Efficiency Of Androgenesis Research Articles

A combination of ANNs and multivariate sensitivity analysis unveils critical factors to increase androgenesis efficiency in tomato (Solanum lycopersicum)

A combination of ANNs and multivariate sensitivity analysis unveils critical factors to increase androgenesis efficiency in tomato (Solanum lycopersicum)

Efficiency of Androgenesis In Vitro in the Culture of Isolated Winter Wheat Anthers: 1. The Influence of the Duration of Low-Temperature Pretreatment of Donor Plants on Respiration, Carbohydrate Content, and ROS Levels in Flowers from Different Parts of the Spike

Efficiency of Androgenesis In Vitro in the Culture of Isolated Winter Wheat Anthers: 1. The Influence of the Duration of Low-Temperature Pretreatment of Donor Plants on Respiration, Carbohydrate Content, and ROS Levels in Flowers from Different Parts of the Spike

Efficiency of Androgenesis In Vitro in the Culture of Isolated Winter Wheat Anthers: 2. The Influence of the Duration of Low-Temperature Pretreatment of Donor Plants on the Content of Lipid Peroxidation Products and Fatty Acid Composition in Flowers from Different Parts of the Spike

Efficiency of Androgenesis In Vitro in the Culture of Isolated Winter Wheat Anthers: 2. The Influence of the Duration of Low-Temperature Pretreatment of Donor Plants on the Content of Lipid Peroxidation Products and Fatty Acid Composition in Flowers from Different Parts of the Spike

The study of the efficiency of anther culture <i>in vitro</i> of bread wheat varieties and hybrids <i>(Triticum aestivum L.)</i>

Anther culture is the most effective and not technically complex tool for obtaining double wheat haploids. The current study is devoted to the research of the influence of the genotype and concentration of the growth regulator 2.4-D on the efficiency of androgenesis in vitro of spring bread wheat. The purpose of the study was to estimate the efficiency of the anther culture of hybrids of the first and second generation and their parental varieties and to study its dependence on different concentrations of the growth regulator 2.4-D. There has been carried out the estimation of the traits of the F1hybrids, the F2population, and the parental varieties ‘Obskaya 2’ and ‘Novosibirskaya 15’ when 2.4-D was added to the induction medium at a concentration of 1 and 2 mg/l. There has been established that the F2population and the parental variety ‘Novosibirskaya 15’ are characterized by the largest indicators. The average values were significantly surpassed by the F2population in the traits ‘number of productive anthers per 100 anthers’ and ‘number of all regenerants per 100 anthers’ at 1 mg/l of 2.4-D. The variety ‘Novosibirskaya 15’ surpassed the average values in traits ‘number of sprouts per 100 anthers’ and ‘number of all regenerants per 100 sprouts’ at 2 mg /l of 2.4-D. The trial has shown that the best responsiveness of the anther culture for the studied genotypes was when the growth regulator 2.4-D was added to the induction medium at a concentration of 1 mg/l. The variety ‘Novosibirskaya 15’ has been characterized by good responsiveness in anther culturein vitroand could be used as a donor of valuable alleles in wheat breeding programs using DH technologies.

Induction of in vitro androgenesis in anther culture of recalcitrant einkorn (Triticum monococcum L.)

Einkorn (Triticum monococcum L.) can be applied as a model species for cereal genomic studies due to its small genome size and high level of polymorphism. The in vitro somatic tissue culture protocol in einkorn was significantly improved recently, however the in vitro androgenesis remained an unresolved research topic. Five different pre-treatments were compared to study the effects of stress pre-treatments on the efficiency of androgenesis in two einkorn genotypes. The long cold pre-treatment (2 weeks, 4 °C) of donor tillers increased significantly the number of microspore derived embryo-like structures (ELS). Green and albino plantlets were regenerated from these structures. The ploidy level of microspore-derived green plantlet was determined as haploid by flow cytometric analyses. This is the first report published on the successful androgenesis induction (ELS production) and green- and albino plantlet regeneration in in vitro anther culture of the recalcitrant einkorn wheat (Triticum monococcum L.).

The influence of combinations of alien translocations on in vitro androgenesis in spring common wheat (Triticum aestivum L.)

Backgr ound.The basic approach to the production of new common wheat genotypes involving introgressive hybridization entails a long-term process. Doubled haploid production could accelerate it. However, this method is not widely used in breeding programs due to its main limitation: the genotype dependence. Due to genetic differences between wheat and related species, it was assumed that alien genetic materials are different in their capacity to affect androgenesis. The effect of alien translocations on androgenesis has been shown earlier. The aim of this study was to develop a set of DH wheat lines containing a wheat-alien translocation in the genome and study the effect of alien translocations on androgenesis of anther culture in such lines.Materials and methods. The plant material included: the spring wheat cultivar ‘Novosibirskaya 16’, line Velut 991 carrying wheat-alien translocations 1RS.1BL from rye and 5BS.5BL-5SL from Aegilops speltoides Tausch, and four hybrid F3 generation lines (10-7, 14-8, 15-8, 15-12) from their crossing, differing in the content of alien translocations.Results.It was shown that parameters of androgenesis such as the number of embryo-like structures per 100 anthers, the number of albino regenerants per 100 anthers, and the number of green regenerants per 100 anthers varied depending on the line. The best -responding lines Velut 991, 10-7 and 14-8 are characterized by the presence of a 1RS.1BL wheat-rye translocation chromosome. Regeneration frequency of green plants was recorded to be 8,6%, 3,6% and 10,1% respectively. The values of the parameters for lines 15-12 (carrying 5BS.5BL-5SL translocation) and 15-8 (without translocations) did not differ significantly.Conclusion.Therefore, it can be concluded that the presence of the introgressive fragment of chromosome 5S did not affect the efficiency of androgenesis and the short shoulder of chromosome 1R carries genes that stimulated androgenesis in anther culture.

IDENTIFICATION OF MICROSATELLITE LOCI ASSOCIATED WITH EMBRYOGENIC POTENTIAL IN WHEAT GENOTYPES IN ANTHER CULTURE IN VITRO

In order to identify microsatellite loci that can be used as markers in screening for responsiveness in the in vitro
 anther culture, the efficiency of androgenesis in vitro was evaluated in 24 wheat genotypes. The forms contrasting in responsiveness in the in vitro anther culture were identified, and four F2 segregating populations were obtained on their basis. The study of interlinear polymorphism by 20 microsatellite loci located on chromosomes 5A and 5B was carried out. It was found that the cleavage by the ability to embryogenesis coincides with the cleavage by the alleles of the Xgwm371 locus: the 169 bp allele is associated with the ability to embryogenesis; the 185 bp allele — with the absence of such an ability or extremely low embryoid yield in the anther culture in vitro.

Effects of temperature pretreatments and carbohydrate type on androgenesis induction in anther culture of Capparis spinosa L.

Caper (Capparis spinosa L.) is a rich source of rutin, plays an essential role in human health. In the present study, the effects of cold (25°C as control, 4°C, and 7°C for 2, 4, and 7 days), heat (25°C as control, 30°C for 14 days, 32°C for 2 and 4 days, and 35°C for 8 hours), and carbohydrate treatments on the androgenesis efficiency were studied in the anther culture of caper. Also, the effects of maltose and sucrose at the concentrations of 30 and 60 g L-1 in combination with two temperature treatments (1- 30°C for 14 days and 2- 7°C for 7 days + azacytidine and 2,4-D pretreatments) on the androgenesis induction was evaluated. The temperature and carbohydrate treatments showed statistically significant differences (p ≤0.01) in terms of callus and embryo formation. The 7°C for 2, 4, and 7 days produced the highest percentage (at the third week: 80, 78.34, and 76.67%, respectively) and callogenesis speed (7.85, 7.75, and 7.60 calli week-1, respectively) and the 7°C for 7 days produced the highest embryo production (0.57 embryo anther-1). The 30°C for 14 days treatment showed the highest percentage (at the third week: 100%) and callogenesis speed (9.44 calli week-1). While the 32°C for 2 and 3 days and also 30°C for 14 days produced the highest number of embryos per anther (0.22, 0.20, and 0.18 embryo, respectively). The use of 30 g L-1 maltose in combination with the 30°C for 14 days produced the highest percentage (at the third week: 91.66%) and callogenesis speed (8.94 calli week-1), while the 30 g L-1 maltose in combination with the 7°C for 7 days + azacytidine and 2,4-D pretreatments produced the highest mean embryo number per anther (0.55 embryo). The results of this research are of great importance for the use in the caper breeding programs.

The testing of haploproduction ability of soft winter wheat different hybrids in anther culture in vitro

Aim. Study the the efficiency of androgenesis in vitro in anther culture of different genotyps winter wheat. Methods. Obtaining of soft winter wheat double haploid lines by anther culture in vitro. The statistical methods. Results The ability to androgenesis in an in vitro anthers culture of 30 soft winter wheat genotypes was tested. The differences in the frequency of callus induction and the ability to regenerate plants in the process of androgenesiss in vitro of winter soft wheat were detected. The range of variation haploproduction activiti (from % of implanted anthers) was broad. The sign of "the formation of callus" was in limited from 0 to 13.2 % and on the sign of "regeneration of green plants" – from 0 to 1.8 %. The 126 green plants-regenerants were received. Conclusions. Genotype-specific of microspores morphogenetic reactions of soft winter wheat in the process of androgenesis in vitro were revealed. It is advisable to use the anthers from the F2 hybrid population plant as a donor material to create an effective technology for producing double haploids of soft winter wheat was shown.
 Keywords: wheat hybrid, anther culture in vitro, callus, regeneration.

Impedance flow cytometry allows the early prediction of embryo yields in wheat (Triticum aestivum L.) microspore cultures

Haplomethods are key biotechnological tools that make it possible to rapidly produce perfectly homozygous lines, speeding up plant breeding programs. Under specific stress conditions, microspores are reprogrammed toward sporophytic pathways, leading to embryo formation. Various endogenous and exogenous factors affect embryo yield in androgenesis, so the improvement of androgenesis efficiency requires the development of early, reliable and robust reactivity markers. During the last decade, numerous cytological, cellular and biochemical approaches were carried out to finely characterize microspore development and fate during androgenesis. However, the different available markers are often species-dependent, and their development and application are time-consuming and cumbersome. In this study, we show the suitable use of impedance flow cytometry (IFC) to develop new robust, reliable and strong markers of androgenesis reactivity in wheat, leading to: (i) routine monitoring of the viability of heterogeneous cell cultures; (ii) quick and simple evaluation of stress treatment efficiency; and (iii) early prediction of embryo yields from microspore suspensions. IFC can therefore provide the fine characterization of all of the microspore developmental pathways that occur in a cell suspension, for embryogenic microspores as well as pollen-like microspores. IFC technology has become a very useful tool to track and characterize wheat microspores in androgenesis, but can also be adapted to other species and other in vitro cell culture systems.

Comparison of the Androgenic Response of Spring and Winter Wheat (Triticum aestivum L.)

Androgenesis is potentially the most effective technique for doubled haploid production of wheat. It is not however widely used in breeding programmes due to its main limitation: the genotype dependence. Due to genetic differences between spring and winter wheat, it was assumed that both phenotypes are different in their capacity to conduct androgenesis. And so, the aim of this investigation was to verify the effectiveness of androgenesis induction and plant regeneration of spring and winter wheat genotypes while considering varying amounts of growth hormones in the induction medium. Fifteen genotypes of spring wheat and fifteen of winter wheat were used in the experiment. Six hundred anthers of each of the 30 genotypes were plated and analysed. Previous studies have allowed selection of the best medium for wheat androgenesis and a combination of growth hormones that are the most effective in stimulating microspore proliferation. Therefore, C17 induction media with two combinations of growth hormones were used: I—supplemented only by auxins (2,4-D and dicamba), and II—supplemented by auxin and cytokinin (2,4-D and kinetin). Data was recorded according to the efficiency of androgenic structure formation (ASF), green plant regeneration (GPR), and albino plant regeneration (APR). The results showed that the induction and regeneration of androgenesis in the spring wheat were more efficient than in the winter ones. The spring genotypes formed more androgenic structures and green plants on anthers plated on the medium supplemented only by auxins, in contrast to the winter genotypes which were better induced and regenerated on the medium supplemented by auxin and cytokinin. The study showed that to increase the efficiency of androgenesis, it is necessary to select appropriate factors such as concentration and type of hormones in medium composition, affecting the course of the culturing procedure according to the winter or spring phenotype of donor plants.

Comparison of methods for obtaining doubled haploids of carrot

Doubled haploid lines of carrot can be obtained through androgenesis in anther cultures and in isolated microspore cultures. The two methods were compared using three carrot cultivars (‘Kazan F1’, ‘Feria F1’, and ‘Narbonne F1’) at the androgenesis induction stage, during plant regeneration from embryos, and during acclimatization of androgenetic plants as well as their characterization. It was found that cultivar was the main factor affecting the efficiency at each stage of plant production in both anther and isolated microspore cultures. The efficiency of androgenesis in anther cultures of ‘Feria F1’ was considerably higher in comparison with isolated microspore cultures, and more plants were obtained from the embryos of androgenesis-cultured plants. In ‘Kazan F1’ and ‘Narbonne F1’, more acclimatized androgenetic plants were produced from anther cultures. Ploidy assessment of acclimatized plants of ‘Narbonne F1’ showed that the majority of the plants in the population derived from anther cultures had a doubled chromosome (DH) set. On the other hand, the majority of plants obtained from isolated microspore cultures were haploids. When assessing homozygosity, it was found among plants obtained in anther cultures that the percentage of homozygotes for phosphoglucose isomerase (PGI) and aspartate aminotransferase (AAT) depended on the cultivar. In contrast, the majority of plants derived from isolated microspore cultures were homozygous regardless of cultivar.

Effect of medium gelatinized component on the effectiveness of androgenesis in vitro Oryza sativa L.

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.

Androgenesis, gynogenesis, and parthenogenesis haploids in cucurbit species

Haploids and doubled haploids are critical components of plant breeding. This review is focused on studies on haploids and double haploids inducted in cucurbits through in vitro pollination with irradiated pollen, unfertilized ovule/ovary culture, and anther/microspore culture during the last 30years, as well as comprehensive analysis of the main factors of each process and comparison between chromosome doubling and ploidy identification methods, with special focus on the application of double haploids in plant breeding and genetics. This review identifies existing problems affecting the efficiency of androgenesis, gynogenesis, and parthenogenesis in cucurbit species. Donor plant genotypes and surrounding environments, developmental stages of explants, culture media, stress factors, and chromosome doubling and ploidy identification are compared at length and discussed as methodologies and protocols for androgenesis, gynogenesis, and parthenogenesis in haploid and double haploid production technologies.

Enhancement in androgenesis efficiency in barley (Hordeum vulgare L.) and bread wheat (Triticum aestivum L.) by the addition of dimethyl sulfoxide to the mannitol pretreatment medium

Dimethyl sulfoxide (DMSO) is a well-known solvent widely used in cell biology owing to its specific physicochemical properties, which allow it to decrease the lipid bilayer’s thickness and membrane fluidity while increasing membrane permeability. To improve doubled haploid production using an anther culture method in barley and bread wheat, the addition of DMSO to the pretreatment medium was tested. The first experiment was carried out on four barley cultivars with varying degrees of androgenetic ability by exposing them to four DMSO concentrations (0, 0.2, 1 and 2 % v/v). The medium with 1 % DMSO was the most effective in increasing the numbers of embryos and plants. The highest concentration tested (2 %) negatively impacted all of the measured variables when compared with the results of the 1 % DMSO addition, probably owing to a toxic effect. The effects caused by this solvent were more remarkable on the most recalcitrant cultivars, in which there was a threefold increase in the number of green plants. Furthermore, a downward trend in the albinism rate was observed as the concentration of DMSO increased. In a second experiment, we compared a 1 % DMSO supplement with the control to determine whether its addition was effective in several cultivars and F1 crosses of bread wheat. As in barley, there was a marked increase in the number of green plants, leading to a twofold to fourfold increase in both cultivars and F1 crosses.

Androgenic capability among genotypes of winter and spring barley

AbstractVariable efficiency of androgenesis remains a serious problem in many species of cereals. It is still unclear what makes certain genotypes more amenable to androgenesis than others. This study was undertaken to quantify the previously suspected advantage of winter barley genotypes over spring ones with regard to regeneration efficiency in anther culture. The material consisted of 40 barley hybrids originating from Polish breeding companies. The number of androgenic structures per 100 anthers did not differ significantly between analysed groups (119 vs. 152 non‐significant), but the average regeneration of green plants per 100 anthers was five times higher in winter genotypes (6.4 vs. 1.3). The incidence of albinism was lower for the winter than for the spring materials (70% vs. 90%), while the rate of spontaneous chromosome doubling was similar in both groups (58% vs. 56%). The results strongly support the notion that winter genotypes are more amenable to androgenesis and this may be a consequence of their better adaptation to stress conditions.

Doubled haploid production in Brassica L.

Doubled haploids (DHs) production through androgenesis is a biotechnological method for genetic improvement of crops. Biotechnological DH line production offers advantages to plant breeders, including the possibility to obtain homozygous lines within a year in contrast to common inbreeding methods, which may take 6–12 years. The greatest success in androgenesis has been achieved in some varieties of rapeseed ( Brassica napus L.). However, the efficiency of androgenesis in other Brassica species is still poor. Induction of microspore embryogenesis is usually induced by many factors such as conditions of donor plant growth, genotype, microspore developmental stage, culture medium composition, and culture conditions. The reprogramming of microspores from the gametophytic to the sporophytic habit of development depends on various stress factors. Certain pretreatments of microspores, such as high temperature and colchicine, can favor androgenesis in Brassica species. Plant regeneration from microspores can be improved by proper application of different growth regulators (ethylene, abscisic acid, and indole acetic acid). Optimal combinations of these factors are mandatory for efficient androgenesis. In this review, we summarize the experience of our colleagues in DH-technology in the Brassica genus. Attention is focused on some factors influencing the development of doubled haploid plants and their impact on enhancing the efficiency of androgenesis in Brassica species.

Effect of polyamines on in vitro anther cultures of carrot (Daucus carota L.).

This study examines the influence of the polyamines putrescine and spermidine on the efficiency of androgenesis in anther cultures of 2 carrot cultivars, Kazan F_1 and Narbonne F_1, and the effect of putrescine on the process of plant regeneration from androgenetic embryos. In the Kazan F_1 variety, an increase in the number of obtained embryos was achieved using each of the 2 polyamines separately and in combination. In contrast, no beneficial effects of polyamines on the efficiency of androgenesis were observed in the Narbonne F_1 variety. Putrescine added to the regeneration medium increased the number of obtained plants. For embryos obtained on induction medium without putrescine, the best concentration was 160 mg putrescine in 1 L of regeneration medium, and for those obtained on the induction medium with putrescine the best concentration was 0.5 mg putrescine per 1 L. All of the resulting plants, both in the experimental and control combinations, had a doubled set of chromosomes. The vast majority of them were homozygous for both isoenzymes, glucose-6-phosphate isomerase and aspartate aminotransferase, and the distribution of homo- and heterozygous received combinations with the polyamines and in the control was very similar.

Quantitative trait loci associated with androgenic responsiveness in triticale (×Triticosecale Wittm.) anther culture

Quantitative trait loci (QTLs) associated with androgenic responsiveness in triticale were analyzed using a population of 90 DH lines derived from the F1 cross between inbred line ‘Saka 3006’ and cv. ‘Modus’, which was used in a number of earlier studies on molecular mapping in this crop. Using Windows QTL Cartographer and MapQTL 5.0, composite interval mapping (CIM) and association studies (Kruskal–Wallis test; K–W) for five androgenesis parameters (androgenic embryo induction, total regeneration and green plant regeneration ability, and two characteristics describing final androgenesis efficiency) were conducted. For the studied components of androgenic response, CIM detected in total 28 QTLs which were localized on 5 chromosomes from A and R genomes. Effects of all QTLs that were identified at 2.0 or above of the LOD score explained 5.1–21.7 % of the phenotypic variation. Androgenesis induction was associated with seven QTLs (LOD between 2.0 and 5.8) detected on chromosomes 5A, 4R, 5R and 7R, all of them confirmed by K–W test as regions containing the markers significantly linked to the studied trait. What is more, K–W test revealed additional markers on chromosomes: 5A, 2BL, 7B and 5R. Both total and green regeneration ability were controlled by genes localized on chromosome 4A. Some of the QTLs that affected final androgenesis efficiency were identical with those associated with androgenic embryo induction efficiency, suggesting that the observed correlation may be either due to tight linkage or to pleiotropy. Key message Five regions of the triticale genome were indicated as revealing significant marker/trait association. Markers located in these regions are potentially useful for triticale breeding through marker-assisted selection.

Enhancement of androgenesis by abiotic stress and other pretreatments in major crop species

Rapid production of doubled haploids (DHs) through androgenesis is an important and promising method for genetic improvement of crop plants. Through androgenesis complete homozygous plants can be produced within a year compared to long inbreeding methods that may take several years and costly. Significant advantage of androgenesis is that it not only speeds up the process to achieve homozygosity, but also increases the selection efficiency. Though success in androgenesis has been achieved in many crop plants, yet there are certain limitations especially, low frequency of embryogenesis and regeneration in few species. In fact in many cereals, induction of embryos and regeneration of green plants is still a hurdle that one needs to overcome to improve the efficiency of androgenesis. Efficient androgenesis is usually induced by the successful application of different stress pretreatment. Since so many stress factors can trigger the reprogramming of microspores and that have been co-related to change the ultrastuctural changes of cells to embryos and finally haploid plants. It has been shown that certain pretreatment such as (i) physical stresses as cold, heat shock, starvation, drought stress, osmotic pressure, gamma irradiation, oxidative stress, reduced atmospheric pressure, and (ii) chemical treatments such as colchicine, heavy metal, ABA, CGA, AEC, Azetidine, 2-NHA, either individual or combined effect of more than one stress factors may positively influence androgenetic efficiency. This review highlights the recent and past work on uses of various abiotic stresses and pretreatments and their impact on enhancing the efficiency of androgenesis on some major crop species for the development of doubled haploid plants.