In Vitro Induction of Callus and Flowers in Immature Oil Palm Inflorescences

This work aimed to investigate the effects of different combinations of auxin (0, 225 and 450 µM Picloram) and Murashige and Skoog salts (50% and 100% MS salt concentrations, respectively named as ½ MS and MS) in mediate the induction of callus and flowers from immature inflorescences (developmental stages I-9, I-12 and I-15) of the oil palm BRS – Manicoré hybrid. In I-9 inflorescence explants, the rate of callus induction was 17% higher in ½ MS than in MS; while in both I-12 and I-15 inflorescence explants, callus induction was over than 95% regardless of the MS salt concentration. The higher rate of callus induction in I-9 inflorescences was mediated by 450 µM Picloram, while the callus induction rates for either I-12 or I-15 inflorescences were higher than 92% regardless of Picloram concentration. Floral induction was lower in I-9 inflorescence explants (45% floral induction rate) than in both I-12 and I-15 inflorescence explants (floral induction rate higher than 90%). Our results suggest that Picloram (450 µM) and MS salts concentrations (½ MS) led to the highest rates of calluses and flowers induction in younger inflorescence explants (I-9); however, the medium composition is indifferent on callus and flower induction in I-12 and I-15 inflorescences.

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In vitro propagation of six selected sugarcane mutant clones through leaf explants

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/883/1/012075 ◽

2021 ◽

Vol 883 (1) ◽

pp. 012075

Author(s):

R Purnamaningsih ◽

D Sukmadjaja ◽

S Suhesti ◽

S Rahayu

Keyword(s):

Callus Induction ◽

Casein Hydrolysate ◽

Leaf Explants ◽

Shoot Proliferation ◽

Medium Composition ◽

Culture Techniques ◽

High Productivity ◽

Media Formulation ◽

Number Of Shoots

Abstract Six mutant clones of sugarcane with high productivity have been produced through tissue culture techniques combined with mutations using gamma-ray irradiation and Ethyl Methane Sulfonate. The six mutant clones have been tested for stability in the field. They are proven to have high productivity and yields, so that they are very potential to be developed as superior varieties. To support the planting material sufficiency of these clones, an efficient propagation method was needed. Media formulations with different physical properties and composition of growth regulators were tested to obtain high seedling propagation rates. The media formulation for callus induction was Murashige dan Skoog (MS) + 3 mg/l 2,4-D + 3 g/l casein hydrolysate + 3% sucrose and for shoot regeneration was MS + 0,5 mg/l BA + 0,1 mg/l IBA + 100 mg/l PVP and 2% sucrose. Shoot proliferation was carried out on MS liquid (1, ½) + (0.3; 0.5 mg/l) BA + 0.1 mg/l IBA + 1 mg/l Kinetin + (0; 0.5 mg/l) GA3+ sucrose 2%. The results showed that callus induction, callus regeneration, and shoot proliferation of sugarcane mutant clones were influenced by the genotype and medium composition. The fastest callus induction was obtained from the MSP-4 clone (5.82 days), and the longest was MSB-7 (8.82 days). The largest callus diameter was obtained from MSB-6 clone on MS medium containing 1 mg/l BA, 100 mg/l PVP, and 2% sucrose. The highest number of shoots was obtained from the MSB-6 clone, while the least number of shoots conducted from the MSB-8 clone. The MSB-8 clones were more difficult to regenerate compared to the others. The best media formulation for shoot proliferation was ½ MS containing 0.5 mg/l BA, 1 mg/l Kinetin, and 0.1 mg/l IBA, while the best formulation for rooting was ½ MS.

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Indirect in vitro Regeneration of the Medicinal Plant, Aspilia africana, and Histological Assessment at Different Developmental Stages

Frontiers in Plant Science ◽

10.3389/fpls.2021.797721 ◽

2021 ◽

Vol 12 ◽

Author(s):

Denis Okello ◽

Sungyu Yang ◽

Richard Komakech ◽

Yuseong Chung ◽

Endang Rahmat ◽

...

Keyword(s):

Medicinal Plant ◽

Shoot Regeneration ◽

Callus Induction ◽

Developmental Stages ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Regeneration Capacity ◽

Root Explants ◽

Indirect Regeneration

The medicinal plant, Aspilia africana, has been traditionally used in several African countries to treat many diseases such as tuberculosis, cough, inflammation, malaria, osteoporosis, and diabetes. In this study, we developed a protocol for in vitro propagation of A. africana using indirect shoot organogenesis from leaf and root explants of in vitro-grown seedlings and assessed the tissues at different developmental stages. The highest callus induction (91.9 ± 2.96%) from leaf explants was in the Murashige and Skoog (MS) medium augmented with 1.0 mg/L 6-Benzylaminopurine (BAP) and 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while from root explants, the highest callus induction (92.6 ± 2.80%) was in the same plant tissue culture medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D. The best shoot regeneration capacity from leaf-derived calli (i.e., 80.0 ± 6.23% regeneration percentage and 12.0 ± 6.23 shoots per callus) was obtained in medium augmented with 1.0 mg/L BAP and 0.05 mg/L α-Naphthaleneacetic acid (NAA); the best regeneration capacity for root-derived calli (i.e., 86.7 ± 6.24% shoot regeneration percentage and 14.7 ± 1.11 shoots per callus) was obtained in the MS medium augmented with 1.0 mg/L BAP, 0.05 mg/L NAA, and 0.1 mg/L Thidiazuron (TDZ). Regenerated plantlets developed a robust root system in 1/2 MS medium augmented with 0.1 mg/L NAA and had a survival rate of 93.6% at acclimatization. The in vitro regenerated stem tissue was fully differentiated, while the young leaf tissue consisted of largely unorganized and poorly differentiated cells with large intercellular airspaces typical of in vitro leaf tissues. Our study established a protocol for the indirect regeneration of A. africana and offers a basis for its domestication, large-scale multiplication, and germplasm preservation. To the best of our knowledge, this is the first study to develop an indirect regeneration protocol for A. africana and conduct anatomical assessment through the different stages of development from callus to a fully developed plantlet.

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Study on the Establishment of a Rapid Propagation System for Succulent Plant Haworthia heidelbergensis

Journal of Biobased Materials and Bioenergy ◽

10.1166/jbmb.2020.2001 ◽

2020 ◽

Vol 14 (5) ◽

pp. 632-638

Author(s):

Zhongyong Cen ◽

Jiang Su ◽

Hongrun Tu ◽

Shiting Lin

Keyword(s):

In Vitro Culture ◽

Callus Induction ◽

Proliferation Rate ◽

Medium Component ◽

Succulent Plant ◽

Induction Rate ◽

Rapid Propagation ◽

Callus Differentiation ◽

Optimal Medium

This study took the inflorescence and leaves of the succulent plant Haworthia heidelbergensis as explants, and explored the effects of different mediums with different hormone ratios on the rapid propagation of Haworthia heidelbergensis. The results showed that the optimal medium component for inflorescence callus induction was MS (Murashige and Skoog)+2.0 mg/L 6-BA+1.0 mg/L 2,4-D+0.2 mg/L NAA, the callus induction rate was 90%; the optimal medium component for leaf callus induction was MS+0.5 mg/L 6-BA+2.0 mg/L KT+0.3 mg/L NAA; the optimal medium for callus differentiation was MS+1.0 mg/L 6-BA+2.0 mg/L KT+0.3 mg/L NAA, the clump bud differentiation rate was 50%; the optimal medium for clump bud proliferation was MS+0.5 mg/L 6-BA+1.0 mg/L KT+0.05 mg/L NAA, the proliferation rate was 600%; the best medium for rooting was 1/2 MS+ 0.2 mg/L NAA+0.3 g/L AC. In conclusion, this study selected the explants and culture mediums, established an aseptic propagation system and provided a reference for the in vitro culture and rapid propagation of Haworthia heidelbergensis.

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In vitro induction of oil palm (Elaeis guineensis Jacq.) shoot roots and their acclimatization in mycorrhiza-enriched media

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/759/1/012014 ◽

2021 ◽

Vol 759 (1) ◽

pp. 012014

Author(s):

Karyanti ◽

H Khairiyah ◽

T Sukarnih ◽

N F Hanifah ◽

Y Rudiyana ◽

...

Keyword(s):

Oil Palm ◽

Elaeis Guineensis ◽

Elaeis Guineensis Jacq ◽

In Vitro Induction

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In Vitro Indirect Plantlet Regeneration from Hypocotyl Segments and Cotyledonary Explant Derived Calli in Lady's Finger (Abelmoschus esculentus L. Monech)

Journal of Bio-Science ◽

10.3329/jbs.v16i0.3741 ◽

1970 ◽

Vol 16 ◽

pp. 49-57

Author(s):

MT Rahman ◽

MJ Hossain ◽

M Khalekuzzaman

Keyword(s):

Callus Induction ◽

In Vitro Regeneration ◽

Abelmoschus Esculentus ◽

Root Induction ◽

Ms Medium ◽

Embryogenic Calli ◽

Induction Rate ◽

Cotyledonary Explants ◽

Hypocotyl Segments

An experiment was conducted to develop an efficient protocol for in vitro regeneration of plantlets from hypocotyl segments and cotyledonary explants of Abelmoschus esculentus L. Monech. In this investigation the cultivar OK-285 of Abelmoschus esculentus L. Monech was used. At first somatic embryogenic calli were induced using MS medium supplemented with different concentrations of auxin and cytokinin singly or in combination. The cotyledonary explants showed the best callus induction rate (87.9%) in MS medium containing 1.5mgl-l 2.4-D + 0.1mgl-l BAP hormonal concentration while hypocotyl segments showed 82.6% callus induction rate in the same medium. For shoot formation, calli were subcultured on MS solid medium and the hypocotyl segments showed the best result (72.1%) in MS medium containing 2.0 mgl-l BAP + 0.1 mgl-l IAA and the mean number of shoots per callus was recorded 4.2. For root induction from shoots MS and ½MS media were used. The highest 63.5% microshoots initiated roots in ½ MS + 0.1 mgl-l IBA medium and the highest mean number of root was 4.8. Rooted shoots were acclimated and successfully established into soil under natural condition with 70% survival. Key words: Abelmoschus esculentus, hypocotyl segments and cotyledon, embryogenic calli, regeneration. DOI:10.3329/jbs.v16i0.3741 J. bio-sci. 16: 49-57, 2008

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INDUKSI KALUS CENGKEH DARI EKSPAN DAUN MENGGUNAKAN 2,4-D SECARA IN VITRO

J-PEN Borneo : Jurnal Ilmu Pertanian ◽

10.35334/jpen.v2i3.1533 ◽

2020 ◽

Vol 2 (2) ◽

Author(s):

Yulianti Rasud ◽

Zainuddin Basri ◽

Nirwan Sahiri

Keyword(s):

Growth Regulators ◽

Callus Induction ◽

Callus Formation ◽

Cell Mass ◽

Medium Composition ◽

Ms Medium ◽

Plant Parts ◽

Randomized Design ◽

Completely Randomized Design

ABSTRACT Callus induction is one method of tissue culture which is done by stimulating cell division continuously from certain plant parts such as leaves, roots, stems, and so on by using growth regulators to form cell mass. The cell mass (callus) will then regenerate through organogenesis or embryogenesis to become a new plant. One of the growth regulators used for callus induction is 2,4-D. The aims of this experiments was to evaluate the best concentration of 2,4-D for callus induction of clove leaves. The experiment used Completely Randomized Design with treatment tested was concentrations of 2,4-D, consisted of six levels, namely 0.5 ppm, 1.0 ppm, 1.5 ppm, 2.0 ppm, 2.5 ppm and 3.0 ppm. Results of this experiments indicated that the best medium composition for callus induction was MS medium supplemented with 0.5 ppm 2,4-D. In the medium composition, the fastest callus formation, namely 6.00 weeks after culture and the percentage of callus formation reached 100% with the color and texture of the resulting callus white and crumb. Keyword : Callus Induction, Clove, 2,4-DABSTRAK Induksi kalus merupakan salah satu metode kultur jaringan yang dilakukan dengan jalan memacu pembelahan sel secara terus menerus dari bagian tanaman tertentu seperti daun, akar, batang, dan sebagainya dengan menggunakan zat pengatur tumbuh hingga terbentuk massa sel. Massa sel (kalus) tersebut selanjutnya akan beregenerasi melalui organogenesis ataupun embriogenesis hingga menjadi tanaman baru. Salah satu zat pengatur tumbuh yang digunakan untuk induksi kalus adalah 2,4-D. Penelitian ini bertujuan menentukan konsentrasi 2,4-D yang lebih baik untuk induksi kalus daun cengkeh. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan tiga kali ulangan. Media dasar yang digunakan adalah media MS yang ditambahkan berbagai konsentrasi 2,4-D yaitu 0,50 ppm, 1,5 ppm, 2 ppm, 2,5 ppm, dan 3 ppm. Hasil penelitian menunjukkan bahwa komposisi media yang terbaik untuk induksi kalus daun cengkeh adalah media MS yang ditambahkan 0,5 ppm 2,4-D. Pada komposisi media tersebut diperoleh saat muncul kalus paling cepat, yaitu rata-rata 6,00 MST dengan persentase pembentukan kalus tertinggi mencapai 100% dengan warna dan tekstur kalus yang dihasilkan putih dan remah. Kata Kunci : Induksi Kalus, Cengkeh, 2,4-D.

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The explant developmental stage profoundly impacts small RNA-mediated regulation at the dedifferentiation step of maize somatic embryogenesis

Scientific Reports ◽

10.1038/s41598-019-50962-y ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 7

Author(s):

Vasti T. Juárez-González ◽

Brenda A. López-Ruiz ◽

Patricia Baldrich ◽

Eduardo Luján-Soto ◽

Blake C. Meyers ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Callus Induction ◽

Molecular Mechanisms ◽

High Throughput Sequencing ◽

Developmental Stages ◽

Immature Embryo ◽

Zygotic Embryos ◽

Embryogenic Potential

Abstract Maize somatic embryogenesis (SE) requires the induction of embryogenic callus and establishment of proliferation before plant regeneration. The molecular mechanisms underlying callus embryogenic potential are not well understood. Here we explored the role of small RNAs (sRNAs) and the accumulation of their target transcripts in maize SE at the dedifferentiation step using VS-535 zygotic embryos collected at distinct developmental stages and displaying contrasting in vitro embryogenic potential and morphology. MicroRNAs (miRNAs), trans-acting siRNAs (tasiRNAs), heterochromatic siRNAs (hc-siRNAs) populations and their RNA targets were analyzed by high-throughput sequencing. Abundances of specific miRNAs, tasiRNAs and targets were validated by qRT-PCR. Unique accumulation patterns were found for immature embryo at 15 Days After Pollination (DAP) and for the callus induction from this explant, as compared to 23 DAP and mature embryos. miR156, miR164, miR166, tasiARFs and the 24 nt hc-siRNAs displayed the most strikingly different patterns between explants and during dedifferentiation. According to their role in auxin responses and developmental cues, we conclude that sRNA-target regulation operating within the 15 DAP immature embryo explant provides key molecular hints as to why this stage is relevant for callus induction with successful proliferation and plant regeneration.

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Investigation of explants sterilization process, effect of light intensity and concentration of agar for callus induction of Kappaphycus alvarezii (Doty) doty (Rhodophyta) in vitro

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/14/3/9868 ◽

2016 ◽

Vol 14 (3) ◽

pp. 515-522

Author(s):

Vũ Thị Mơ ◽

CRK Reddy

Keyword(s):

Light Intensity ◽

Callus Induction ◽

Kappaphycus Alvarezii ◽

Agar Concentration ◽

Induction Rate ◽

Callus Production ◽

Sterilization Process ◽

Different Levels ◽

Effect Of Light

Objective of this study was to ascertain the optimum condition for callus induction of K. alvarezii (Doty) in vitro such as to determine the explants sterilization process, the effect of intensity of light and the concentration of agar. Fresh thalli treated with 0.5% - 1% detergent for 5 mins followed by 0.5% - 1% betadine for 2 – 3 mins and incubated with 0.5% - 1% broad spectrum antibiotic mixture in PES medium for 1 day produced 95 – 98% bacteria free healthy explants. Two independent experiments with light intensity and agar contentration of the environment were carried out at 5 different levels of 0, 5, 25, 50, 70 µmol photon.m2.s-1 and 9 agar concentrations of 0.5%, 0.75%; 1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.5%, 3.0%. The highest callus induction rate was (96 ± 3.5 – 98 ± 2.1%) at 5 - 25 µmol photon.m-2.s-1 and (87 ± 5.8% – 90 ± 5.0%) in 1% - 3% agar concentration after 2 weeks of explants. The highest callus living rate was 98% at the light intensity of 25 µmol photon.m-2.s-1 and (75 ± 5.7 – 84 ± 1.1%) in 0.75 – 1.5% agar concentration after 2 months of explants. The highest callus re-induction rate was 50 – 55% at the light intensity of 5 – 25 µmol photon.m-2.s-1 and 60 – 65% in 1 – 1.5% agar concentration. Callus was not observed in dark condition (0 µmol photon.m-2.s-1). These calluses, that were strong, big and had filamentous type, will be a good material for the next production stage of embryonic callus production and seedling regeneration from micropropagules.

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In vitro induction of callus from cotyledon and hypocotyl explants of Glycine wightii (Wight & Arn.) Verdc.

Ciência e Agrotecnologia ◽

10.1590/s1413-70542003000600011 ◽

2003 ◽

Vol 27 (6) ◽

pp. 1277-1284 ◽

Cited By ~ 2

Author(s):

André Luis Coelho da Silva ◽

Cecília Sulzbacher Caruso ◽

Renato de Azevedo Moreira ◽

Ana Cecília Góes Horta

Keyword(s):

Callus Induction ◽

Basal Medium ◽

Average Weight ◽

Dichlorophenoxyacetic Acid ◽

Fresh Matter ◽

Growth Room ◽

High Concentrations ◽

2,4 Dichlorophenoxyacetic Acid ◽

In Vitro Induction

With the objective to promote in vitro callus induction, cotyledon and hypocotyl segments of "perennial soybean" (Glycine wightii (Wight & Arn.) Verdc.) were inoculated in basal medium MS supplemented with sucrose (1.5 e 3%) and 0.8% agar and different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-furfurylaminopurine (kinetin). The explants were maintained in a dark growth room at 28ºC. The best callus induction was observed in explants (cotyledon and hypocotyl) maintained in medium containing the combination of 2,4-D (1 mg.L-1), kinetin (0.1 mg.L-1) and 3% sucrose. To promote callus subculture, the MS medium was supplemented with different combinations of 2,4-D (0.5 to 4.0 mg.L-1), with or without kinetin (0.1 mg.L-1) and sucrose (1.5 e 3%). The calli were maintained 35 days in a dark growth room at 28ºC. The results indicated that the use of 2,4-D 1.0 mg.L-1 + kinetin 0.1 mg.L-1 + sucrose 3% provided the highest average weight of cotyledons calli fresh matter, whereas the use of 2,4-D 2.0 mg.L-1 + kinetin 0.1 mg.L-1 + sucrose 3% provided the highest average weight of hypocotyl calli fresh matter. High concentrations of 2,4-D, independent of kinetin and sucrose concentrations, promoted oxidation and reduction in fresh weight from calli of cotyledon and hypocotyls.

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In vitro host-free seed culture, callus development and organogenesis of an obligatory root-parasite Striga hermonthica (Del.) Benth: the witch-weed and medicinal plant

International Journal of Plant Biology ◽

10.4081/pb.2011.e9 ◽

2011 ◽

Vol 2 (1) ◽

pp. 9 ◽

Cited By ~ 1

Author(s):

Faisal Hammad Mekky Koua ◽

Fatima Misbah Abbas ◽

Eisa Ibrahim Elgaali ◽

Mutasim Mohammed Khalafallah ◽

Hind Ahmed Ali Babiker

Keyword(s):

Acetic Acid ◽

Callus Induction ◽

Sucrose Concentration ◽

Folk Medicine ◽

Striga Hermonthica ◽

Seed Culture ◽

Induction Rate ◽

Root Parasite ◽

The Witch

Striga hermonthica (Del.) Benth a well-known hemi-parasitic weed, it also has been used widely in African folk medicine to remedy broad spectra of diseases. The current contribution is an attempt to establish reproducible in vitro callusing system. In vitro seedling’s stem segments were used as an explant for callus induction, in 1.5% or 3.0% sucrose added into Murashig and Skoog medium (MS) and supplemented with different auxins, α-Naphthalene-3-acetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), Indole-3-acetic acid (IAA), or Indole-3-butryic acid (IBA) at different concentrations each alone or in combination with cytokinin 0.5 mgl-1 6-benzyl aminopurine. The most effective auxin was NAA with maximum 79% callus induction rate. All auxin treatments induced callus in all concentration when used alone or combined with BAP, except 2,4-D, which induced the callus only when combined with BAP. A high sucrose concentration was required for high callus induction rate by all auxin treatments. IAA and IBA auxins induced direct shoot regeneration and had low callus induction rates. NAA, IAA and IBA induced organogenic calli, whereas 2,4-D in combination with BAP induced non-organogenic callus. We further screened preliminarily the phytochemical contents of the callus and intact plant, which was revealed the presence of flavonoids, terpenes, saponins, cardiac glycosides, alkaloids, tannins and coumarins. Experimental data of both seed culture and callus induction could provide a route to further enhance the efficiency of callus initiation of S. hermonthica for medicinal purposes and understanding the infection mechanism of the witch-weed plant.

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