An efficient in vitro propagation protocol for direct organogenesis from root explants of a multi-purpose plant, Broussonetia papyrifera (L.) L’Hér. ex Vent.

2021 ◽  
Vol 170 ◽  
pp. 113686
Author(s):  
Jiana Lin ◽  
Jintuo Zou ◽  
Bingnan Zhang ◽  
Qingmin Que ◽  
Junjie Zhang ◽  
...  
Biologia ◽  
2014 ◽  
Vol 69 (5) ◽  
Author(s):  
Ziba Bakhtiar ◽  
Mohammad Mirjalili ◽  
Ali Sonboli ◽  
Mahdi Farimani ◽  
Mahdi Ayyari

AbstractThymus persicus (Ronniger ex Rech. f.) Jalas is a valuable and endangered natural source of antitumor pentacyclic triterpenoids, i.e., betulinic acid, oleanolic acid and ursolic acid, which grows in northwest Iran. As the plant has a low propagation rate in nature, a suitable method for in vitro-propagation is needed. With the aim of identifying a suitable system for regenerating T. persicus via direct organogenesis, Murashige & Skoog (MS) medium supplemented with different plant growth regulators (PGRs) was tested. In vitro-grown shoot tips were exposed to the cytokinins 6-benzylaminopurine (BAP), kinetin (KN), and thidiazuron (TDZ), alone or in combination with the auxins 1-naphthalene-acetic acid (NAA), 2,4-dichlorophenoxyacetic (2,4-D), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). The highest shoot formation (7.1 ± 0.9) was obtained with a medium fortified with 8.9 μM BAP plus 2.7 μM NAA. Regenerated shoots were easily rooted on the different tested media, with the most abundant (16.6 ± 1.4) and strongest roots obtained on half-strength MS medium containing 2.5 μM IBA. The rooted plantlets were successfully acclimatized (76.6%) in a greenhouse before transference to natural conditions. Homogeneity and phytochemical productivity of the in vitro regenerated plantlets were confirmed by random amplified polymorphic DNA (RAPD) profiles and high performance liquid chromatography (HPLC), respectively.


2007 ◽  
Vol 8 (1) ◽  
pp. 22 ◽  
Author(s):  
Fabiola Ocampo ◽  
Víctor Manuel Núñez

<p>Se indujeron múltiples brotes mediante organogénesis directa a partir de segmentos nodales de 10 genotipos diferentes de guayaba. Para ello se estableció un sistema de propagación clonal <em>in vitro </em>combinado con inducción rápida de brotes <em>ex vitro </em>para propagar árboles élite. La utilización de segmentos nodales permitió obtener en poco tiempo brotes adventicios adecuados para multiplicación masiva. La respuesta <em>in vitro </em>de los genotipos fue evaluada usando los medios de cultivo MS (Murashige y Skoog, 1962), Mc (Mascarenhas) y WPM (<em>Woody Plant Medium</em>) suplementados con 0,1 mg•L-1 de ácido indolácetico (AIA) y 0,25 mg•L<sup>-1</sup> de bencilaminopurina (BAP). El procedimiento de desinfección con hipoclorito de sodio previno eficientemente la contaminación de los explantes después de la inoculación en el medio de cultivo. El mayor porcentaje en la inducción de brotes se logró con 0,25 mg•L<sup>-1</sup> de BAP. Los segmentos nodales presentaron de 1 a 2 brotes adventicios por explante después de 15 días de inoculados y de 3 a 7 brotes a los 30 días después del inicio del cultivo. Una vez individualizados los brotes se usaron en una nueva fase de multiplicación masiva en la que se probaron cuatro sustratos diferentes durante el enraizamiento y el endurecimiento. Esta metodología permitió la propagación <em>in vitro </em>de guayaba cuatro semanas después del inicio del cultivo. Los mejores resultados se lograron con el medio WPM que permitió obtener las primeras plántulas enraizadas dos semanas después de la transferencia al sustrato de enraizamiento.</p><p> </p><p><strong>In vitro propagation of Psidium guajaba using direct organogenesis from nodal segments</strong></p><p>Multiple shoots were induced using direct organogenesis from nodal segments of 10 different genotypes of guayaba. For this an in vitro clonal propagation system combined with rapid ex vitro induction of shoots was established in order to propagate elite trees. The use of nodal segments resulted in adventitious shoots adequate for mass multiplication in less time. The in vitro response of the genotypes was evaluated using the culture media MS (Murashige and Skoog, 1962), Mc (Mascarenhas) and WPM (Woody Plant Medium) supplemented with 0.1 mg·L-1 of indole acetic acid (IAA) and 0.25 mg·L-1 of benzoaminopurine (BAP). The procedure for sterilizing with sodium hypochlorite effectively prevented the contamination of the explants after the inoculation of the culture medium. The greatest percentage of shoot induction was achieved with 0.25 mg·L-1 of BAP. The nodal segments showed between 1-2 adventitious shoots per explant 15 days post-inoculation and 3-7 shoots 30 days post-inoculation. Once individualized, the shoots were used in a new mass multiplication phase in which four different substrates were tested during rooting and hardening. This methodology permitted the in vitro propagation of guayaba four weeks post-inoculation. The best results were achieved with the WPM medium that resulted in the first rooted plantlets two weeks after the transfer to the rooting substrate.</p>


Agronomy ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 2164
Author(s):  
Anamica Upadhyay ◽  
Anwar Shahzad ◽  
Zishan Ahmad ◽  
Abdulrahman A. Alatar ◽  
Gea Guerriero ◽  
...  

Diplocyclos palmatus (L.) C. Jeffrey, commonly referred to as “Shivalingi” or “Lollipop climber” is a valuable medicinal plant with a climbing growth habit used in traditional medicine. It is reputed to have antiarthritic, anti-diabetic properties and to be useful in various skin and reproductive problems. Overexploitation of wild plants and low seed germination have resulted in the decline of the species in the wild. Thus, the present investigation was aimed to establish an effective in vitro propagation procedure for its large-scale production and conservation. Nodal explants, obtained from an established mother plant were grown on MS basal medium augmented with various cytokinins, alone or in combination with auxins, to study the morphogenic response. A maximum of 8.3 shoots/explants with an average shoot length of 7.2 cm were produced after six weeks on MS containing benzylaminopurine 5.0 µM + 1-naphthaleneacetic acid 2.0 µM. After 4 weeks of transfer, microshoots rooted well on a low nutrient medium of ½ MS + 1.0 µM indole-3-butyric acid, with a maximum of 11.0 roots/microshoot and an average root length of 7.4 cm. With an 80% survival rate, the regenerated plantlets were effectively acclimatized to natural conditions. DNA-based molecular markers were used to investigate the genetic uniformity. Scanning Electron Microscopic examination of leaves indicated the adaptation of the plantlets to natural, as evidenced by the formation of normal stomata. Gas chromatography-mass spectrometry analyses of mother and micropropagated plants were performed to identify essential secondary metabolites. The results obtained show that the in vitro propagation system can be adopted for preservation, large-scale production and secondary metabolites’ production in D. palmatus.


2019 ◽  
Vol 49 ◽  
Author(s):  
Lilian Marcia Santana Mascarenhas ◽  
José Raniere Ferreira de Santana ◽  
Alone Lima Brito

ABSTRACT Physalis peruviana L. (Solanaceae) is an herbaceous fruit-bearing species that has been gaining market acceptance due to its nutritional and medicinal potential. The main limitations to its cultivation are the short reproductive cycle, the susceptibility of the fruits to pests and the lack of information about the crop management. Hence, studies are necessary to develop strategies for its propagation. This study aimed to evaluate the effects of 6-benzylaminopurine (BAP) and explants on the morphogenetic potential of P. peruviana, as well as to establish a protocol for the micropropagation of the species via direct organogenesis. To evaluate the morphogenesis, cotyledonary node, cotyledon, leaf, epicotyl, hypocotyl and root explants were inoculated in Murashige & Skoog culture medium with half the normal concentration of salts and supplemented with cytokinin BAP (0.00 µM, 2.22 µM, 4.44 µM, 6.66 µM or 8.88 µM), plus 30 g L-1 of sucrose and 7 g L-1 of agar. Aiming at a direct production of shoots, the cotyledonary node explant was submitted to 0.00 µM, 2.22 µM, 4.44 µM, 6.66 µM, 8.88 µM, 13.32 µM, 17.76 µM or 22.20 µM of BAP. The obtained shoots were tested regarding their rooting potential in media with and without the addition of activated charcoal and then were transferred for acclimatation. The cotyledonary node and leaf explants were the most efficient sources for the regeneration of shoots via direct and indirect organogenesis, respectively. The most significant results for direct shoot production were obtained with 12.50 µM of BAP. These shoots were successfully rooted in vitro in medium without activated charcoal, and the microplants acclimated in vegetable earth attained 100 % of survival after 90 days of acclimatation.


2019 ◽  
Vol 48 (3) ◽  
pp. 575-581
Author(s):  
Li-Juan Zou ◽  
Jin-Yao Hu ◽  
Ming-Hua Luo ◽  
Qing-Gui Wu

Three explants such as stem tips, leaves and petioles of Heracleum scabridum were compared for their shoot development/differentiation ability by using different plant growth regulators (PGRs). The most effective PGRs combination for callus induction and organogenesis was determined. TDZ, Kn, Zn and IAA were used to induce multiple shoots. For indirect organogenesis (from the calli), the best response (27.25 ± 4.19 shoot per stem tips) and (18.23 ± 2.12 per leaves) were obtained in Murashige and Skoog (MS) medium fortified with 0.5 mg/l IAA and 3.0 mg/l Zn with 100, 97.3% induction rate, respectively. MS medium containing 0.5 mg/l IAA and 3.0 mg/l Zn was also found to be optimal for shoot regeneration from petioles. The highest percentage of regeneration (94.6) with mean number of shoots 17.83 ± 4.87 from petioles were produced. For direct organogenesis (from axillary bud shoot clumps), 0.1 mg/l IAA and 1.5 mg/l TDZ were found to be optimal for shoot regeneration of stem tips, with mean numbers of axillary bud shoot clumps 7.12 ± 1.23 were produced. Plantlets were transferred to soil with 95% of plants acclimatized recovered successfully.  


2014 ◽  
Vol 24 (1) ◽  
pp. 65-76
Author(s):  
Sayantani Nath ◽  
Anusree Das

Solanum pseudocapsicum L. is an unexplored plant of Solanaceae having horticultural and medicinal importance. Earlier the in vitro morphogenic potential of this plant species has not been explored. Induction of direct organogenesis was obtained from internodes, leaf bases and root explants from in vitro grown seedlings and plants  in MS supplemented with different concentrations and combinations of BAP (0.44 - 8.88 µM/l) and IAA (0.29 - 2.85 µM/l). Direct organogenesis was achieved in 90% of internodes, 65% of leaf bases and 75% of root explants. The optimum regeneration medium was MS supplemented with 4.44 µM/l BAP and 1.43 µM/l IAA. The regeneration rate observed six weeks after culture was approximately 20, 9 and 12 shoot buds/explants from internodes, leaf bases and root explants, respectively. cDNA RAPD analysis from organogenic stages was carried out and revealed distinct gene expression pattern. In vitro rooting in 100% shoots was achieved on MS within ten days of culture. Successfully hardened rooted plants (R0) on transfer  to the field  produced flowers, fruits and viable seeds (R1). Genetic characterization of R0 and R1 plants revealed stability and uniformity at the chromosomal and genomic DNA level. Plant Tissue Cult. & Biotech. 24(1): 65-76, 2014 (June)  D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19201


Planta Medica ◽  
2007 ◽  
Vol 73 (09) ◽  
Author(s):  
M Gangopadhyay ◽  
R Bhattacharya ◽  
D Chakraborty ◽  
S Bhattacharya ◽  
A Mitra ◽  
...  

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