scholarly journals Suppression of Metacaspase- and Autophagy-Dependent Cell Death Improves Stress-Induced Microspore Embryogenesis in Brassica napus

2020 ◽  
Author(s):  
Eduardo Berenguer ◽  
Elena A Minina ◽  
Elena Carneros ◽  
Ivett Bárány ◽  
Peter V Bozhkov ◽  
...  
Keyword(s):  
Cell Death ◽  
Brassica Napus ◽  
Embryonic Development ◽  
Doubled Haploid ◽  
Microspore Embryogenesis ◽  
Brassica Species ◽  
Cysteine Proteases ◽  
Plant Production ◽  
Haploid Plant

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.

scholarly journals Proteases with caspase 3-like activity participate in cell death during stress-induced microspore embryogenesis of Brassica napus

2019 ◽  
Vol 3 (3) ◽  
pp. 152-159 ◽  
Author(s):  
Eduardo Berenguer ◽  
María-Teresa Solís ◽  
Yolanda Pérez-Pérez ◽  
Pilar S. Testillano
Keyword(s):  
Cell Death ◽  
Brassica Napus ◽  
Caspase 3 ◽  
Specific Inhibitor ◽  
Microspore Embryogenesis ◽  
Plant Production ◽  
Haploid Plant ◽  
Stress Treatment ◽  
Proteolytic Activities

Abstract Microspore embryogenesis is a model system of plant cell reprogramming, totipotency acquisition, stress response and embryogenesis initiation. This in vitro system constitutes an important biotechnological tool for haploid and doubled-haploid plant production, very useful for crop breeding. In this process, microspores (cells that produce pollen grains in planta) are reprogrammed toward embryogenesis by specific stress treatment, but many microspores die after the stress. The occurrence of cell death is a serious limiting problem that greatly reduces microspore embryogenesis yield. In animals, increasing evidence has revealed caspase proteolytic activities as essential executioners of programmed cell death (PCD) processes, however, less is known in plants. Although plant genomes do not contain caspase homologues, caspase-like proteolytic activities have been detected in many plant PCD processes. In the present study, we have analysed caspase 3-like activity and its involvement in stress-induced cell death during initial stages of microspore embryogenesis of Brassica napus. After stress treatment to induce embryogenesis, isolated microspore cultures showed high levels of cell death and caspase 3-like proteolytic activity was induced. Treatments with specific inhibitor of caspase 3-like activity reduced cell death and increased embryogenesis induction efficiency. Our findings indicate the involvement of proteases with caspase 3-like activity in the initiation and/or execution of cell death at early microspore embryogenesis in B. napus, giving new insights into the pathways of stress-induced cell death in plants and opening a new way to improve in vitro embryogenesis efficiency by using chemical modulators of cell death proteases.

scholarly journals High Frequency of Doubled Haploid Plant Production in Spelt Wheat

2016 ◽  
Vol 58 (2) ◽  
pp. 107-112 ◽  
Author(s):  
Csaba Lantos ◽  
Barnabás Jenes ◽  
Lajos Bóna ◽  
Mátyás Cserháti ◽  
János Pauk
Keyword(s):  
Anther Culture ◽  
Ploidy Level ◽  
Doubled Haploid ◽  
High Frequency ◽  
Applied Research ◽  
Plant Production ◽  
Haploid Plant ◽  
Spelt Wheat ◽  
Regenerated Plantlets

AbstractThis is the first study to report an efficient anther culture (AC) method for spelt wheat, which has an increasing importance not only in applied research but also in organic farming and changing nutritional standards. In this study, an efficient AC protocol has been described for ‘GK Fehér’ spelt wheat. The number of AC-derived embryo-like structures (ELS) was 62.2/100 anthers, from which we were able to regenerate 30.6 green plantlets per 100 anthers. The percentage of green plantlets production was 89.0% among the regenerated plantlets, while the phenomenon of albinism was restricted (3.8/100 anthers). Altogether, from AC of ‘GK Fehér’ 306 green plantlets were producedin vitroand 241 plants were acclimatized to the greenhouse conditions. Based on ploidy level analyses, 83 spontaneous doubled haploid (DH) plants were produced (8.3 DH plants/100 anthers), so the percentage of spontaneous rediploidization was 34.4%. The spontaneous DH plants produced fertile spikes, while a few seeds were harvested from seven partially fertile plants.

scholarly journals In Vitro Gynogenesis and Flow Cytometry Analysis of the Regenerated Haploids of Black Cumin (Nigella sativa)

HortScience ◽  
2018 ◽  
Vol 53 (5) ◽  
pp. 681-686 ◽  
Author(s):  
Mohammed Elsayed El-Mahrouk ◽  
Mossad K. Maamoun ◽  
Antar Nasr EL-Banna ◽  
Soliman A. Omran ◽  
Yaser Hassan Dewir ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nigella Sativa ◽  
Plant Production ◽  
Haploid Plant ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Flow Cytometry Analysis ◽  
Black Cumin ◽  
Haploid Plants

In vitro ovule culture could be used to generate homozygous lines through the production of haploid plants. The present study reports on in vitro regeneration and production of haploid plants through ovule cultures and identification of the regenerated haploids using flow cytometry. The ovules were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kin), 2,4-dichlorophenoxyacetic acid (2,4-D), and naphthalene acetic acid (NAA) at 0, 0.5, 1, and 2 mg·L−1 for their gynogenesis. Among different plant growth regulators (PGRs) tested, 2,4-D at 2 mg·L−1 produced direct gynogenesis. The highest callogenesis percentage (100%) was obtained on MS medium containing 1 mg·L−1 2,4-D and 2 mg·L−1 NAA. Flow cytometry analysis was used to identify the regenerated haploids. It also confirmed gynogenic occurrence at 1 and 2 mg·L−1 2,4-D with percentages of 21.7% and 41%, respectively. Therefore, 2,4-D proved effective for the induction of haploids in black cumin. The regenerated haploids were developed on MS medium without PGRs. The obtained results of in vitro gynogenesis and haploid plant production can tremendously facilitate breeding programs of black cumin.

scholarly journals Factors Affecting Doubled Haploid Plant Production Via Maize Technique in Bread Wheat

2015 ◽  
Vol 56 (2) ◽  
pp. 67-73
Author(s):  
Ioannis Xynias ◽  
Antonios Koufalis ◽  
Evdokia Gouli-Vavdinoudi ◽  
Demetrios Roupakias
Keyword(s):  
Bread Wheat ◽  
Doubled Haploid ◽  
Embryo Rescue ◽  
Plant Production ◽  
Haploid Plant ◽  
In Planta ◽  
Maize Pollen ◽  
Doubled Haploid Plants ◽  
Haploid Embryos ◽  
Haploid Plants

Abstract The effect of two in planta factors (growth conditions, genotype) and two in vitro factors (time of embryo rescue, embryo rescue medium) on doubled haploid (DH) plant production in bread wheat via maize technique was investigated in nine F1 hybrids produced after crossing four bread wheat cultivars. During the first year one group of F1 plants was grown in a field and at the proper stage pollinated with maize pollen (sweet corn popu-lation). In parallel, a second group of F1 plants was grown in a growth chamber and pollinated as in the former group. In the second growing season the experiment was repeated but only field-grown plants were used. All the produced haploid embryos were cultured in three different media and the resulting 146 haploid plants were sub-sequently treated with aqueous solution of colchicine. Finally, 86 doubled haploid plants were obtained. We noted that the growing conditions of the parental plants and the intervening time between day of pollination and day of embryo rescue influenced the percentage of haploid embryo production. Culture medium also influenced haploid and doubled haploid plant production. The two media (MS/2, B5) were found equally effective. Most of the haploid embryos originated from the Penios × Acheloos cross, whereas most of the doubled haploid plants were produced from the KVZ × Penios cross. Doubled haploid plants were produced from all crosses.

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