scholarly journals In Vitro Regeneration of Grass Pea (Lathyrus sativus L.) from Cotyledonary Node Explants

2017 ◽  
Vol 5 (2) ◽  
pp. 57 ◽  
Author(s):  
Diriba Tesfaye ◽  
Kassahun Bantte ◽  
Tewodros Tadesse

Full potential of grass pea has not been utilized because of the presence of the neurotoxin amino acid β-N-oxalyl-L-αβ -diaminopropionic acid (ODAP/BOAA). Conventional breeding and other approaches have not been successful in reducing the toxin. Integration of in vitro techniques can contribute significantly to meet the challenge. Therefore, this study was carried out to evaluate the in vitro regeneration capacity of grass pea genotypes. Shoot initiation, multiplication and rooting of IVAT-LS-690 were conducted using completely randomized design with five replications. Genotypes were treated with BAP and NAA for shoot initiation while BAP and Kn Combination were used for multiplication. Different concentrations of IBA and IAA were used for rooting. Shoot proliferation percentage was the highest (100%) for IVAT-LS-690,on Murashige and Skoog (MS) medium augmented with 2.0 mg/l BAP +0.1 mg/lNAA.For in vitro shoot multiplication, best results were obtained on concentrations of 3mg/l BAP+1mg/l Kn with maximum shoot number per explants (11.5). High number of roots per shoot (6) and percent of rooted shoot (86.66%) were obtained from ½ MS medium supplemented with 0.5 mg/l IBA. This study inferred that both genotype and BAP levels play a crucial role for shoot regeneration capacity and the optimum hormonal combination for grass pea is genotype specific.

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sharone gladies E ◽  
Chithra Devi B. S

We can see Orchids come in a wide variety of shapes, sizes, colours, and textures far beyond the human mind’s imagination. They emerge from seeds in nature, but in the absence of suitable hosts, they do not germinate in sufficient numbers. This problem was solved by using the tissue culture technique for its germination. One of the successful method used for mass propogation of orchid plantlets is in vitro techniques. Therefore, an initial analysis was conducted in order to establish an appropriate procedure for mass multiplication of Arundina graminifolia. MS (Murashige and Skoog) medium was found to be suitable for the asymbiotic seed germination of Arundina graminifolia. Direct protocorm like bodies were induced by using combinations and individual supplement of MS medium with IAA (Indole-3-acetic acid), IBA (Indole-3- butyric acid), BAP (6-Benzylaminopurine) and KIN (Kinetin). Hormone-free MS basal medium was found suitable for the conversion of PLBs (protocorm-like bodies) into complete plantlets


1970 ◽  
Vol 8 (2) ◽  
pp. 227-232 ◽  
Author(s):  
Sonia B Shahid ◽  
SZ Khan ◽  
L Hassan

The experiment was conducted to optimize a suitable protocol for in vitro regeneration in cole crops. Callus initiation was excellent in the variety Early Tropical. Highest percentage of callus proliferation was observed in Early Tropical (75.0%) followed by Tangail Special Pauslali (55.0%) and the lowest in Tara (40.0% ). Maximum callus proliferation (68.5%) was observed in MS + 3.0 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3. Callus proliferation was lowest (40.0%) in MS + 2.5 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3. MS medium supplemented with 3.0 mgL-1 BAP + 1.0 mgL-1 2,4-D + 2.0 mgL-1 AgNO3 was the best for shoot initiation & plantlet regeneration. The highest number of shoots per vial was 7.20 and the lowest number of shoots per vial was 4.40. Among the concentration MS + 3.0 mgL-1 BAP + 0.1 mgL-1 2,4-D + 2.0 mgL-1 AgNO3 showed the highest performance of shoots per vial. The variety Tangail Special Pauslali was the best for root initiation. Keywords: Brassica; Cole crops; Callus; In vitro; Regeneration DOI: 10.3329/jbau.v8i2.7930 J. Bangladesh Agril. Univ. 8(2): 227-232, 2010


2020 ◽  
Vol 100 (2) ◽  
pp. 202-208
Author(s):  
Mengting Wang ◽  
Guiliang Zhang ◽  
Peiyao Xin ◽  
Yun Liu ◽  
Bin Li ◽  
...  

Camellia fascicularis is an endangered evergreen ornamental plant with pale yellow flowers. An efficient and reproducible in vitro regeneration method is required for its large-scale propagation and germplasm conservation. In this study, one axillary bud per nodal stem was obtained from C. fascicularis cultured on Murashige & Skoog (MS) medium containing 0.1 mg L−1 indole-3-acetic acid (IAA) combined with 1.0 mg L−1 6-benzylaminopurine (BA). Axillary buds from the stem segments were transferred to modified woody plant medium (WPM) supplemented with 3.0 mg L−1 BA in combination with 0.3 mg L−1 IAA for multiplication, thereby resulting in a high shoot multiplication rate of 6.8. Multiple shoots were divided into nodal stems and shoot tips and were induced to root. The shoot tips were induced to root by culturing on one-half MS medium supplemented with 2.0 mg L−1 indole-3-butyric acid (IBA) in combination with 0.3 mg L−1 α-naphthalene acetic acid (NAA), which resulted in 76.0% rooting efficiency with 2.3 roots per shoot. The optimal hormone ratio for inducing rooting of nodal stems was 1.0 mg L−1 IBA in combination with 2.0 mg L−1 NAA, which resulted in 72.7% rooting efficiency with 1.7 roots per nodal stem. These two rooted plantlets were successfully acclimatized and established in a greenhouse.


2019 ◽  
Vol 1 (4) ◽  
pp. 261-269
Author(s):  
Thuy Linh Thi Nguyen ◽  
Ngoc Thi Pham ◽  
Thao Thi Ninh ◽  
Phuong Thao Thi Nguyen

This study was conducted to establish the procedure for in vitro propagation of Japanese cherry (Prunus sp.) to produce large quantity of plantlets and initial planting materials for climate adaptation research of this plant in Hanoi. Single nodal stems were used as the primary explants and initially produced shoots on MS medium supplemented with 1 mg L-1 BA. The highest shoot multiplication rate (9.57 times) was obtained on MS medium containing 1 mg L-1 BA and 0.25 mg L-1 a-NAA after 8 weeks of culture. 100% of the shoots produced roots with a mean of 10.10 roots per plant within 4 weeks on ½ MSM medium with 4 mg L-1 IBA. The survival rate of in vitro derived plantlets after a 6 to 7-week-period of rooting during acclimatization using a soil: coco peat: smoked rice husks (2:2:1, v/v/v) substrate was 100% and acclimatized plantlets showed good growth and development. This is the first report on a practical and efficient in vitro multiplication protocol for Japanese cherry in Vietnam, starting from shoot initiation to establishment of plants under greenhouse conditions.


HortScience ◽  
1999 ◽  
Vol 34 (2) ◽  
pp. 353-354 ◽  
Author(s):  
James R. Ault ◽  
Kayri Havens

Shoot explants from actively growing, greenhouse-maintained plants of Baptisia `Purple Smoke' were cultured in vitro for shoot initiation on Murashige and Skoog (MS) basal medium containing vitamins and supplemented with 30 g·L–1 sucrose, 8.87 μm BA, and 4.14 μm K-IBA. All subsequent media were supplemented with 2.47 mm NaH2PO4 to enhance shoot growth. Single-node explants were subcultured for shoot multiplication on MS medium with either no plant growth regulator or with 2.22, 4.44, 8.87, 17.74, or 35.48 mm BA in combination with 0.0 or 4.14 μm K-IBA. Explants produced a maximum of 4.1 shoots on the medium with 2.22 μm BA. Shoots rooted on all concentrations of K-IBA (2.07, 4.14, 10.36, or 20.72 μm) and K-NAA (2.23, 4.46, 11.15, or 22.29 μm) tested. Maximum rooting was 100% on MS medium with 11.15 μm K-NAA; however, this treatment induced copious stem callusing. Rooted shoots were greenhouse-acclimatized for 2.5 weeks. Overall survival was 86%. For optimal rooting and subsequent acclimatization, treatment with 2.23 μm K-NAA is recommended; this resulted in 83% rooting and 87% acclimatization. Chemical names used: N6 benzyladenine (BA); potassium salt of indole-3-butyric acid (K-IBA); potassium salt of 1-naphthalene acetic acid (K-NAA).


HortScience ◽  
2019 ◽  
Vol 54 (9) ◽  
pp. 1558-1564 ◽  
Author(s):  
Georgia Vlachou ◽  
Maria Papafotiou ◽  
Konstantinos F. Bertsouklis

Seed ecophysiology and micropropagation of Clinopodium nepeta, an aromatic Mediterranean plant with pharmaceutical and horticultural uses was investigated. The optimum germination temperature of seeds stored at room temperature for 0, 6, or 12 months was 15 to 20 °C (100% germination completed in 10 to14 days) and cardinal temperatures were defined at 10 and 30 °C (80% to 82% and 62% to 76% germination, respectively). Six or 12 months of storage did not seem to affect germination, although 12-month-old seeds germinated at higher percentage and completed germination earlier at 15 °C than at 20 °C. Concerning micropropagation, shoot multiplication at subcultures of both adult plant- and seedling-origin nodal explants was tested on Murashige and Skoog (MS) medium supplemented with various cytokinin types, i.e., zeatin (ZEA), 6-benzyladenine (BA), kinetin (KIN), and 6-γ-γ-(dimethylallylamino)-purine (2IP), at various concentrations from 0.0 to 8.0 mg·L−1. Both explant types presented a rather similar response during in vitro culture. Increasing concentration of all cytokinin types resulted in an increase in shoot number per responding explant (1.1–5.3) and in most cases a decrease in shoot length (0.6–3.4 cm). Increasing cytokinin concentration induced hyperhydricity to a number of shoots (0.1–6.5) per explant, mostly when ZEA and BA were used. Supplementing the MS medium with 8.0 mg·L−1 BA combined with 0.1 mg·L−1 1-naphthaleneacetic acid (NAA) led to almost elimination of hyperhydricity and very satisfactory shoot production (80%/88% explant response and 6.5/7.5 shoot number per responding explant for seedling- / adult-origin explants, respectively). Alternatively, increasing the agar concentration to 12.0 g·L−1 and supplementing the medium with 8.0 mg·L−1 BA only, resulted in the same effect on eliminating hyperhydricity, such as the addition of NAA, and in the best shoot multiplication response achieved in this study (100% explant response, 9.4/9.9 shoots per explant for seedling-/adult-origin explants, respectively). Microshoots rooted abundantly (92% to 100%) on half-strength MS medium, either Hf or supplemented with 0.5 mg·L−1 to 4.0 mg·L−1 indole-3-butyric acid (IBA). The addition of IBA to the rooting medium, regardless of its concentration, affected only the root length by increasing it 2- to 3-fold. Microshoot clusters produced on multiplication media rooted at 96% when cultured on Hf half-strength MS medium. Rooted microshoots and shoot clusters survived at 80% to 100%, respectively, after ex vitro acclimatization in peat:perlite 1:1 (v/v).


2016 ◽  
Vol 1 (3) ◽  
pp. 589-599
Author(s):  
Meherunnesa Papry ◽  
SM Ahsan ◽  
Sayeed Shahriyar

The experiment was conducted on in vitro regeneration of tomato at the Plant Biotechnology Laboratory, Department of Horticulture, Patuakhali Science and Technology University, Patuakhali. The objective was to develop an efficient regeneration protocol in tomato through callus induction for subsequent plantlet regeneration. Seeds were inoculated on MS medium where germination rate was 78.4%. The stems of in vitrocultured seedlings were used as explants. Different concentrations and combinations of growth regulators were added to MS medium to observe their efficacy on callus induction, shoot initiation and root formation. Stem explants cultured on MS medium fortified with 2 mg/L BAPgave the highest number of shoots (3.0) at 45 DAC. Among the concentrations of PGRs, 0.25 mg/L IAA produced the highest length (4.064 cm) of plantlets, number (5.0) of leaves and fresh weight (0.663 g) of plantlets with the stem explants at 45 DAC. The concentration of 0.5 mg/L IAA produced the highest number (21.00) of roots/plantlet, length (7.676 cm) of roots at 45 DAC, from the same explants. The highest survival rate of in vitro regenerated plantlets in the pot was 70.00 % with the stem explants. The results of the current study showed significant increase in the growth of callus of Solanumlycopersicon Mill. Indicating a good efficiency of the optimized media composition and the experimental model used in comparison to other studies of similar nature.Asian J. Med. Biol. Res. December 2015, 1(3): 589-599


Author(s):  
Manu Pant ◽  
Prabha Bisht ◽  
Manju P Gusain

Objective: The present study was performed to investigate the role of varying concentrations of carbohydrate source and strengths of nutrient medium in growth and development of in vitro shoots of Swertia chirata-an endangered medicinal plant.Methods: MS medium supplemented with 6-Benzylaminopurine (4.44 µM), Indole-3 acetic acid (2.85 µM) and Adenine sulphate (271.45 µM) was used to test the efficiency of of sucrose at concentrations of 1-5% and of media strength varying from full to one-fourth. The data were analysed using analysis of variance (ANOVA) of Completely Randomized Design (CRD) in GenStat 5 Edition 3.2 for PC/Windows NT (Copyright 1995, Lawes Agricultural Trust (Rothamsted Experimental Station))Results: Observations on axillary shoot multiplication indicated that sucrose at a concentration of 3% and MS medium in its full strength proved to be most optimal for in vitro culture multiplication. On this medium combination mean number of 11.80 shoots (after 4 w) and 18.50 shoots (after 8 w) could be obtained On sucrose free medium the shoots exhibited necrosis while at lower concentrations of 1-2% sucrose, the shoots developed were thin and unsuitable for further growth in vitro. At higher levels of sucrose in the medium, the shoots became thick and stunted. Similarly, reduction in medium strength resulted in a decline in shoot number and shoot length to an average of 6.50 shoots (1.33 cm mean length) on half strength medium and 5.60 shoots (0.88 cm mean length) on one-fourth strength; as observed after 4 w.Conclusion: The experimental findings suggest that any decline from the standard had a significant effect on the number, size and overall health of shoots developed in vitro. The conditions so standardized augment the production of healthy shoots that shall aid in subsequent rooting and survival after transplantation of tissue-culture raised plantlets.


2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Sunil Tulshiram Hajare ◽  
Nitin Mahendra Chauhan ◽  
Girum Kassa

Aim. Potato (Solanum tuberosum L.) is one of the important crops in Ethiopia which has a crucial role in nutritional security, poverty alleviation, and income generation. The aim of the present investigation is to develop an efficient in vitro propagation protocol for Belete and Gudiene potato varieties by using lateral bud as explants. Materials and Methods. Shoot initiation was achieved by inoculating buds on full-strength MS Murashige and Skoog medium (MS) fortified with variable concentrations of BAP and NAA. Basal MS was used as control throughout the experiment. Results. Results of our study showed that best shoot initiation was obtained on MS medium supplemented with 1.5 mg/l BAP + 3.0 mg/l NAA for Gudiene variety, whereas 1.0 mg/l BAP and 2.0 mg/l NAA produced more shoots in Belete variety. The initiated shoots increased two- to three-fold upon subculture on the MS medium fortified with varying concentrations of BAP and Kinetin. The highest numbers of multiple shoots were obtained in the MS medium containing 2.5 mg/l Kinetin. The combined effect of BAP and Kinetin did not produce any additional positive effect for shoot multiplication. Rooting percentage and number of roots/shoot were found best on the MS medium fortified with 1.0 mg/l IBA + 0.5 IAA. Conclusions. The variety Gudiene was found best for shoot initiation and root formation, while Belete variety proved its superiority for multiple shoot formation. A total number of 82.66% of plantlets were acclimatized under field conditions. This work indicates the practical applicability of plant tissue culture using lateral bud as explants is effective for micropropagation of potato in vitro.


Author(s):  
Girmay Mekonen ◽  
Meseret Chimdessa Egigu ◽  
Manikandan Muthsuwamy

Banana is a fruit crop which has high demand in Ethiopia, but its production is constrained by lack of disease free planting material with conventional propagation methods. For shoot initiation, shoot tip explants were cultured on MS medium supplemented with 0.5, 1.0, 1.5 and 2.0 mg/L BAP. Similarly, MS medium supplemented with BAP at 1.0, 1.5, 2.0 mg/L in combination with IBA at 0.25 and 0.50 mg/L were used for shoot multiplication. Half- strength MS medium augmented with IBA at 1.0, 2.0, 3.0 and 4.0 mg/l were used for root induction. MS medium without PGRs were used as controls. Finally, hardening of the in vitro derived plantlets was carried out in green house both in the primary and secondary acclimatization stages. Results showed that the highest shoot initiation percent (93.40%), highest mean number of shoots per explant (4.67) and lesser day for shoot induction (11.00) were observed in explant cultured on MS + 1.0 mg/L BAP. With shoot multiplication, highest shooting percent (92.60%), maximum number of shoots (7.67) and highest shoot length (5.27 cm) were recorded on MS + 1.5 mg/L BAP + 0.5 mg/L IBA. The highest rooting percent (93.40%), maximum root number per shoot (7.67) and highest root length (11.00 cm) were found on a half strength MS medium + 2.0 mg/L IBA. The survival rate of plantlets were 96.00% in coco peat substrate in primary acclimatization and 97.92% in forest soil, sand and manure substrates mixed at 3:2:1 ratio in secondary acclimatization. Overall, the result showed that the PGRs type, concentrations and combinations used are effective for mass propagation of banana variety studied in this experiment.


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