Microdissection and painting of the Y chromosome in spinach (Spinacia oleracea)

Deng, Chuan‐Liang; Qin, Ruiyun; Jian, Gao; Li, Shufen; Gao, Wu‐Jun; Longdou, Lu

doi:10.1007/s10265-013-0549-3

Cited by 19 publications

(17 citation statements)

References 57 publications

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“…In addition, the DOP-PCR products of 1R and 1RS yielded weaker signals in the terminals of chromosomes than in the other regions (Deng et al, 2014). The similar phenomemon could also be found in the chromosome painting of spinach (Spinacia oleracea L.) using DOP-PCR products of microdissected Y chromosome as a probe (Deng et al, 2013b). Although the signals of DOP-PCR products from microdissected 6PS and 6PL in the chromosome ends were weaker, the signals seemed to be still stronger than those of the previous studies.…”

Section: Discussionsupporting

confidence: 60%

“…A DNA library of Gossypium arboreum chromosome 5 containing approximately 173,000 clones was created by chromosome microdissection and microcloning (Peng et al, 2012). In addition, there were chromosomes microdissected from wheat, Thinopyrum intermedium , and spinach ( Spinacia oleracea L.) (Deng et al, 2013a, 2013b; Vega et al, 1994). The microdissected DNA would facilitate specific probe screening, molecular mapping, and gene cloning.…”

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confidence: 99%

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Degenerate Oligonucleotide Primed–Polymerase Chain Reaction‐Based Chromosome Painting of P Genome Chromosomes in Agropyron cristatum and Wheat–A. cristatum Addition Lines

Zhang

Liu

et al. 2015

Crop Science

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Agropyron cristatum (L.) Gaertn. (2n = 4x = 28, PPPP), a wild relative of wheat, could provide many desirable genes for wheat improvement. Microdissection and degenerate oligonucleotide primed –polymerase chain reaction (DOP–PCR) is an effective way to isolate specific sequences. The development of specific sequences and functional markers of P genome could lay the foundation for gene mapping and cloning. In this study, chromosomes 6PS and 6PL were microdissected from wheat–A. cristatum 6PS and 6PL addition lines, and DOP–PCR products from both microdissected chromosomes were used as probes to hybridize with mitotic metaphase chromosomes of diploid A. cristatum, wheat–A. cristatum addition lines 6PS, 6PL, and 6P. The results showed that the distribution patterns on A. cristatum chromosomes using DOP–PCR products of microdissected 6PS as the probe were similar to those using DOP–PCR products of microdissected 6PL as the probe. They both distributed along all chromosomes, and chromosome 6P and other chromosomes were similar according to the hybridization pattern. There were no signals on wheat chromosomes indicating that DOP–PCR products contain P‐genome specific sequences. Actually, we obtained a P‐genome specific sequence from the DOP–PCR products of 6PS, which could be used as a fluoresence in situ hybridization (FISH) probe to identify P genome chromatin in wheat–A. cristatum introgression lines. These results could provide a basis for the study of structure and evolution of P genome chromosomes. Furthermore, this study may lead to mapping and cloning of the desirable genes on 6P chromosome.

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Section: Discussionsupporting

confidence: 60%

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confidence: 99%

Degenerate Oligonucleotide Primed–Polymerase Chain Reaction‐Based Chromosome Painting of P Genome Chromosomes in Agropyron cristatum and Wheat–A. cristatum Addition Lines

Zhang

Liu

et al. 2015

Crop Science

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“…varies substantially among species and even within species (for example, Hough et al, 2014). In species with homomorphic sex chromosomes, sex is likely controlled by a single locus, or several tightly linked loci, with the heterogametic sex being either male (for example, Asparagus officinalis- (Telgmann-Rauber et al, 2007); Sagittaria latifolia- (Dorken and Barrett, 2004) and Spinacia oleracea - (Deng et al, 2013)) or female (for example, Fragaria virginiana- (Spigler et al, 2008) and Populus trichocarpa- (Tuskan et al, 2012)). …”

Section: Introductionmentioning

confidence: 99%

Sex determination in dioecious Mercurialis annua and its close diploid and polyploid relatives

Russell

Pannell

2014

Heredity

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Separate sexes have evolved on numerous independent occasions from hermaphroditic ancestors in flowering plants. The mechanisms of sex determination is known for only a handful of such species, but, in those that have been investigated, it usually involves alleles segregating at a single locus, sometimes on heteromorphic sex chromosomes. In the genus Mercurialis, transitions between combined (hermaphroditism) and separate sexes (dioecy or androdioecy, where males co-occur with hermaphrodites rather than females) have occurred more than once in association with hybridisation and shifts in ploidy. Previous work has pointed to an unusual 3-locus system of sex determination in dioecious populations. Here, we use crosses and genotyping for a sex-linked marker to reject this model: sex in diploid dioecious M. annua is determined at a single locus with a dominant male-determining allele (an XY system). We also crossed individuals among lineages of Mercurialis that differ in their ploidy and sexual system to ascertain the extent to which the same sex-determination system has been conserved following genome duplication, hybridisation and transitions between dioecy and hermaphroditism. Our results indicate that the maledetermining element is fully capable of determining gender in the progeny of hybrids between different lineages. Specifically, males crossed with females or hermaphrodites always generate 1:1 male:female or male:hermaphrodite sex ratios, respectively, regardless of the ploidy levels involved (diploid, tetraploid or hexaploid). Our results throw further light on the genetics of the remarkable variation in sexual systems in the genus Mercurialis. They also illustrate the almost identical expression of sexdetermining alleles in terms of sexual phenotypes across multiple divergent backgrounds, including those that have lost separate sexes altogether.

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“…A reciprocal translocation proved that the sex-determining genes are located on the short arm of chromosome [7]. In 2013, the Y chromosome in spinach was isolated and analyzed by microdissection for the first time; the largest spinach chromosome was confirmed to be a sex chromosome at the molecular level [8]. Some molecular markers linked to the X/Y gene have been found and used for the study of molecular markers of spinach sex.…”

Section: Introductionmentioning

confidence: 99%

Construction of a high-density genetic map and the X/Y sex-determining gene mapping in spinach based on large-scale markers developed by specific-locus amplified fragment sequencing (SLAF-seq)

et al. 2017

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BackgroundCultivated spinach (Spinacia oleracea L.) is one of the most widely cultivated types of leafy vegetable in the world, and it has a high nutritional value. Spinach is also an ideal plant for investigating the mechanism of sex determination because it is a dioecious species with separate male and female plants. Some reports on the sex labeling and localization of spinach in the study of molecular markers have surfaced. However, there have only been two reports completed on the genetic map of spinach. The lack of rich and reliable molecular markers and the shortage of high-density linkage maps are important constraints in spinach research work. In this study, a high-density genetic map of spinach based on the Specific-locus Amplified Fragment Sequencing (SLAF-seq) technique was constructed; the sex-determining gene was also finely mapped.ResultsThrough bio-information analysis, 50.75 Gb of data in total was obtained, including 207.58 million paired-end reads. Finally, 145,456 high-quality SLAF markers were obtained, with 27,800 polymorphic markers and 4080 SLAF markers were finally mapped onto the genetic map after linkage analysis. The map spanned 1,125.97 cM with an average distance of 0.31 cM between the adjacent marker loci. It was divided into 6 linkage groups corresponding to the number of spinach chromosomes. Besides, the combination of Bulked Segregation Analysis (BSA) with SLAF-seq technology(super-BSA) was employed to generate the linkage markers with the sex-determining gene. Combined with the high-density genetic map of spinach, the sex-determining gene X/Y was located at the position of the linkage group (LG) 4 (66.98 cM–69.72 cM and 75.48 cM–92.96 cM), which may be the ideal region for the sex-determining gene.ConclusionsA high-density genetic map of spinach based on the SLAF-seq technique was constructed with a backcross (BC1) population (which is the highest density genetic map of spinach reported at present). At the same time, the sex-determining gene X/Y was mapped to LG4 with super-BSA. This map will offer a suitable basis for further study of spinach, such as gene mapping, map-based cloning of Specific genes, quantitative trait locus (QTL) mapping and marker-assisted selection (MAS). It will also provide an efficient reference for studies on the mechanism of sex determination in other dioecious plants.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3659-9) contains supplementary material, which is available to authorized users.

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Microdissection and painting of the Y chromosome in spinach (Spinacia oleracea)

Cited by 19 publications

References 57 publications

Degenerate Oligonucleotide Primed–Polymerase Chain Reaction‐Based Chromosome Painting of P Genome Chromosomes in Agropyron cristatum and Wheat–A. cristatum Addition Lines

Degenerate Oligonucleotide Primed–Polymerase Chain Reaction‐Based Chromosome Painting of P Genome Chromosomes in Agropyron cristatum and Wheat–A. cristatum Addition Lines

Sex determination in dioecious Mercurialis annua and its close diploid and polyploid relatives

Construction of a high-density genetic map and the X/Y sex-determining gene mapping in spinach based on large-scale markers developed by specific-locus amplified fragment sequencing (SLAF-seq)

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