scholarly journals An efficient protocol for somatic embryogenesis of garlic (Allium sativum L.) using root tip as explant

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

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.


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 14 (2) ◽  
pp. 73-81
Author(s):  
Yelnititis Yelnititis ◽  

Kulim is one of woody plant that have multifunction as wood source and for spice and medicinal. Generative propagation of this plant have trouble because seed use limited. The use of leaf segment through somatic embryogenesis to solve the problem. The objective of this study is to obtain the best treatment to embryogenic callus induction. The modification of basal medium of Murashige and Skoog was used as growth medium. The experiment was conducted in three stages are callus induction, embryogenic callus and somatic embryo induction. The treatment of 2,4-D (3,0 – 12 mg/l) used for callus induction. For embriogenic callus induction used 2,4-D (3,0 – 12,0 mg/l) combined with NAA 0,5 mg/l. The treatment of thidiazuron (0,1 – 0,7 mg/l) used for somatic embryo induction. The result showed that the treatment of 2,4-D 6,0 mg/l is the best for callus induction with compact of texture, green, dry and non embryogenic. The treatment of combination 2,4-D 12.0 mg/l with NAA 0.5 mg/l is the best for friable callus induction. The treatment of 2,4-D 6.0 mg/l combined with NAA 0,5 mg/l is the best for embryogenic callus induction with very friable of texture, easy to separate, dry, smooth and glossy. Thidiazuron of 0,1 mg/l treatment is the best for somatic embryos induction with the average number of 7,8 somatic embryos.


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


2016 ◽  
Vol 3 (2) ◽  
pp. 71
Author(s):  
Nur Ajijah ◽  
RR. Sri Hartati

<p><em>Information on the effect of cytokinins on cacao (</em>Theobroma cacao<em> L.) primary somatic embryogenesis and its interaction with explant types and genotypes is not yet known. This study aimed to evaluate the effect of cytokinins and its interaction with explant types and genotypes on cacao somatic embryogenesis. The study was conducted at tissue culture laboratory of IAARD, Bogor from April until December 2012 and October 2014 until February 2016. Three types of cytokinins i.e. kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M), thidiazuron (0.01, 0.02, and 0.04 </em><em>μ</em><em>M) and benzylaminopurine (0.55, 1.11, and 2.22 </em><em>μ</em><em>M) in combination with 9 </em><em>μ</em><em>M 2,4-D were tested for their effectiveness in inducing somatic embryogenesis from petals and staminoid explants of Cimanggu 1 genotype. Furthermore, three levels of kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M</em><em>) also in combination with 9 </em><em>μ</em><em>M 2,4-D were evaluated for their influences on the somatic embryogenesis from petals and staminoid explants of three cacao genotypes i.e. Sulawesi 02, ICCRI 04 and Cimanggu 3. The result demonstrated that 2.32 </em><em>μ</em><em>M kinetin and staminoids explant were more effective to induce cacao somatic embryogenesis of Cimanggu 1 genotype (7%, 0.23 embryos/explant). Additionally, there were interaction effects between the level of kinetin with explant types and genotype on the percentage of explants forming embryo at 12 weeks after culture. The highest percentage of somatic embryo formation was shown by ICCRI 04 genotype with the use of petals explant and a kinetin level of 1.16 </em><em>μ</em><em>M (31.85%), but not significantly different from the level of kinetin 2.23 </em><em>μ</em><em>M (25.55%). The formation of primary somatic embryos of cacao is largely determined by the type and level of cytokinins, type of explant, and genotype.</em></p>


Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2620
Author(s):  
Dmitry Miroshnichenko ◽  
Anna Klementyeva ◽  
Sergey Dolgov

Triticum timopheevii Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, T. timopheevii exhibits recalcitrance and albinism in tissue cultures, making this species of little use for manipulation through genetic engineering and genome editing. This study tested various approaches to increasing in vitro somatic embryogenesis and plant regeneration, while reducing the portion of albinos in cultures derived from immature embryos (IEs) of T. timopheevii. They included (i) adjusting the balance between 2,4-D and daminozide in callus induction medium; (ii) cultivation using various darkness/illumination schedules; and (iii) inclusion of additional concentrations of copper ions in the tissue culture medium. We achieved a 2.5-fold increase in somatic embryogenesis (up to 80%) when 50 mg L−1 daminozide was included in the callus induction medium together with 3 mg L−1 2,4-D. It was found that the dark cultivation for 20–30 days was superior in terms of achieving maximum culture efficiency; moreover, switching to light in under 2 weeks from culture initiation significantly increased the number of albino plants, suppressed somatic embryogenesis, and decreased the regeneration of green plants. Media containing higher levels of copper ions did not have a positive effect on the regeneration of green plants; contrarily, the elevated concentrations caused albinism in plantlets. The results and relevant conclusions of the present study might be valuable for establishing an improved protocol for the regeneration of green plants in tissue cultures of T. timopheevii.


1970 ◽  
Vol 14 ◽  
pp. 31-38 ◽  
Author(s):  
M Rahman ◽  
M Asaduzzaman ◽  
N Nahar ◽  
MA Bari

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83-85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl-1NAA + 0.05 mgl-1BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA3, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl-1 Zeatin + 1.0 mgl-1 BAP. For root induction, MS + 3.0 mgl-1 IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments. Key words: Eggplant; cotyledon; midrib; callus induction; somatic embryo J. bio-sci. 14: 1-9, 2006


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

2013 ◽  
Vol 18 (2) ◽  
pp. 1-9
Author(s):  
MAZ Al Munsur ◽  
MS Haque ◽  
KM Nasiruddin ◽  
MJ Hasan

Experiments were carried out to study the callus induction and subsequent plantlet regeneration ability of leaf segments and root tips of bitter gourd. A combination of 1.0 mg L-1 2,4-D and 1.0 mg L-1 BAP produced the highest percentage of callus in leaf segments (78.75%) and 1.0 mg L-1 2,4-D combined with 0.5 mg L-1 BAP produced the highest callus frequency in root tips (72.5%). Using the combination of BAP at 2.0 mg L-1 and NAA at 0.3 mg L-1 improved the callus induction frequency (80.00%) in root tips. Root tips showed better response than leaf segments in callus induction. Leaf segments and root tips had equal percentage (65.0) of shoot regeneration upon culture of the calli on medium with 2.0 or 2.5 mg L-1 BAP with 0.2 mg L-1 IAA. The highest length (3.95 cm) of shoot from leaf segments was recorded with 2.5 mg L-1 BAP, 0.5 mg L-1 IBA and 0.2 mg L-1 GA3. The highest number of roots (6.75) and root length (2.45 cm) were found in 3.0 mg L-1 BAP with 0.1 mg L-1 NAA in root tips. The callus formation and regeneration ability of the explants have the potential for use in future biochemical studies and genetic improvement of bitter gourd.DOI: http://dx.doi.org/10.3329/pa.v18i2.17457 Progress. Agric. 18(2): 1 - 9, 2007


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