Improvement of a Genetic Transformation System and Preliminary Study on the Function of LpABCB21 and LpPILS7 Based on Somatic Embryogenesis in Lilium pumilum DC. Fisch

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.

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

Journal of the Bangladesh Agricultural University ◽

10.3329/jbau.v12i1.20747 ◽

2014 ◽

Vol 12 (1) ◽

pp. 1-6 ◽

Cited By ~ 2

Author(s):

MN Hassan ◽

MS Haque ◽

MM Hassan ◽

MS Haque

Keyword(s):

Tissue Culture ◽

Somatic Embryogenesis ◽

Somatic Embryo ◽

Callus Induction ◽

Genetic Improvement ◽

In Vitro Regeneration ◽

Root Tip ◽

Root Tips ◽

Embryo Induction ◽

Somatic Embryo Induction

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

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

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha4129295 ◽

2013 ◽

Vol 41 (2) ◽

pp. 385

Author(s):

Adrian Ioan TIMOFTE ◽

Simona VICAȘ

Keyword(s):

Somatic Embryogenesis ◽

Developmental Stage ◽

Somatic Embryo ◽

Culture Medium ◽

Culture Media ◽

Quercus Petraea ◽

Ex Situ ◽

Zygotic Embryos ◽

Embryo Induction ◽

Somatic Embryo Induction

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.

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EFFECT OF 2,4-D AND NAA ON SOMATIC EMBRYOGENESIS FROM APICAL PORTION OF GROUNDNUT (ARACHIS HYPOGAEA L.)

International journal of multidisciplinary advanced scientific research and innovation ◽

10.53633/ijmasri.2021.1.7.01 ◽

2021 ◽

Vol 1 (7) ◽

pp. 119-124

Author(s):

Muniappan V ◽

Manivel P ◽

Prabakaran V ◽

Palanivel S ◽

Parvathi S

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryo ◽

Arachis Hypogaea ◽

Somatic Embryos ◽

Mature Embryo ◽

Induction Medium ◽

Embryogenic Cultures ◽

Apical Portion ◽

Arachis Hypogaea L ◽

Somatic Embryo Induction

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

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