ and 22 from JMS2 × ‘Jiao5’, respectively, were identified to be authentic hybrids using SSR markers. Thus, JMS2 is a promising male-sterile female parent that possesses a good cross-compatibility and is free from the need for emasculation, embryo rescue, and hybrid identification. The largest hybrid population of 602 progeny was obtained in Chinese jujube using controlled hybridization. This study demonstrated the feasibility of a highly efficient crossbreeding approach using male-sterile germplasm as the female parent in Chinese jujube.

A successful application of the embryo rescue technique as a model for studying crosses between Salix viminalis and Populus species

2011 ◽  
Vol 59 (4) ◽  
pp. 382 ◽  
Author(s):  
Agnieszka Bagniewska-Zadworna ◽  
Maciej Zenkteler ◽  
Elzbieta Zenkteler ◽  
Maria K. Wojciechowicz ◽  
Abdelali Barakat ◽  
...  
Keyword(s):  
Embryo Rescue ◽  
Female Parent ◽  
Botanical Garden ◽  
Intergeneric Hybrids ◽  
Salix Viminalis ◽  
Putative Hybrid ◽  
Ms Medium ◽  
Hybrid Nature ◽  
Rescue Technique

Embryos and plants from the crosses Salix viminalis L. as the female parent and Populus alba L., P. violascens Dode or P. tremula L. as the male parent were obtained by in vitro embryo rescue technique. F1 intergeneric progeny were obtained by overcoming post-zygotic barriers caused by the deficiency of endosperm as nutritive tissue during the first stage of embryo development. Ovules containing immature heart-stage embryos as well as early cotyledonary embryos were isolated and rescued in modified 1/2 MS medium supplemented with 3% sucrose to ensure maturity. Seedlings were cultured under in vitro conditions in 1/2 MS medium that contained 0.2 mg l–1 naphtaleneacetic acid and plantlets after rooting were transferred first into pots and subsequently to the experimental field in the Botanical Garden in Poznań, Poland. After 2 years, the putative hybrid nature of the plants was checked by analysing morphological characters and molecular markers. Scanning electron microscopy, flow cytometry and random amplification of polymorphic DNA screening of individuals confirmed the hybrid nature of the S. viminalis × P. alba and S. viminalis × P. violascens progeny and of 25% of the analysed plants from the cross S. viminalis × P. tremula. This study showed the feasibility of overcoming pre- and post-fertilisation barriers to achieve intergeneric hybrids between species from two genera of Salicaceae. The in vitro system to produce hybrids with characters of both parents could be of great importance for increasing biomass production. This study also opens new opportunities to improve other traits in trees, such as resistance to pathogens inherited from one parent.

Sign in / Sign up

Export Citation Format

Share Document