scholarly journals The explant developmental stage profoundly impacts small RNA-mediated regulation at the dedifferentiation step of maize somatic embryogenesis

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Vasti T. Juárez-González ◽  
Brenda A. López-Ruiz ◽  
Patricia Baldrich ◽  
Eduardo Luján-Soto ◽  
Blake C. Meyers ◽  
...  

Abstract Maize somatic embryogenesis (SE) requires the induction of embryogenic callus and establishment of proliferation before plant regeneration. The molecular mechanisms underlying callus embryogenic potential are not well understood. Here we explored the role of small RNAs (sRNAs) and the accumulation of their target transcripts in maize SE at the dedifferentiation step using VS-535 zygotic embryos collected at distinct developmental stages and displaying contrasting in vitro embryogenic potential and morphology. MicroRNAs (miRNAs), trans-acting siRNAs (tasiRNAs), heterochromatic siRNAs (hc-siRNAs) populations and their RNA targets were analyzed by high-throughput sequencing. Abundances of specific miRNAs, tasiRNAs and targets were validated by qRT-PCR. Unique accumulation patterns were found for immature embryo at 15 Days After Pollination (DAP) and for the callus induction from this explant, as compared to 23 DAP and mature embryos. miR156, miR164, miR166, tasiARFs and the 24 nt hc-siRNAs displayed the most strikingly different patterns between explants and during dedifferentiation. According to their role in auxin responses and developmental cues, we conclude that sRNA-target regulation operating within the 15 DAP immature embryo explant provides key molecular hints as to why this stage is relevant for callus induction with successful proliferation and plant regeneration.

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 461G-462
Author(s):  
S.N. Talhouk ◽  
M. Shmoury ◽  
R. Baalbaki ◽  
S. Khuri

Somatic embryogenesis offers a great potential for large-scale production of Cedrus libani, which is important not only as a forest tree, but also for the development of a timber industry. In an attempt to optimize conditions for embryogenic callus induction, we used zygotic embryos at different developmental stages as explants, compared different media, and used several hormone levels and combinations. Results indicated that post-cotyledonary immature embryos had highest induction efficiency. Four different media namely 1/2 MS, Durzan, Litvay's, and Von Arnold supplemented with similar hormone levels showed no significant difference in efficiency of callus induction. Induction frequencies of embryogenic callus from explants subjected to different hormone levels and combinations were dependent on the developmental stage of the explant.


2021 ◽  
Author(s):  
Thiago Sanches Ornellas ◽  
Yohan Fritsche ◽  
Edison Cardona Medina ◽  
Miguel Pedro Guerra

Abstract Bamboos are an important worldwide non-timber forest product with current rising interest due to their environmentally friendly applications. Besides the consolidated uses of the sweet shoots and culms for structural uses, Dendrocalamus asper is an imposing ornamental bamboo for horticulture. The present work aimed to establish in vitro calli culture and plant regeneration through somatic embryogenesis starting from young inflorescences of the giant bamboo, D. asper. Pre-anthesis inflorescences were collected, disinfested, and subjected to callus induction on MS basal medium supplemented by 0 µM, 9 µM, 18 µM, 27 µM, and 36 µM of 2,4-D in combination with 9 µM of 2-iP or 9 µM Kin. The different obtained calli types were characterized and subcultured in 0 µM, 4.5 µM, 9 µM, and 18 µM of 2,4-D in combination with 9 µM of both cytokinins for multiplication and differentiation. Additionally, the explant incision and its inoculation orientation onto culture media were tested for callus induction improvement. The 2,4-D was essential for callus induction, and its combination with both cytokinins resulted in embryogenic callus induction and further somatic embryos regeneration. The subsequent reduction of this auxin to 4.5 µM resulted in somatic embryo maturation. Somatic embryos transferred to a plant growth regulator-free medium resulted in plantlet conversion. The present work showed the feasibility of using inflorescences as explants and the efficiency of using the 2-iP in combination with 2,4-D to callus induction and in vitro bamboo plant regeneration through somatic embryogenesis.


2021 ◽  
Vol 42 (5) ◽  
pp. 1232-1238
Author(s):  
D.S. Sparjanbabu ◽  
◽  
P.N. Kumar ◽  
S.R.K. Motukuri ◽  
D. Ramajayam ◽  
...  

Aim: This study evaluated efficient culture media for the regeneration of elite material through somatic embryogenesis from oil palm zygotic embryos. Methodology: For callus induction, zygotic embryos of four elite genotypes (G1-264T, G2-238DX17P, G3-37DX17P and G4-237T) were cultured on three basal media (Y3, MS and N6) with different auxin 2 mg l-1 (Picloram, 2,4-D and Dicamba) combinations. Subculture was made every month for three passages. It evaluated various callus characters. The embryogenic calli from callus induction media were transferred to the embryo maturation medium and subcultured until the polyembryoids formed. For shoot and root formation, somatic embryo clumps were transferred into regeneration media. In-vitro plantlets with well-grown roots were hardened in pots for six weeks and assessed clonal fidelity using polymorphic SSR primers. Results: Among the treatments, calli from N6+2,4-D, Y3+2,4-D and N6+Picloram showed the highest embryogenic callus potential. G4-237T induced more embryogenic calli (32.982) among genotypes, which was on par with G1-264T (24.196). Embryogenic calli grown on N6 media with Dicamba showed the highest proliferation rate (1.141). After 60 days of culture on regeneration media, the highest number of plantlets per somatic embryogenic clump was obtained from G1-264T on N6 media supplemented with Dicamba. Interpretation: Culture media salt concentration showed a significant difference among media by causing perturbations of auxin flow during somatic embryogenesis affecting callus induction, proliferation and plantlet regeneration. This may be useful for standardizing the genotype-specific regeneration media in oil palm.


2015 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Kee Hwa Bae ◽  
Eui Soo Yoon

Lychnis cognate Maxim and Lychnis fulgens Fish. Ex Spreng are two valued ornamental plants in Korea. Soaking of seeds in GA3 solution remarkably promoted germination up to 60%, but the control (0 mg/l) was not effective (> 5%). To select an adequate temperature for seed germination, seeds, previously soaked in a 1000 mg/l GA3 for 24 hrs, were incubated at 15, 20, 25, and 30°C. Seed germination of over 20% was obtained at 15, 20, and 25°C, but only 10% at 30°C. These results indicate that the seeds of L. cognate and L. fulgens are in a such dormant state that they hardly germinate even by dormancy breaker (GA3) and low (15 ? 25°C) temperature treatment. The highest callus induction was observed in the leaf explants of the seedlings on MS containing specific concentrations of 3.0 mg/l BA and 1.0 mg/l NAA. The adventitious shoot was formed < 90% of calli on 1/2 WPM medium. The height of in vitro propagated plantlet was no different media used for regeneration. This in vitro propagation protocol should be useful for conservation of endangered and ornamental plant.Plant Tissue Cult. & Biotech. 25(1): 1-12, 2015 (June)


1970 ◽  
Vol 35 (2) ◽  
pp. 331-341 ◽  
Author(s):  
MA Sayem ◽  
M Maniruzzaman ◽  
SS Siddique ◽  
M Al-Amin

The experiment was conducted to investigate the performance of three different genotypes (BARI Sarisha-6, BARI Sarisha-8, and BARI Sarisha-11) in two different media viz., MS and B5 with different concentrations of phytohormone (2, 4-D) for callus induction from uninucleate stage anthers of Brassica and subsequent plant regeneration in MS media with different concentrations of phytohormone (BAP and NAA). Among the genotypes, BARI Sarisha-8 showed the best performance for all the parameters of callus induction. The performance of BARI Sarisha-6 was poor compared to others. Maximum rate of callus induction (%) was observed in MS + 0.5 mg/L 2, 4-D followed by B5 + 0.5 mg/L 2,4-D. The media combination MS + 1.0 mg/L BAP 0.3 mg/L 2,4-D showed the best performance for maintenance of calli. Significant variations were observed among the genotypes and media composition for shoot regeneration. Among the genotypes, BARI Sarisha-8 showed the best performance for shoot regeneration followed by BARJ Sarisha-l1. The genotype BARI Sarisha-8 produced higher percent of shoots/calli and required minimum days for shoot initiation. Higher percent calli without shoot were produced by the genotype BARI Sarisha-6. The media combination MS + 2.0 mg/L BAP + 0.5 mg/L NAA showed the best performance for shoot regeneration and required maximum days for shoot initiation. Keywords: Regeneration; BARI Sarisha-6; BARI Sarisha-8; BARI Sarisha-11; anther culture; phytohormone  DOI: 10.3329/bjar.v35i2.5896Bangladesh J. Agril. Res. 35(2) : 331-341, June 2010


2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Dwi Hapsoro ◽  
Rahmadyah Hamiranti ◽  
Yusnita Yusnita

Abstract. Hapsoro D, Hamiranti R, Yusnita Y. 2020. In vitro somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media. Biodiversitas 21: 3811-3817. This study aimed to investigate the effects of genotypes and primary callus induction media on somatic embryogenesis of superior robusta coffee clones of Lampung. Leaf explants of clones Tugusari, Komari, Tugino, and Wanto were cultured on two types of primary callus induction media (PCIM). PCIM1 consisted of half-strength MS salts, 30 gL-1 sucrose, added with (mgL-1) 0.1 thiamine-HCl, 0.5 nicotinic acids, 0.5 pyridoxine-HCl, 100 Myo-inositol, 200 ascorbic acids, 150 citric acids, and 1 benzyl adenine. PCIM2 consisted of NPCM salts, 30 gL-1 sucrose, added with (mgL-1) 15 thiamine-HCl, 1 nicotinic acid, 1 pyridoxine-HCl, 2 glycines, 130 Myo-inositol, 200 ascorbic acids, 150 citric acids, 1 2,4-dichlorophenoxyacetic acid, and 2 thidiazuron. The highest percentage (100%) of primary callus formation was found in Komari and Wanto clones. PCIM2 resulted in more primary calli than PCIM1. When subcultured to embryogenic callus induction medium, primary calli of clone Komari and Wanto developed into a high percentage of embryogenic calli, while those of the other two turned brown and died. PCIM2-derived primary calli developed into more embryogenic calli. When subcultured on somatic embryo (SE) regeneration medium, these calli underwent the formation of SE of various stages. When subcultured to plant regeneration medium, these SEs developed into plantlets.


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