Genetic Fidelity Testing Using SSR Marker Assay Confirms Trueness to Type of Micropropagated Coconut (Cocos nucifera L.) Plantlets Derived from Unfertilized Ovaries

2017 ◽  
Vol 10 (1) ◽  
pp. 46-54 ◽  
Author(s):  
H.D.D. Bandupriya ◽  
W.W.M.A. Iroshini ◽  
S A C N Perera ◽  
V.R.M. Vidhanaarachchi ◽  
S.C. Fernando ◽  
...  

Background: In vitro culture techniques provide an excellent platform for the multiplication of recalcitrant species such as coconut and thereby increase the homogeneity of the plantations. Clonal fidelity is one of the most important pre-requisites in a micropropagation protocol of crop species especially those with long life spans. Objective: The present study was conducted in order to determine the genetic homogeneity of coconut plantlets derived from unfertilized ovaries through somatic embryogenesis. Method: Twenty randomly selected plantlets at acclimatization stage, from two mother palms were subjected to Simple Sequence Repeats analysis. Thirteen highly polymorphic microsatellite primers were used for the detection of genetic fidelity in the clonal plantlets and their respective parent. Results: These plantlets showed no apparent differences among themselves and were comparable with the respective mother palm in the Simple Sequence Repeats analysis. The results obtained from this study suggest that there is no somaclonal variation or genetic instability occurring in plantlets that are regenerated from ovary explants. Conclusion: The absence of any sign of somaclonal variation suggests that somatic embryogenesis protocol did not induce the changes in gene structure, which had remained stable throughout the period that had been maintained in vitro. Determination of genetic fidelity of in vitro plants proved the suitability of regeneration protocol for large scale micropropagation applications for coconut.

2012 ◽  
Vol 60 (5) ◽  
pp. 396 ◽  
Author(s):  
Andrea Kodym ◽  
Eva M. Temsch ◽  
Eric Bunn ◽  
John Delpratt

We report on the development of a somatic embryogenesis system for Lepidosperma concavum R.Br. and L. laterale R.Br. and the determination of ploidy stability of plants derived from somatic embryos. These keystone Lepidosperma species cannot currently be returned to restoration sites because of propagation difficulties (i.e. seed dormancy, low seed fill and recalcitrance to vegetative propagation). Three explant types (in vitro-germinated seedlings, immature seed and immature inflorescences) were used for the assessment of callus production potential. Embryogenic callus was induced and multiplied on 1/2MS medium with 2,4-D either alone, or in combination with zeatin. Over 90% of seedling explants of L. laterale produced regenerative calli after 6 weeks and 53% of seedling explants of L. concavum produced calli after 16 weeks on media containing 2,4-D and zeatin. Inflorescence material appeared to be least responsive. High rates of conversion to plants were achieved on medium containing activated charcoal, followed by thidiazuron medium. Acclimatisation success of plants ranged from 86% to 95%. Acclimatised plants grew vigorously under standard nursery conditions. The DNA ploidy level of 486 somatic embryogenesis-derived plantlets was analysed by flow cytometry. Only one plant (=0.2% of all plantlets tested) was found mixoploid. All other plants showed a stable ploidy level and stable C-values within the species. There was a small but significant C-value difference between the two Lepidosperma species. Five variegated plants (=0.3%) were observed among a total of ~1600 plants acclimatised. The application of tissue culture techniques such as somatic embryogenesis brings large-scale production of Lepidosperma plants for revegetation and horticultural purposes closer to commercial feasibility.


Plants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2050
Author(s):  
Sundaravelpandian Kalaipandian ◽  
Zhihua Mu ◽  
Eveline Yee Yan Kong ◽  
Julianne Biddle ◽  
Robyn Cave ◽  
...  

Coconut [Cocos nucifera L.] is often called “the tree of life” because of its many uses in the food, beverage, medicinal, and cosmetic industries. Currently, more than 50% of the palms grown throughout the world are senile and need to be replanted immediately to ensure production levels meet the present and increasing demand for coconut products. Mass replanting will not be possible using traditional propagation methods from seed. Recent studies have indicated that in vitro cloning via somatic embryogenesis is the most promising alternative for the large-scale production of new coconut palms. This paper provides a review on the status and prospects for the application of somatic embryogenesis to mass clonal propagation of coconut.


Rodriguésia ◽  
2021 ◽  
Vol 72 ◽  
Author(s):  
Claudia Simões-Gurgel ◽  
Tatiana Carvalho de Castro ◽  
Cátia Henriques Callado ◽  
Lívia da Silva Cordeiro ◽  
Norma Albarello

Abstract In vitro culture techniques are recognized as efficient strategies for large-scale plant production, as well as providing alternatives for plant conservation. In this study the micropropagation of Tarenaya rosea was established using petiole and foliar blade segments cultivated on MS medium with 6-benzyladenine (BA) and/or 6-furfurylaminopurine (KIN). The regeneration rate from explants was evaluated after 30-days in culture, as well as the proliferation rate from explant-derived shoots, reached after four subcultures performed at 30-days in culture. In vitro propagation occurred by both direct (DO) and indirect (IO) organogenesis. The highest regeneration rates by DO (50% to 100%) were reached on media containing only BA, while morphogenic calluses (IO) were mainly formed with BA+KIN. Explants on media with BA showed the presence of small black nodules on their surface, and histological analysis revealed the presence of trichomes with anthocyanin content. Elongation and rooting were reached on growth regulator-free MS. Acclimatization rates around 80% were achieved and the in vitro-regenerated plants were successfully maintained under field conditions. Results show significant morphogenetic potential of T. rosea from leaf explants, mainly when cultivated in the presence of 4.4 µM BA, providing a new alternative source of plant material for biotechnological and in vitro conservation studies.


2018 ◽  
Vol 41 (4) ◽  
Author(s):  
Marlúcia Souza Pádua ◽  
Raíssa Silveira Santos ◽  
Luciano Vilela Paiva ◽  
Vanessa Cristina Stein ◽  
Luciano Coutinho Silva

ABSTRACT Oil palm is a woody monocot of economic importance due to high oil production from its fruits. Currently, the conventional method most used to propagate oil palm is seed germination, but success is limited by long time requirements and low germination percentage. An alternative for large-scale propagation of oil palm is the biotechnological technique of somatic embryogenesis. The rooting of plants germinated from somatic embryos is a difficult step, yet it is of great importance for later acclimatization and success in propagation. The aim of this study was to evaluate the effect of the auxins indole acetic acid (IAA) and indole butyric acid (IBA) on the rooting of somatic embryos of Tenera hybrid oil palm. Plants obtained by somatic embryogenesis were inoculated in modified MS medium with 10% sucrose and 0.6% agar and supplemented with IAA or IBA at concentrations of 5 µM, 10 µM, and 15 µM, and the absence of growth regulators. After 120 days, the presence of roots, root type, length of the longest root, number of roots, number of leaves, and shoot length were analyzed. Growth regulators were favorable to rooting; plants cultivated with IBA growth regulator at 15 µM showed higher rooting percentage (87%) and better results for the parameters of number of roots (1.33) and shoot length (9.83).


2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Jingli Yang ◽  
Songquan Wu ◽  
Chenghao Li

Embryogenic callus was obtained from mature seed explants on medium supplemented with 2,4-dichlorophenoxyacetic acid. Primary somatic embryos (SEs) can only develop into abnormal plants. Well-developed SEs could be obtained through secondary somatic embryogenesis both in solid and liquid cultures. Temperature strongly affected induction frequency of secondary embryogenesis. Relatively high temperature (30∘C) and germinated SEs explants were effective for induction of secondary somatic embryos, and low temperature (20∘C) was more suitable for further embryo development, plantlet conversion, and transplant survival. Somatic embryos formed on agar medium had larger cotyledons than those of embryos formed in liquid medium. Supplementing 0.1 mg L−16-benzyladenine (BA) was effective for plant conversion; the rate of plant conversion was 43.3% in somatic embryos from solid culture and 36.5% in embryos from liquid culture.In vitroplants were successfully acclimatized in the greenhouse. The protocol established in this study will be helpful for large-scale vegetative propagation of this medicinal tree.


2017 ◽  
Vol 41 (4) ◽  
pp. 347-358 ◽  
Author(s):  
Jameel Mohammed Al-Khayri ◽  
Poornananda Madhava Naik

ABSTRACT Date palm (Phoenix dactylifera L.) is a fruit tree resilient to adverse climatic conditions predominating in hot arid regions of the Middle East and North Africa. The date fruit contains numerous chemical components that possess high nutritional and medicinal values. Traditional propagation by offshoots is inefficient to satisfy current demands for date palm trees. Alternatively, micropropagation provides an efficient means for large-scale propagation of date palm cultivars. Both somatic embryogenesis and organogenesis, either directly or indirectly though the callus phase, have been demonstrated in date palm in vitro regeneration. Culture initiation commonly utilizes shoot-tip explants isolated from young offshoots. Recently, the immature inflorescences of adult trees were utilized as an alternative nondestructive source of explants. In addition to the nature of the explant used, successful plant regeneration depends on the cultivar, composition of the culture medium and physical status. Challenges of date palm micropropagation include long in vitro cycle, latent contamination, browning, somaclonal variation as well as ex vitro acclimatization and transplanting. A remarkable amount of research investigating these factors has led to optimized protocols for the micropropagation of numerous commercially important cultivars. This has encouraged the development of several international commercial tissue culture laboratories. Molecular characterization provides an assurance of genetic conformity of regenerated plantlets, a key feature for commercial production. This article describes date palm micropropagation protocols and also discusses recent achievements with respect to somaclonal variation, molecular markers, cryopreservation and future prospects.


2005 ◽  
Vol 130 (6) ◽  
pp. 912-917 ◽  
Author(s):  
Jernej Jakse ◽  
William Martin ◽  
John McCallum ◽  
Michael J. Havey

The commercial production of onion (Allium cepa L.) inbreds, hybrids, and open-pollinated (OP) cultivars would benefit from a robust set of molecular markers that confidently distinguish among elite germplasms. Large-scale DNA sequencing has revealed that single nucleotide polymorphisms (SNPs), short insertion-deletion (indel) events, and simple sequence repeats (SSRs) are relatively abundant classes of codominant DNA markers. We identified 398 SNPs, indels, and SSRs among 35 elite onion ulations and observed that all populations could be distinguished. Phylogenetic analyses of simple-matching and Jaccard's coefficients for SSRs produced essentially identical trees and relationships were consistent with known pedigrees and previous marker evaluations. The SSRs revealed that elite germplasms from specific companies or breeding programs were often closely related. In contrast, phylogenetic analyses of SNPs and indels did not reveal clear relationships among elite onion populations and there was no agreement among trees generated using SNPs and indels vs. SSRs. This discrepancy was likely due to SNPs and indels occurring among amplicons from duplicated regions (paralogs) of the onion genome. Nevertheless, these PCR-based markers will be useful in the quality control of inbred, hybrid, and OP onion seed lots.


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