scholarly journals EMBRYO CULTURE AND IN VITRO CLONAL PROPAGATION OF OAK (Quercus aegilops L.)

2020 ◽  
Vol 51 (1) ◽  
Author(s):  
Fadladeen & Toma

An attempt was done to develop a micropropagation protocol for oak using embryo culture. Oak is considered a hard-to-root woody plant by conventional propagation methods, that’s why using tissue culture techniques is a very suitable alternative method. For oak embryo culture, WPM was used and found to be better than MS medium for embryo germination which gave 66.13%. As well as adding of GA3 to the medium improved the germination rate of embryos (43.25% and 82.25 %). At initiation stage, WPM was used and found to be the best medium by giving the highest number of shoots/ explant which was 1.80, the highest number of leaves (15.17 leaves/ explant) and the longest shoots (1.42 cm) followed by MS medium then GD which gave the lowest parameters which gave 0.98 shoots/ explant, 7.20 leaves/ explant and 1.06 cm shoot length. At shoot multiplication stage, BA was better than Kinetin for multiplication of oak explants. The addition of BA at 3 mg.l-l gave the highest number of shoot and leaves which were 3.33 and 26.11 respectively. The longest shoots were achieved when 4.5 mg.l-l of BA was used. Furthermore, kinetin at 3 and 4.5 mg-l gave the lowest parameters which were 1 cm in length and 1.54 leaves/ explant. For rooting stage, NAA was better than IAA in giving better parameters and rooting percentage. The highest number of roots and rooting percentage were achieved when 1 mg.l-l was added by giving 6 roots/ explant and 100% rooting percentage. While the longest roots were achieved when 0.5 mg.l-l of NAA was used (3.67 cm) followed by 1.5 mg.l-l IAA which gave 3.55 roots/ explant with rooting percentage 90%. The produced plantlets were successfully acclimatized and transferred to the open-air conditions with a rate reached 85%.

2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Efah FITRAMALA ◽  
Eva KHAERUNNISA ◽  
Nina Ratna Djuita Ratna DJUITA ◽  
Hadi SUNARSO ◽  
Diah RATNADEWI

AbstractBanana (Musa paradisiaca L) cv. Kepok Merah has a high commercial value as it is used in food industries such as banana chip. Besides, Kepok Merah contains high B-complex vitamins that serve in energy metabolism and are important in the development of infant brain. The establishment of industrial plantations of this plant has been restricted by the lack of planting materials. This research aimed at ameliorating the capacity of plantlets multiplication up to rooting of this banana in a rapid way through in vitro multiplication techniques. Murashige and Skoog (MS) and Woody Plant (WP) were used as the basic media. For the initiation stage, the media was fortified with 0.2 mg/L IAA and two levels of BA at 3 and 5 mg/L.  For shoot multiplication, the concentrations of IAA as well as BA were increased. For rooting, 1 mg/L NAA or IBA was applied. The observations demonstrated that for shoots initiation, both basic media performed good results when enriched with 0.2 mg/L IAA and 5 mg/L BA. The highest rate of shoots multiplication at 6 – 17 shoots per explant, was obtained on MS medium added with 0.5 mg/L IAA and 5 mg/L BA.  NAA at 1 mg/L in MS medium produced more rooted plantlets, 3 – 16 roots per plantlet, than those of other treatments. Keywords: Musa paradisiaca cv. Kepok Merah, in vitro micropropagation, scalps.AbstrakPisang (Musa paradisiaca L.) kultivar Kepok Merah memiliki nilai komersial yang cukup tinggi yaitu sebagai bahan dalam industri pembuatan keripik pisang. Selain itu, pisang Kepok Merah memiliki kandungan vitamin B kompleks cukup tinggi untuk membantu produksi energi dan pembentukan sel-sel otak pada bayi. Pertanaman pisang ini dalam skala industri terkendala oleh kurangnya ketersediaan sumber benih. Teknik kultur jaringan diharapkan dapat menghasilkan benih secara massal dalam waktu yang relatif singkat. Tujuan dari penelitian ini adalah meningkatkan keberhasilan multiplikasi tunas in vitro hingga pengakaran tanaman pisang Kepok Merah secara cepat. Pada tahap inisiasi tunas digunakan media dasar Murashige and Skoog (MS) dan media Woody Plant (WP); ke dalam media dasar tersebut ditambahkan IAA 0,2 mg/Ldan 2 taraf BA yaitu 3 dan 5 mg/L. Multiplikasi tunas dilakukan pada media dasar yang sama namun dengan taraf konsentrasi IAA serta BA yang ditingkatkan. Tahap perakaran menggunakan media dasar MS dan WP dengan auksin NAA 1 mg/L atau IBA 1 mg/L. Hasil penelitian menunjukkan bahwa untuk inisiasi tunas, media MS dan WP yang diperkaya dengan IAA 0,2 mg/L dan BA 5 mg/L   sama baiknya. Untuk multiplikasi tunas, media MS dengan IAA 0.5 mg/L   yang dikombinasikan dengan BA 5 mg/L   memberikan jumlah tunas paling banyak, yaitu 6 – 17 tunas per eksplan, dan pertumbuhannyapun lebih baik. Pemberian NAA 1 mg/L   pada media MS dapat memberikan lebih banyak tunas yang berakar, dengan jumlah akar 3 – 16 per planlet.  Kata kunci: Musa paradisiaca cv. Kepok Merah, mikropropagasi in vitro, nodul meristematik


2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Efah FITRAMALA ◽  
Eva KHAERUNNISA ◽  
Nina Ratna Djuita Ratna DJUITA ◽  
Hadi SUNARSO ◽  
Diah RATNADEWI

 Banana (Musa paradisiaca L) cv. Kepok Merah has a high commercial value as it is used in food industries such as banana chip. Besides, Kepok Merah contains high B-complex vitamins that serve in energy metabolism and  in the development of infant brain. The establishment of industrial plantations of this plant has been restricted by the lack of planting materials. This research aimed at ameliorating the capacity of plantlets multiplication up to rooting of this banana in a rapid way through in vitro multiplication techniques. Murashige and Skoog (MS) and Woody Plant (WP) media were used as the basic media. For the initiation stage, the media was fortified with 0.2 mg/L IAA and two levels of BA at 3 and 5 mg/L.  For shoot multiplication, the concentrations of IAA as well as BA were increased. For rooting, 1 mg/L NAA or IBA was applied. The observations demonstrated that for shoots initiation, both basic media performed good results when enriched with 0.2 mg/L IAA and 5 mg/L BA. The highest rate of shoots multiplication at 6 – 17 shoots per explant, was obtained on MS medium added with 0.5 mg/L IAA and 5 mg/L BA.  NAA at 1 mg/L in MS medium produced more rooted plantlets, 3 – 16 roots per plantlet, than those of other treatments. [Keywords: Musa paradisiaca cv. Kepok Merah, in vitro micropropagation, scalps.]AbstrakPisang (Musa paradisiaca L.) kultivar Kepok Merah memiliki nilai komersial yang cukup tinggi yaitu sebagai bahan dalam industri pembuatan keripik pisang. Selain itu, pisang Kepok Merah memiliki kandungan vitamin B kompleks cukup tinggi untuk membantu produksi energi dan pembentukan sel-sel otak pada bayi. Pertanaman pisang ini dalam skala industri terkendala oleh kurangnya ketersediaan sumber benih. Teknik kultur jaringan diharapkan dapat menghasilkan benih secara massal dalam waktu yang relatif singkat. Tujuan dari penelitian ini adalah meningkatkan keberhasilan multiplikasi tunas in vitro hingga pengakaran tanaman pisang Kepok Merah secara cepat. Pada tahap inisiasi tunas digunakan media dasar Murashige and Skoog (MS) dan Woody Plant (WP), ke dalam media dasar tersebut ditambahkan IAA 0,2 mg/L dan 2 taraf BA yaitu 3 dan 5 mg/L. Multiplikasi tunas dilakukan pada media dasar yang sama namun dengan taraf konsentrasi IAA serta BA yang ditingkatkan. Tahap perakaran menggunakan media dasar MS dan WP dengan auksin NAA 1 mg/L atau IBA 1 mg/L. Hasil penelitian menunjukkan bahwa untuk inisiasi tunas, media MS dan WP yang diperkaya dengan IAA 0,2 mg/L dan BA 5 mg/L   sama baiknya. Untuk  multiplikasi  tunas,   media  MS dengan IAA 0,5 mg/L   yang dikombinasikan dengan BA 5 mg/L   memberikan jumlah tunas paling banyak, yaitu 6 – 17 tunas per eksplan, dan pertumbuhannyapun lebih baik. Pemberian  NAA 1 mg/L pada media MS dapat memberikan lebih banyak tunas yang berakar, dengan jumlah akar 3 – 16 per planlet.  [Kata kunci: Musa paradisiaca cv. Kepok Merah, mikropropagasi in vitro, nodul meristematik.]


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.


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1050A-1050
Author(s):  
Natalie Anderson ◽  
David H. Byrne ◽  
Maria B. Raseira

A major obstacle faced by programs that breed early-ripening peach cultivars [Prunus persica (L.) Batsch] is the low viability of the embryos from the early-ripening parents that are used as females. Embryo culture techniques have been developed to allow embryos to mature in vitro, thus increasing the chances of germination and survivability. Several media types exist for Prunus embryo culture. Two types, Woody Plant Medium (WPM) and Smith, Bailey, and Hough (SBH) were investigated for this report. The WPM type was studied in two forms, one made from scratch and the other in a prepackaged form. The SBH type was studied with the addition of vitamins and without vitamins. Eight peach genotypes with embryo lengths ranging from 9.6 to 12.7 mm were used. Surprisingly enough, it was found that WPM from scratch performed better than WPM from a prepackaged mix. For all eight genotypes studied, WPM from scratch resulted in as good as or better germination than SBH with or without vitamins. A large media by genotype effect was found, which is partially attributed to the embryo size. The genotypes with larger embryos (>11 mm) tended to perform equally on all media tested whereas the embryos <10.5 mm germinated better on WPM as compared to SBH.


2016 ◽  
Vol 75 (2) ◽  
pp. 244-252 ◽  
Author(s):  
Jana Ambrožič-Dolinšek ◽  
Terezija Ciringer ◽  
Mitja Kaligarič

Abstract The monotypic Hladnikia pastinacifolia Rchb. is a narrow endemic species, with an extremely small distribution area in Slovenia, prone to any kind of threat that could lead to species extinction. Tissue culture techniques are proposed as a conservation measure for rapid propagation and ex-situ conservation. Tissue culture was initiated from seeds and juvenile plants obtained from natural sites on a solid Murashige and Skoog (MS) medium, with and without growth regulators. We tested various combinations and concentrations of growth regulators, and the best proliferation of axillary shoots, on average 14, was obtained on MS medium with 5 μM BAP and 3 μM IBA and 3% sucrose. Rooting was achieved after transferral of the shoots to an MS medium with 2 μM IBA and 3% sucrose. The rooted plants were acclimatized on a mixture of limestone sand, potting soil and vermiculite in a ratio of 10:2:2, with pH in the range of 7.5–8.0. In vitro propagation methods provide an important opportunity for the propagation and preservation of H. pastinacifolia by rapidly increasing the number of plants, without disturbing the wild population.


2020 ◽  
Author(s):  
Adugnaw Admas ◽  
Berhane Kidane ◽  
Melaku Admasu ◽  
Tesaka Misga

ABSTRACTIn Ethiopia, Oxytenanthera abyssinica A.Rich. Munro has varies economic importance. However, conventional propagation methods of O. abyssinica are generally inefficient due to their low multiplication rate, time consuming, labor intensive, and too costly. The objective of this study was to develop a protocol for mass micropropagation of O. abyssinica through seed culture. Murashige and Skoog (MS) medium augmented with 6-Benzylaminopurine (BAP) was used for shoot initiation and multiplication. For in vitro rooting, MS medium supplemented with 3-Indole –butric acid (IBA) was used.In shoot initiation experiment all viable seeds were proliferated in 5-7 days of culturing. In shoot multiplication at 0.004 g/L BAP was Sucssefuly shoot multiplied, also best root responding were found at 0.005 g/l IBA.The present optimized protocol enables for any acters who needs large numbers of low land bamboo seedling for industery, small and micro enterprize or for reafforestation programms.


2018 ◽  
Vol 24 (2) ◽  
Author(s):  
J. D. BARSHILE

Present investigation was undertaken to standardize technique for in vitro micro-propagation of chickpea( Cicer arietinum ) cultivar Vishwas (Phule G 12). Micropropagation method for chickpea was established and this method enabled much more efficient propagation of plants. The present work was aimed at evolving a protocol for rapid multiplication of chickpea using micropropagation technique. Explants from shoot tip and node segment were cultured on MS medium supplemented with different concentrations of BAP and Kinetin (1.0 to 2.5 mg/l) and their growth responses like shooting were elucidated. The maximum multiple response was observed with 2 mg/l concentration of BAP from both types of explant. The highest number of shoots (12.5 ± 0.3) was achieved on MS medium with 2 mg/l BAP using node segments. The medium supplemented with 2 mg/l of BAP was found better than all other concentrations. Individual shoots were transferred to IBA and IAA (1.0-1.5 mg/l) for root induction. MS medium supplemented with 2 mg/l of IBA proved better for rooting. Rooted plantlets were successfully hardened in greenhouse and established in the pot.


Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3229
Author(s):  
Mat Yunus Najhah ◽  
Hawa Z. E. Jaafar ◽  
Jaafar Juju Nakasha ◽  
Mansor Hakiman

This study aims to investigate whether the in vitro-cultured L. pumila var. alata has higher antioxidant activity than its wild plant. An 8-week-old L. pumila var. alata nodal segment and leaf explants were cultured onto Murashige and Skoog (MS) medium supplemented with various cytokinins (zeatin, kinetin, and 6-benzylaminopurine (BAP)) for shoot multiplication and auxins (2,4-dichlorophenoxyacetic acid (2,4-D) and picloram) for callus induction, respectively. The results showed that 2 mg/L zeatin produced the optimal results for shoot and leaf development, and 0.5 mg/L 2,4-D produced the highest callus induction results (60%). After this, 0.5 mg/L 2,4-D was combined with 0.25 mg/L cytokinins and supplemented to the MS medium. The optimal results for callus induction (100%) with yellowish to greenish and compact texture were obtained using 0.5 mg/L 2,4-D combined with 0.25 mg/L zeatin. Leaves obtained from in vitro plantlets and wild plants as well as callus were extracted and analyzed for their antioxidant activities (DPPH and FRAP methods) and polyphenolic properties (total flavonoid and total phenolic content). When compared with leaf extracts of in vitro plantlets and wild plants of L. pumila var. alata, the callus extract displayed significantly higher antioxidant activities and total phenolic and flavonoid content. Hence, callus culture potentially can be adapted for antioxidant and polyphenolic production to satisfy pharmaceutical and nutraceutical needs while conserving wild L. pumila var. alata.


2019 ◽  
Vol 11 (2) ◽  
Author(s):  
POPY HARTATIE HARDJO ◽  
DANNY PUTRA SENTOSA SUSANTO ◽  
WINA DIAN SAVITRI ◽  
MARIA GORETTI MARIANTI PURWANTO

Abstract. Hardjo PH, Susanto DPS, Savitri WD, Purwanto MGM. 2019. Shoot multiplication of Pogostemon cablin var. Sidikalang and patchouli oil profile. Nusantara Bioscience 11: 123-127. Pogostemon cablin Benth. is a plant producing patchouli oil, which mostly consists of patchouli alcohol compound. Patchouli oil has great potentials in the world market because of its stability and high price. In this study, in vitro multiplication of Sidikalang variety of Acehnese patchouli shoots was done on solid and liquid Murashige & Skoog (MS) medium. This study aimed to determine the effect of cytokinins in various combinations of shoot multiplication and to compare the patchouli oil yield of in vitro and ex vitro culture. In vitro multiplication of Acehnese patchouli shoots by using solid MS medium with addition of 0.2 ppm benzyl aminopurine (BAP) and 0.2 ppm Kinetin resulted in shoot explants with an average growth index of 82.198 ± 0.690. Patchouli oil extraction was done on 7 weeks old in vitro shoot explants cultured on solid MS medium + 0.2 ppm BAP + 0.2 ppm Kinetin using water distillation method. In vitro shoots yielded 2.5% patchouli oil and contained ± 35% patchouli alcohol compound, whereas ex vitro shoots produced 4% patchouli oil and contained ± 25% patchouli alcohol compound. The qualitative analysis by using thin layer chromatography (TLC) showed that there were similarities in the number of spot and Rf value for each spot of ex vitro and in vitro patchouli oil.


2015 ◽  
Vol 48 ◽  
pp. 23-31 ◽  
Author(s):  
S. Kaveri ◽  
Rao Srinath

In vitro seed germination and embryo culture have been achieved in Nothapodytes foetida, this plant is known for its rich source of anticancer drug i. e., Camptothecin. In present study both normal and decoated seeds were subjected to different treatments viz., H2O, GA3, H2O2, H2SO4, chlorine water and mechanical scarification, further these were germinated on water agar medium (WA), filter paper bridge (FB), half strength MS (HMS) and full strength MS (FMS) medium. The highest percentage (69%) of germination was achieved from decoated seeds treated with 10mg/L GA3 and germinated on Filter Paper Bridge. And for embryo culture mature embryos were inoculated on MS medium containing various combination and concentrations of cytokinins (BAP, Kn and TDZ) and auxin (IAA and NAA) for rapid conversion into a plantlet. Among the different combinations of growth regulators; highest frequency (100%) of plantlet conversion was obtained on MS medium containing Kn (1.0mg/L) and NAA (0.2mg/L).


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