In vitro propagation of Syzygium travancoricum Gamble – an endangered tree species

Rapid multiplication in Syzygium travancoricum, an endangered tree species was achieved from nodal explants on Murashige and Skoog's (MS) medium supplemented with Woody Plant Medium (WPM) micronutrients and plant growth regulators. Multiple shoot induction from young 1–2-year-old seedlings was observed on basal medium supplemented with 17.7 M benzyladenine and 1.3 M -naphthalene acetic acid. A high number of multiple shoots (25 shoots/nodal explant) was observed by the third subculture on multiplication medium. The dark brown nodular callus obtained from in vitro grown shoots through axillary buds regenerated at a high frequency when transferred to 1/2 strength basal medium. The plants rooted on 1/2 strength basal medium supplemented with 1.1 M indole-3-acetic acid and were transferred to the field with 40% survival.     

Micropropagation of the Mediterranean species Viburnum tinus

In vitro propagation of the Mediterranean species Viburnum tinus L. was established from an outdoor-grown shrub. Two standard macrosalt formulations (Margara N30K and Murashige and Skoog), a range of benzyladenine and sucrose concentrations were tested for their effect on shoot multiplication. The cytokinin concentration was the most important factor affecting shoot multiplication. The highest shoot multiplication rate was obtained from single-node explants cultured on Murashige and Skoog medium supplemented with 4.4 M benzyladenine. Cytokinin concentration and an interaction of macrosalts and benzyladenine influenced shoot length on the multiplication stage: best shoot growth was observed on MS medium containing 1.1 M benzyladenine. In addition, sucrose concentrations of 87.6–146.0 mM gave the highest multiplication rates and improved shoot growth. Following a shoot ellongation stage, single shoots were rooted on media containing naphtaleneacetic acid (1.3–5.4 M). Although enhanced in vitro rooting was obtained on media containing 5.4 M naphtaleneacetic acid, reducing the auxin concentration to 1.3 M during the in vitro rooting stage improved acclimatisation frequency and further plant growth in a horticultural substrate.     

Micropropagation of safed musli (Chlorophytum borivilianum), a rare Indian medicinal herb

In vitro clonal multiplication of safed musli (Chlorophytum borivilianum Sant. et. Fernand.), a rare Indian medicinal herb, has been achieved on Murashige and Skoog's (MS) medium supplemented with 22.2 M benzyladenine using young shoot bases as explants. Shoots multiplied at a rate of four-fold every 3 weeks. All shoots rooted when transferred to MS medium with 3/4-strength inorganic and organic constituents and 9.8 M indolebutyric acid and 67% of the micropropagated plants were successfully established in pots. Such plants produced normal fasciculated storage roots as in wild plants.Abbreviations BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - IAA indoleacetic acid - IBA indolebutyric acid - MS Murashige and Skoog (1962) - NAA -naphthaleneacetic acid     

Micropropagation of parsley through axillary shoot proliferation

A micropropagation protocol of parsley,Petroselinum crispum (Mill.) Nyman (curled type) has been developed. Surface-sterilized axillary buds cultured on Murashige and Skoog medium supplemented with benzyladenine, kinetin, or thidiazuron developed axillary shoots (rosettes). Kinetin resulted in only a low proliferation rate. The concentrations of thidiazuron or benzyladenine that were optimal for shoot proliferation, resulted in shoots with a low capability to root. During the rooting treatment, these shoots showed wilting signs. Rooting was increased significantly by using a two-week inductive stage with 2.5 M naphthaleneacetic acid directly followed by acclimatization. Two proliferation media (5 M benzyladenine and 0.5 M naphthaleneacetic acid or 5 M kinetin and 2.5 M naphthaleneacetic acid) resulted in moderate proliferation but produced shoots that were easy-to-root. These media have been tested by repeated axillary proliferation on the same medium. The medium with 5 M benzyladenine and 0.5 M naphthaleneacetic acid was optimal.Abbreviations BA 6-Benzyladenine - IBA Indole-3-butyric acid - MS Murashige & Skoog - NAA -Naphthaleneacetic acid     

In vitro organogenesis of elephant apple (Feronia limonia)

Elephant apple (Feronia limonia L.). was micropropagated on MS medium containing 4.4 M benzyladenine and 4.6 M kinetin using cotyledon explants taken from in vitro-grown seedlings. Adventitious buds formed on the cotyledon developed into shoots that were rooted in half-strength MS medium containing 0.57 M indoleacetic acid and 0.49 M indolebutyric acid. Plants were successfully established in soil.Abbreviations BA 6-benzyladenine - IAA 3-indoleacetic acid - IBA 3-indolebutyric acid - MS Murashige & Skoog     

Clonal propagation of mature elite trees of Commiphora wightii

Elite trees of Commiphora wightii (Arnott) Bhandari were selected from the wild on the basis of their content of guggul, an oleoresin. The selected tree was micropropagated through forced axillary branching on Murashige and Skoog's (MS) medium supplemented with benzyladenine (BA) and kinetin. Highest frequency of shoot formation was achieved on MS medium supplemented with 17.8 M BA, 18.6 M kinetin, 100 mg l^-1 glutamine, 10 mg l^-1 thiamine HCL and 0.3% activated charcoal. Seasonal changes affected the shoot proliferating potential of the initial explants in vitro. Transfer of shoots to a medium containing a lower concentration of BA (1.8 M) and kinetin (1.9 M) before rooting markedly stimulated shoot elongation. Shoots could be rooted by treating them with both indoleacetic acid and indolebutryic acid for 24 h in darkness and transferring them to a low-salt basal medium with activated charcoal. After rooting, transfer to a half-strength White's (modified) medium was necessary for further development of the plantlet. Regenerated plantlets were successfully established in soil.Abbreviations AC activated charcoal - BA benzyladenine - IAA indoleacetic acid - IBA indolebutyric acid - 2-iP isopentenyladenine - MS Murashige and Skoog's medium - NAA -naphthaleneacetic acid - PG phloroglucinol     

Plant regeneration from cotyledons of niger

Callus initiation from seedling explants of niger (Guizotia abyssinica Cass) cv. Ootacamund was found to be better on LS medium containing kinetin (1.4 M) plus 2,4-dichlorophenoxyacetic acid (9 M) than its analogues. Embryoids were induced directly from cotyledons on LS medium supplemented with 2,4,5-trichlorophenoxyacetic acid and 2,4,5-trichlorophenoxypropionic acid. When cotyledon-derived callus was subcultured onto medium with 10.7 M naphthalene-acetic acid and 2.3 M kinetin, embryogenesis was observed. Multiple shoots were obtained from cotyledonary explants in presence of MS medium containing 4.4 M benzyladenine and 11.4 M indoleacetic acid. Regenerated plants that were transferred to pots and grown to maturity were morphologically normal and fertile.Abbreviations NAA naphthaleneacetic acid - IAA indoleacetic acid - BA benzyladenine - GA₃ gibberellic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - 2,4,5-T 2,4,5-trichlorophenoxypropionic acid - 2,4,5-TP 2,4,5-trichlorophenoxypropionic acid - ABA abscisic acid     

Axillary shoot induction and plant regeneration in Plantago ovata Forssk

Axillary shoot induction and plant regeneration were obtained in Plantago ovata. The optimum medium for inducing axillary shoots was Murashige & Skoog (MS) medium [5] supplemented with 4.6 M kinetin and 0.05 M NAA. Rooting of shoots was best on half-strength MS medium containing 5.0 M IBA and 0.05 M kinetin. The regenerated plants were similar to the control plants in karyotypic and phenotypic details.     

In vitro clonal propagation of Capsicum spp.

The effect of various concentrations of benzyladenine (BA 4.4–177.5 M) or kinetin (4.7–185.9 M) on shoot proliferation from shoot-tip explants was investigated in C. praetermissum Heiser & Smith and C. annuum L. Maximum number of shoots were obtained on Murashige & Skoog's medium with 66.6 M BA or 92.9 M kinetin in C. praetermissum, and 88.8 M BA or 116.2 M kinetin in C. annuum after 4 weeks of culture. Combining 1 M 2, 3, 5-triiodobenzoic acid (TIBA) with low levels of BA or kinetin significantly increased shoot number as compared to using either cytokinin alone. Rooting of regenerated shoots was achieved on MS medium containing 5.7 M indoleacetic acid. Best rooting (80–100%) was observed in shoots from TIBA plus BA or kinetin media while only 40–50% of shoots from the BA or kinetin treatments were rootable. Plantlets obtained from TIBA plus BA or kinetin were normal diploids while those from BA or kinetin alone revealed distinct chromosomal aberrations in their root tip squashes. Regenerants from TIBA plus BA or kinetin media were successfully established in the soil (86% survival rate), where they flowered and showed normal meiotic behaviour with 100% pollen viability.Abbreviations BA benzyladenine - IAA indole-3-acetic acid - MS Murashige & Skoog's medium - TIBA 2, 3, 5-triiodobenzoic acid     

Micropropagation of Tamarix gallica from nodal explants of mature trees

Tamarix gallica L. was micropropagated from four-to six-node explants taken from mature trees. Shoot proliferation was induced on Linsmaier and Skoog medium containing 30 g l^-1 sucrose, 7 g l^-1 agar, 200 mg l^-1 reduced glutathione (basal medium) and supplemented with 3.3 M benzyladenine. Adding 0.5 or 1.0 M indole-3-butyric acid (IBA) to the basal medium increased lateral shoot formation and ease of rooting. Microcuttings repeatedly subcultured on 1.0 M IBA produced well-developed roots, a high number of axillary shoots and could be acclimatized in the greenhouse.