In Vitro Culture of Zygotic Embryos: Its Use in Soya and Sunflower Improvement

Author(s):  
H. Serieys
HortScience ◽  
2001 ◽  
Vol 36 (2) ◽  
pp. 351-352 ◽  
Author(s):  
Pedro A. Sansberro ◽  
Hebe Y. Rey ◽  
Luis A. Mroginski

Plants of Ilex argentina L., I. brasiliensis (S.) L., I. brevicuspis R., I. dumosa R., I. integerrima (V.C.) L., I. microdonta R., I. pseudoboxus R., and I. theezans C.M. were obtained by immature embryo culture. Heart-stage zygotic embryos were removed from immature fruits and cultured aseptically on quarter-strength Murashige and Skoog medium with 3% sucrose, 0.65% agar, and 0.1 mg·L-1 zeatin. Cultures were incubated at 27±2°C for 4 weeks, in darkness and subsequently transferred to a culture room with a 14-hour photoperiod (116 μmol·m-2·s-1) for another 4 weeks. Seedlings with two leaves, derived from germinated embryos, were successfully transplanted to pots containing 1 peat: 1 perlite: 1 sand (v/v) and were maintained in greenhouse conditions. From 95% to 100% of transplanted seedlings survived. Chemical name used: 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).


2020 ◽  
Vol 9 (9) ◽  
pp. e231997031
Author(s):  
João Bosco de Oliveira Júnior ◽  
Camilla Mendes Pedroza Pessoa ◽  
Hendril da Silva Lopes ◽  
Frederico Henrique da Silva Costa ◽  
Jonny Everson Scherwinski Pereira

Dragon's blood, native species from South America, has therapeutic properties scientifically proved. This study had the objective of developing a protocol for in vitro establishment and plantlets acclimatization from zygotic embryos, since researches with species are incipient. Culture media (MS, WPM, QL and N6) were assessed, without and with activated charcoal (2 g L-1). It was observed the percentage of developed embryos from 83% to 100% and the effect of medium composition for plant height and leaf number in vitro. The plantlets were acclimatized with 100% of survival rate, and for treatments from in vitro culture only the culture medium influenced diameter and plant height. The WPM and QL formulations without activated charcoal are indicated for in vitro culture of embryos and acclimatization of Dragon blood. The suggested methodology represents an important strategy for reproduction, physiological study and preservation of species.


HortScience ◽  
1994 ◽  
Vol 29 (6) ◽  
pp. 695-697 ◽  
Author(s):  
Paula P. Chee

An embryo culture method overcomes the lengthy dormancy requirement of Taxus L. spp. (yew) seeds. When zygotic embryos excised from mature T. brevifolia L. seeds were cultured in darkness for 4 weeks on one of three basal salt media (B5, Litvay, and Murashige and Skoog), radicle emergence and seedling development was highest on B5 basal salt medium. After 1 to 2 weeks on B5 basal salt medium, seedling development of T. brevifolia, T. cuspidata L., T. baccata L., and T. baccata stricta L. ranged from 2% to 36%. BA at 2.25 μm had no effect on radicle emergence; 22.5 μm prevented it. Embryos excised from mature or nearly mature seeds had the highest frequency of radicle emergence and seedling development. Cultured embryos developed seedlings in only 8 to 10 weeks. Chemical name used: N6-benzyladenine (BA).


2018 ◽  
Vol 36 (2) ◽  
pp. 229-234
Author(s):  
Cláudia Ulisses ◽  
Marciana Morais ◽  
Marta R Barbosa ◽  
Cynthia C Albuquerque ◽  
Lilia Willadino ◽  
...  

ABSTRACT The aim of this study is to compare physiological development of Heliconia bihai cv. Lobster Claw Two plants derived from in vitro culture of zygotic embryos and conventional propagation. Heliconias obtained from rhizomes and from in vitro multiplication were evaluated every 30 days during ten months under greenhouse conditions. The experimental design was completely randomized, with ten repetitions, and the 2x10 factorial arrangement consisted of two plant multiplication methods and ten evaluations performed at different times. The analyzed biometric parameters were plant height, number of leaves, number of tillers, leaf area, and color intensity in the bracts. Plants derived from in vitro culture showed significant differences in the development of the evaluated physiological parameters in comparison to plants derived from rhizomes, and they also showed early flowering. Although the in vitro cultured plants were derived from zygotic embryos, no morphological changes were found in the vegetative and reproductive parts (inflorescence) of the plants or in the colorimetry. It shows that the in vitro cultures of zygotic embryos may be used as a technique to produce seedlings on a large-scale, thus allowing the floriculture sector to grow in the region and all over the country.


Author(s):  
Iraida N. Tretyakova ◽  
◽  
Maria E. Park ◽  
Angelica P. Pakhomova ◽  
Irina S. Sheveleva ◽  
...  

The biotechnology of somatic embryogenesis in in vitro culture is the most promising direction in the reproduction of conifers. The use of this technology makes it possible not only to massively propagate the best genotypes of trees, but also serves a model for studying the structural, physiological and molecular and genetic mechanisms of both somatic and zygotic embryogenesis in conifers. The main aim of this research was to obtain embryogenic cultures (ECs) producing somatic embryos and embryonic suspension mass (ESM) of Picea obovata. The studies were carried out in 2014-2019 on 30 Siberian spruce trees growing in the vicinity of the city of Krasnoyarsk. To detect genotypes competent for somatic embryogenesis, new donor trees were selected every year for the experiment. 3-10 cones were collected from each tree at different stages of embryo development: globular embryo (the first decade of July), the initiation stage cotyledons (second decade of July), the stage of developed cotyledons (third decade of July) and mature embryos (August). Sterilized explants (zygotic embryos at different stages of development) were introduced into in vitro culture on basic media DCR (Gupta PK and Durzan DJ, 1985), ½LV (Litvay JD et al., 1985), MS (Murashige T and Skoog F, 1962) and AI (Tretyakova IN, 2012). All media were supplemented with myo-inositol - 100 mg/L, casein hydrolyzate - 500-1000 mg/L, L-glutamine - 500 mg/L, sucrose - 30 g/L and agar - 7 g/L. Ascorbic acid at a concentration of 400 mg/L was used as an antioxidant. The level of growth regulators was: 2,4-dichlorophenoxyacetic acid (2,4-D) - 2 mg/L and N6 -benzoaminopurine (BAP) - 1 mg/L. For the proliferation of the ESM, DCR and AI basic media containing 2,4-D (2 mg/L), BAP (0.5 mg/L) and sucrose (20 g/L) were used. The pH was adjusted to pH = 5.8. All culture medium and components were sterilized depending on their termolabile properties. Under aseptic conditions, embryos were removed from megagametophytes and inoculated into nutrient media, 10 explants per flask in 25 replicates. The cultures were incubated in the dark at 24 ± 1 °C. Subcultivation to fresh nutrient medium was carried out every 14 days. To control the quality of cell lines (CL) during subculturing, we performed cytological analyzes using temporary preparations (3-5 preparations for each CL). We evaluated the quality of the embryogenicity of the cultures by the presence of even single structures with pronounced polarity - a globular embryo with a suspensor. The results of the study showed that the induction of callus cultures of Siberian spruce is influenced by such factors as the development stage of the explant, the nutrient medium and the genotype of the donor tree. The introduction of P. obovata immature zygotic embryos into in vitro culture at the stage of the globular embryo, both with megagametophytes and extracted from them, turned out to be ineffective. The induction of callus cultures in Siberian spruce was significantly reduced when mature zygotic embryos were introduced into the culture in vitro. The highest response of explants of Siberian spruce was at the stage of developed cotyledons (See Table 1). In the DCR medium, 90% of explants formed callus (See Table 2). The mineral composition of the media did not significantly affect the induction of callus formation in Siberian spruce. The exception was the MS medium, in which callus cultures were formed only in 41% of explants (See Table 2). The growth of callus cultures was most active in the DCR medium. After 6 months of cultivation, 15-32% of calli remained viable (See Table 2). Cytological analysis of callus cultures showed that they include cells of different types (See Fig. 1 and 2). The first type of cells consisted of elongated cells reaching a length of 10 ± 3 μm, others consisted of isodiametric cells with a diameter of 60 ± 3.5 μm. The somatic embryo globule and embryonic tubes were formed from elongated cells. Isodiametric cells were actively dividing and forming callus. Only 3 cell lines (out of 300 cell lines) belonging to two donor trees had an active ability to proliferate. Globular somatic embryos were actively forming in these cell lines (See Fig. 3). An actively proliferating ESM was formed. Thus, we carried out a comprehensive assessment of the factors influencing the induction of somatic embryogenesis in Siberian spruce. The results obtained indicate that for the successful formation of somatic embryos, the determining factor is not only the choice of donor plants, but also the development stage of the explant. We found that the best stage in the development of zygotic embryos when introduced into in vitro culture of Siberian spruce is the stage of immature embryos with formed cotyledons, while the DCR, ½LV and AI nutrient medium supplemented with growth regulators (2.4-D and BAP) is optimal.


2008 ◽  
Vol 4 (4) ◽  
pp. 450-455 ◽  
Author(s):  
A. W. Grootbo ◽  
M. M. O`Kenne ◽  
N. L. Mkhonza ◽  
K. Kunert ◽  
E. Chakauya ◽  
...  

Hoehnea ◽  
2018 ◽  
Vol 45 (4) ◽  
pp. 663-668
Author(s):  
Daniel da Silva ◽  
Angela Maria Imakawa ◽  
Suely de Souza Costa ◽  
Paulo de Tarso Barbosa Sampaio

ABSTRACT The aim of this study was to evaluate the in vitro germination of zygotic embryos and seeds of Caesalpinia ferrea Martius and the morphogenetic responses of the explants to different concentrations of growth regulators. Seeds and zygotic embryos were inoculated in MS culture medium and kept in a growth room at a temperature of 25 ± 2 ºC for 16 hours of photoperiod for 30 days. The seeds had a higher in vitro germination rate than the explants from zygotic embryos. However, zygotic embryos in MS medium supplemented with 0.9 mg L-1 BAP had the highest percentage of regeneration (50%), number of shoots (3.25), buds (2.85) and leaves (3.15), multiplication rate (27.75), and length of shoots (1.96 cm). The in vitro culture of zygotic embryos and seeds made possible the multiplication of a higher number of healthy seedlings. Thus, it can be used as an alternative technique for the propagation of this species.


2013 ◽  
Vol 34 (5) ◽  
pp. 2179
Author(s):  
Daniel Arthur Gaglik Waldow ◽  
Lia Rejane Silveira Reiniger ◽  
Diego Pascoal Golle ◽  
Aline Ritter Curti

Sign in / Sign up

Export Citation Format

Share Document