Number Of Green Plants Research Articles

Genetic analysis of anther culture response and identification of QTLs associated with response traits in wheat (Triticum aestivum L.).

Anther culture is the most effective tool for doubled haploid production of wheat. This investigation was conducted to estimate genetic parameters of anther culture response in wheat and identification of putative Quantitative trait loci (QTLs) associated with response traits. Two varieties of wheat, namely ICR-DH (P1) and Sle 1 × 15 (P2) and their F1 and F2 progenies were used in the present investigation to estimate genetic parameters of anther culture response. Two molecular marker systems, SRAP and SSR markers were used to detect the polymorphism between two anther donor parents. Single marker analysis (SMA) and Composite interval mapping (CIM) were used to localize the putative QTL associated with four anther culture response in wheat using 100 plants of F2 population derived from F1 cross 'ICR-DH' × 'Sel 1 × 15'. Analyses of variance indicated significant differences between four populations (P1, P2, F1 and F2) for callus induction (CAL), number of green plants per 100 anther (GR), number of albino plant per 100 anther (AR) and total regenerated plants per 100 anther (TR). The additive effects were more important than dominance effects in controlling these traits. The two molecular marker systems were sufficient in detecting polymorphism between two parents. Thirty two putative QTLs were detected on eight linkage groups. Our study indicated that the additive effects of genes and detection of new QTLs permit marker-assisted selection of genotypes with high green plantlet regeneration efficiency in anther culture, and therefore favor efficient use of anther culture in wheat breeding programs.

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.

Effects of n-butanol on barley microspore embryogenesis

Doubled haploid (DH) production is an efficient tool in barley breeding, but efficiency of DH methods is not consistent. Hence, the aim of this study was to study the effect of n-butanol application on DH barley plant production efficiency. Five elite cultivars of barley and thirteen breeding crosses with different microspore embryogenesis capacities were selected for n-butanol application in anther and isolated microspore cultures. Application of 0.1 % n-butanol after a mannitol stress treatment in anther culture significantly increased the number of embryos (up to almost twice) and green plants (from 1.7 to 3 times) in three low-responding cultivars: Albacete, Astoria and Majestic. No significant differences on microspore embryogenesis efficiency were observed in medium and high responding cultivars. The application of n-butanol treatment to isolated microspores from cold treated spikes in thirteen spring breeding crosses with a low or very low androgenetic response did not have a significant effect on the overall number of green plants. Nevertheless, an increase in the number of green plants was observed when 0.2 % n-butanol was applied in four out of seven low-responding crosses. Therefore, application of n-butanol could be routinely applied to anther cultures using mannitol treatment, in low-responding material. However, further studies are needed to determine optimal conditions in protocols using cold treatment and isolated microspore cultures.

The effect of liquid and solid medium on production of winter triticale (×TriticosecaleWittm.) anther-derived embryos and plants

The efficiency of anther culture induction in solid and liquid medium C17 was compared for nine triticale genotypes. For all genotypes androgenic embryos and green plants efficiency was higher on liquid medium. On liquid medium 22.2–135.9 and on solid medium 27.5–121.3 androgenic embryos/100 anthers were produced. The embryos were transferred to a regeneration medium 190-2 and also a higher number of green plants (ranging from 3.0 to 31.4/100 anthers) were obtained in liquid culture compared with the yield of plants (1.6–21.0) on solid medium.

The effect of osmotic potential on anther culture in spring wheat ( Triticum aestivum )

This study was conducted to determine the effect of osmotic potential in a modified 85D12 medium on both callus induction and plant regeneration in the anther culture of two wheat genotypes, cv. Chris and cv. Pavon. Altering the medium osmotic potential by changing the carbohydrate source and concentration or by adding a non-metabolized osmoticum appeared to have the greatest potential for improving anther-derived green plant production. The medium osmotic potentials were varied (-0.67 to −2.30 MPa) by altering sucrose and PEG concentration. Both osmotica affected callus production, with −0.9 to −1.4 MPa media producing the most calluses. Callus production depended on genotype and osmoticum. Only PEG concentration affected green plant regeneration. The greatest number of green plants (11.5 plants per 100 anthers in cv. Chris) was obtained with 0.0125 M of PEG. This experiment suggested that a low level of PEG in the medium was beneficial for producing green plants from wheat anthers.

Effect of genotype, medium and sugar on triticale (× Triticosecale Wittm.) anther culture response.

The response of sixteen triticale genotypes to the anther culture induction medium composition was investigated. Two sugar versions of P2 and C17 media were tested, containing 9 % of either maltose or sucrose. Subsequently obtained embryo-like structures were regenerated on 190-2 medium. The results showed the significant differences between genotypes for all the investigated traits. Green plants were obtained at frequency ranging from 0 to 2.3 per 100 plated anthers. Maltose was superior to sucrose (on average respectively 0.69 and 0.03 green plants per 100 plated anthers). P2 with maltose yielded the highest number of green plants in more genotypes than either of remaining media. The genotype × medium, genotype × sugar and medium × sugar interaction appeared significant. Both, P2 and CI7 media supplemented with maltose are advisable for triticale haploid production.

Variations in green plant regeneration response from anthers of indica rice and their hybrids with japonica cv. Taipei 309

The green plant regeneration ability from anthers of BR-7, a high yielding indica cultivar, Binnatoa (BA), a salt tolerant indica land race and IR-43 was tested in N6, M8, He2 and R2 media. The response was calculated on the basis of number of anthers producing green plants. The number of green plants per responding anther was also recorded. The response of BR-7 and BA was poor compared to the indica cultivar IR-43 in three of the media that were tested. In N6 medium, green plant regeneration of BA and BR-7 was respectively 10-fold and 100-fold less than the japonica cultivar Taipei 309 (T-309). No anther-derived green regenerant was obtained from another salt tolerant indica land race, Rajashail (RAJ). The N6 medium was selected to test green plant regeneration frequency from anthers obtained from the F1 crosses of T-309 × BR-7, T-309 × BA, T-309 × RAJ and T-309 × BR-7 AC regenerants backcrossed with BA. Our objective was to combine the salt tolerant trait of BA and the high yield of BR-7 in a single line. The intermediate crossing step with T-309 was performed to increase the green plant regenerability of the anthers. All F1 progeny from the crosses with T-309 showed significantly increased callus induction compared to the indica parent although the values were lower than the midparent means. Green plant regeneration compared to their respective indica parents either increased or decreased but never approached the level of T-309.

Regeneration of green plants from embryogenic suspension on cultures of kentucky blue grass ( Poa pratensis L.)

Summary Plants were regenerated from embryogenic suspension cultures of Kentucky blue grass ( Poa pratensis L.) cv. «Geronimo». Suspension cultures capable of regeneration of green plants were established from embryogenic callus derived from either entire mature seeds or isolated mature embryos. From both donor materials suspensions were established with a frequency of 100%. Green plants were regenerated from two out of 17 isolated-embryo-derived suspension cultures. One of the cultures yielded exclusively green plants and the other culture gave rise to both green and albino plantlets. Out of three seed-derived cultures, two possessed regeneration capacity, one of these also regenerating exclusively green plants and one culture regenerating albinos only. The capacity for regeneration of green plants remained constant in the two embryo-derived suspension cultures for at least 1 year. The number of green plants from 200mg fresh weight suspension cells varied from one in the least responsive culture up to 246 in the most responsive one. More than 500 green regenerants have been potted in soil and 400 are in the process of field testing. Evaluation of the morphology and fertility of the plants is in progress and the first three plants have undergone seed setting. This is the first report on regeneration of green plants from embryogenic suspension cultures in Kentucky blue grass.

Anther Culture in Rye: Improved Plant Regeneration Using Modified MS‐Media

AbstractThe use of androgenetic doubled haploids in rye breeding is still limited by low regeneration rates. In this study we tested the influence of two modified MS‐media on the number of reacting anthers and on plant regeneration. Picloram and 2,4‐D were used as auxin components in the induction media. The highest induction rates (reacting anthers per 100 cultivated anthers) and the highest regeneration rates (number of green plants per 100 cultivated anthers) were reached on a modified MS‐medium containing Picloram as auxin source, regardless of the donor plant material. The comparison of the results obtained with the single cross ‘SC 35’ which contains genes of Secale vavilovii and the true Secale cereale donor forms clearly show the genotypic influence of donor plant material on induction and regeneration ability.

Fertile indica and japonica rice plants regenerated from protoplasts isolated from embryogenic haploid suspension cultures

A system to regenerate fertile rice (Oryza sativa L.) plants (both indica and japonica varieties) from protoplasts isolated from anther-derived embryogenic haploid suspension cultures has been established. Green plants were regenerated from protoplast-derived cell clusters five months after suspension culture initiation. Protoplast yields and subsequent growth of the protoplast-derived microcalli were enhanced by transferring suspension cells into AA medium (Muller et al. 1978) three to four days prior to protoplast isolation. Protoplasts were cultured initially in Kao medium (Kao et al. 1977) and in association with nurse cells for four weeks. Protoplast-derived microcalli were transferred onto N6 (Chu et al. 1975) or MS (Murashige and Skoog 1962) media for callus proliferation. Callus growth was more rapid and the calli were more enbryogenic when grown on N6 medium. The 2,4-D concentration used to develop the suspension culture was important. Cell cultures grown in medium containing 0.5 mg/l 2,4-D released protoplasts whose plating efficiency was higher than for protoplasts obtained from suspension cultures grown in 2.0 mg/l 2,4-D. However, suspension cells grown in 2.0 mg/l 2,4-D were superior with regard to the ability of protoplast-derived calli to regenerate green plants. Amongst several hormone treatments evaluated, a combination of 0.5 mg/l NAA + 5.0 mg/l BAP resulted in the largest number of green plants regenerated. There were no significant differences between BAP or kinetin regarding total number of plants regenerated. More than 200 green plants have been produced form six independently initiated suspension cell lines. The number of regenerated plants per 10(6) protoplats plated anged from 0.4 to 20.0, and the average seed fertility of single panicles of these RO plants was about 40%.