scholarly journals Regeneration and Transformation of Taro (Colocasia esculenta) with a Rice Chitinase Gene Enhances Resistance to Sclerotium rolfsii

HortScience ◽  
2010 ◽  
Vol 45 (7) ◽  
pp. 1014-1020 ◽  
Author(s):  
Xiaoling He ◽  
Susan C. Miyasaka ◽  
Yi Zou ◽  
Maureen M.M. Fitch ◽  
Yun J. Zhu
Keyword(s):  
Transgenic Line ◽  
Sclerotium Rolfsii ◽  
Multiple Shoots ◽  
Chitinase Gene ◽  
Colocasia Esculenta ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Reverse Transcription Pcr ◽  
Rice Chitinase

Genetic engineering has the potential to improve disease resistance in taro [Colocasia esculenta (L.) Schott]. To develop a method to produce highly regenerable calluses of taro, more than 40 combinations of Murashige and Skoog (MS) media at full- or half-strength with varying concentrations of auxin [α-naphthaleneacetic acid (NAA) or 2, 4-dichlorophenoxyacetic acid (2, 4-D)], cytokinin [benzyladenine (BA) or kinetin], and taro extract were tested for callus initiation and plant regeneration. The best combination, MS medium with 2 mg·L−1 BA and 1 mg·L−1 NAA (M5 medium), was used to produce regenerable calluses from taro cv. Bun Long initiated from shoot tip explants. After 8 weeks of growth, multiple shoots from these calluses could be induced on MS medium with 4 mg·L−1 BA (M15 medium). The rice chitinase gene (ricchi11) along with the neomycin phosphotransferase (npt II) selectable marker and β-glucuronidase (gus) genes were introduced into these taro calluses through particle bombardment. Transformed calluses were selected on M5 medium containing 50 mg·L−1 geneticin (G418). Histochemical assays for beta-glucuronidase (GUS), polymerase chain reaction (PCR), reverse transcription–PCR, and Southern blot analyses confirmed the presence, integration, and expression of the rice chitinase gene in one transgenic line (efficiency less than 0.1%). Growth and morphology of the transgenic plants appeared normal and similar to non-transformed controls. In pathogenicity tests, the transgenic line exhibited improved resistance to the fungal pathogen, Sclerotium rolfsii, but not to the oomycete pathogen, Phytophthora colocasiae.

scholarly journals Shoot Proliferation, Callus Induction And Plant Regeneration In Tripsacum Laxum Nash

2021 ◽  
Author(s):  
Yuping Xiong ◽  
Jinhui Pang ◽  
Kunlin Wu ◽  
Jaime A. Teixeira Silva ◽  
Xinhua Zhang ◽  
...  
Keyword(s):  
Peat Soil ◽  
Shoot Proliferation ◽  
Multiple Shoots ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Rooting Medium ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract The peduncles of Tripsacum laxum Nash were used as explants to induce axillary shoots. Multiple shoots were proliferated on Murashige and Skoog (MS) medium to establish, for the first time, efficient shoot proliferation and plant in vitro regeneration systems. Optimal shoot proliferation medium was MS with 3.0 mg/L 6-benzyladenine (BA) and 0.2 mg/L α-naphthaleneacetic acid (NAA), resulting in a shoot proliferation coefficient of 11.0 within 45 d. Optimal rooting medium was MS with 0.1 mg/L NAA and/or 0.1 mg/L indole-3-butyric acid (IBA), inducing 100% root formation from shoots within 30 d. When young roots, leaf sheaths and shoot bases were used as explants, MS medium with 1.0 mg/L thidiazuron (TDZ) and 0.2 mg/L BA induced most shoots, with the least callus. Shoot bases induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), while leaf sheaths induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L BA. Rooted plantlets showed 99.3% survival when transplanted into a substrate of vermiculite: peat soil (1:3, v/v).

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 Micropropagation, Callus Induction and Regeneration of Ginger (Zingiber officinale Rosc.)

2020 ◽  
Vol 5 (1) ◽  
pp. 75-84
Author(s):  
Seied Mehdi Miri
Keyword(s):  
Acetic Acid ◽  
Callus Induction ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Zingiber Officinale ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Shoot Number ◽  
Micro Propagation ◽  
Dichlorophenoxy Acetic Acid

AbstractThe present study describes a protocol for micro-propagation, callus induction, and shoot regeneration of ginger (Zingiber officinale). The rhizomes were surface-sterilized with ethanol (70%) for 45 s, sodium hypochlorite (2.5%) for 10 min, and mercuric chloride (0.1%) for 10 min. Multiple shoots were induced from sprouting bud explants cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA) combined with kinetin (Kin). The maximum shoot number was obtained from MS medium containing 10 mg/l BA with a mean of 20.6 shoots per explant. The leaf explants were cultured on MS medium supplemented with indole-3-acetic acid (IAA), naphthaleneacetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), Dicamba, or BA for callus culture. Green-red compact calli were induced using 2,4-D, Dicamba or BA. Also, BA successfully induced plant regeneration. The multiplied shoots that were transferred to the rooting medium (½MS supplemented with 0, 1 and 2 mg/l IAA, indole-3-butyric acid (IBA) or NAA) showed development of roots (100%). The rooted plantlets were transferred to pots containing a 1:1 mixture of cocopeat and perlite, and acclimatization was successful, resulting in 85% survival of the plantlets in the greenhouse.

scholarly journals The Effects of 2,4-Dichlorophenoxyacetic Acid and α-Naphthaleneacetic Acid on Biomass Increment, Rhizogenesis and Somatic Embryogenesis of Suspension-cultured Dinh Lang Cells [Polyscias fruticosa (L.) Harms]

2021 ◽  
Author(s):  
T.T.B. Phuong ◽  
V.P. Trung ◽  
N.H. An ◽  
N.D. Tuan ◽  
P.T.T. Nguyen
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Bioactive Compound ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Triterpenoid Saponins ◽  
Cell Biomass ◽  
2,4 Dichlorophenoxyacetic Acid

Background: Dinh Lang [Polyscias fruticosa (L.) Harms] is a medicinal plant widely grown in Vietnam, with proven note-worthy health benefits. However, Dinh Lang’s amounts of triterpenoid saponins could not meet the need of the pharmaceutical industry. Thus, this study’s purpose is to figure out the optimal condition for raising Dinh Lang’s cell biomass, rhizogenesis and somatic embryogenesis to provide materials for bioactive compound productions. Methods: Different 2,4-dichlorophenoxyacetic acid and α-naphthaleneacetic acid concentrations (0.5, 1.0, 1.5 and 2.0 mg/L) were examined to determine the best amount of each plant growth regulator for raising cells’ biomass, rhizogenesis and somatic embryogenesis. In each treatment, two grams of eight-week-old calli were cultured in 50 mL of liquid MS medium. Result: It is demonstrated by the results that liquid MS medium containing 1.5 mg/L α-naphthaleneacetic acid has the capacity of producing the highest numbers of somatic embryos (489 embryos per flask) and rooted cells (259.5 cells per flask), while the fresh weight of cells cultured in the medium given 1.5 mg/L 2,4-dichlorophenoxyacetic acid reached its peak of 5.7 g.

scholarly journals ENHANCED REGENERATION OF SHOOTS FROM PEACH CELLS INFECTED WITH A SHOOTY MUTANT STRAIN OF AGROBACTERIUM.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1150d-1150
Author(s):  
A. Smigocki ◽  
F. Hammerschlag
Keyword(s):  
Prunus Persica ◽  
Northern Analysis ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Gene Transcripts ◽  
Half Strength ◽  
Low Levels ◽  
2,4 Dichlorophenoxyacetic Acid

Immature `Redhaven' peach (Prunus persica L. Batsch) embryos were infected with Agrobacterium tumefaciens strain tms328::Tn5 carrying the functional cytokinin gene. Shoots were regenerated from callus grown on MS medium without added phytohormones and subsequently rooted on half-strength MS medium with 2.8 -naphthaleneacetic acid. These plants exhibited an increased frequency of branching in vitro. Low levels of cytokinin gene transcripts were detected in these cells by Northern analysis, and using an ELISA assay, the cytokinins zeatin and zeatinriboside were determined to be on the average 30-fold higher. From these results, the expression of the cytokinin gene appears to promote growth of cells in the absence of phytohormones thus serving as a marker for transformation and a promoter of morphogenesis without a 2,4-dichlorophenoxyacetic acid inductive step.

scholarly journals Regeneration of Hemidesmus indicus (L.) R. Br. using in vitro nodes: an alternative method for efficient multiplication of shoots

2021 ◽  
Vol 13 (2) ◽  
pp. 10831
Author(s):  
Ashutosh R. PATHAK ◽  
Aruna G. JOSHI
Keyword(s):  
Multiple Shoots ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Nodal Explant ◽  
Mass Propagation ◽  
Hemidesmus Indicus ◽  
Full Strength ◽  
Alternative Method

In vivo nodes of Hemidesmus indicus (L.) R. Br. induced healthy multiple shoots with branching in our earlier studies and thus in the present study, potency of in vitro nodes to regenerate shoots was evaluated. In vitro nodes were excised from eight-week-old shoots and placed in Murashige and Skoog’s (MS) medium fortified with sucrose (3%) and different concentrations of 6-benzyladenine (BA) and kinetin (Kn). After eight weeks, optimum of 5.42 ± 0.36 shoots with 100% response were regenerated in medium supplemented with BA (10 µM) and Kn (5 µM). These healthy shoots were placed in full, half and quarter strengths of liquid MS medium fortified with sucrose (1%) and α-naphthaleneacetic acid (NAA, 1-25 µM) for rooting. Among all the strengths of MS medium, full strength MS medium having 8 µM NAA formed maximum of 3.42 ± 0.55 roots (91.67% response) within four weeks. The protocol is in continuation with earlier study and it was confirmed that a single in vivo nodal explant can regenerate around 385 healthy elongated shoots within 4 months, which will help in mass-propagation of the species.

scholarly journals IN VITRO PROPAGATION OF GUAVA (PSIDIUM GUAJAVA L.) FROM SEEDLING EXPLANTS

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 696e-696
Author(s):  
Yaseen Mohamed-Yaseen ◽  
Raymond J. Schnell ◽  
Robert J. Knight ◽  
T. L. Davenport
Keyword(s):  
Shoot Growth ◽  
Multiple Shoots ◽  
Psidium Guajava ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Indolebutyric Acid ◽  
Seedling Explants ◽  
Cultivated Species ◽  
Regenerated Plantlets

Guava (Psidium guajava L.) is an exceptional source of vitamin. C. It is also considered to be the most important cultivated species of the Myrtel family. Shoot tip and stem node were taken from seedling germinated in Murashige and Skoog medium (MS) and cultured in the same medium supplemented with 1-3mg/l benzylaminopurine (BA) and 0.1mg/l naphthaleneacetic acid (NAA) or 0.2-2mg/l thidiazuron (TDZ) and 0.1mg/l NAA. Multiple shoots (4-6) were obtained in 4-5 weeks from culture in 1-2mg/l BA and 0.1mg/l NAA, while TDZ caused abnormal shoot growth. Shoots were rooted successfully with 100% frequency in MS medium containing 2mg/l indolebutyric acid and further elongation of shoots was achieved in MS medium, supplemented with lg/l activated charcoal. Regenerated plantlets were successfully established in soil.

scholarly journals Production of callus mediated gynogenic haploids in winter squash (Cucurbita maxima Duch.) and pumpkin (Cucurbita moschata Duch.)

2018 ◽  
Vol 54 (No. 1) ◽  
pp. 9-16 ◽  
Author(s):  
E.S. Kurtar ◽  
A. Balkaya ◽  
M. Ozbakir Ozer
Keyword(s):  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Cucurbita Moschata ◽  
Cucurbita Maxima ◽  
Culture Time ◽  
Irradiated Pollen ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Winter Squash ◽  
Indole 3 Acetic Acid

Although haploids were successfully produced via irradiated pollen technique and anther culture in Cucurbita maxima and Cucurbita moschata, the haploidization efficiency is still low due to genotype dependence. Thus, as an alternative technique, the efficacy of the ovule culture was investigated. Ovules were extracted at different flowering time and then cultured on a solid MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), benzylaminopurine (BAP), thidiazuron (TDZ), and naphthaleneacetic acid (NAA) to induce callogenesis and plant regeneration. The gynogenic response was influenced by the combination of plant growth regulators, genotype and culture time. The medium containing of 4.0 mg/l BAP + 0.05 mg/l NAA + 0.1 mg/l TDZ provided the highest response at anthesis time. Plantlets were rooted and elongated on a solid MS medium supplemented with 0.01 mg/l indole-3-acetic acid (IAA) + 1.0 mg/l BAP. The ploidy observations of 122 plants revealed that 70 plants were haploid, 46 plants were diploid and the others were mixoploid.  

scholarly journals Somatic embryogenesis and plant regeneration from callus cultures of Cleome rosea Vahl

2010 ◽  
Vol 53 (3) ◽  
pp. 679-686 ◽  
Author(s):  
Claudia Simões ◽  
Norma Albarello ◽  
Cátia Henriques Callado ◽  
Tatiana Carvalho de Castro ◽  
Elisabeth Mansur
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Formation ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Fold Reduction ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Callus Formation

This paper describes a protocol for the efficient vegetative propagation of Cleome rosea by somatic embryogenesis. Leaf and stem explants from nursery-grown seedlings of C. rosea were cultivated on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA), a -naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram) or 2,4-dichlorophenoxyacetic acid (2,4-D). Nodular calli were produced from both explant types in the presence of 4.5 and 9.0 µM 2,4-D. Embryo development and maturation were achieved when calli from stem explants were transferred to media containing a ten-fold reduction of 2,4-D concentration initially used (0.45 and 0.90 µM). Leaf-derived calli did not form embryos with the same treatments. The highest frequency of embryogenic callus formation (85%) and number of embryo per callus (13.45 ± 2.8) were achieved during the first subculture on medium supplemented with 0.90 µM 2,4-D. Embryo conversion into plantlets was achieved following transfer to growth regulator-free MS medium solidified with 2 g.L-1 phytagel. An acclimatization rate of 53% was found three months after transfer to ex vitro conditions and the recovered plants presented a normal phenotypic aspect.

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