Perbandingan Metode Sterilisasi Untuk Perbanyakan Rubus rosifolius Secara in Vitro

2021 ◽  
Vol 14 (1) ◽  
pp. 127-137
Author(s):  
Muhammad Imam Surya ◽  
Lily Ismaini

AbstrakRubus rosifolius adalah salah satu jenis rasberi liar yang memiliki potensi cukup tinggi untuk dikembangkan sebagai tanaman buah. Selain itu, metode perbanyakan tanaman merupakan salah satu faktor yang berpengaruh dalam pembudidayaan. Lebih lanjut, informasi terkait upaya perbanyakan R. rosifolius secara in vitro masih sangat terbatas. Percobaan ini ditujukan untuk mengetahui metode sterilisasi yang tepat pada eksplan R. rosifolius. Sebanyak 17 metode sterilisasi telah diujicobakan di Laboratorium Kultur Jaringan BKT Kebun Raya Cibodas-LIPI. Bahan sterilisasi yang digunakan, yaitu detergen, tween 80, bakterisida, fungisida, clorox/pemutih (NaClO), alkohol 70%, larutan Plant Preservative Mixture (PPM), vitamin C/asam askorbat, dan povidone iodine/antiseptik. Hasil percobaan menunjukkan bahwa metode sembilan merupakan metode sterilisasi yang cukup optimum untuk sterilisasi eksplan R. rosifolius. Metode sembilan mampu menghambat munculnya mikroorganisme endofitik hingga 8 hari dan tidak menyebabkan warna eksplan menjadi cokelat/browning. Tahapan sterilisasi pada metode sembilan meliputi pencucian dengan detergen, perendaman dengan bakterisida + fungisida selama +30 menit, perendaman dengan clorox 10% + tween 80 selama +15 menit, pencucian dengan larutan PPM selama +15 menit.  AbstractRubus rosifolius is one of the species from wild raspberries, which is has high potential to develop as a fruit crops. In the other hand, the technique of plant propagation became an important factor for cultivation. Moreover, the information related to the in vitro propagation of R. rosifolius is very limited. This experiment was aimed to determine the best method to sterilize an explants of R. rosifolius. About 17 methods of sterilization have been tried in the laboratorium of tissue culture at Cibodas Botanical Garden-Indonesian Institute of Sciences. The combination of detergent, tween 80, bactericide, fungicide, sodium hypochlorite (NaClO), alcohol 70%, plant preservative mixture (PPM), ascorbic acid, and povidone iodine were used during the experiment. The results show that the method of sterilization number nine could be inhibit the emergence of endophytic organisms for eight days and keep an explant in green with a little brownish compared by the others methods. The method of sterilization number nine was consist of several steps i.e. wash by detergent, soak in bactericide + fungicide for +30 minutes, soak in sodium hypoclorite 10% + tween 80 for +15 minutes, wash by PPM solution for +15 minutes.

Lankesteriana ◽  
2015 ◽  
Vol 7 (1-2) ◽  
Author(s):  
Alla Lavrentyeva ◽  
Roman Ivannikov

Nowadays many wild species of South American’s orchids are under threat of extinction from over-col- lection and habitat destruction. Many tropical native orchid species were propagated in the National Botanical Garden of National Academy of Sciences of Ukraine through a range of asymbiotic seed germi- nation techniques and tissue culture procedures aimed to preserve a number of individuals under artificial conditions in glasshouses in the temperate zone, with the aim to protect these species from complete extinc- tion. Our orchid collection includes plants of Cattleya and Laelia species. Some of these species are rare in the wild. To protect them from extinction the meth- ods of propagation should be developed. 


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Helena Abramovič ◽  
Blaž Grobin ◽  
Nataša Poklar Ulrih ◽  
Blaž Cigić

Trolox, gallic acid, chlorogenic acid, caffeic acid, catechin, epigallocatechin gallate, and ascorbic acid are antioxidants used as standards for reaction with chromogenic radicals, 2,2-diphenyl-1-picrylhydrazyl (DPPH⋅) and 2,2′-azino-bis-3-ethylbenzotiazolin-6-sulfonic acid (ABTS⋅+), and Folin–Ciocalteu (FC) reagent. The number of exchanged electrons has been analyzed as function of method and solvent. A majority of compounds exchange more electrons in FC assay than in ABTS and DPPH assays. In reaction with chromogenic radicals, the largest number of electrons was exchanged in buffer (pH 7.4) and the lowest reactivity was in methanol (DPPH) and water (ABTS). At physiological pH, the number of exchanged electrons of polyphenols exceeded the number of OH groups, pointing to the important contribution of partially oxidized antioxidants, formed in the course of reaction, to the antioxidant potential. For Trolox, small impact on the number of exchanged electrons was observed, confirming that it is more suitable as a standard compound than the other antioxidants.


1957 ◽  
Vol 106 (1) ◽  
pp. 159-177 ◽  
Author(s):  
Dorothy M. Horstmann ◽  
J. R. Paul ◽  
J. L. Melnick ◽  
Joyce V. Deutsch

Four of five individuals possessing homotypic antibody in titers of 8 to 64 were infected on being fed Type III (Leon KP-34) poliovirus attenuated by Sabin by passage through tissue culture. None of the infected subjects or controls showed any evidence of illness which could be attributed to virus infection. There was no evidence of spread of infection to any of the control adult wardmates of the experimental subjects, although the two groups were in close contact: none of the controls excreted virus, none showed any antibody shift. One control who had no Type III antibodies at the start of the experiment was still antibody-negative on the 63rd day of the experiment. Three of the four individuals who became infected had naturally acquired-Type III antibodies; the other had antibodies induced by formalinized vaccine. Virus excretion in the stool was of short duration (7 to 13 days) in the three with natural antibodies, and lasted at least 6 weeks after feeding in the vaccinated child. Virus in the throat was detected only in the two persons receiving the larger virus dose (107.5 TCD50). In them it was present in small amounts between the 2nd and 6th day after feeding. No virus was detected in the blood of any of the infected individuals. The antibody responses of the four infected individuals were variable. There was no clear correlation with virus dosage, amount of virus excretion in the stools, or presence of virus in the throat. Only the child whose neutralizing antibodies were "Salk" vaccine induced showed a marked CF response. The virus excreted by two of the individuals who became infected, as tested in the 2nd tissue culture passage by monkey inoculation, was slightly more neurotropic than the virus which was ingested. Virus excreted by one of these individuals behaved as a virulent strain when tested by the in vitro plaque virulence test, while that isolated from the other had the characteristics of an attenuated strain in this test.


1986 ◽  
Vol 112 (2) ◽  
pp. 271-277 ◽  
Author(s):  
Mervi Julkunen

Abstract. Human decidua was found to synthesize and secrete placental protein 14 (PP14). The presence of PP14 in tissue explants of decidua, foetal membranes and placenta from term pregnancy was demonstrated by radioimmunoassay. The PP14 content in decidua (1300 ±410 ng/100 mg tissue) was higher than in chorion (570 ± 120 ng/100 mg; P < 0.01), amnion (240 ± 100 ng/100 mg; P < 0.001) or placenta (300 ± 130 ng/100 mg; P< 0.001). Three to 36 times more PP14 was released into culture medium by decidual explants compared with the other tissues, and cycloheximide decreased this release by 39%. Placental tissues released hardly any PP14. Decidual synthesis of PP14 was demonstrated by incorporation of [35S]methionine into immunoprecipitable PP14 in tissue culture.


1985 ◽  
Vol 65 (4) ◽  
pp. 1025-1032 ◽  
Author(s):  
BRIAN W. DYKEMAN ◽  
BRUCE G. CUMMING

Methods were developed for the successful in vitro propagation of ostrich fern (Matteuccia struthiopteris (L.) Todaro) clones utilizing shoot tips derived by forcing lateral buds on the rhizome. Maximum shoot proliferation was attained with 6-furfurylaminopurine (kinetin) at 1.0 mg/L with half-strength Murashige and Skoog (MS) inorganic salts and sucrose, agar, NaH2PO4, adenine sulphate, i-inositol and thiamine∙HCl at 30 000, 4000, 85, 40, 100, 0.4 mg/L, respectively. Excellent frond and root development was achieved with half-strength MS salts and sucrose, agar, i-inositol and thiamine∙HCl at 7500, 4000, 100 and 0.4 mg/L, respectively. The methods developed were satisfactory for a cross section of clones. Morphogenesis in vitro was dependent on medium osmotic potential.Key words: Matteuccia struthiopteris, in vitro propagation, tissue culture, morphogenesis, fern (ostrich)


2020 ◽  
Vol 8 (10) ◽  
pp. 237-244
Author(s):  
Duru Maduabuchi ◽  
◽  
Mbata Ikechukwu ◽  
Osikwe Keziah ◽  
Ukaoma Adamma ◽  
...  

The study investigated an in vitro propagation of Manihot esculenta Crantz in a substituted substrate regime. The aim was to proffer and affordable alternative to the expensive high tech media formulations usually employed in tissue culture protocol. The experiment was conducted on laboratory bench, using standard tissue culture and micropropagation methods under aseptic conditions. The morphogenesis effect of the substrate was determined based on the integer number of explants’ callus and adventitious shoot regeneration. Results showed that MS + Agar, supported embryogenic callus formation with 38% viability, NH4NO3 + KH2PO4 + Agar, supported same with 29%. MS + 2, 4-D + BAP +Agar supported shoot establishment with 32%. While NH4NO3 + KH2PO4 + Zea mays extracts + Agar, did same with 43.26%. MS + Soil, supported callugenesis with 27% viability while NH4NO3 + KH2PO4 + Soil supported the callus establishment with 25%. MS + 2,4 - D + BAP + Soil, supported shoot establishment with 38.41% viability while NH4NO3 + KH2PO4 + Zea mays Extracts + Soil supported same with 36%. The application of crude Zea mays seedling extracts can serve as potent alternative to the synthetic 2, 4 – D and BAP, in in vitro somatic cell morphogenesis. NH4NO3 + KH2 + PO4 can substitute for the MS salt in the same protocol. Loamy top soil can be a good alternative to agar powder as gelling agent in cassava somatic cell embryogenesis and shoot regeneration. Keywords: Ammonium nitrate, Potassium biphosphate, MS salt, axillary meristem, morphogenesis.


2020 ◽  
Vol 17 (3) ◽  
pp. 473-481
Author(s):  
Do Hong Gam ◽  
Duong Huong Huynh ◽  
Phan Thi Lan Anh ◽  
Nguyen Hoang Duong ◽  
Do Thi Kim Hoa

In this study, the effects of various agricultural LED lights (LED NN), including single red LED (R), single blue LED (B), and four combinations of blue, red, and warm white (W) LED (BR, BRW1, BRW2, BRW3) on the growth and development of C. militaris (Link.) Fries were evaluated in vitro. After 7 days, samples subjected to LED NN showed shorter sporocarp sprouting time and higher sprouting ratio than the control, which was subjected to T5 fluorescent light. After 2 months, LED lights with high red ratio, such as single red LED and LED BR, had suppressing effect on the growth and development of C. militaris (Link.) Fries. On the other hand, combinations of red, blue, and warm white such as LED BRW1, LED BRW2, and LED BRW3 had the positive impact on the growth and development of this fungus. Notably, samples subjected to LED BRW2 reached 5.79 cm in height, fresh biomass of 3.67 g/20 samples. Cordycepin and Adenosine levels were 64.2 and 6.37 mg/100 g fresh mass, respectively. All of studied  indicators were the higher compared to those of the control and other LED lighting schemes. Therefore, it can be conlcuded that LED lighting combination with BRW2 ratio of 1:5:1 and luminous intensity of 45±2 µmol.m-2.s-1 (511,59 Lux) was suitable for the growth and development of C. militaris (Link.) Friesand a potential replacement of fluorescent light for C. militaris (Link.) Friesin vitro propagation.


2014 ◽  
Vol 22 (2) ◽  
pp. 47-52 ◽  
Author(s):  
Jaime A. Teixeira da Silva ◽  
Kamal P. Acharya

AbstractNepalese orchids are made up of 458 taxa. Despite a ban on the collection and trade of all orchid species in Nepal, numerous anthropogenic factors are leading to the rapid loss of natural stands of germplasm. Biotechnology, specifically in vitro propagation, may be the only viable solution for preserving and reintroducing endangered germplasm back into the wild. Despite the large germplasm base, only tissue culture studies have been conducted, and most have focused almost exclusively on in vitro seed germination, the bulk of which have been conducted in the past few years. No other biotechnological advances have yet been made. This brief review provides a short synopsis of the advances made thus far in the in vitro propagation of Nepalese orchids.


2017 ◽  
Vol 47 (6) ◽  
Author(s):  
Aline Meneguzzi ◽  
Mayra Juline Gonçalves ◽  
Samila Silva Camargo ◽  
Fernanda Grimaldi ◽  
Gabriela Candido Weber ◽  
...  

ABSTRACT: International breeding programs launched new genetic material of apple rootstocks that in addition to precocity and great yield are resistant to major diseases and soil pests encountered in the largest apple producing regions in Brazil. Given this, there is a necessity for vegetative propagation of these materials for study and possible replacement of existing rootstocks. The objective was to adapt a micropropagation protocol for new apple rootstock ‘G. 814’. In the multiplication phase were evaluated BAP concentrations: 0; 0.5; 1; 2 and 4mg L-1 and in the rooting phase were evaluated IBA concentrations: 0; 0.25; 0.50; 1; 1.5 and 2.5mg L-1. These new results demonstrated that this new rootstock selection can be propagated with this tissue culture adapted protocol. For the successful in vitro propagation of apple rootstock ‘G. 814’ it is indicated the use of 1mg L-1 BAP at multiplication phase and 1.5mg L-1 IBA at rooting phase.


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