scholarly journals The Influence of the Developmental Stage and Culture Medium upon the Somatic Embryo Induction in Quercus petraea

2013 ◽  
Vol 41 (2) ◽  
pp. 385
Author(s):  
Adrian Ioan TIMOFTE ◽  
Simona VICAȘ

The somatic embryogenesis is an advanced method for clonal propagation and a useful tool for ex situ conservation of genetic resources. In this paper an experimental device was designed, composed of: one oak species (Quercus petraea), one provenance, four types of explants (developmental stages of the zygotic embryo) and four variants of culture medium. It was released the two-way statistical analyses of the two analyzed factors affecting the embryo induction efficiency: the stage of explant and culture medium (4 stages x 4 culture media). The most important factor affecting the efficiency of somatic embryogenesis was found to be the developmental stage of the zygotic embryos used as juvenile explants. The high embryogenic ability of young zygotic embryos and low embryogenic ability of maturing zygotic ones showed the strong positive relationship between somatic embryo induction and accumulation of reserve nutrients in cotyledons. The best results were obtained with the youngest embryos (stage 1), with a linear decrement towards the stage 4. The embryo induction efficiency ranged between 44% and 10% in Q. petraea. Two nutritive culture media and two growth hormone combinations were tested. The effect of culture medium proved to be totally insignificant, besides the differences observed among the variants cultivated on different media. The situation was the same as well for the nutrients as for growth regulators.

2020 ◽  
Vol 21 (18) ◽  
pp. 6784
Author(s):  
Shengli Song ◽  
Rui Yan ◽  
Chunxia Wang ◽  
Jinxia Wang ◽  
Hongmei Sun

Auxin transport mediates the asymmetric distribution of auxin that determines the fate of cell development. Agrobacterium-mediated genetic transformation is an important technical means to study gene function. Our previous study showed that the expression levels of LpABCB21 and LpPILS7 are significantly up-regulated in the somatic embryogenesis (SE) of Lilium pumilum DC. Fisch. (L. pumilum), but the functions of both genes remain unclear. Here, the genetic transformation technology previously developed by our team based on the L.pumilum system was improved, and the genetic transformation efficiency increased by 5.7–13.0%. Use of overexpression and CRISPR/Cas9 technology produced three overexpression and seven mutant lines of LpABCB21, and seven overexpression and six mutant lines of LpPILS7. Analysis of the differences in somatic embryo induction of transgenic lines confirmed that LpABCB21 regulates the early formation of the somatic embryo; however, excessive expression level of LpABCB21 inhibits somatic embryo induction efficiency. LpPILS7 mainly regulates somatic embryo induction efficiency. This study provides a more efficient method of genetic transformation of L. pumilum. LpABCB21 and LpPILS7 are confirmed to have important regulatory roles in L. pumilum SE thus laying the foundation for subsequent studies of the molecular mechanism of Lilium SE.


2014 ◽  
Vol 12 (1) ◽  
pp. 1-6 ◽  
Author(s):  
MN Hassan ◽  
MS Haque ◽  
MM Hassan ◽  
MS Haque

Genetic improvement of garlic through conventional breeding is very difficult due to sterile nature of its flower. Hence, an alternative system is desirable to induce genetic variation. Tissue culture could be a good opportunities and somatic embryogenesis is one of the potential techniques of tissue culture for in vitro regeneration of garlic plant. The successes and production of somatic embryo depends on several factors such as optimization of media components, genotypes and explant type. Therefore, in the present investigation, garlic root tips were used as explant for callus and somatic embryo induction under different plant growth regulator combination. It was found that MS+1.0 mg l-1 2,4-D was the most favorable (86.10% regeneration with 2.19 cm callus diameter) for callus induction. This concentration also induced and produced good quality somatic embryo. In addition, MS+2.0 mg l-1 Kinetin gave better regeneration of somatic embryo and yielded the highest number (4.670) and longest length (7.0 cm) of shoots per callus. The procedure used a single hormonal signal for callus and somatic embryo induction as well as hormone free medium for further development of plantlet. Besides, maximum duration for callus induction and somatic embryo production was 17 and 10.67 days respectively. Thus, it appears that the protocol is cheap and time bound and particularly useful for conducting experiment for genetic improvement of garlic. Furthermore, as the protocol is cost effective, it can be further tested for commercial feasibility. DOI: http://dx.doi.org/10.3329/jbau.v12i1.20747 J. Bangladesh Agril. Univ. 12(1): 1-6, June 2014


2005 ◽  
Vol 35 (5) ◽  
pp. 1053-1060 ◽  
Author(s):  
Harald Kvaalen ◽  
Ola Gram Daehlen ◽  
Anne Tove Rognstad ◽  
Borgny Grønstad ◽  
Ulrika Egertsdotter

Seeds of Abies lasiocarpa (Hook.) Nutt. (subalpine fir) were dissected, and the different parts were analyzed for elemental composition. The data were used to design a novel growth medium for initiation of somatic embryogenesis. Embryogenic cultures were initiated from immature zygotic embryos from six open-pollinated families of A. lasiocarpa on three different media. The frequency of initiation was the highest in early to mid-July when the zygotic embryo explants were ca. 0.8 mm long. Thereafter the response declined rapidly. The culture media did not significantly affect the initiation frequencies, but the subsequent growth and culture survival was dependent on the culture medium. On the Schenk Hildebrandt medium, many cultures ceased to grow and died. Several of the decaying cultures were rescued after transfer to one of the new media. Proliferating cultures could be stimulated to produce mature embryos. Of 2510 mature somatic embryos, 212 (8.4%) converted to plants, and 35 plants have grown over two periods.


Author(s):  
Muniappan V ◽  
Manivel P ◽  
Prabakaran V ◽  
Palanivel S ◽  
Parvathi S

Somatic embryogenesis was carried out epicotyl portion of the mature embryo/apical portion. The somatic embryo induction medium containing 2,4-D or NAA (10.0 to 50.0 mg/l). Of the two concentrations tested 2,4-D (30.0mg/l) recorded the highest percentage of response followed by NAA (30.0mg/l). But the highest number of somatic embryo were recorded in 30.0mg/l of 2,4-D followed by NAA. The apical portion of the mature embryo formed direct embryos without any intervention of callus. The maximum percentage of embryogenic cultures were noticed in 30.0mg/l of 2,4-D followed by NAA at 30.0mg/l. for the differentiation of somatic embryos, the embryogenic masses were transferred to medium without any growth regulator. The maximum number of somatic embryos per culture was recorded in 30 mg/l of 2,4-D followed by 30.0 mg/l of NAA. Keywords: Arachis hypogaea L.,Somatic Embryogenesis, 2,4-D and NAA


2014 ◽  
Vol 23 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Dinesh Giri ◽  
Sushma Tamta

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013  (December)


2010 ◽  
Vol 140 (2) ◽  
pp. 225-243 ◽  
Author(s):  
Priyanka Saha ◽  
Subhendu Bandyopadhyay ◽  
Sarmistha Sen Raychaudhuri

Forests ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1335
Author(s):  
Shuaifei Jiang ◽  
Xiaoyi Chen ◽  
Ying Gao ◽  
Ying Cui ◽  
Lisheng Kong ◽  
...  

Prince Rupprecht’s larch (Larix principis-rupprechtii Mayr) is a native conifer in North China with great economic and ecological values. Somatic embryogenesis (SE) is a powerful tool for the mass clonal propagation in plants. In this study, we described a high-efficiency SE system via indirect pathways and investigated the effect of genotype, culture conditions and phytohormones on SE. Immature zygotic embryos (IZEs) of L. principis-rupprechtii Mayr were used as explant materials. In the induction stage, embryogenic tissues (ETs) were induced on mLV medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzylaminopurine (6-BA). The initiation frequencies showed significant differences (p < 0.05) among 20 genotypes of open-pollinated mother trees with the highest induction frequency reaching 30%. For tissue proliferation, proliferation in liquid medium was more efficient compared with proliferation in semi-solid medium, providing a multiplication rate of 3.12 in an 8-day subculture period. As a necessary exogenous plant growth regulator (PGR) for somatic embryo maturation in conifers, abscisic acid (ABA) was optimized at 16 mg L−1 in this system. Next, an orthogonal test on osmotic pressure factors showed 50 g L−1 sucrose, 7 g L−1 phytagel and 75 g L−1 polyethylene glycol (PEG) was the optimal combination for somatic embryo maturation in L. principis-rupprechtii Mayr. Moreover, the dispersion culture method provided a more efficient somatic embryo maturation, up to 545 per gram of fresh weight (FW). Finally, 2 g L−1 of active charcoal (AC) was found to increase the somatic embryo germination rate to 63.46%. The improved protocol of SE will serve as a foundation for establishing mass propagation and genetic transformation of L. principis-rupprechtii Mayr.


Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 3 ◽  
Author(s):  
Xiuxia Ren ◽  
Ya Liu ◽  
Byoung Ryong Jeong

Somatic embryogenesis is a preferred method for vegetative propagation due to its high propagation efficiency. In this study, zygotic embryos, cotyledons, and hypocotyls of Paeonia ostii ‘Fengdan’ were used as the explant to induce somatic embryogenesis. The results showed that a combination of 0.5 mg·L−1 thidiazuron (TDZ) and 0.5 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D) was effective in inducing somatic embryos from the zygotic embryo and cotyledon explants. Hypocotyls only formed somatic embryos on Murashige and Skoog (MS) medium supplemented with both 0.5 mg·L−1 TDZ and 0.5 mg·L−1 1-naphthylacetic acid (NAA). Moreover, the compact callus was effectively produced from zygotic embryo, cotyledon, and hypocotyl explants in medium supplemented with a combination of 3.0 mg·L−1 6-benzylaminopurine (BA) and 1.0 mg·L−1 NAA, and then converted into somatic embryos in the same medium, and the ratio of the explants with embryo induction and number of embryos induced per explant were much higher than those induced by 0.5 mg·L−1 TDZ and either 0.5 mg·L−1 2,4-D or 0.5 mg·L−1 NAA. The MS medium was better than the woody plant medium (WPM) for inducing somatic embryos from zygotic embryo and hypocotyl explants, whereas the WPM was better than the MS medium for somatic embryogenesis induction from cotyledon explants. All of the somatic embryos developed well into mature embryos on their respective media supplemented with both 3.0 mg·L−1 BA and 1.0 mg·L−1 NAA. Overall, the protocols for indirect somatic embryogenesis from zygotic embryo, cotyledon, and hypocotyl of P. ostii ‘Fengdan’ were successfully established, which can greatly facilitate their propagation and breeding processes.


Sign in / Sign up

Export Citation Format

Share Document