Callus formation and organogenesis by explants of six Lycopersicon species

1983 ◽  
Vol 61 (4) ◽  
pp. 1072-1079 ◽  
Author(s):  
R. D. Locy

Stem and hypocotyl explants of six Lycopersicon species, L. esculentum var. cerasiformae, L. cheesmanii, L. hirsutum, L. pimpinellifolium, L. peruvianum, and L. glandulosum, were examined for ability to form callus, adventitious shoots, and adventitious roots on media containing varying levels of indole acetic acid (IAA) and kinetin. Callus growth of L. peruvianum and L. glandulosum explants was better than that of the other species tested. Stem explants tended to form more callus than hypocotyl explants. Shoot formation by hypocotyl explants was superior to stem explants for all species and at nearly all hormone levels. Hypocotyl explants of L. peruvianum and L. glandulosum showed the highest levels of shoot formation, and L. cheesmanii and L. esculentum var. cerasiformae hypocotyl explants showed good shoot formation on media containing 3 mg/L kinetin and 0.3 or 1 mg/L indole acetic acid. No shoot formation was observed by L. hirsutum explants, and L. pimpinellifolium explants gave only a poor response. Shoot formation by stem explants was obtained only with L.. esculentum var. cerasiformae and L. glandulosum. When callus of the six species, subcultured for 63 days on a medium optimal for callus growth, was subcultured on shoot-forming media, only callus of L. peruvianum and L. glandulosum subsequently formed shoots. Callus of L. glandulosum has retained its morphogenic potential for 1 year in culture and can be used to initiate liquid suspension cultures.

2021 ◽  
Vol 5 (1) ◽  
pp. 9-20
Author(s):  
Khairan Khairan ◽  
Betty Mauliya Bustam ◽  
Yunita ◽  
Riska Meilinda ◽  
Raudhatul Muna

This study aims to determine the effect of 2,4-Diclorophenoxy Acetic Acid (2,4-D) and Benzyl Amino Purin (BAP) on the formation of callus of patchouli (Pogostemon cablin Benth.) shoot explants by horizontal and vertical cutting methods. The parameters that observed in this study were the percentage growth of callus, time appearance of callus, weight of callus and the morphology of callus. The results showed that horizontal cutting method was able to induce callus growth with the percentages growth of callus were 18,75%, with the time appearance of callus was at 16 days at P1; P10; P12; P13 dan P14. The highest weight of callus obtained was 0.19 grams at P8. The results also showed that the callus yielded had a yellow and cream color, with a compact and crumb textures. Meanwhile, the vertical cutting method was able to induce callus formation with the percentage growth of callus were 12,5%. The fastest time of callus appearance was obtained in P6 and P8, which was 12 day after planting with the highest weight of callus obtained was 0.05 grams at P12.   The results also showed that vertical cutting method had brown and dark-brown of callus with a compact and crumb textures.


HortScience ◽  
2011 ◽  
Vol 46 (3) ◽  
pp. 466-469
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Jianjun Chen ◽  
Richard J. Henny

Chlorophytum amaniense Engl. ‘Fire Flash’ is a popular exotic ornamental foliage plant as a result of its unique coral-colored midribs and petioles and tolerance to interior low light levels. Currently, demand for propagative materials exceeds the availability of seeds. This study was intended to develop an in vitro culture method for rapid propagation of this cultivar. Leaf and sprouted seed explants were cultured on a Murashige and Skoog basal medium supplemented with different cytokinins with 1.1 μM α-naphthalene acetic acid (NAA) or 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Leaf explants showed poor responses in callus production and no adventitious shoots were obtained. Callus formation frequencies from sprouted seeds were 71% and 85% when induced by 9.8 μM N6-(2-isopentyl) adenine (2iP) with 1.1 μM NAA and 9.1 μM N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with 1.1 μM NAA, respectively. Adventitious shoots occurred after the induced calluses were subcultured on the same concentrations of TDZ or 2iP with NAA. Shoot formation frequencies from calluses cultured on TDZ with NAA and 2iP with NAA were 92% and 85%, and the corresponding mean shoot numbers were 37 and 31 per piece of callus (1 cm3), respectively. Adventitious shoots rooted at 100% after transferring to the basal medium containing 4.4 μM 6-benzylaminopurine (BA) with 2.7 μM NAA. Plantlets, after transplanting to a soilless substrate were easily acclimatized in a shaded greenhouse under a photosynthetic photon flux (PPF) density of 200 μmol·m−2·s−1. Regenerated plants grew vigorously without undesirable basal branching or distorted leaves. This newly established regeneration method can provide the foliage plant industry with a means for rapidly propagating ‘Fire Flash’ liners in a year-round fashion.


HortScience ◽  
2010 ◽  
Vol 45 (8) ◽  
pp. 1250-1254 ◽  
Author(s):  
Juanxu Liu ◽  
Min Deng ◽  
Richard J. Henny ◽  
Jianjun Chen ◽  
Jiahua Xie

This study established a method of regenerating Dracaena surculosa Lindl. ‘Florida Beauty’ through indirect shoot organogenesis. Bud, leaf, and stem explants were cultured on a Murashige and Skoog basal medium supplemented with N6-(2-isopentyl) adenine (2iP) at 12.3 and 24.6 μM with 3-indoleacetic acid (IAA) at 0, 1.1, and 2.3 μM, respectively, and 2iP at 36.9, 49.2, 61.5, and 73.8 μM with IAA at 1.1 and 2.3 μM, respectively. Calluses were induced from leaf explants but failed to produce adventitious shoots. Calluses were also induced from stem and bud explants cultured on the basal medium containing 12.3 μM 2iP and 2.3 μM IAA, 24.6 μM 2iP or higher with either 1.1 or 2.3 μM IAA. The highest callus induction frequency was 63.2% from stem explants and 69.6% from bud explants when they were cultured on the basal medium supplemented with 49.2 μM 2iP and 2.3 μM IAA. The highest shoot formation frequency was 65.7% from stem-derived callus cultured on the basal medium containing 61.5 μM 2iP and 1.1 μM IAA and 88% from bud-derived callus cultured with 49.2 μM 2iP and 1.1 μM IAA. The highest number of shoots per piece of stem- and bud-derived calluses was 3.8 and 6.7, respectively. Adventitious shoots developed better root systems in the basal medium supplemented with 2.0 μM IAA. Plantlets after transplantation into a soilless substrate grew vigorously in a shaded greenhouse under a maximum photosynthetic photon flux density of 300 μmol·m−2·s−1. Neither disease incidence nor somaclonal variants were observed in the regenerated population. This established method could be used for efficient micropropagation of D. surculosa, and the availability of tissue-cultured liners could reduce the dependency on imported cuttings, which often bring new or invasive pests into the United States.


Genetika ◽  
2021 ◽  
Vol 53 (1) ◽  
pp. 295-303
Author(s):  
Hüseyin Uysal

This study was carried out to determine in vitro development using Black cumin leaf and stem explants. ?ameli black cumin variety was used as plant material. Five different nutrient mediums (1. LS2.5, 2. MS, 3. MS + 0.5 mg.l-1 IAA, 4. MS + 0.5 mg.l-1 BAP, 5. MS + 0.5 mg.l-1 IAA + 0.5 mg.l-1 BAP) containing 30 g sugar were used in this study. As a result of the research, 100% callus formation was detected in the stem explants cultured in the number 1 and number 5 mediums. These were followed by stem explants cultured in medium 4 with a success rate of 96%. Of this rate, 66% was shoot formation, and 30% was callus formation. Direct shoot regeneration was performed only on stem explants cultured in mediums 4 and 3, with a 66% success rate in medium four and a 36% success rate in medium 3. The highest plant regenerations from calluses were gained from stem explants (273.3%) in medium 4, followed by calluses gained from leaf explants (262.5%) in the same medium. These were followed by cultures in medium 3, with calluses derived from stem explants (255%) and leaf explants (150%). No plant regeneration was determined from calluses gained in the medium 1. Thus it is evident that high auxin content and auxin-cytokinin balanced mediums encouraged callus formation in the black cumin plants. The addition of only IAA or BAP to the medium promoted shoot formation in the stem explants, but direct shoot regeneration was not thereby achieved from the leaf explants. These results show that, for in vitro clonal propagation studies done on black cumin plants, a high auxin containing medium is preferable if the aim is callus formation. If the aim is direct shoot regeneration, BAP or other cytokinin-containing medium is preferred.


1999 ◽  
Vol 77 (2) ◽  
pp. 318-322
Author(s):  
Jacintha Miranda ◽  
Michele N Konschuh ◽  
E C Yeung ◽  
C C Chinnappa

An in vitro regeneration protocol for Stellaria longipes Goldie was developed using young hypocotyl explants. Optimal regeneration was obtained using Murashige and Skoog (MS) basal medium supplemented with 0.5 µM N6-benzyladenine and 1 µM indole-3-butyric acid. Three different patterns of shoot regeneration were observed: (i) "direct shoot" formation within 3-5 days of inoculation, (ii) nodular structures appeared followed by shoot formation, and (iii) callus formation followed by the appearance of shoots. Histological observation revealed that cells within the central vascular cylinder of the hypocotyl were responsible for shoot organogenesis. Shoot production was not synchronous or uniform among explants. A more synchronous shoot production was obtained by excising the direct shoots or by wounding the nodular structures. Excision and wounding increased the regeneration capability of the explants. Regenerated shoots were readily rooted in MS medium lacking growth regulators and were successfully transferred to greenhouse conditions. These showed morphology consistence with greenhouse-grown plants.Key words: hypocotyl, organogenesis, regeneration, Stellaria longipes.


2014 ◽  
Vol 49 (1-2) ◽  
pp. 5-20 ◽  
Author(s):  
Janina H. Rogozińska ◽  
Lucyna Drozdowska

Cotyledon explants of rape were excised from aseptically germinated seedlings and cultured during 2 weeks on M u r a s h i g e and S k o o g medium supplemented with auxins, cytokinins, auxin-cytokinin combinations and abscisic acid. Callus formation occurred on medium with 2,4-dichlorophenoxyacetic acid (2,4-D), naphthalene-l-acetic acid (NAA), indole-3-acetic acid (IAA) and on their combinations with kinetin (K) or 6-benzylaminopurine (BAP). Regeneration of roots was achieved on media with NAA, IAA and indole-3-butyric acid (IBA) and on combinations of these auxins with cytokinins. The presence of 2,4-D in the medium, though it promoted compact callus growth, had an inhibitory effect on root formation. Callus derived from the cotyledons had somewhat different requirements for growth in subculture and the root formation ability diminished in the course of the culture. Lower ABA concentrations stimulated callus growth whereas higher concentrations inhibited it similary as in the case of cotyledons. Shoot buds regenerated from the cotyledons after ca. 3 weeks on media supplemented with NAA + BAP. The 9-week-old plantlets transferred to the soil developed into complete plants. The plants which underwent vernalization formed flowers and normal seeds.


HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 477B-477
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart

One-centimeter hypocotyl explants from 2-week-old Antirrhinum majus L. (snapdragon) seedlings germinated and grown in vitro under 12-h cool-white fluorescent light and 12 h dark or 24 h dark were placed on Murashige and Skoog (MS) medium containing 0, 0.44, 2.22, 4.44, 8.88, or 44.4 μM N6-benzyladenine (BA). Cultures were maintained under the light/dark regime at 25°C. After 2 weeks, adventitious shoots were counted. A shoot was considered adventitious and counted if a stem and leaf developed. Shoots developed along the entire length of the hypocotyl sections. Mean shoot production per hypocotyl explant ranged from 2.4 to 6.1 shoots when seedlings were germinated and grown in 24 h darkness and 2.2 to 10.9 shoots when started in the light/dark regime. Highest shoot counts were attained /from hypocotyl explants when seedlings were germinated and grown under the light/dark regime for 2 weeks and transferred to 2.22, 4.44, or 8.88 μM BA. Shoot development appeared normal at the 2.22 and 4.44 μM level, while at 8.88 μM BA, development was slightly abnormal along with slightly more callus production.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 478e-479
Author(s):  
M.M. Jenderek ◽  
A.J. Olney

Hibiscus syriacus is a difficult species in micropropagation due to its endogenous contamination and recalcitrant shoot formation; therefore, studies on using explants other than shoot tip or axillary buds of growing shrubs were initiated. Three different seedling fragments (root, hypocotyl, and leaf petiole) from aseptically germinated seedlings of hibiscus (var. Aphrodite) were evaluated for adventitious bud formation, shoot and leaf development. The explants were cultured on McCown's woody plant basal salt medium supplemented with KNO3 (800 mg/L), adenine sulfate (80 mg/L) and MS vitamins containing BA or 2iP or TDZ at 0.5, 1.0, 2.2, 4.4 and 10 mM. Adventitious buds were present on all of the three different explants grown on medium containing TDZ; however, the most abundant bud formation, with many small leaves originating from callus was observed on hypocotyl explants cultured on medium with 1 mM of TDZ. Petiole explants were the most frequent to develop short shoots (≈15 mm) and one to nine leaves without callus formation, where 70% of hypocotyl and the root explants formed leaves originating from callus. Callus was induced on all explant types regardless of the level or type of cytokinin used. However, the number of shoots produced by any explant type was low, petioles cultured on 0.5 and 1mM of TDZ were the most suitable material for non-callus shoot development in H. syriacus. Hypocotyl explants proved to be an excellent source for adventitious bud formation but their ability to develop shoots needs to be investigated.


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