Tissue culture and genetic analysis of somaclonal variations of Solanum melongena L. cv. Nirrala

2014 ◽  
Vol 9 (12) ◽  
pp. 1182-1195
Author(s):  
Samar Naseer ◽  
Tariq Mahmood
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Soluble Protein ◽  
Solanum Melongena ◽  
Total Soluble Protein ◽  
Naphthalene Acetic Acid ◽  
Fresh Tissue ◽  
Somaclonal Variations ◽  
Random Amplification ◽  
Plant Grown

AbstractThe present study was designed to analyze genetically somaclonal variants using biochemical and molecular markers. Efficient tissue culture protocol for Solanum melongena L. cv. Nirrala was developed. Maximum callus induction (100%) was observed for Murashige and Skoog (MS) media supplemented with 2.0 mg L−1 naphthalene acetic acid +0.5 mg L−1 6-benzylaminopurine; and nodal explants gave best callusing response (88.8%) as compared to internodes (88.3%) and leaves (87.7%). The best shooting was induced on nodal and internodal callus in the presence of 2.0 mg L−1 6-benzylaminopurine. Total soluble protein content of callus and regenerated variant plants was estimated for biochemical analysis, and largest amount of soluble protein was found in callus (6.54 mg g−1 fresh tissue) followed by variant plant grown on 2.0 mg L−1 6-benzylaminopurine (5.96 mg g−1 fresh tissue). Random amplification of polymorphic DNA technique was done with five decamer primers (OPC1-OPC5) and maximum polymorphism was detected by OPC 2 (26.99%) among all samples, whereas nodal callus on media containing 1.0 mg L−1 naphthalene acetic acid +1.0 mg L−1 6-benzylaminopurine showed highest polymorphism producing 22 bands, out of which 8 bands were polymorphic. The study shows that this marker system can provide better evaluation of genetic variation induced by tissue culture.

scholarly journals Multiplikasi Eksplan Tunas Anggrek Hitam (Coelogyne pandurata Lindl.) dengan Penambahan NAA (Naphthalene Acetic Acid) dan Ekstrak Biji Jagung (Zea mays) Secara In Vitro

2021 ◽  
Vol 11 (2) ◽  
pp. 114
Author(s):  
Nurkapita Nurkapita ◽  
Riza Linda ◽  
Zulfa Zakiah
Keyword(s):  
Tissue Culture ◽  
Zea Mays ◽  
Acetic Acid ◽  
Seed Extract ◽  
Naphthalene Acetic Acid ◽  
Shoot Height ◽  
Culture Techniques ◽  
Corn Seed ◽  
Randomized Design

(Article History: Received February 18, 2021; Revised April 27, 2021; Accepted May 19, 2021) ABSTRAKPerkembangbiakan anggrek secara generatif alami membutuhkan bantuan jamur mikoriza untuk perkecambahan biji, sedangkan usaha perbanyakan konvensional memerlukan waktu lama untuk memperoleh tanaman dalam jumlah banyak. Salah satu alternatif untuk perbanyakan anggrek hitam (Coelogyne pandurata Lindl.) adalah melalui multiplikasi tunas anggrek secara in vitro. Tujuan penelitian adalah untuk membuktikan pengaruh pemberian NAA (Naphthalene Acetic Acid) dan ekstrak biji jagung (Zea mays) terhadap multiplikasi tunas anggrek hitam. Penelitian dilakukan di Laboratorium Kultur Jaringan Jurusan Biologi Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Tanjungpura Pontianak. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) pola faktorial dengan dua faktor perlakuan. Faktor pertama adalah NAA terdiri dari 5 taraf konsentrasi yaitu A0 (0 M/ kontrol) A1 (10-7 M), A2 (10-6 M), A3 (5x10-7 M) dan A4 (5x10-6 M ) dan faktor ekstrak biji jagung (B) dengan 5 taraf konsentrasi yaitu B0 (0%), B1 (2,5%), B2 (5%); B3 (7,5%) dan B4 (10%). Pemberian kombinasi NAA dan ekstrak biji jagung berpengaruh nyata terhadap semua parameter yaitu jumlah tunas, jumlah daun, dan tinggi tunas. Hasil terbaik rerata jumlah tunas pada perlakuan A4+B4 yaitu 5x10-6M NAA+10% ekstrak biji jagung. Hasil terbaik pada rerata jumlah daun pada perlakuan A2+B2 yaitu 5x10-7M NAA+5% ekstrak biji jagung dan hasil terbaik pada rerata tinggi tunas pada perlakuan A1+B1 yaitu 10-7M NAA+2,5% ekstrak biji jagung.Kata Kunci: multiplikasi; tunas anggrek hitam; ekstrak biji jagung; NAA. ABSTRACTGenerative reproduction of orchid plants it takes a requires the help of mycorriza mushrooms for seed germination, whereas conventional propagation business takes a long time to obtain large quantities of plants. One alternative to the propagation black orchids (Coelogyne pandurata Lindl.) is required through tissue culture techniques. The purpose of this study is to find the influence and concentration corn seed extract (Zea mays) and NAA (Naphthalene Acetic Acid) on the multiplication black orchids. This research was conducted in the tissue culture laboratory Biology Department Faculty of Mathematics and Natural Sciences Tanjungpura University Pontianak. The study used a Complete Randomized Design (RAL) of factorial patterns with two treatment factors. The first factor is that the NAA consists of 5 concentration levels  A0 (0 M) A1 (10-7 M), A2 (10-6 M), A3 (5x10-7 M) and A4 (5x10-6 M ) and the second factor is that corn seed extract of 5 levels concentratio B0(0%), B1 (2,5%), B2 (5%); B3 (7,5%) and B4 (10%). The administration NAA and corn seed extract in combination has a real effect on all parameters namely the number shoots, the number leaves, and the height shoots. The best results where the average number of shoots in the treatment of A2+B2 namely 5x10-6M NAA + 10% corn seed extract. The best results average number of leaves in the treatment  A2+B2 namely 5x10-7M NAA + 5% corn seed extract and in the best results for shoot height in the treatment of A1+B1 namely 10-7M NAA + 2.5% corn seed extract.Keywords: Multiplication; black orchid’s shoot; corn  seed extract; NAA

scholarly journals Variation of axillary growth as respond of Morus spp. micropropagation using various concentration of Indonesian local solid substance

2019 ◽  
Vol 21 (1) ◽  
Author(s):  
Yasinta Ratna Esti Wulandari ◽  
Laurensia Danis Anggradita
Keyword(s):  
Growth Rate ◽  
Tissue Culture ◽  
Acetic Acid ◽  
Callus Formation ◽  
Culture Technique ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Solid Substance ◽  
Local Plant ◽  
Morus Spp

Abstract. Wulandari YRE, Anggradita LD. 2020. Variation of axillary growth as respond of Morus spp. micropropagation using various concentrations of Indonesian local solid substance. Biodiversitas 21: 80-85. The difficulties of growing Morus spp. makes it become one local plant that hard to cultivate conventionally even though it’s a beneficiary plant. Hence cultivation Morus spp. through tissue culture technique could help growing this plant. This research is aimed to design the optimal condition for micropropagation of local Morus spp. (Morus bombycis var. lembang, M. cathayana, M. multicaulis, and M. alba var. kanva-2) using agar-agar as Indonesian local solid substance. This solid substance is used as its cheap and easy to find compared to other solid substances. This research used MS medium supplemented with 0.1 ppm naphthalene acetic acid + 1.0 ppm benzyl aminopurine and various concentration of agar-agar (0.6%, 0.8%, 1.0%). Growth rate, axillary bud length and number, leaf number, callus formation and contamination were observed in this research. All those concentrations could be used for micropropagation of Morus spp. Agar concentrations of 0.8 and 1.0% showed better results than 0.6% because it showed the highest results.

scholarly journals Effect of synthetic auxins on in vitro and ex vitro bromeliad rooting

2013 ◽  
Vol 43 (2) ◽  
pp. 138-146 ◽  
Author(s):  
João Paulo Rodrigues Martins ◽  
Edilson Romais Schimildt ◽  
Rodrigo Sobreira Alexandre ◽  
Breno Régis Santos ◽  
Gizele Cristina Magevski
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
In Vitro Growth ◽  
Free Medium ◽  
Ex Vitro ◽  
Synthetic Auxins ◽  
Parameters Analysis

The tissue culture can contribute to the propagation of several economic species, such as the bromeliads. This research aimed at evaluating the auxins type and concentration in the in vitro and ex vitro rhizogenesis of Neoregelia concentrica bromeliad. N. concentrica shoots were induced in a growth medium with 15.0 µM of 6-benzylaminopurine, for 80 days, followed by sub-cultivation in phytoregulator-free medium, for 45 days. In the in vitro rhizogenesis, the shoots grew in a medium supplemented with indole-3-butyric acid (IBA) or naphthalene-acetic acid (NAA), at the concentrations of 0.0 µM, 1.0 µM, 2.0 µM, 3.0 µM and 4.0 µM. In the ex vitro rhizogenesis, the bases of shoots were immersed, for 60 minutes, in IBA or NAA solutions, at the concentrations of 0.0 µM, 5.0 µM, 10.0 µM and 15.0 µM. After immersion, the shoots were planted in plastic trays with vermiculite. At the end of each rhizogenesis method, the phytotechnical parameters analysis was carried out. For the in vitro rhizogenesis, a higher number of roots were observed when the shoots were cultivated in concentrations higher than 1.0 µM of NAA, when compared to the IBA. However, the rooting rate differed only at 30 days after the in vitro growth, with a higher root induction in the shoots grown with NAA. At 60 days, the rooting rate was higher than 90% and statistically similar in all treatments. In the ex vitro rhizogenesis, a better formation of the rooting system was observed when 5.0 µM of IBA was applied, with higher rooting averages and number of roots.

scholarly journals Tissue culture of Cecropia glaziovii Sneth (urticaceae): vegetative micropropagation and plant regeneration from callus

2010 ◽  
Vol 34 (5) ◽  
pp. 1245-1252 ◽  
Author(s):  
Marcos Nopper Alves
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Shoot Regeneration ◽  
Culture Media ◽  
Adventitious Shoot ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Adventitious Shoot Regeneration ◽  
Popular Medicine ◽  
Superior Genotype

Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS) medium supplemented with 6-benzylaminopurine alone (BAP) (1.0, 5.0 or 10.0 mg L-1) or combined with -naphthalene acetic acid (NAA) (1.0 or 2.0 mg L-1), after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D) (5.0 mg L-1) combined with BAP (1.0 mg L-1). Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA) (0.1 mg L-1) alone or combined with 2,4-D (1.0 or 5.0 mg L-1) was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA) (1.0 mg L-1). Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.

scholarly journals Propagation Techniques for a Spineless Acacia wrightii

HortScience ◽  
2005 ◽  
Vol 40 (6) ◽  
pp. 1832-1837 ◽  
Author(s):  
Donita L. Bryan ◽  
Michael A. Arnold ◽  
R. Daniel Lineberger ◽  
W. Todd Watson
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Butyric Acid ◽  
Shoot Proliferation ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Root Number ◽  
Indole Butyric Acid ◽  
Tissue Culture Propagation

Three spineless phenotypes of Acacia wrightii G. Bentham ex A. Gray were identified with aesthetic landscape potential. Experiments in seed, cutting, grafting, and tissue culture propagation were undertaken to perpetuate this desired spineless phenotype. Germination percentages for mechanically scarified seeds ranged from 33% to 94%, however yield of spineless seedlings was low (0% to 34%). Sulfuric acid scarification for 10, 20, 30, or 60 minutes hastened and unified germination compared to nontreated seeds by 7 to 8 days. Vegetative propagation was successful for softwood cuttings. Rooting measures increased with auxin (2:1 indole butyric acid to naphthalene acetic acid) concentrations from 0 to 15000 mg·L–1, with maximum rooting percentage (70%), root number (9.2), and root length (12.4 cm) per softwood cutting at 15000 mg·L–1 auxin 8 weeks after treatment. Rooting was not successful for semi-hardwood or hardwood cuttings. Whip-and-tongue or T-bud grafting was not successful. Tissue culture of shoots from in vitro germinated seedlings indicated that shoot proliferation was greatest in Murashige and Skoog (MS) medium with 15 μm zeatin. The number of shoots that rooted in vitro increased with increasing concentrations of indole-3-butyric acid from 0 to 25 μm.

scholarly journals Response of In Vitro Strawberry to Silver Nitrate (AgNO3)

HortScience ◽  
2005 ◽  
Vol 40 (3) ◽  
pp. 747-751 ◽  
Author(s):  
Yonghua Qin ◽  
Shanglong Zhang ◽  
Lingxiao Zhang ◽  
Daoyu Zhu ◽  
Asghar Syed
Keyword(s):  
Acetic Acid ◽  
Silver Nitrate ◽  
Shoot Regeneration ◽  
Soluble Protein ◽  
Dry Weight ◽  
Naphthalene Acetic Acid ◽  
Antioxidant Enzyme Activities ◽  
Tetrazolium Chloride ◽  
Regeneration Efficiency

Response of Toyonoka strawberry to AgNO3 was studied. Types and combinations of plant growth regulators had significant effects on shoot regeneration efficiency. Explants cultured for 10 days in shoot regeneration medium in the presence of AgNO3 not only enhanced shoot regeneration efficiency but also expedited the initiation of adventitious buds. Highest regeneration (87.38%) and number of shoots per explant (11.67) were achieved in shoot regeneration media containing 1.5 mg·L–1 TDZ, 0.4 mg·L–1 IBA and 1.0 mg·L–1 AgNO3. Half-strength MS containing 1.0 mg·L–1 AgNO3 was an optimum medium for rooting. AgNO3 advanced root emergence and increased percent rooting, root length, dry weight and activity. Lower concentrations of AgNO3 inhibited ethylene production and promoted shoot regeneration and growth. It had a significant stimulatory effect on chlorophyll, soluble protein contents and antioxidant enzyme activities. Chlorophyll and soluble protein contents, superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities were increased in the presence of AgNO3 and reached maximum at 1.0 mg·L–1 AgNO3. Root water content, superoxide free radicals (O2.-), malondialdehyde (MDA) content, proline accumulation and IAA-oxidase activity in leaves were increased while (IAA) content was decreased in the presence of AgNO3. Chemical names used: indole-3-butyric acid (IBA); silver nitrate (AgNO3); thidiazuron (TDZ); N6-benzyladenine (BA); 2,4-dichlorophenoxy acetic acid (2,4-D); indole-3-acetic acid (IAA); α-naphthalene acetic acid (NAA); gibberellic acid (GA3); bovine serum albumin (BSA); 2,3,5-triphenyl-2H-tetrazolium chloride (TTC).

scholarly journals INFLUNCE OF AUXIN AND POLYAMINEES ON ROOTING OF SHOOTS OF CITRUS VOLKAMERIANA ROOTSTOCK IN VITRO

2016 ◽  
Vol 47 (3) ◽  
Author(s):  
Al- Khazali & Hamad
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Plant Tissue Culture ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Root Number ◽  
College Of Agriculture ◽  
Indole Butyric Acid ◽  
Citrus Volkameriana

This  research  was  conducted  in  the  plant  tissue  culture  Lab. College  of Agriculture / University  of  Baghdad  from  February to  October  2015. The aim  of  the  study  was  investigating  the  influence  of  combinations  of Indole  butyric  acid (IBA) ,  Naphthalene  acetic  acid (NAA) and  polyamine Spermidine (Spd.) on rooting of shoots of  citrus volkameriana rootstock cultured  on 1\2  MS medium. The Results indicated that 1/2 MS medium supplemented  with 1.0 mg L-1 (IBA)  gave the highest  percentage  of  rooting  (67 %) which differed significantly  from the MS medium with free auxin IBA  that gave (22%) while  the  MS medium  supplemented with 0.5 mg L-1  spermidine  gave the highest percentage of rooting (63%) that was not significantly different  than other concentrations of Spd . MS medium supplemented with 1.0 mg L-1 IBA and 0.5 mg L-1 Spd. gave the highest percentage of rooting (83%) and the highest root number / shoot (3.17) and highest length of root (3.15 cm) while the MS medium with free auxin IBA and spd. did not give percentage of rooting (0%) for citrus volkameriana rootstock plantlets . The MS medium supplemented with 1.0 mg L-1 NAA gave the highest percentage of rooting (56%) which differed significantly from MS medium with free auxin NAA that gave (22%)  while MS medium supplemented with 0.5mg L-1 spd. gave the highest percentage of rooting (50%) that was not significantly different from other concentration of Spd . MS medium supplemented with 1.0 mg L-1 NAA and 0.5 mg L-1 Spd. gave the highest percentage of rooting (68%) and the highest root number /shoot (2.5) and highest length of root (2.65 cm) while the MS medium with free auxin NAA and Spd. did not gave percentage of rooting (0%) .

scholarly journals Tissue Culture and Plant Regeneration of Cowpea (Vigna unguiculata)

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 627f-627
Author(s):  
Lurline Marsh
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Plant Regeneration ◽  
Vigna Unguiculata ◽  
Nutrient Medium ◽  
Cotyledonary Node ◽  
Naphthalene Acetic Acid ◽  
Leaf Segments ◽  
Dichlorophenoxy Acetic Acid ◽  
Cotyledonary Node Explants

Explants (cotyledon, cotyledonary node, second node, hypocotyl, epicotyl, and leaf) of cowpea (Vigna unguiculata) genotypes MN13 and Pinkeye Purple Hull were cultured on Murashige and Skoog basal nutrient medium. The medium was supplemented with 1 mg·L–1 benzyladenine (BA) or 1 mg·L–1 benzyladenine plus naphthalene acetic acid (BA + NAA) or 2 mg·L–1 2,4-dichlorophenoxy acetic acid (2,4-D). Cultures were maintained at 22°C for 1 month, after which they were transferred to 1 mg·L–1 BA + NAA. Cotyledons, hypocotyl, epicotyl, and leaf segments produced only calli after subculturing in BA + NAA. The second node and cotyledonary node explants cultured on the BA or BA + NAA followed by subculture on BA + NAA produced calli, shoots, and roots. The plants were then transplanted to promix but later died.

Media for growth of flax tissue culture

1971 ◽  
Vol 49 (2) ◽  
pp. 295-298 ◽  
Author(s):  
R. K. Ibrahim
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Root Formation ◽  
Callus Tissue ◽  
Maximum Growth ◽  
Shoot Formation ◽  
Tissue Differentiation ◽  
Naphthalene Acetic Acid ◽  
Mineral Salts ◽  
Callus Tissue Culture

A callus tissue culture from flax cotyledons reached maximum growth on a medium rich in mineral salts and supplemented with 2 mg/l α-naphthalene acetic acid, 0.05 mg/l kinetin, 15% coconut milk, and 3% sucrose. While tissue differentiation was evident and root formation was induced with increasing auxin concentrations, higher kinetin levels or the addition of L-tyrosine failed to promote shoot formation.

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