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

2016 ◽  
Vol 14 (3) ◽  
pp. 515-522
Author(s):  
Vũ Thị Mơ ◽  
CRK Reddy

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.

2014 ◽  
Vol 8 (2) ◽  
pp. 27-34
Author(s):  
Emad H. Jassim ◽  
Sami K. M. Ameen

An experiment on the effect of sucrose and mannitol on leave induced callus of Catharanthus roseus was conducted from February 2011 to May 2012. Callus induction was achieved by culturing leaves of the plant on MS medium supplemented with 0.5 mg /L 2,4-D and 1mg / L Kin, The best medium to maintain callus was on MS medium modified with 0.5 mg /L 2,4-D and 1.5 mg / L Kin. when different levels were added to MS medium for each Mannitol 0, 6000 ,8000 ,10000 mg /L and Sucrose 40, 60 ,80, 100 g /L in split experimental and control treatment was MS medium supplemented with 30 g/L sucrose. The results showed that medium supplemented with 100 gL of sucrose gave the highest quantity of Ajmalicine 32.27 µg/100 mg fresh weight of callus,as well as medium supplemented with 8000 mgL of Mannitol gave the highest value of Ajmalicine 120.19 µg/100 mg fresh weight of callus. The concentrations of Ajmalicine, derived from the leaves of the plants grown in pot, were lowest than the concentrations produced by the callus grown in vitro it was 0.047 µg/100 fresh weight of the leaves.


2020 ◽  
Vol 14 (5) ◽  
pp. 632-638
Author(s):  
Zhongyong Cen ◽  
Jiang Su ◽  
Hongrun Tu ◽  
Shiting Lin

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.


2021 ◽  
pp. 27-34
Author(s):  
O.V. Mitrofanova ◽  
N.N. Ivanova ◽  
N.P. Lesnikova-Sedoshenko ◽  
V.A. Brailko ◽  
I.V. Zhdanova ◽  
...  

2015 ◽  
Vol 10 (1) ◽  
pp. 13
Author(s):  
Emma Suryati ◽  
Rosmiati Rosmiati ◽  
Andi Parenrengi ◽  
Andi Tenriulo

The development of micropropagule and embryo of seaweed depend on nutrient and fertilizer used. Seaweed has been reported contain hormone regulators such as auxine, cytokinine, gibbereline, and various minerals applied in stimulating the growth ocra plant and wheat culture. The objectives of this study were to determine the potential of Kappaphycus alvarezii extract and its optimal concentration in accelerating of Kappaphycus alvarezii micropropagule and embryo growth. Micropropagule and embryo produced through callus induction were planted into PES 1/20 liquid medium supplemented with seaweed extract at the concentrations of 0 (control), 25, 50, 75, and 100 μL in 10 mL of medium. The results showed that medium enrichment with 50 μL of seaweed extract had the highest survival rate and growth of thallus. In addition, this concentration was also resulted in a good performance of K. alvarezii thallus with the lighter color. The advantage of this study for seaweed cultivation in Indonesia, among others, seaweed can be used as fertilizer, especially in the maintenance of seaweed seed, so that cultivation can be better develop.


2016 ◽  
Vol 5 (2) ◽  
pp. 211 ◽  
Author(s):  
Muh. Alias L. Rajamuddin ◽  
Andi Asdar Jaya ◽  
Ridwan Ridwan ◽  
Emma Suryati

Untuk mendukung program transgenesis pada rumput laut, embrio somatik dapat digunakan sebagai material untuk transfer gen baik secara individu sel ataupun kluster sel embriogenik, sehingga mempercepat keberhasilan dengan peluang transformasi yang lebih tinggi. Penelitian ini bertujuan untuk mengkaji induksi kalus rumput laut K. alvarezii untuk produksi sel embrio somatik (e.s.) dengan beberapa rasio zat pengatur tumbuh (ZPT) dan konsentrasi agar media induksi, sampai sel menjadi filamen. Penelitian terdiri atas dua tahap: Tahap (1) induksi kalus, dengan rasio ZPT asam indol asetat (IAA):kinetin = 0,5:0,0 mg/L; 1,0:1,0 mg/L; dan 2,0:0,2 mg/L dengan konsentrasi agar media induksi = 0,6%; 0,8%; 1,0%; dan 1,5%. Tahap (2) regenerasi massa sel e.s., dengan rasio IAA:kinetin = 0,1:1,0 mg/L; 0,0:0,1 mg/L dan tanpa ZPT dengan konsentrasi agar media = 0,4%; 0,6%; dan 0,8%. Untuk perkembangan sel-sel e.s. lebih lanjut dipelihara pada kultur cair. Hasil penelitian menunjukkan pada tahap induksi kalus, rasio IAA: kinetin = 1:1 mg/L dengan konsentrasi agar media 0,8% dan 1,0% menghasilkan persentase induksi kalus tertinggi (90%). Pada tahap regenerasi massa sel e.s., ZPT tidak berpengaruh terhadap perkembangan massa sel e.s., di mana tanpa ZPT dengan konsentrasi agar 0,6% memperlihatkan perkembangan tertinggi (rata-rata diameter massa sel 5 mm). Pada media cair, perkembangan sel e.s. dari single cell ukuran 3-4 mm menjadi filamen-filamen ukuran rata-rata 0,5 mm dapat dicapai dalam satu bulan kultur. Keberhasilan produksi sel e.s. K. alvarezii, selain sebagai material untuk transfer gen juga dapat dijadikan acuan dalam produksi benih rumput laut kultur jaringan.To support the program of seaweed transgenesis, somatic embryo can be used as a materials for gene transfer purpose either by individual or cluster of cells in accelerating the higher rate of transformation. This research aims to study the callus induction of seaweed K. alvarezii for production of somatic embrio (s.e) cell by different ratio of growth regulators (GR) and agar media concentrations. The study consists of two stages: stage (1) callus induction to the cells filament using GR of indol acetic acid (IAA):kinetin in ratio of  0.5:0.0 mg/L; 1.0:1.0 mg/L; and 2.0:0.2 mg/L on the media agar concentration of 0.6%; 0.8%; 1.0%; and 1.5%, and stage (2) regeneration of the cell mass of s.e, using GR of IAA:kinetin in ratio of 0.1:1.0 mg/L; 0.0:0.1 mg/L on the media agar concentration of 0.4%; 0.6%; and 0.8%. For further maintenance, the s.e cells were cultured in liquid media. The results of callus induction showed that the ratio of IAA:kinetin (1:1) on the media agar concentration of 0.8% and 1.0% produced the highest callus induction (90%). The study of mass cell regeneration of s.e showed did not different of GR IAA:kinetin ratio of the cell mass development, but the media agar concentration of 0.6% and 0.4% showed the higher growth of cell mass (in diameter size of 4-5 mm). The development of cells culture of s.e from the size of 3-4 mm to filament of 0.5 mm could be reached in one month of culture period. The success of K. alvarezii s.e production will be helpful not only as a material for gene transfer but also as a reference on tissue culture for seed production of seaweed.


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