scholarly journals Optimization of in vitro rooting protocol for tomato (Lycopersicon esculentum mill.) varieties

2020 ◽  
Vol 12 (3) ◽  
pp. 365-371
Author(s):  
Endalkachew Baye ◽  
Temesgen Matewos ◽  
Derbew Belew

In vitro rooting of micropropagated shoots were carried out with the aim of evaluating the root induction responses of two tomatoes (Lycopersicon esculentum MILL) varieties (Gelilema and Chali) using Indole -3- butyric acid (IBA). Seven levels of IBA (0.0, 0.25, 0.5, 0.75, 1.0, 1.25 and 1.5 mg L-1) were used in a completely randomized design (CRD) in factorial combinations (seven level of IBA*two varieties) with three replications.  After the plantlets were kept in the rooting media for three weeks, data on rooting percentage, number of roots/shoots and root length in cm were collected. The analysis of variance showed that the interaction of IBA*Var was highly significantly different for rooting percentage, a number of roots/shoot and root length at p<0.01. The highest rooting percentage (100.00±0.00), number of roots/shoot (14.20±0.35) and root length (10.7±0.29) were received from Chali on free Murashige and Skoog medium (MS). At the same time, the lowest percentage of rooting (11.11±0.00), number of roots/shoot (0.887±0.19) and root length (1.00±0.00 cm) were obtained from Gelilema on MS+1.5mg/l IBA. For acclimatization, the in vitro rooted shoots were transplanted into plastic pots containing a mixture of oven sterilized soil and sand at a ratio of 2:1. After three weeks, a survival rate of 67.7% for Chali and 58.1% for Gelilema was obtained. From the above result, it can be concluded that free MS medium was the best for in vitro rooting of the two tomato varieties. The optimized protocol will be useful for rapid in vitro multiplication of the two tomato varieties.

2021 ◽  
Vol 58 (1) ◽  
pp. 1640-1643
Author(s):  
Thanwamas Kassanuk Et al.

Kluai Nam Thai’ (Musa AA group) are rare in Thailand and it is at risk of extinction because of environmental changes and human behavior. Hence, the plant tissue culture technique is used for propagation. This research aimed to study the effect of coconut water on root induction of Kluai Nam Thai in vitro. The shootlets from laboratory were cultured on Murashige and Skoog (MS) medium supplemented with coconut water at concentrations 10, 15, 20, and 25% for 1 month. The experimental design was Completely Randomized Design (CRD). The results indicated that MS medium supplemented with 20% coconut water gave the highest average number of roots at 1.85 while MS medium supplemented with 10% coconut water gave the highest length of roots at 2.34 cm which were significantly different (p<0.01) from the other concentrations


2021 ◽  
Vol 3 (2) ◽  
Author(s):  
Wulan Dari Neng Gumiwang ◽  
Tintrim Rahayu ◽  
Ari Hayati

The purpose of this research is to determine the concentration of young coconut water that is appropriate for the growth of orchid plantlets (Dendrobium sp.) In vitro. This study used an experimental method, descriptive data analysis to compare several different concentrations of coconut water. The design of this study uses a completely randomized design (CRD). The treatments consist of 0% coconut water concentration (as a control), 15%, 30% and 60%. Each concentration was carried out 5 replications and each repetition consisted of 5 Dendrobium sp plantlets in each culture bottle conducted for 40 HST, for observing the root length carried out for 50 HST. The highest number of shoots and leaves were produced at the same concentration, namely 150 ml / L coconut water treatment (15% concentration) with an average of 2.8 shoots and the average number of leaves 10.8 leaves. The average number of roots and the longest root length was produced at a concentration of 600 ml / L coconut water (60% concentration) with an average of 6 roots, and the longest root length was 0.5 cm.Keywords: Young coconut water, (Cocos nucifera L.), Dendrobium sp., in vitro, growth.ABSTRAKTujuan penelitian ini ialah menentukan konsentrasi air kelapa muda yang tepat untuk pertumbuhan planlet anggrek (Dendrobium sp.) secara in vitro. Penelitian ini menggunakan metode eksperimen, analisis data secara deskriptif untuk membandingan beberapa konsentrasi air kelapa yang berbeda. Rancangan penelitian ini menggunakan Rancangan Acak Lengkap (RAL). Perlakukan terdiri dari konsentrasi air kelapa 0 % (sebagai kontrol), 15% , 30% dan 60%. Masing-masing konsentrasi dilakukan 5 kali ulangan dan setiap ulangan terdiri dari 5 planlet Dendrobium sp dalam setiap botol kultur yang dilakukan selama 40 HST, untuk pengamatan panjang akar dilakukan selama 50 HST. Jumlah tunas dan jumlah daun terbanyak dihasilkan pada konsentrasi yang sama, yaitu perlakuan air kelapa 150 ml/L (konsentrasi 15%)  dengan rata-rata jumlah tunas terbanyak 2,8 tunas dan rata-rata jumlah daun terbanyak 10,8 helai daun. Rata-rata jumlah akar terbanyak dan panjang akar terpanjang dihasilkan pada konsentrasi air kelapa 600 ml/L (Konsentrasi 60%) dengan rata-rata jumlah akar terbanyak sebanyak 6 akar, dan rata-rata panjang akar terpanjang 0,5 cm.Kata kunci : Air kelapa Muda (Cocos nucifera L.), Dendrobium sp., in vitro, pertumbuhan 


1970 ◽  
Vol 19 (1) ◽  
pp. 101-111 ◽  
Author(s):  
Rakha Hari Sarker ◽  
Khaleda Islam ◽  
M.I. Hoque

Agrobacterium-mediated genetic transformation system has been developed for two tomato (Lycopersicon esculentum Mill.) varieties, namely Pusa Ruby (PR) and BARI Tomato-3 (BT-3). Prior to the establishment of transformation protocol cotyledonary leaf explants from the two varieties were cultured to obtain genotype independent in vitro regeneration. Healthy multiple shoot regeneration was obtained from the cut ends of cotyledonary leaf segments for both the varieties on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. The maximum root induction from the regenerated shoots was achieved on half the strength of MS medium supplemented with 0.2 mg/l IAA. The in vitro grown plantlets were successfully transplanted into soil where they flowered and produced fruits identical to those developed by control plants. Transformation ability of cotyledonary leaf explants was tested with Agrobacterium tumefaciens strain LBA4404 harboring binary plasmid pBI121, containing GUS and npt II genes. Transformed cotyledonary leaf explants were found to produce multiple shoots on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin to 200 mg/l since kanamycin resistant gene was used for transformation experiments. Shoots that survived under selection pressure were subjected to rooting. Transformed rooted plantlets were transferred to soil. Stable expression of GUS gene was detected in the various tissues from putatively transformed plantlets using GUS histochemical assay.  Key words: In vitro regeneration, transformation, tomato D.O.I. 10.3329/ptcb.v19i1.5004 Plant Tissue Cult. & Biotech. 19(1): 101-111, 2009 (June)


Author(s):  
Talib Khashan Kareem ◽  
Abbas Tikki Karrar

This experiment was conducted in faculty of Science labs, Kufa University, carried out during 2015 to applied methods for extraction, purification and Quantitative of Lycopene red pigments, from callus tissue and tomato fruits mother plant (Lycopersicon esculentum Mill).This study include of three parts, Firstly; Tomato seeds(Supper queen) hybrid were germinated in free MS medium and callus induction from shoot tip (3cmpieces) by using MS medium supplemented with Dichlorophenoxiactic acid (2,4-D) at different concentration (0.5,1, 1.5mg/l)with benzyl adenine (BA) at concentration of (0.3 mg/l). Secondly; identically callus fresh weight re-cultured in the same MS medium supplemented with high molecular weight polyethylene glycol (PEG) was used as selective agent at level of (5,10,15 and 25%). Thirdly; comparisons study were made between in vitro and in vivo grown plant. Powder of control lycopene used as standard solution. The content of lycopene was done by using high performance liquid chromatography (HPLC), and compare of the quantitatively of lycopene with these content in fruits of mother plant, and callus tissue. Also, include alcohol extraction of Lycopene from tomato fruit by using acetone and hexane mixture. The result showed significant increased (P< 0.05) of lycopene production and the superiority of lycopene content in callus than the content in fruits of mother plant. Antioxidant enzymes activity like Catalase (CAT),Guaiacol peroxidise (POX) and Superoxide dismutase(SOD) were high in callus under drought stress than in fruit of mother plant. However, Proline and total sugar content were at higher levels in callus under drought stress than in fruit of mother plant.


2016 ◽  
Vol 74 (2) ◽  
Author(s):  
Nurita TORUAN-MATHIUS ◽  
. LUKMAN ◽  
. AGUS-PURWITO

Summary In vitro micrografting is a technique for grafting scions to rootstocks of plantlets from tissue culture. In vitro micrografting of Cinchona plant has never been carried out. The objective of this research was to obtain the best method of in vitro micrografting, medium for micrografted plantlets, and acclimatization  for Cinchona plantlets from  micrografting. The research consisted of (i) optimization of micrografting method, (ii) optimization of medium for growing plantlets, and (iii) acclimatization of micrografted plantlet. Plantlets of four-month-old of  C. ledgeriana  QRC clone were used as  scions, while of C. succirubra as  rootstocks. Each of experiments was arranged according to Completely Randomized Design, consisted of  combination of scion and rootstock and type of micro-grafting with 10 replicates. Parameters measured were  the percentage of survived plantlet, leaf number, and callus productions on union area, and percentage of survived  plantlet. The results show that V type of micrografting was the best for Cinchona micrografting. MS medium with the addition of 3 mg/L IBA was the best medium for growing of micrografted plantlet. Husk charcoal mixed with top soil (1 : 1) was the best medium for acclimatization.  Acclimatization  consisted  of two steps: preaclimatization in a culture room with 12- hour photoperiod at temperature 25 – 27oC  for two weeks,  followed by aclimatization in a plastic house with  70% reduced light intensity for one month. Using this method, 90% of the seedlings were survived. It is concluded that in vitro micrografting can be used as a technique for clonal propagation of Cinchona sp.Ringkasan  Teknik sambung mikro (mikrografting) in vitro adalah teknik penyambungan potongan batang atas pada batang bawah dalam kultur jaringan.  Pada tanaman kina teknik sambung mikro  in vitro belum pernah dilakukan. Tujuan penelitian ini adalah  menetapkan tipe sambung mikro, medium terbaik untuk planlet hasil sambung  mikro, dan perbanyakan tanaman kina dengan sambung mikro. Pelaksanaan percobaan meliputi (i) optimasi tipe sambung, (ii) optimasi  medium, dan (iii) aklimatisasi planlet hasil sambung mikro. Bahan tanaman yang digunakan sebagai batang atas adalah planlet Cinchona ledgeriana klon QRC, sedangkan sebagai batang bawah digunakan planlet  C. succirubra, berumur empat bulan. Masing- masing percobaan disusun dengan Rancangan Acak Lengkap terdiri dari dua taraf yaitu  kombinasi batang bawah dengan batang atas bentuk sambung tipe V dan L dilakukan  dengan 10 ulangan. Peubah yang diukur meliputi persentase planlet yang bertahan hidup,  jumlah daun,  berkalus atau tidak berkalus pada daerah pertautan, dan persentase planlet yang bertahan hidup. Hasil yang diperoleh menunjukkan bahwa tipe V merupakan cara sambung  mikro  yang terbaik. Medium MS dengan penambahan 3 mg/L IBA adalah medium terbaik untuk pertumbuhan dan perakaran planlet hasil sambung mikro.  Aklimatisasi planlet dilakukan dengan medium tumbuh arang sekam : top soil (1 : 1) yang disterilkan. Tahapan aklimatisasi adalah pre-aklimatisasi dalam ruang kultur  suhu 25 -     27 oCdengan pencahayaan 12 jam per hari dan diikuti dengan aklimatisasi di rumah plastik bernaungan 70% paranet. Dengan metode aklimatisasi ini  90% dari bibit mampu bertahan hidup. Kesimpulan dari penelitian ini menunjukkan bahwa teknik sambung mikro dapat digunakan untuk perbanyakan klonal   Cinchona sp..


2019 ◽  
Vol 11 (6) ◽  
pp. 171
Author(s):  
Yuri Lima Melo ◽  
Isabele Aragão Gomes Trindade ◽  
Monique Cristina Simão Lopes ◽  
Cibelley Vanúcia Santana Dantas ◽  
Josemir Moura Maia ◽  
...  

The aim of this study was to determine the concentration and exposure time to NaCl suitable for the micropropagation of banana, through the analysis of growth traits. Banana propagules were inoculated in MS medium with different concentrations of NaCl (0; 50; 75 and 100 mM) for 120 days (multiplication and rooting, 60 days each), with monthly subcultures. These propagules were measured for plant height, number of leaves, sprouting rate, average number of formed propagules, rooting rate, root length and survival rate. After 30 days, NaCl reduced sprouting rate at multiplication; the number of leaves, rooting rate and root length in rooting; and the height and propagules number in both phases. After 60 days, the NaCl affected the sprouting rate and propagules number in the multiplication; length of root in rooting; and the height and number of leaves in both phases. After 120 days, the reduction in the survival rate was proportional to the increase of NaCl in the medium. Thus, it is concluded that NaCl reduces most of the growth traits and the treatments with 75 and 100 mM NaCl affected multiplication and in vitro rooting more intensely.


2019 ◽  
Vol 24 (2) ◽  
pp. 143-152
Author(s):  
Marhan Nurullia ◽  
Erni Suminar Suminar ◽  
Anne Nurani

This study was aimed at determining the response of turmeric shoot explants after the provision of various types and concentrations of cytokinins in vitro. This experiment was conducted at the Tissue Culture Laboratory, Faculty of Agriculture, Padjadjaran University from January to April 2018. The data were analyzed using T-Test. The experimental method used in this research was Completely Randomized Design (CRD). Explant planting was carried out in Laminar Air Flow. The experiment consisted of 7 treatments consisting of 4 replications and each test consisted of 4 units. Observation of this experiment was carried out for 12 MST. The main observations were made on the data that were tested statistically namely the percentage of explant growing shoots, percentage of explant growing roots, shoot height, number of tuns, number of roots and root length. The treatments consisted of Control, 2.5 mg L-1 BAP, 5 mg L-1 BAP, 0.5 mg L-1 TDZ, 1 mg L-1 TDZ, 0.01 mg L-1 Zeatin and 0.1 mg L Zeatin -1. The results show that the treatment of 1 mg L-1 TDZ shows the best response to the growth of turmeric explants by increasing the number of turmeric shoot explants than the others.RESPONS EKSPLAN TUNAS KUNYIT SETELAH SITOKININ SECARA IN VITROTujuan dari penelitian ini yaitu untuk melihat respons eksplan tunas kunyit terhadap pemberian berbagai jenis dan konsentrasi sitokinin secara in vitro. Percobaan ini dilakukan di Laboratorium Kultur Jaringan, Fakultas Pertanian, Universitas Padjadjaran dari bulan Januari sampai April 2018. Hasil percobaan dianalisis dengan Sample T-Test. Metode percobaan yang digunakan dalam penelitian ini yaitu Rancangan Acak Lengkap (RAL). Penanaman eksplan dilakukan di dalam Laminar Air Flow. Percobaan terdiri dari 7 perlakuan sebanyak 4 ulangan dan setiap ulangan terdiri dari 4 unit. Pengamatan percobaan ini dilakukan selama 12 MST. Pengamatan utama dilakukan terhadap data-data yang diuji secara statistik yakni persentase eksplan tumbuh tunas, persentse eksplan tumbuh akar, tinggi tunas, jumlah tunas, jumlah akar dan panjang akar. Perlakuan terdiri dari Kontrol; 2,5 mg L-1 BAP; 5 mg L-1 BAP; 0,5 mg L-1 TDZ; 1 mg L-1 TDZ; 0,01 mg L-1 Zeatin; dan 0,1 mg L-1 Zeatin. Hasil penelitian menunjukkan bahwa perlakuan 1 mg L-1 TDZ menunjukkan respons yang lebih baik terhadap pertumbuhan eksplan kunyit dengan meningkatkan jumlah tunas eksplan tanaman kunyit daripada yang lainnya.


2015 ◽  
Vol 7 (2) ◽  
pp. 47-52 ◽  
Author(s):  
S Mojumder ◽  
MD Hossain ◽  
MS Haque ◽  
KM Nasiruddin

The experiment was conducted to develop an efficient protocol for in vitro regeneration of mungbean (Vignaradiata) on the aspect of regeneration potentiality of two mungbean varieties (BINA mung 5 and BINA mung 7) as influenced by different combinations of growth regulators supplemented with MS medium. Cotyledon explant of both varieties was used for the present study. Data were collected for various characters of callus initiation, shoot regeneration and root proliferation. Initiation of callus (%) and required days for its initiation and weight of callus were influenced significantly due to the effect of varieties where BINA mung 5 produced more callus induction (40.36%) at minimum requiring time (18.27 days) including heavier sizes of callus (1.54 g) than BINA mung 7 when BINA mung 5 further recorded the longest root (2.92 cm) compare to BINA mung 7. Effect of treatments of the present study were significantly influenced the whole characters regarding callus culture, shoot regeneration and root proliferation. The highest percentage of callus (88.44%) within minimum time (12.53 days) including larger sizes callus (3.521 g) were produced in 1.0 mg L–1 BAP + 2.5 mg L–1 NAA among the treatments while the highest percentage of regenerated shoot (83.44%) at minimum requiring time (17.59 days) and more shoots (7.69 callus–1) were obtained in 1.0 mg L–1 BAP + 2.0 mg L– 1 NAA. Root induction (82.50%), number of roots plantlet–1 (8.469) with minimum requiring time for initiation (14.13 days) and root length (5.250 cm) were the highest in 0.2 mg L–1 IAA + 1.0 mg L–1 kinetin + 0.2 mg L–1 BAP. Incase of interaction, percentage of callus initiation (89.38 %) was the highest in BINA mung 5 treated by 1.0 mg L–1 BAP + 2.5 mg L–1 NAA at requiring minimum time (12.38 days) while same treatment produced the larger callus (3.581 g) among the interactions. The highest percentage (84.38%) and number (7.813 callus–1) of shoot with minimum requiring time (17.50 days) were found from BINA mung 5 treated by 1.0 mg L–1 BAP + 2.0 mg L–1 NAA. Similarly, the longest shoot (5.58 cm) was produced from the BINA mung 5 treated by 1.0 mg L–1 BAP + 2.0 mg L–1 NAA. However, root induction (%), roots plantlet–1, days required for root initiation and root length were statistically similar among the whole interaction treatments due to non significant variation. This result mentioned that the variety BINA mung 5 was better than BINA mung 7 for callus induction, shoot regeneration and root initiation while 1.0 mg L–1 BAP + 2.5 mg L–1 NAA, 1.0 mg L–1 BAP + 2.0 mg L–1 NAA and 0.2 mg L–1 IAA + 1.0 mg L–1 kinetin + 0.2 mg L–1 BAP supplemented with MS medium were the best combinations for better callusing, higher ability of shoot regeneration and root proliferation.DOI: http://dx.doi.org/10.3329/jesnr.v7i2.22203 J. Environ. Sci. & Natural Resources, 7(2): 47-52 2014


2021 ◽  
Vol 49 (2) ◽  
pp. 12038
Author(s):  
Afsaneh TAVANGAR ◽  
Leila KARAMI ◽  
Mohammad HEDAYAT ◽  
Gholamreza ABDI

In this study, micro propagation of two Iranian fenugreek populations and their morphological and biochemical responses to salinity and drought stresses in in vitro culture condition were conducted using factorial experiment in a completely randomized design in three replications. Different explant type (terminal bud, cotyledon and epicotyledon explant) were cultured in MS medium contain different concentration of plant growth regulators such as kin (0, 0.5 and 1 mg / l) and 2,4-D (0.5, 1 and 2 mg / l). Murashige and Skoog (MS) medium supplemented with 1 mg/l kinetin and 2 mg/l 2,4-D showed the highest callus proliferation rate per explants in both populations. The highest amount of callus volume was obtained from the explants of the terminal bud. Proliferated calli from terminal bud explant were green and yellowish, from cotyledon were yellowish to white with soft texture, and the cotyledons were greenish and compact. The results of salinity stresses with different concentrations of sodium chloride (0, 70 and 120 mM) and drought stress with polyethylene glycol (0, 5 and 10%) showed that both stresses decreased callus growth and increased total protein, proline, catalase, peroxidase and trigonelin content in both populations. Trigonelin measurement showed that ‘Borazjan’ papulation had higher trigonelin content, in vitro, than ‘Ardestan’ papulation. 


2020 ◽  
Vol 10 (5) ◽  
pp. 1-7
Author(s):  
H. Mehreen ◽  
J. Zafar ◽  
G. Zishan

Current Status of Saussurea heteromalla was investigated in Galiyat areas of District Abbottabad, viz., Jahaffar, Seri, Beeran Gali, Banj, Haryala, Daryala Gali, Sarbhanna, Barriyan, Akhreela and Broangiala. Saussurea heteromalla was found common in Seri, Sarbhanna and Barriyan; endangered in Haryala, Jahaffar, Banj, Daryala Gali and Beeran Gali and absent in Akhreela and Broangiala. In vitro regeneration of Saussurea heteromalla (D. Don.) Hand-Mazz on MS media was conducted and shoots were developed on full strength MS medium supplemented with 1 mgL-1 GA3. The developed shoots were transferred for root induction to half strength MS medium fortified with various concentrations of Indole butyric acid (IBA) and α-Naphthalene acetic acid (NAA) i.e. T1 (control), T2 (2 mgL-1 IBA), T3 (3 mgL-1 IBA), T4 (4 mgL-1 IBA), T5 (1 mgL-1 NAA), T6 (2 mgL-1 NAA) and T7 (3 mgL-1 NAA). Maximum mean shoot length (6.3 cm), mean number of leaves (7), mean number of nodes (5.25); highest root emergence percentage (71%), means root length (1.5 cm), mean number of roots (3) and highest survival rate (100%) was recorded in treatment T4. However, treatments T5 and T6 also seem to be effective for in vitro rooting of Saussurea heteromalla. Treatments T2, T3 and T7 showed minimum root growth. It was concluded that IBA at higher concentration is more effective for in vitro rooting and better shoot growth of Saussurea heteromalla whereas NAA also initiate rooting but at lower concentration.


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