Microspore Embryogenesis

Author(s):  
A. Olmedilla
HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 484b-484
Author(s):  
Tatiana Boluarte ◽  
R.E. Veilleux

Genes for anther culture response (ACR) need to be mapped to enable efficient transfer of the trait to unresponsive but agronomically desirable clones. The objective of this study was to find extremes for ACR in a segregating population to target genes controlling this trait using bulk segregant analysis. Populations resulting from backcrosses (BC) of a diploid interspecific clone [CP2: Solanum chacoense (chc: low ACR) and S. phureja (phu: high ACR)] to both parents were used to characterize two phases of androgenesis in a preliminary study: microspore embryogenesis and embryo regeneration. Among 24 plants from each BC, consistently high- and low-responding genotypes with regard to embryo production were identified. Low ACR clones in each BC produced from 0.0–0.1 embryos per anther (EPA), whereas high ACR clones in the chc and phu BC produced from 1.4–2.9 and 2.8–7.5 EPA, respectively. The frequency of embryos regenerating into plants ranged from 3% to 20% for the chc high EPA clones and 12% to 64% for the phu high EPA clones. Ploidy of regenerated plants was determined by flow cytometry. The frequency of monoploid regenerants among the high EPA clones ranged from 3.6% to 68%. Since these populations showed sufficient genetic variation for ACR, a series of statistically designed experiments were conducted to develop high and low ACR bulks. From four groups of phenotypically characterized PBC plants, a high ACR bulk (–x = 4.9 EPA; n = 14) and a low ACR bulk (–x = 0.12 EPA; n =9) were constructed.


2016 ◽  
Vol 128 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Giuseppe Cimò ◽  
Annalisa Marchese ◽  
Maria Antonietta Germanà

Author(s):  
Eduardo Berenguer ◽  
Elena A Minina ◽  
Elena Carneros ◽  
Ivett Bárány ◽  
Peter V Bozhkov ◽  
...  

Abstract Microspore embryogenesis is a biotechnological process that allows us to rapidly obtain doubled-haploid plants for breeding programs. The process is initiated by the application of stress treatment, which reprograms microspores to embark on embryonic development. Typically, a part of the microspores undergoes cell death that reduces the efficiency of the process. Metacaspases (MCAs), a phylogenetically broad group of cysteine proteases, and autophagy, the major catabolic process in eukaryotes, are critical regulators of the balance between cell death and survival in various organisms. In this study, we analyzed the role of MCAs and autophagy in cell death during stress-induced microspore embryogenesis in Brassica napus. We demonstrate that this cell death is accompanied by the transcriptional upregulation of three BnMCA genes (BnMCA-Ia, BnMCA-IIa and BnMCA-IIi), an increase in MCA proteolytic activity and the activation of autophagy. Accordingly, inhibition of autophagy and MCA activity, either individually or in combination, suppressed cell death and increased the number of proembryos, indicating that both components play a pro-cell death role and account for decreased efficiency of early embryonic development. Therefore, MCAs and/or autophagy can be used as new biotechnological targets to improve in vitro embryogenesis in Brassica species and doubled-haploid plant production in crop breeding and propagation programs.


2011 ◽  
Vol 47 (No. 3) ◽  
pp. 114-122 ◽  
Author(s):  
S.H. Ardebili ◽  
M.E. Shariatpanahi ◽  
R. Amiri ◽  
M. Emamifar ◽  
M. Oroojloo ◽  
...  

The effect of 2,4-dichlorophenoxyacetic acid (2,4-D) applied at high concentrations for a short time was investigated as a novel stress for induction of microspore embryogenesis for the first time. Brassica napus L. cvs. Topas and Hyola 420 were used as model plants for testing this hypothesis. Microspores were subjected to 2,4-D at 4 concentrations (15, 25, 35 and 45 mg/l) for 15–45 min while the classical heat shock was used as the control treatment. Among 2,4-D treatments in Topas, the highest yield of torpedo-stage embryos was achieved at 15 mg/l 2,4-D for 30 min while more normal plantlets were produced when 2,4-D (25 mg/l for 30 min) was applied to the microspores. In Hyola 420 the results showed a lower number of embryos and normal plantlets at all concentrations of 2,4-D. Although Hyola 420 was almost equally embryogenic as Topas after heat shock treatment, large differences between genotypes (concerning embryogenic response) occurred after 2,4-D treatment. However, the mean number of embryos and regenerants was higher in heat shock as compared to 2,4-D induced stress (one magnitude of order). According to the results obtained, 2,4-D can be introduced as a new stress for induction of embryogenesis in microspores similarly like in zygotic and somatic cells. This novel stress is very important for plant species whose microspores are extremely sensitive to classical stresses.


2013 ◽  
Vol 5 (4) ◽  
pp. 485-489 ◽  
Author(s):  
Tina Oana CRISTEA

In vitro microspore culture is one of the top techniques utilised now-a-days for the obtaining of double haploid plants in many plant species, including Brassica. The pH of the medium is a critical factor for the success of In vitro microspore culture as it influences the invertase enzyme activity, translated at cellular level through an acceleration or reduction of sucrose cleavage. The results published until now shows rather contradictory findings, as the response of microspores have been proved to be highly depending on genotypes, most of them being focused on Brassica napus. Thus, in the present study, the effect of different NLN liquid medium pH, ranging between 5.0 to 7.0 were tested in order to establish the most suitable pH for the expression of embryogenic competences of microspores cultivated on medium In vitro and ultimately for the obtaining of microspore-derived embryos. Among the 11 values of pH tested, the best results were obtained on variants with pH 5.8 and 6.0, both in what concern the maintaining of microspores viability and the number of microspore-derived embryos. The findings of the present study provide a strong base for the establishment of an efficient protocol for the In vitro culture of microspore at Brassica oleracea L. genotypes with Romanian origin.


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