scholarly journals High efficient de novo root-to-shoot organogenesis in Citrus jambhiri Lush.: Gene expression, genetic stability and virus indexing

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0246971
Author(s):  
Tongbram Roshni Devi ◽  
Madhumita Dasgupta ◽  
Manas Ranjan Sahoo ◽  
Paresh Chandra Kole ◽  
Narendra Prakash
Keyword(s):  
High Frequency ◽  
De Novo ◽  
Random Amplified Polymorphic Dna ◽  
Basal Medium ◽  
Direct Regeneration ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Citrus Jambhiri ◽  
High Efficient ◽  
Root Explant

A protocol for high-frequency direct organogenesis from root explants of Kachai lemon (Citrus jambhiri Lush.) was developed. Full-length roots (~3 cm) were isolated from the in vitro grown seedlings and cultured on Murashige and Skoog basal medium supplemented with Nitsch vitamin (MSN) with different concentrations of cytokinin [6-benzylaminopurine, (BAP)] and gibberellic acid (GA3). The frequency of multiple shoot proliferation was very high, with an average of 34.3 shoots per root explant when inoculated on the MSN medium supplemented with BAP (1.0 mg L–1) and GA3 (1.0 mg L–1). Optimal rooting was induced in the plantlets under half strength MSN medium supplemented with indole-3-acetic acid (IAA, 0.5–1.0 mg L–1). IAA induced better root structure than 1-naphthaleneacetic acid (NAA), which was evident from the scanning electron microscopy (SEM). The expressions of growth regulating factor genes (GRF1 and GRF5) and GA3 signaling genes (GA2OX1 and KO1) were elevated in the regenerants obtained from MSN+BAP (1.0 mg L-1)+GA3 (1.0 mg L-1). The expressions of auxin regulating genes were high in roots obtained in ½ MSN+IAA 1.0 mg L-1. Furthermore, indexing of the regenerants confirmed that there was no amplicons detected for Huanglongbing bacterium and Citrus tristeza virus. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers detected no polymorphic bands amongst the regenerated plants. This is the first report that describes direct organogenesis from the root explant of Citrus jambhiri Lush. The high-frequency direct regeneration protocol in the present study provides an enormous significance in Citrus organogenesis, its commercial cultivation and genetic conservation.

Micropropagation and assessment of genetic stability in Celastrus paniculatus: An endangered medicinal plant

Biologia ◽  
2013 ◽  
Vol 68 (4) ◽  
Author(s):  
Sunil Senapati ◽  
Subhashree Aparajita ◽  
Gyana Rout
Keyword(s):  
Medicinal Plant ◽  
Genetic Stability ◽  
In Vitro Regeneration ◽  
Random Amplified Polymorphic Dna ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Endangered Medicinal Plant ◽  
Celastrus Paniculatus

AbstractA highly efficient protocol for in vitro regeneration of an indigenous, endangered medicinal plant Celastrus paniculatus was achieved using nodal explants. Murashige and Skoog (MS) basal medium supplemented with 0.5 mg/L 6-benzylaminopurine (BAP) and 0.1 mg/L naphthaleneacetic acid (NAA) showed maximum percentage of shoot multiplication (83.4%) with 8.2 shoots/explants. Maximum rooting of 73.3% with 4.8 roots/shoot was achieved on half-strength MS media supplemented with 0.5 mg/L indole-3-acetic acid (IAA) and the percentage of survival was 91% after acclimatization. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) marker study confirmed genetic stability for in vitro raised explants by showing 100% monomorphism. High multiplication rate associated with genetic stability ensure the efficacy of the present in vitro clonal propagation protocol of this important medicinal plant species.

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 New Basal Media for Protocorm-Like Body and Callus Induction of Hybrid Cymbidium

2012 ◽  
Vol 20 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Jaime A. Teixeira da Silva
Keyword(s):  
Callus Induction ◽  
High Frequency ◽  
Callus Formation ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Control Medium ◽  
Industry Standard ◽  
Basal Media ◽  
Bacto Agar

Abstract High frequency protocorm-like body (PLB) production from hybrid Cymbidium Twilight Moon ‘Day Light’ has been developed through a new medium, Teixeira Cymbidium (TC) medium. Two new TC media containing variable amounts of macroand micronutrients and other additives, inspired by Winarto and Teixeira (WT) medium for Anthurium and Murashige and Skoog (MS) basal medium were used to induce PLBs and callus. Control medium was research- and industry-standard Vacin and Went (VW) medium. The first TC medium, TCPLB, could induce significantly more PLBs than on VW while high levels of macronutrients in the second TC medium, TCCALLUS, and MS were required to induce callus. All PLB induction media contained 0.1 mg/l α-naphthaleneacetic acid (NAA) and 0.1 mg/l kinetin (KIN), 2 g/l tryptone and 20 g/l sucrose, and solidified with 8 g/l Bacto agar while callusinduction media were identical, except that KIN was substituted by thidiazuron (TDZ). Basal medium had a significant effect on PLB and callus formation. This protocol could be used to induce PLBs and callus from other Cymbidium species or cultivars.

scholarly journals Agrobacterium Mediated Transformation Optimizations for Sugarcane (Saccharum Officinarum L.) Cultivar SPF-234 with Direct Organogenesis

2021 ◽  
Vol 8 (2) ◽  
pp. 61-71
Author(s):  
Muhammad Nawaz ◽  
Naeem Iqbal ◽  
Rabia Hameed ◽  
Mehwish Mehwish ◽  
Shakra Jamil
Keyword(s):  
Shoot Elongation ◽  
Saccharum Officinarum ◽  
Energy Crop ◽  
Direct Regeneration ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Equal Concentration ◽  
Sugarcane Cultivar ◽  
Agrobacterium Mediated Transformation

Sugarcane (Saccharum officinarum L.) is the most important food and energy crop worldwide. In the present study, an efficient Agrobacterium mediated transformation and regeneration system for sugarcane cultivar SPF-234 was established. Agrobacterium tumefaciens strains EHA101and LBA4404 using vector pIG121 Hm, having GUS, HPTII and NPTII genes were used. Polymerase chain reaction (PCR) and histochemical assays confirmed the GUS gene expression. A 620 bp fragment from GUS positive plants was amplified. The GUS expressing putative transformants were 35% of the total plants formed under 30 minute immersion time and 72 hr of incubation period. The co-cultivation media having 60 µM acetosyringone produced 66% GUS expressing plants for LBA4404 and 58% for EHA101. The maximum average number of directly produced shoot (59.5%) from leaf explant was in M6 media having 1.00 mg/l 6-Benzylaminopurine (BAP) and 2.5 mg/l Naphthaleneacetic acid (NAA). A significant decrease (17%) was observed when auxin (NAA) concentration was increased to 4.0 mg/l. The best response of shoot elongation was observed in SE4 media having equal concentration (2.00 mg/l) of both kinetin and BAP. Increased concentrations of kinetin significantly decreased shoot elongation of the subject cultivar. Agrobacterium strain LBA4404 performed better for genetic transformation of the said sugarcane cultivar.This quick and less expensive transformation and direct regeneration system could be exploited for sugarcane on commercial scale in general, and for this elite cultivar in particular.

scholarly journals Optimizing Turmeric Tissue Culture, Testing Different Media and a Plant Growth Regulator Matrix

2021 ◽  
pp. 1-13
Author(s):  
Toktam Taghavi ◽  
Alireza Rahemi ◽  
Reza Rafie ◽  
Maru K. Kering
Keyword(s):  
Tissue Culture ◽  
Plant Growth ◽  
Growth Regulator ◽  
Plant Growth Regulator ◽  
Woody Plant ◽  
Curcuma Longa ◽  
Basal Medium ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate

Rapid multiplication of turmeric (Curcuma longa) by micropropagation is needed to produce a continuous source of uniformly sized, high-quality, and disease-free plantlets. Three in vitro experiments were conducted to optimize the medium by evaluating nine media and a full factorial combination (matrix) of two plant growth regulators for direct organogenesis of ‘Hawaiian Red’ turmeric. Two experiments evaluated the media, and the third studied the plant growth regulator matrix. As a result, Driver and Kuniyuki walnut (DKW), Murashige and Skoog (MS), and broadleaf tree basal (BLT) media performed better than woody plant media [Lloyd & McCown woody plant basal medium (L&M), and McCown’s woody plant basal salt mixture (McCown)] for shoot and root formation. The multiplication rate was 18 plants per explant in DKW with 1 mg⋅L−1 6-benzylaminopurine (BAP) and 0.1 mg⋅L−1 1-naphthaleneacetic acid (NAA). After transferring the plants to an ex vitro environment, the survival rate was 97%, and 30% higher than previously reported. DKW produced the highest number of plantlets (with shoots and roots), and BLT produced fewer plants with higher biomass. In the MS media, higher BAP to NAA ratio (2.5 to 0.1 mg⋅L−1) produced the most significant number of shoots; however, the lowest concentration of BAP and NAA (0.1 mg⋅L−1 of both) produced the highest number of rooted plantlets. There are two recommendations for tissue culture of ‘Hawaiian Red’ turmeric. To produce the highest number of plantlets, one should use the higher BAP to NAA ratio (2.5 mg⋅L−1 BAP and 0.1 mg⋅L−1 NAA) for shoot proliferation and then transfer the explants to the root initiation media. However, to reduce the number of subcultures, the explants can be grown in the lowest concentration of both BAP and NAA (0.1 mg⋅L−1) to induce both shoot and root. Although, the number of plantlets (with roots and shoots) will decrease in this method, there is no need for subsequent subcultures and changing of the plant growth regulator combinations.

scholarly journals High Frequency Direct Organogenesis, Genetic Homogeneity, Chemical Characterization and Leaf Ultra-Structural Study of Regenerants in Diplocyclos palmatus (L.) C. Jeffrey

Agronomy ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 2164
Author(s):  
Anamica Upadhyay ◽  
Anwar Shahzad ◽  
Zishan Ahmad ◽  
Abdulrahman A. Alatar ◽  
Gea Guerriero ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
In Vitro Propagation ◽  
Large Scale ◽  
Basal Medium ◽  
Wild Plants ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Scale Production ◽  
Large Scale Production

Diplocyclos palmatus (L.) C. Jeffrey, commonly referred to as “Shivalingi” or “Lollipop climber” is a valuable medicinal plant with a climbing growth habit used in traditional medicine. It is reputed to have antiarthritic, anti-diabetic properties and to be useful in various skin and reproductive problems. Overexploitation of wild plants and low seed germination have resulted in the decline of the species in the wild. Thus, the present investigation was aimed to establish an effective in vitro propagation procedure for its large-scale production and conservation. Nodal explants, obtained from an established mother plant were grown on MS basal medium augmented with various cytokinins, alone or in combination with auxins, to study the morphogenic response. A maximum of 8.3 shoots/explants with an average shoot length of 7.2 cm were produced after six weeks on MS containing benzylaminopurine 5.0 µM + 1-naphthaleneacetic acid 2.0 µM. After 4 weeks of transfer, microshoots rooted well on a low nutrient medium of ½ MS + 1.0 µM indole-3-butyric acid, with a maximum of 11.0 roots/microshoot and an average root length of 7.4 cm. With an 80% survival rate, the regenerated plantlets were effectively acclimatized to natural conditions. DNA-based molecular markers were used to investigate the genetic uniformity. Scanning Electron Microscopic examination of leaves indicated the adaptation of the plantlets to natural, as evidenced by the formation of normal stomata. Gas chromatography-mass spectrometry analyses of mother and micropropagated plants were performed to identify essential secondary metabolites. The results obtained show that the in vitro propagation system can be adopted for preservation, large-scale production and secondary metabolites’ production in D. palmatus.

Plant regeneration from sugar beet leaf protoplasts: analysis of shoots by DNA fingerprinting and restriction fragment length polymorphism

2000 ◽  
Vol 78 (1) ◽  
pp. 10-18 ◽  
Author(s):  
E Jazdzewska ◽  
Z Sadoch ◽  
A Niklas ◽  
A Majewska-Sawka
Keyword(s):  
Sugar Beet ◽  
Beta Vulgaris ◽  
Fragment Length Polymorphism ◽  
Random Amplified Polymorphic Dna ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Chromosome Counting ◽  
Male Sterile ◽  
Somaclonal Variant ◽  
Leaf Protoplasts

Shoots were regenerated from leaf protoplasts of cytoplasmic male sterile and male fertile diploid sugar beet (Beta vulgaris L.) genotypes. Protoplasts cultured in Murashige and Skoog medium supplemented with 5 µM naphthaleneacetic acid, 2 µM 6-benzylaminopurine, 100 µM n-propyl gallate, and diamine putrescine at concentrations of 50, 100, or 500 µM were able to synthesize a new cell wall and entered successive mitotic divisions leading to the formation of callus colonies. Shoots were obtained via organogenesis by continuous culture of calli on the same basal medium supplemented with either cytokinin alone, or with a combination of cytokinin and auxin. The regenerants of both lines were characterized with regard to ploidy, and the regenerants of the male sterile line were further characterized with regard to possible somaclonal variation and organization of two mitochondrial genes: atpA and atp6. Chromosome counting revealed that tetra-, hexa-, and octa-ploids were present among regenerants. Random amplified polymorphic DNA (RAPD) analysis identified one somaclonal variant among 31 shoots tested, whereas hybridization with both mitochondrial probes showed no notable changes in the organization of mtDNA within these loci.Key words: Beta vulgaris L., protoplasts, regeneration, random amplified polymorphic DNA (RAPD), atpA, atp6.

scholarly journals In vitro Regeneration of Alstroemeria cv. ‘Balance’ Based On Direct Organogenesis

2017 ◽  
Vol 14 (2) ◽  
pp. 557-566
Author(s):  
Maryam Beigi Harchegani ◽  
Hossein Nazarian ◽  
Mahmoud Otroshy ◽  
Mohammad Ali Ebrahimi ◽  
Ali Motamedi
Keyword(s):  
Shoot Regeneration ◽  
Culture Medium ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Direct Regeneration ◽  
Direct Organogenesis ◽  
Root Induction ◽  
Factorial Experiments ◽  
Apical Bud

ABSTRACT: The present study aimed at improving the efficiency of the in vitro regeneration of Alstroemeria cv. ‘Balance’ protocol through direct organogenesis technique using three different origins of explants (nodal stem, rhizome apical bud, rhizome segments) as two separate factorial experiments with completely randomized design with six replications which were implemented in three stages. Firstly, Direct regeneration, including two factorial experiments to induce regeneration in organogenesis media by utilizing the four NAA concentrations (0, 0.5, 1, 2 mg/l) combined with five BAP concentrations (0, 0.5, 1, 1.5, 2 mg/l) in the first experiment, and TDZ concentrations (0, 0.5, 10 mg/l) in combination with three IBA concentrations (0, 1, 2 mg/l) in the second experiment. Secondly, shoot regeneration and elongation of stems in regenerated buds in the MS basal medium and finally, rooting of the regenerated shoots in the root induction mediums by NAA and IBA. Results of regeneration experiment revealed that, in the culture medium containing 10 mg/l TDZ in combination with 2 mg/l IBA and rhizome apical bud explants, the maximum rates of regeneration percentage, the highest number and length of the shoots were produced. Resulted shoots produced the greatest number of roots and rhizomes in the culture medium comprising 1 mg/l NAA in combination with 2 mg/l IBA as auxins. From the two tested explants, rhizome apical bud, was known as the best explants for shoot regeneration obtained plantlets successfully adapted to environmental conditions and were transferred to the greenhouse. Results of current study indicated that Alstroemeria can be produced through direct organogenesis.

scholarly journals FAST DIRECT REGENERATION OF PLANTS FROM NODAL EXPLANTS OF Stevia rebaudiana Bert.

2019 ◽  
Vol 18 (5) ◽  
pp. 95-103
Author(s):  
Romuald Doliński ◽  
Krzysztof Kowalczyk
Keyword(s):  
Basal Medium ◽  
Stevia Rebaudiana ◽  
Production Costs ◽  
Nodal Explants ◽  
Direct Regeneration ◽  
Naphthaleneacetic Acid ◽  
Production Scale ◽  
Plant Weight ◽  
The Media

In the preceding research, stevia has been typically cloned in vitro using two media, on which the shoots were formed (3–6 weeks), and on the other they were rooted (3–5 weeks). This study aimed at finding the possibility for rapid stevia propagation from large nodal explants using the MS basal medium [Murashige and Skoog 1962], with low auxin concentrations (0.5, 1 and 2 mg‧dm–3). The plants were obtained as soon as after three weeks. The best results were obtained from media with various concentrations of the indole-3-acetic acid (IAA) and the highest concentration of phenylacetic acid (PAA). Plants were formed by 83.9-86.0% of explants, they had high weight (234−253 mg), two shoots measuring 2.07−2.37 cm and 5.8−8.3 roots measuring 1.00–1.24 cm. Mean plant weight was lowest on the media with indole-3-butyric acid (IBA) (185–192 mg). Both explant buds formed single shoots, but their development was typically uneven. The differences in the length and weight of shoots were lowest on the media with IAA and at lower PAA concentrations. Plants from the media with IAA and the control medium were distinguished by a higher number of nodes. The percentage share of shoots in the total plant weight was highest on the media with PAA (62.1–62.7%), and lowest at higher concentrations of α-naphthaleneacetic acid (NAA) (47.9 and 48.9%). Parts of explants immersed in media developed callus, and the highest amounts of this tissue were found in the media with NAA. 92.3% of plants survived the acclimatization. The applied procedure may be used for rapid in vitro cloning of selected stevia genotypes. The use of one medium enables reduction of seedling production costs. Moreover, cyclical cloning and extending the production scale is possible.

RUNX1 DNA-binding mutations and RUNX1-PRDM16 cryptic fusions in BCR-ABL+ leukemias are frequently associated with secondary trisomy 21 and may contribute to clonal evolution and imatinib resistance

Blood ◽  
2008 ◽  
Vol 111 (7) ◽  
pp. 3735-3741 ◽  
Author(s):  
Catherine Roche-Lestienne ◽  
Lauréline Deluche ◽  
Sélim Corm ◽  
Isabelle Tigaud ◽  
Sami Joha ◽  
...  
Keyword(s):  
Dna Binding ◽  
High Frequency ◽  
Trisomy 21 ◽  
De Novo ◽  
Blast Crisis ◽  
Clonal Evolution ◽  
Chronic Phase ◽  
Chromosome 21 ◽  
Imatinib Resistance ◽  
Molecular Abnormalities

Abstract Acquired molecular abnormalities (mutations or chromosomal translocations) of the RUNX1 transcription factor gene are frequent in acute myeloblastic leukemias (AMLs) and in therapy-related myelodysplastic syndromes, but rarely in acute lymphoblastic leukemias (ALLs) and chronic myelogenous leukemias (CMLs). Among 18 BCR-ABL+ leukemias presenting acquired trisomy of chromosome 21, we report a high frequency (33%) of recurrent point mutations (4 in myeloid blast crisis [BC] CML and one in chronic phase CML) within the DNA-binding region of RUNX1. We did not found any mutation in de novo BCR-ABL+ ALLs or lymphoid BC CML. Emergence of the RUNX1 mutations was detected at diagnosis or before the acquisition of trisomy 21 during disease progression. In addition, we also report a high frequency of cryptic chromosomal RUNX1 translocation to a novel recently described gene partner, PRDM16 on chromosome 1p36, for 3 (21.4%) of 14 investigated patients: 2 myeloid BC CMLs and, for the first time, 1 therapy-related BCR-ABL+ ALL. Two patients presented both RUNX1 mutations and RUNX1-PRDM16 fusion. These events are associated with a short survival and support the concept of a cooperative effect of BCR-ABL with molecular RUNX1 abnormalities on the differentiation arrest phenotype observed during progression of CML and in BCR-ABL+ ALL.

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