scholarly journals Optimization of Culture Media Formulations for Micropropagation of Lepisanthes fruticosa

2018 ◽  
Vol 15 (1) ◽  
pp. 51-58 ◽  
Author(s):  
Zuraida Ab Rahman ◽  
Mohd Shukri Mat Ali ◽  
Mohd Norfaizal Ghazalli ◽  
Khadijah Awang ◽  
Ayu Nazreena Othman

Tissue culture provides an avenue for the production of high quality clonal plants in large numbers within a short time. Here, we describe the development of protocols for reproducible in vitro micropropagation of Lepisanthes fruticosa via direct organogenesis. Shoots were initiated from two types of explants, nodes and young shoots, to establish in vitro cultures on Murashige and Skoog’s (MS) medium or Woody Plant Medium (WPM) supplemented with different concentrations of benzylaminopurine (BAP). Semi-solid WPM media containing 1 mg/L BAP was most effective in shoot initiation in both node and young shoot explants, giving 40% and 20% shoot induction, respectively. The highest rate of shoot proliferation from young shoot explants was obtained using BAP at 3.0 mg/L in combination with NAA at 1.0 mg/L in WPM culture medium. This combination of growth regulators in the medium was also suited to root initiation.

2021 ◽  
Vol 31 (1) ◽  
pp. 1-12
Author(s):  
PV Chaithanya Lakshmi ◽  
CM Narendra Reddy ◽  
B Srinivas

In general, antimicrobial agents are often used in micropropagation techniques to obtain contaminant free clones. The objective of the present study was to evaluate the effects of bavistin and cefotaxime on producing contaminant free plants of Ruellia tuberosa cultured on MS supplemented with phytohormones. Field grown nodal explants of Ruellia tuberosa was used to regenerate entire plants via direct organogenesis. Among the decontaminants tested, the fungicide bavistin along with higher concentration of BAP (2.0 mg/l) and lower concentration of NAA (1.0 mg/l) was the most effective in regeneration and producing contaminant free shoots from cultured explants. This fungicide at 300 mg/l minimised fungal contamination with survival rate of 54%. While the addition of decontaminant cefotaxime at low concentration (200 mg/l) along with same concentration of BAP and NAA stimulated the bud formation and controlled the bacterial contamination. However, its increasing concentration adversely affected the survival rate of Ruellia tuberosa. These findings clearly showed that low concentrations of bavistin and cefotaxime were not only non-toxic but also facilitated bud regeneration. The results achieved showed the decisive role not only of the use of successful fungicides and antibiotics, but also of their sufficient doses were very important in reducing contamination and helping multiple shoot proliferation. Plant Tissue Cult. & Biotech. 31(1): 1-12, 2021 (June)


2018 ◽  
Vol 30 (2) ◽  
pp. 283-294 ◽  
Author(s):  
Mani Manokari ◽  
Mahipal S. Shekhawat

Abstract The present study reports an efficient in vitro propagation system for Turnera ulmifolia using nodal segments as explants. Turnera ulmifolia (Passifloraceae) is an important garden plant with multipotent medicinal values. Effective shoot proliferation was achieved on agar gelled MS medium (Murashige and Skoog, 1962). The maximum number of shoots (8.3 ± 0.57) per initial explant was obtained on MS medium supplemented with 8.88 mM of 6-benzylaminopurine (BAP) and 0.54 mM of α-naphthalene acetic acid (NAA). The highest number of shoots (59.5 ± 2.10) proliferated on semi-solid MS medium (with agar) augmented with 2.22 mM of BAP and 2.32 mM of kinetin (Kin) along with 0.54 mM of NAA. Longer (4-5 cm) and healthy shoots were rooted (12.0 ± 0.10 roots per shoot) on half-strength MS medium fortified with 9.84 mM of indole-3 butyric acid (IBA). The in vitro regenerated plantlets were hardened in the greenhouse and transferred to the field. Significant developmental changes were observed in the foliar micromorphology of in vitro raised plantlets when these were transferred to the field. The stomatal index was gradually reduced (26.72 to 21.25) in the leaves from in vitro to field environments. But, vein-islets and veinlet terminations (13.4 and 7.6) were increased (39.7 and 18.4) respectively from in vitro to in vivo grown plants. Simple, unicellular, less frequent and underdeveloped trichomes were observed with the leaves of in vitro plants but fully developed trichomes recorded in the field transferred plants. The study could help in understanding the response and adaptation of tissue culture raised plantlets towards changed environmental conditions.


2014 ◽  
Vol 23 (2) ◽  
pp. 211-219
Author(s):  
Md. Abul Kalam Azad ◽  
Hasnatul Arefin ◽  
Md. Amzad Hossain

After inoculation of young leaves of date palm offshoot required about six months to complete the morphogenesis process. Fourteen weeks were required for embryogenic callus formation under continuous dark condition and nine weeks for shoot initiation (under 16/8 h light/dark). The highest number of explants (80%) produced callus in modified MS containing 5 mg/l 2,4-D + 2 mg/l 2ip. Sixty per cent of explants produced callus in the modified medium containing 5 mg/l 2,4-D + 5 mg/l NAA. while only 50 per cent of the explants formed callus in the same medium when supplemented with only 5 mg/l 2,4-D. The induced calli were transferred to modified MS for shoot proliferation. A combination of two cytokines showed better performance than single ones in shoot induction. The highest percentage (70) of shoot developed in modified MS containing 2 mg/l BAP + 1 mg/l Kn. Forty per cent shoot induction was found in the same medium supplemented with 2 mg/l of BAP. Thirty per cent shoot formed in MS containing 1 mg/l of Kn. The shoots were subcultured at three- four week intervals throughout culture duration. D. O. I. http://dx.doi.org/10.3329/ptcb.v23i2.17522 Plant Tissue Cult. & Biotech. 23(2): 211-219, 2013  (December)


1970 ◽  
Vol 20 (2) ◽  
pp. 225-231 ◽  
Author(s):  
Meenakshi Banerjee ◽  
Priyanka Modi

Hot extract of Aulosira fertilissima (cyanobacterium) added in different proportions to MS as a liquid culture media for the in vitro propagation of Bacopa monnieri (L.) Pennell. Maximum numbers of shoots were induced from axillary node in MS media (40 ml) + Aulosira extract (60 ml) and maximum shoot multiplication was observed when Kn (1.0 mg/l) was added in the shoot initiation media (mentioned above). Surprisingly rooting was also found to be best in the same combination of MS + cyanobacterial extract that was used for initiation and multiplication of shoots. On an average within a period of three subcultures (2 - 3 months) the nodal explants generated 400 shoots.  Rooted plantlets were successfully transferred to the field, after acclimation in the net house.   Key words: Baccopa monnieri, Cyanobacterial extract, Regeneration, Acclimation   D.O.I. 10.3329/ptcb.v20i2.6917   Plant Tissue Cult. & Biotech. 20(2): 225-231, 2010 (December)


CORD ◽  
2017 ◽  
Vol 33 (2) ◽  
pp. 11
Author(s):  
Anitha Karun

Coconut is one of the principal crops of India cultivated in over 35 districts mainly in the southern states. The productivity of the crop is declining in many of the traditionally cultivated regions owing to ageing plantations as well as biotic and abiotic stresses. These plantations are to be replanted with high yielding varieties/hybrids for which adequate quantity of quality planting material is not available. Even though tissue culture research was initiated in many laboratories in the country, the work was eventually phased out in most of the laboratories for want of a repeatable protocol.  At ICAR-CPCRI, coconut tissue culture programs have been continuing for the past three decades. The attempts made include experimentation with different explants viz., immature inflorescence, plumular tissues, mature palm shoot meristem, ovary and anthers and different culture media supplemented with varying levels and types of hormones. Some of the successful protocols developed at the Institute include coconut zygotic embryo culture for collection and exchange of germplasm, cryopreservation and retrieval of zygotic embryos and pollen and plantlet regeneration from plumular tissues. Even though ICAR-CPCRI has succeeded in obtaining plantlets via direct organogenesis from inflorescence explants, the absence of friable calli formation from explants, the low rate of somatic embryo formation, large number of cultures turning to abnormal shoot development, non conversion of somatic embryos into plantlets, and formation of abnormal somatic embryos remain the major bottlenecks. Gene expression studies are being currently undertaken to decipher the molecular basis of in vitro recalcitrance.


HortScience ◽  
2002 ◽  
Vol 37 (1) ◽  
pp. 200-201 ◽  
Author(s):  
James R. Ault

Shoot tip and stem segment explants collected from greenhouse-maintained plants of Hymenoxys acaulis var. glabra were cultured in vitro for shoot initiation on a Murashige and Skoog (MS) medium supplemented with 30 g·L-1 sucrose, 2.5 μm BA, and 7 g·L-1 agar at a pH of 5.7. Unbranched shoot explants were subcultured to MS medium with 0.0, 0.5, 1, 2, 4 or 8 μm BA for shoot proliferation. A maximum of 10.3 shoots per explant was produced on the medium with 2.0 μm BA. Nonrooted shoots were subcultured to MS medium with 0.0, 0.5, 2, or 8 μm K-IBA for rooting. Maximum rooting was 90% on MS medium with 0.5 μm K-IBA. Rooted shoots were greenhouse-acclimatized for 10 days. Overall survival was 75%. Chemical names used: 6-benzyl adenine (BA); potassium salt of indole-3-butyric acid (K-IBA).


2007 ◽  
pp. 79-90 ◽  
Author(s):  
Tessie Nuñez

Using a suitable medium for specific plant genotype greatly improves the efficiency of the in vitro culture method. The Visayas State University (VSU)-based National Coconut Research Center -Visayas (NCRC-V) evaluated the comparative suitability of the COGENT medium and VSU-modified Y3 (mY3) as in vitro culture media for coconut embryos using Albuera Dwarf (ALD), Baybay Tall (BAYT),and the VSU-developed Coconiño x Makapuno (VMAC1) hybrid. These two media differ in vitamin components, iron concentration and state during the first two stages of culture, namely germination and first subculture. Results showed that mY3 was more suitable for the in vitro germination and development of the coconut embryos than the COGENT medium. Significantly higher germination rates were observed in BAYT, ALD, and VMAC1 cultured in the semi-solid mY3 than those in the liquid COGENT medium from the first week until the fourth week of initial culture. Germination rates of 100%, 85.8% and 84.5% were obtained from CÑO x MAC, ALD, and BAYT, respectively. Furthermore, significantly higher percentages of germinating embryos with developing shoot and root were observed in the semi-solid mY3 than in the liquid COGENT. Likewise, better growth of plantlets in liquid mY3 was noted during the fourth and fifth months of culture. Among coconut types, VMAC1 had the highest germination rates in the two media and the best growth in mY3. BAYT had better growth in the COGENT medium while ALD had better shoot development in mY3.


2007 ◽  
Vol 44 (07) ◽  
pp. 514-519 ◽  
Author(s):  
Ved Prakash Pandey ◽  
Jose Kudakasseril ◽  
Elizabeth Cherian ◽  
George Patani ◽  

Two different methods of in vitro multiplication of Rauwolfia serpentina from nodal explants were compared viz. multiplication via callus morphogenesis and that via shoot proliferation from axillary buds. The second method was found to be far better. The optimum shoot proliferation occurred on Murashige and Skoog (MS) medium supplemented with 1 mg/L naphthalene acetic acid (NAA) and 2 mg/L of benzyl aminopurine (BAP). The best rooting of shoots occurred on MS medium containing 4% sucrose and 1 mg/L of NAA. Solid and liquid MS media were found to be similar in supporting shoot proliferation. The plants produced were successfully hardened and established in soil. An easy, reliable and reproducible protocol was developed for in vitro micropropagation of Rauwolfia serpentina from nodal explants.


2018 ◽  
Vol 28 (1) ◽  
pp. 69-76
Author(s):  
H Reshmi Singha ◽  
Sangram Sinha ◽  
Rabindra Kumar Sinha

An efficient method of clonal propagation through nodal culture of Solanum torvum Swartz. is described. Different concentrations of BAP/Kn alone or in combination with IAA were tested for direct shoot bud induction and proliferation. Lower concentration of BAP/Kn alone produced better shoot proliferation and elongation. Maximum number of shoot proliferation was achieved from MS supplemented with Kn 0.5 mg/l with an average 4.0 ± 1.41 shoots during 28 days of culture. Addition of IAA to the culture media in combination with BAP/ Kn significantly reduced the number of shoot formation. Regenerated plants also produced roots during subsequent culture in the same media supplemented with BAP/Kn alone or in combination with IAA. The easy nature of in vitro rooting of S. torvum was recorded without any separate root induction phase. Regenerated plants were successfully transferred to the field condition. Clonal feature was cytologically confirmed through the study of mitotic metaphase chromosomes of regenerated plants which reveals 2n = 24 somatic chromosomes. Comparative karyomorphological details between the mother and regenerated plants of S. torvum revealed close similarity in their chromosomal complements and falls under the category of "1B" Stebbin’s symmetric index suggesting true to type nature of the regenerated plant.Plant Tissue Cult. & Biotech. 28(1): 69-76, 2018 (June)


2013 ◽  
Vol 2013 ◽  
pp. 1-3 ◽  
Author(s):  
Mehmet Ugur Yildirim

Origanum acutidens(HAND.-MAZZ.) IETSWAART is a promising ornamental plant that can be widely used in landscape management. It is endemic to Eastern Anatolian region of Turkey. Tissue culture has not been used to micropropagate it. The study reports stem node explants from one-week-old seedlings of the plant for successful micropropagation. The stem nodes were cultured on MS medium containing 0.6, 1.2, 1.8, and 2.4 mg/L BAP with 0.2 mg/L NAA. Visible effects of culture media on shoot proliferation were recorded. Shoot regeneration rate was maximum on MS medium containing 1.80 mg/L BAP-0.2 mg/L NAA. The micropropagated shoots were rooted on MS medium containing 0.2 mg/L NAA. All microrooted plantlets survived during acclimatisation on peat moss. It was concluded thatO. acutidenscan be successfully micropropagated underin vitroconditions.


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