Genetic transformation of Stella De Oro daylily by particle bombardment

2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou
Keyword(s):  
Southern Blotting ◽  
Basal Medium ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Semi Solid ◽  
Independent Transformation

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis

scholarly journals Total Polyphenol Content, in vitro Antifungal and Antioxidant Activities of Callus Cultures from Inula crithmoides

2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Anahi Bucchini ◽  
Laura Giamperi ◽  
Donata Ricci
Keyword(s):  
Methanol Extract ◽  
Antioxidant Activities ◽  
Leaf Explants ◽  
Basal Medium ◽  
Alternaria Solani ◽  
Antimicrobial Activities ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Stable Free Radical

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.

scholarly journals Resistance of in Vitro-derived Cucumber Plants to Pythium aphanidermatum

HortScience ◽  
1990 ◽  
Vol 25 (8) ◽  
pp. 967-969 ◽  
Author(s):  
W. Msikita ◽  
H.T. Wilkinson ◽  
R.M. Skirvin
Keyword(s):  
Cucumis Sativus ◽  
Pythium Aphanidermatum ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Embryonic Axes ◽  
Rate Of Development ◽  
Primary Roots ◽  
Solid Agar ◽  
Semi Solid

`Embryonic axes-derived `Burpless Hybrid' cucumber (Cucumis sativus L.) plantlets germinated on Murashige and Skoog (MS) medium supplemented with 16 combinations of BAP and NAA and seedlings derived from whole seeds cultured on semi-solid agar were inoculated in vitro with two isolates (WFU3 and WFM13) of Pythium aphanidermatum. All axes-derived plantlets and whole seedlings inoculated with WFM13 isolates were susceptible to blight and died 2 days after inoculation. Similarly, all seedlings inoculated with WFU3 isolates were killed within 2 days after inoculation; however, the rate of development and severity of blight varied among the axes-derived plantlets. Blight on axes-derived plantlets, regenerated on MS medium supplemented with 2 mg BAP/liter and 0.2 mg NAA/liter, was significantly less than on regenerants cultured on all other amended MS media. On some media, callus developed on crowns and/or primary roots. The presence of callus influenced resistance to Pythium. In a second experiment, axes-derived cucumber regenerants from five genotypes, cultured on MS medium supplemented with 2 mg BAP/liter and 0.2 mg N&A/liter, were compared for their resistance to P. aphanidermatum isolate WFU3. Resistance was significantly greater for `Burpless Hybrid' and `Sweetslice' than for three other genotypes. Chemical names used: 6-benzylaminopurine (BAP); α -naphthaleneacetic acid (NAA).

scholarly journals In vitro Multiple Shoot Regeneration from Petunia hybrida

2019 ◽  
Vol 7 (10) ◽  
pp. 1554
Author(s):  
Rebaz Rasul Habas ◽  
Musa Turker ◽  
Fethi Ahmet Ozdemir
Keyword(s):  
Petunia Hybrida ◽  
Survival Rates ◽  
Basal Medium ◽  
Multiple Shoot ◽  
Shoot Length ◽  
Peat Moss ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Shoot Number

An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.

scholarly journals In Vitro Propagation of Eriostemon myoporoides and Eriostemon `Stardust'

HortScience ◽  
1994 ◽  
Vol 29 (6) ◽  
pp. 686-688 ◽  
Author(s):  
James R. Ault
Keyword(s):  
Root Length ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Shoot Length ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Axillary Shoot Proliferation

Optimal axillary shoot proliferation was obtained from stem explants of a clone of Eriostemon myoporoides DC. on Murashige and Skoog (MS) basal medium containing 0.1 mg BA/liter, and of Eriostemon `Stardust' on MS medium containing 0.5 mg BA/liter. Overall average number of shoots and shoot lengths for all treatments was greater for E. `Stardust' (22.4 shoots and 12.1-mm shoot length) than for E. myoporoides (4.5 shoots and 8.3-mm shoot length). Maximum percent rooting of E. myoporoides (42%) and E. `Stardust' (95%) was obtained on MS medium supplemented with 1.0 mg K-IBA/liter for E. myoporoides and 0.1 mg NAA/liter for E. `Stardust'. Overall average percent rooting and root lengths were greater for E. `Stardust' (42% rooting and 11.0-mm root length) than for E. myoporoides (27% rooting and 2.3-mm root length). For E. `Stardust', reducing sucrose in the rooting medium from 50 to 25 g·liter-1 significantly decreased overall average percent rooting to 1670 and root length to 6.8 mm. Plantlets of both clones were acclimatized in the greenhouse and transferred successfully to soil, although survival was <7070. Chemical names used: N -(phenylmethyl) -l H -purine-6-amine (BA); potassium-l H -indole-3-butyric acid (K-IBA); l-naphthaleneacetic acid (NAA).

Micropropagation and In Vitro Inflorescence of Pentas lanceolata

2022 ◽  
Author(s):  
Kitti Bodhipadma ◽  
Sompoch Noichinda ◽  
Chutikarn Tangtivaporn ◽  
Saowaros Phanomchai ◽  
David W. M. Leung
Keyword(s):  
Basal Medium ◽  
Tubular Structure ◽  
Nodal Explants ◽  
Ms Medium ◽  
Water Accumulation ◽  
Semi Solid ◽  
Pentas Lanceolata

In this study, different concentrations of 6-benzyladenine (BA) on in vitro shoot and inflorescence inductions of P. lanceolata were investigated. The in vivo and in vitro floral characteristics of this plant were also compared. Nodal explants of P. lanceolata were cultured vertically with the cut ends inserted into semi-solid Murashige and Skoog (MS) medium supplemented with 0, 0.5, 1, 2, 4, and 8 mg L–1 BA. The results showed that the explants formed the highest numbers of shoots even when cultured in MS basal medium without any addition of BA, while the shoots formed in the explants cultured in MS medium supplemented with 1 mg L–1 BA were the longest. No inflorescence was found in the shoots cultured in MS medium supplemented with 8 mg L–1 BA, while the highest percentage of inflorescence induction was found in the shoots cultured in the medium supplemented with 0.5 mg L–1 BA. The apperances of in vivo and in vitro flowers of P. lanceolata were the same in many aspects except that the number of flower/inflorescence formed was different. In addition, water accumulation was observed only inside the in vitro flowers. Water deposit in the long tubular structure of P. lanceolata flower could cause anther injury, suggesting that flowers developed in vitro may not always produce pollen.

Embryo induction and plant regeneration of Callerya speciosa (Fabaceae) through anther culture

2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li
Keyword(s):  
Anther Culture ◽  
Culture Medium ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Anther Wall ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.

scholarly journals Fast protocol for high frequency in vitro cloning of Banana (Musa acuminata) cv. Grande Naine

2017 ◽  
Vol 9 (1) ◽  
pp. 72-79
Author(s):  
S. R. Parida ◽  
S. Beura ◽  
S. Rout ◽  
R. Beura ◽  
P. N. Jagadev
Keyword(s):  
High Frequency ◽  
Basal Medium ◽  
Shoot Multiplication ◽  
Musa Acuminata ◽  
Future Research ◽  
Naphthaleneacetic Acid ◽  
Maximum Height ◽  
Ms Medium ◽  
Number Of Leaves

An investigation was conducted on Fast Protocol for High Frequency in vitro cloning of Banana (Musa acuminata) cv. Grande Naine at the Biotechnology-cum-Tissue Culture Center, OUAT, Bhubaneswar, during the year 2012. This has helped to determine the best media compositions for shoot multiplication and rooting of cv. Grande Naine, so as to get optimum results with a minimized cost of production. MS medium supplemented with 4.0 mg/1 Benzylaminopurine (BAP) and 2.0 mg/1 Kinetin gave the highest number of shoot/explants (11.33) in 30 days. However, MS medium when supplemented with 6.0 mg/1 BAP produced a maximum number of leaves (19.07) with a maximum height 2.73 cm. Among various concentrations of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and naphthaleneacetic acid (NAA) for rooting. Half MS medium supplemented with 1.0 mg/1 IBA was found to be ideal for early rooting and producing more number of roots in 21 days. However, MS basal medium was found to be the best treatment to support the formation of long roots. This protocol can be very useful to the future research worker and as well as entrepreneurs for mass production of banana (Musa acuminata) cv. Grande Naine.

scholarly journals Kultur akar rambut Cinchona ledgeriana dan C. succirubra dalam kultur in vitro Hairy root culture of Cinchona ledgeriana and C. succirubra by in vitro culture

2016 ◽  
Vol 72 (2) ◽  
Author(s):  
Nurita TORUAN-MATHIUS ◽  
. REFLINI ◽  
. NURHAIMI-HARIS ◽  
. JOKO-SANTOSO ◽  
A PRIANGANI-ROSWIEM
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Basal Medium ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Chain Reaction ◽  
Ri Plasmid ◽  
Polymerase Chain ◽  
Cinchona Ledgeriana

Summary Problems encountered in hairy root culture  of  C. ledgeriana and C. succirubra are low percentage of transformation of explants by Agrobacterium rhizogenes and slow growth of hairy root. The objective of this research was to evaluate the potential of several A. rhizogenes strains for initiation  hairy roots of  C. succirubra and C. ledgeriana, and to obtain the best medium for hairy root culture of Cinchona spesies. Axenic shoot and leaves explants of eight-month-old of C. ledgeriana and  C. succirubra seedlings were inoculated with A. rhizogenes strain ATCC-15834, ATCC-8196,    R-20001, 07-20001, A4, R-MAFFA, TISTR509, TISTR510 and LBA9457. Inoculated explants were cultured in solid MS medium with the addition of 100 mg/L amphicylin. Subculture of the hairy root was performed by transferred of root pieces into fresh liquid basal medium MS, B5, White and Heller. Hairy roots from the best of basal medium were subcultured on the same medium with the addition of 50  and 100 mg/L   L-tryptophane, three or five times concentration of MS vitamins. The integration of T-DNA of   A. rhizogenes in hairy root was confirmed with specific primer for TL and TR-DNA of plasmid by Polymerase Chain Reaction analysis. The results showed that only A. rhizogenes strain  LBA 9457 were effective for  transformation of explants from both Cinchona species. The fastest hairy roots growth were found  in MS medium, while growth in others medium was poor. Hairy roots of  C. ledgeriana has vigor and growth better than hairy roots of C. succirubra. MS with the addition of 50 mg/L  L-tryptophane and  three times the concen-trations of vitamin  is the best medium for hairy root growth and vigor. Hairy roots of  C. succirubra and C. ledgeriana used in this studies were confirmed that hairy roots  contained TL and TR-DNA region of Ri plasmid with molecular weight 780 and 1600 bp.  The results showed that strain of A. rhizogenes, plant species, source of explant and composition of medium affect the initiation, growth, development  and vigor of hairy roots.Ringkasan Masalah dalam kultur akar rambut  C. ledgeriana dan C. succirubra adalah rendahnya tingkat keberhasilan transformasi eksplan dengan Agrobacterium rhizogenesdan pertumbuhannya yang lambat. Penelitian ini bertujuan untuk mengevaluasi  potensi dari beberapa galur A. Rhizogenes untuk inisiasi, mendapatkan komposisi medium terbaik untuk pertumbuhan akar rambut C. ledgeriana dan C. succirubra, serta konfirmasi terintegrasinya TR dan TL-DNA Ri plasmid ke dalam jaringan eksplan.  Eksplan batang  dan  daun  berasal  dari kecambah aksenik C. ledgeriana dan C. succirubra berumur delapan bulan diinokulasi dengan A. rhizogenes galur 15834, 8196, R-20001, 07-20001, A4, R.MAFFA,TISTR 509, TISTR 510 dan LBA 9457. Eksplan yang sudah diinokulasi dikulturkan dalam medium MS padat. Subkultur dilakukan dengan cara mentransfer potongan ujung akar rambut ke dalam medium cair MS, B5, White dan Heller. Akar rambut dari medium kultur yang terbaik kemudian disubkultur ke dalam medium yang sama dengan penambahan 50 dan 100 mg/L L-triptofan dengan konsentrasi vitamin sebanyak tiga kali dan lima kali dari konsentrasi normal MS. Integrasi T-DNA dalam akar rambut dikonfirmasi meng-gunakan Polymerase Chain Reaction  dengan primer spesifik untuk TL dan TR-DNA plasmid. Hasil yang diperoleh menunjukkan bahwa hanya A.rhizogenes galur LB9457 yang efektif menginfeksi eksplan baik batang maupun daun dari kedua spesies kina. Induksi, pertumbuhan dan vigor akar rambut yang terbaik diperoleh dari medium MS dengan penambahan 50 mg/L L-triptofan dan tiga kali konsentrasi vitamin. Hasil konfirmasi akar rambut baik dari batang maupun daun menggunakan PCR, menunjukkan bahwa TL dan TR-DNA dari Ri plasmid  A. rhizogenes mampu menghasilkan pita-pita DNA dengan BM780 dan 1600 pb. Hasil yang diperoleh menunjukkan bahwa galur  A. rhizogenes, spesies tanaman, sumber eksplan dan komposisi medium berpengaruh terhadap inisiasi, pertumbuhan,  perkembangan dan vigor akar rambut.

scholarly journals Plant Regeneration from Leaf Explants of the Medicinal Herb Wedelia chinensis

Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To
Keyword(s):  
Plant Regeneration ◽  
In Vitro Propagation ◽  
Liver Toxicity ◽  
Leaf Explants ◽  
Basal Medium ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Average Percentage

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.

scholarly journals Polyphenols profiles and antioxidant activities of extracts from Capsicum chinense in vitro plants and callus cultures

2019 ◽  
Vol 2 (1) ◽  
pp. 30 ◽  
Author(s):  
Gergana Sherova ◽  
Atanas Pavlov ◽  
Vasil Georgiev
Keyword(s):  
Callus Culture ◽  
Antioxidant Activities ◽  
Sinapic Acid ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Capsicum Chinense ◽  
Practical Applications ◽  
In Vitro Plants

In vitro plants of Capsicum chinense cv. Carolina Reaper have been obtained by planting seeds on sterile MS medium. The plants were used to initiate callus culture on half strength MS medium, supplied with 2 mg/l 1-Naphthaleneacetic acid and 0.5 mg/L 6-Benzylaminopurine. The polyphenol profiles (phenolic acids and flavonoids) of methanol extracts from obtained callus and in vitro plants have been analyzed by HPLC. The main constituents in plant extract were protocatechuic acid, sinapic acid, rutin, hesperetin and myricetin, whereas in callus extract the major compounds were found to be chlorogenic acid, ferulic acid, rutin, hyperoside, myricetin and hesperetin. The antioxidant capacity of both extracts have been evaluated by using DPPH, TEAC, FRAP and CUPRAC assays. In our knowledge, this is the first report for obtaining of callus culture from Capsicum chinense cv. Carolina Reaper and evaluation of phytochemical profiles and antioxidant activities of its extract. Practical applications: The research outlines the potential of Capsicum in vitro systems as a renewable source of active ingredients for application in cosmetic and food products.

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