Combination Of BAP Research Articles

Use of biodegradable polyester-based microvessels for micropropagation of mature Eucalyptus microcorys

Background: Micropropagation, an in vitro vegetative propagation technique using small propagules is one of the main applications of plant tissue culture. It can be used to clone specific plants with desired traits and reduce the cost of plant propagation. In this study, we developed a protocol for micropropagation of Eucalyptus microcorys F.Muell using a selected mature tree, in which we tested various combinations of different culture media and evaluated the use of biodegradable polyester-based microvessels during the adventitious rooting and acclimatisation phases. Methods: Epicormic shoots were used as an explant source. After the in vitro explant establishment and multiplication, we tested 8 combinations of BAP, NAA and IBA in the elongation phase. Three types of microvessels were tested in the adventitious rooting phase and acclimatisation of the microcuttings. Results: Epicormic shoots had an establishment percentage of 40.6% and a total of 820 explants were generated by the 11th subculture, with an average of 12 buds per explant. Best shoot elongation results were achieved with BAP (0.05 mg L-1) + NAA (1 mg L-1) and BAP (0.05 mg L-1) + NAA (1 mg L-1) + IBA (1 mg L-1) combinations, whereas microvessel types M2 and M3 provided higher rooting and acclimatisation. According to the results of ISSR markers, at the end of 535 days of in vitro cultivation, cloning was successful between acclimatised micro-plantlets and the parent plant. Conclusions: The micropropagation protocol using microvessels was efficient in producing E. microcorys clonal micro-plantlets and is recommended for further studies with this species, and for testing in the micropropagation of other species.

In Vitro Multiple Shoot Induction From Embryonic Axes of Annual Herbaceous Legume Faba Bean (Vicia faba L.)

The faba bean (Vicia faba L.) is cultivated in the Mediterranean and Aegean regions only. It is a rich source of protein and an important source of food and feed for human and animal consumption. Faba beans have a narrow genetic base and their improvement through hybridization is not feasible because of high pollen self-incompatibility. This study was carried out using the embryonic axis of cv. Filiz99 and Eresen87 regenerated on MS medium containing 11 different combinations of BAP and NAA. The results showed 100% shoot regeneration frequency with maximum number of 3.3 and 3.5 shoots per explant on cv. Filiz99 and Eresen87 respectively. Regenerating shoots were rooted on 1 mg L–1 IAA. The in vitro regenerated shoots were continuously cultured for 3 weeks to acclimatize them. This approach could improve broad bean seed germination and subsequently regeneration. The results could also facilitate genetic transformation studies.

Callus Induction of Fenugreek Trigonella Foenum-Graecum via Auxin Combined with Cytokinins Hormones, and Assessment of Toxicity via Brine Shrimp Assay

Trigonella foenum-graecum (TFG) is a significant leguminous plant with diverse pharmacological effects. However, the resistant character of this plant accounts for significant difficulties in vitro multiplication, justifying the necessity to try new techniques for in vitro propagation of this plant. Hence, this study reports the effects of BAP, NAA, and 2,4-D on in vitro callus formation of seeds. The toxicity properties of seeds, plant, and callus aqueous extracts of TFG were measured by brine shrimp assay (BSA). Callus index, frequency of callus, callus weight, and morphology of callus were recorded after 30 days of culture. No callus formation was observed in the absence of plant growth regulators. The maximum callus formation observed in the MS media containing 1.0 mg/l 2,4-D, the highest mean of the callus index (52±9.5) with 100% frequency and callus yield (0.52±0.08 g) in 30 days of culture. The highest mean of callus index (37±0 4.05) for combination hormones with 100% callusing and yield (0.37±0.02 g) in 30 days of culture by 1.0 mg/l BAA with 0.5 mg/l NAA. Seeds extract of TFG showed the highest toxicity (954.99 µg/ml), aqueous plant extract (1237.98 µg/ml), and aqueous callus extract (1801 µg/ml) from BSA. Comparing individual hormones, the highest amount of callus in TFG can be yielded 2,4-D hormone alone, and a combination of BAP and NAA can yield 100% callus.

Effect of IAA and BAP application in varying concentration on seed yield and oil quality of Guizotia abyssinica (L.f.) Cass

A low production of oil seed crops, paired with their increased demand, has become a challenge for the edible oil industry in India. Guizotia abyssinica (L.f.) Cass. (niger seed plant) is an annual, dicotyledonous, multipurpose oil seed crop belonging to the family of Asteraceae. Although this plant produces abundant flowers, it fails to produce mature seeds with vigor due to poor vascularization. We investigated the effects of the plant growth-promoting hormone auxin, indole-3-acetic acid (IAA), and the cytokinin, 6-benzyl aminopurine (BAP) individually and together on the growth and oil quality of niger in lateritic soil. Pot experiments were conducted during the wet seasons of 2014 and 2015 in subtropical eastern India. The plants were sprayed with 25, 50, 75, and 100 mg L−1 each of IAA and BAP, 50, 75, and 100 mg L−1 of IAA + BAP, or water (control). When applied individually—IAA or BAP at 75 mg L−1 (I75B0 or I0B75)—the biomass production was maximized. The combination of IAA (50 mg L−1) and BAP (100 mg L−1; I50B100) yielded significantly high biomass (38 and 40 g plant−1) and seed yield (13.24 and 12.67 g plant−1) in 2014 and 2015, respectively. The most effective combinations for increasing niger seed oleic acid content were I50B100 and I50B75 as compared to other combinations of growth hormone. These results suggest that exogenous phytohormones can improve the seed yield and oil quality of niger seed plants in acid lateritic soil.

Effect of Plant Growth Regulators on in Vitro Propagation of Paris Polyphylla

ABSTRACT In vitro propagation using plant growth regulators (PGRs) was evaluated in Paris polyphylla. Thin cell layers (0.5–1 mm) of rhizomes (explants) were sliced and inoculated on half-strength MS (Murashige and Skoog) medium with 3% (w/v) sucrose and solidified with 0.3% (w/v) clerigel. In this medium, 50% of the explants were treated with (0.5 µg mL−1) activated charcoal (AC) and 50% without AC. The half strength MS medium was supplemented with different concentrations of 6-benzylaminopurine (BAP) either individually or in combinations with auxins. The optimal percentage (80%) of shoot response was induced by the synergistic action of BAP (0.5 µg mL−1) and α-naphthalene acetic acid (NAA) (0.2 µg mL−1) in the medium containing AC. After sub-culturing, the highest number (4.00 ± 1.00) of shoot buds per explant was achieved in the MS medium with AC, which was augmented with a combination of BAP (0.5 µg mL−1) and NAA (0.2 µg mL−1). The medium containing AC showed greater shoot induction than the non-AC medium. Up to 94.4% of the regenerated shoots successfully established in the greenhouse soil.

In vitro regeneration and Agrobacterium-mediated genetic transformation of Dragon\u2019s Head plant (Lallemantia iberica)

Dragon’s head plant (Lallemantia iberica), is a flowering species belongs to the mint family (Lamiaceae). The species contains valuable essential oils, mucilage and oil which are used in pharmaceutical and food industries. Tissue culture is a feasible strategy to attain large‐scale production of plantlets with a huge potential to produce plants with superior quality. The objective of this study was to develop a simple and efficient method for regeneration and transformation of L. iberica. To reach this goal, the regeneration ability of various explants including leaf, cotyledonary node, hypocotyl and cotyledon segments was investigated in MS medium supplemented with diverse concentrations of NAA (Naphthalene acetic acid) and BAP (6-Benzyl Amino Purine). According to the results, cotyledonary nodes showed the best regeneration response. The maximum rate of regeneration (and number of induced shoots was achieved in 1 mg l−1 BAP in combination with 0.05 mg l−1 NAA from the cotyledonary nodes. Additionally, through the optimized regeneration technique Agrobacterium-mediated transformation of L. iberica was successfully accomplished. Gene transfer was assessed on leaf samples from regenerated plantlets under a fluorescent microscope to detect the GFP signals. Moreover, transgene integration and its expression were confirmed by PCR and RT-PCR analysis, respectively. The establishment of these efficient regeneration and genetic transformation methods paved the way for further application such as plant improvement, functional analysis and gene editing.

ANALISIS KANDUNGAN SULFORAFAN PADA FASE KECAMBAH BEBERAPA JENIS BROKOLI DALAM MEDIA MS YANG DIBERIKAN NAA DAN BAP SECARA IN VITRO

This study aims to: (1) determine the content of sulforaphane in several types of broccoli sprouts. (2) Knowing the difference in sulforaphane content in the combination of NAA, BAP and types of broccoli grown on MS media; (3) knowing the difference in sulforaphane content in the combination of NAA and types of broccoli sprouts grown on MS media; (4) knowing the difference in sulforaphane content in the combination of BAP and the type of broccoli sprouts grown on MS media. This study used a completely randomized design (CRD) arranged in a factorial manner, namely factor A: NAA 0, 0,5, and 1 ppm. B: BAP 0, 1, 2 ppm and C : 2 Types of Broccoli. The variables observed were: germination time, germination weight, and sulforaphane content. The data were analyzed by analysis of variance and continued with the 5% BNT test. The results showed that germination time began to appear on day 3 and all seeds germinated. Sprout weight varied greatly, and the combination of treatments N1, B0 and BR2 gave the highest average weight and calculated concentration (ENT) of sulforaphane, followed by treatments N0B2BR2 and N0,5B0BR1.

OPTIMIZATION OF GROWTH REGULATORS ON In vitro PROPAGATION OF Moringa oleifera Lam. AND PERFORMANCE EVALUATION OF FIELD GROWN TISSUE CULTURED PLANTS

Moringa oleifera is the most widely cultivated species of the Moringaceae family attributed to several medicinal uses and a high nutritional value. To obtain plantlets of drumstick tree (Moringa oleifera Lam.) with identical genetic make up for propagation or field production, in vitro cultures of drumstick tree were produced. For multiple shoot induction, the stem explants of drumstick tree plantlets were cultured on Murashige and Skoog (MS) agar medium supplemented with different concentrations of 6-benzyl adenine purine (BAP) and maintained at 25 ± 2°C for 5 weeks. The results showed that multiple shoot induction was present in the stem explants that were cultured on the MS medium supplemented with BAP at the concentration range of 0.5 - 3.0 mg/l or the combination of BAP at 1.5 mg/l, TDZ at the range of 0.2 - 1.0 mg/l and NAA at the range of 0.2 -1.0 mg/l. The MS medium containing BAP at 1.5 mg/l was found to produce 95.3% shoot formation with the highest average number of 8.4 shoots per explants. For plant regeneration, the shoot explants were cultured on ½ MS + 7 g/l agar + 15 g/l sucrose supplemented with IBA and IAA at different concentration root induction. The results showed that ½ MS + 7 g/l agar + 15 g/l sucrose supplemented with IBA at 0.3 mg/l and IAA at 0.2 mg/l produced 100% root formation with the highest average number of 4.2 roots per explants and 3.5 cm in length. The medium consisting of (40% soil + 50% sawdust + 10% worm compost) gave the highest average survival rate (88.9%) after 5 weeks under greenhouse conditions. Tissue culture protocol reported in this study is an alternative means of micro propagation of drumstick plantlets with uniform genotypes for breeding selection and field production.

In Vitro Regeneration of Korarima (Aframomum corrorima (Braun) P. C. M. Jansen): A Threatened Spice and Medicinal Herb from Ethiopia.

Developing an in vitro regeneration system is very important to increase production and productivity of plants as well as for the conservation of rare and threatened medicinal plants like korarima (Aframomum corrorima (Braun) P. C. M. Jansen). To date, no study dealing with in vitro indirect regeneration system of korarima has been reported. Thus, in this study, we developed an efficient and reproducible protocol for in vitro regeneration of korarima via callus. The procedure involved soaking seeds in 50% H2SO4 for 16 h that resulted in 92.5% germination on plant growth regulators (PGRs)-free half-strength Murashige and Skoog (MS) basal medium after a month. Shoot and rhizome induction rate of 93.75% was obtained on the MS medium containing 1.5 mg/l BAP in combination with 0.1 mg/l IBA after five weeks. Whitish yellow friable callus was obtained from rhizome culture taken from in vitro grown plantlets. The MS medium containing 2.0 mg/l 2, 4D in combination with 0.5 mg/l kinetin, resulted in 77.5% callus induction. The shoot regeneration rate of 45% was obtained from callus on the MS medium containing 2.0 mg/l TDZ in combination with 0.5 mg/l IBA. The mean shoot number of 10.83 per explant was obtained upon multiplication on the MS medium containing 1.5 mg/l BAP with a mean shoot height of 5.37 cm. The best rooting responses were obtained on half MS medium supplemented with 0.5 mg/l IAA resulting in a mean number of root of 18.59, mean root length of 9.71 cm, and mean shoot height of 7.32 cm. The plantlets showed 75% survival efficiency after acclimatization. The present regeneration protocol offers a conceivable system towards effective conservation and genetic improvement of the crop by increasing the efficiency of genetic transformation.

PENGARUH KOMBINASI IAA (Indole-3-Acetic Acid) DAN BAP (6-Benzylaminopurine) TERHADAP INISIASI TANAMAN VANILI (Vanilla planifolia Andrews)

Provision of Vanilla (Vanilla planifolia Andrews) with traditional cultivation often has problems, including the availability of unhealthy seeds due to disease caused by Fusarium oxysporum. This disease can thwart vanilla plantations up to 85% because the pathogen can infect all parts of the vanilla plant making it difficult to control efforts. Alternative efforts were made to overcome this problem, namely through the initiation process in tissue culture by combining the hormones IAA (indole-3-acetic acid) and BAP (6-benzylaminopurine). This study aims to determine the effect and the maximum concentration of the combination of IAA and BAP on the initiation of vanilla plants. This study was conducted based on a completely randomized design (CRD) with 5 treatments and 3 repetitions. This treatment consisted of: A (MS + 0.1 ppm IAA + 1 ppm BAP), B (MS + 0.2 ppm IAA + 1 ppm BAP), C (MS + 0.3 ppm IAA + 1 ppm BAP), D (MS + 0.4 ppm IAA + 1 ppm BAP), and E (MS + 0.5 ppm IAA + 1 ppm BAP). Observations were made after 40 days from the planting process. The results showed that the concentration of the combination of IAA and BAP can affect the growth and organogenesis of the initiation of vanilla plants (Vanilla planifolia Andrews). Treatment C (MS + 0.3 ppm IAA + 1 ppm BAP) was the best concentration from this study, with average values: number of shoots (1), shoot length (1.73 cm), number of roots (1), and number of leaves (1.33).

Development of an Efficient In vitro Regeneration Protocol for Chrysanthemum (Chrysanthemum morifolium Ramat)

An efficient and rapid in vitro regeneration protocol was developed for chrysanthemum (Chrysanthemum morifolium Ramat) using two local varieties of Bangladesh namely, BARI Chrysanthemum-2 (BARI Chry-2) and local yellow (Y). MS medium supplemented with nine different concentrations and combinations of BAP and IAA was employed to optimize regeneration protocol using young in vitro derived leaf explants. Direct organogenesis was observed from the leaf explants on MS medium supplemented with 0.5 mg/l BAP and 2.0 mg/l IAA (T6) for both the varieties. This treatment (T6) induced shoot buds directly on the adaxial surface of the leaf providing the highest regeneration percentage (90% for BARI Chry-2 and 94.73% for Y), the highest number of shoot/explant (7.6 for BARI Chry-2 and 8.6 for Y) and maximum length of the shoot after six weeks (3 cm for BARI Chry-2 and 2.9 cm for Y) of culture. Explants with initially regenerated shoots were subculture on hormone free MS medium for shoot elongation after 4 weeks of their inoculation. During elongation of shoots, 90-95% of the regenerated shoots produced roots spontaneously in hormone free MS medium within 7-8 weeks of their inoculation. Rooted plantlets were transplanted to the field following hardening where 100% plantlets were survived and produced flower without any variation. Plant Tissue Cult. & Biotech. 31(2): 161-171, 2021 (December)

Vegetative rescue and in vitro propagation of Persea willdenovii

Persea willdenovii is a native species popularly known for its high potential for medicinal use. Due to problems in low seed production and germination, vegetative propagation appears promising. Thus, we seek with this work to evaluate techniques of vegetative rescue and the potential use of shoots obtained in tissue culture. Adult trees of the species were used to apply total and partial girdling treatments and obtain pruned branches as vegetative rescue techniques. Shoot production was monitored over time (90 to 240 days). Sprouts were used for in vitro propagation, and disinfestation treatments were carried out using biocides such as NaClO and Plant Preservative Mixture PPMTM (PPMTM), changing the time and concentrations of the products. Multiplication was also tested using doses of 6-benzylaminopurine (BAP) and media and the potential for callogenesis under combinations of BAP and Naphthalene Acetic Acid (ANA). The vegetative rescue by complete or semi-girdling has the potential to produce shoots, though pruned branches proved to be one of the best techniques for rescue. In the micropropagation, the use of biocides such NaOCl for a long period of time (2 % for 20 min) with PPMTM added to the culture medium shows a potential for disinfestation in the in vitro establishment. For in vitro multiplication, the usage of WPM and BAP (2 a 4 g L-1) promotes higher shoot length, shoot number and leaf number. For indirect organogenesis by leaf segments, the cultivation with BAP and ANA was not responsive to the induction of callogenesis.

The effect of 2,4-dichlorophenoxyacetic acid, benzyl amino purin and cupric sulphate on in vitro propagation system from shoot apices of shoot tiller of hybrid Napier grass (Pennisetum purpureum Schum)

An efficient micropropagation method of hybrid Napier grass (Pennisetum purpureumn Schum) for in vitro plant production and material breeding was established from multiple-shoot clumps (MCS) regeneration system. This system was important for forage breeding system. Shoot apices from shoot-tillers produced MSC on Murashige-Skoog (MS) induction medium containing several combinations of BAP and 2,4-D in induction stage. The addition of 5 μM (v/v) and 50 μM (v/v) CuSO4 were added in best medium for inoculation to proliferate the clump in proliferation/multiplication stage. Plant regeneration was achieved by culturing on solid MS with several combination of medium containing NAA and BAP in regeneration stage. The best results for induction were Murashige-Skoog (MS) induction medium containing 2 mgL−1 BAP and 0.1 mgL−1 2,4-D. The proliferation stage on MS medium containing 5 μM CuSÜ4 effective for proliferation (50% multiple shoot formation). The regeneration stage using 0.1 mgL−1 NAA and 2.0 mgL−1 BAP (51.6% number of shoot can regenerate). All plantlets were successfully grown up in an acclimatization stage. Based on the results, the hybrid Napier grass regeneration via MSC was a stable tissue culture system (no albino plats), which could be applied either for further genetic transformation assay or for alternative supply of nursery plant in the future.

Growth regulators and their reflection on different hop genotypes cultivated under in vitro conditions

Abstract Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.

Propagation of Coccinia cordifolia (L.) Cogn. from shoot tip and nodal segment through micropropagation techniques

The present work was undertaken to develop a reproducible protocol for the micropropagation of an important medicinal plant of the Cucurbitaceae family, Coccinia cordifolia (L.) Cogn., by using shoot tips and nodal segments to overcome the impediment in seed settings and seed germination of conventional reproduction. To develop an efficient protocol, 0.1% HgCl2 treatment for 6 minutes was found effective for surface sterilization of field-grown explants to eliminate microbes and fungus and to get healthy tissues. Throughout the study, different concentrations of auxin, cytokine, and gibberellin were used either alone or in combination as supplemented in MS medium to find suitable conditions and suitable explant (nodal segment). BAP was the best cytokine source as 2.0 mg/l BAP produced 4.0±0.20 and 4.8±0.09 shoots per culture and gained 5.8±0.11 cm and 6.7±0.32 cm length with regeneration rate of 90 and 100 percent in shoot tips and node, respectively. The highest percentage (90%) of regeneration for axillary shoot proliferation was obtained in node explants at MS medium containing 2.0 mg/l BAP + 0.1 mg/l NAA. The highest number of shoots regenerated per culture was 3.0±0.41, with a length of 6.0±0.16 cm. GA3 was also found effective in producing longer shoots with the combination of BAP, but the average shoot number was reduced drastically. Although full-strength MS medium was found to be ideal for shoot regeneration and used in shoots proliferated experiments, half and full-strength of MS medium with auxins supplements of 0.5, 1.0, and 2.0 mg/l either of IBA, NAA, and IAA) were used for roots growth and half strength nutrients supplemented with 0.5 mg/l IBA was found most compelling. The highest number of roots regenerated per shoot was 3.1±0.30, and the average root length was 1.8±0.30 cm. Rooted plantlets were finally transplanted into small plastic pots containing sun sterilized sand, soil and humus (1: 2: 1) to adapt the plantlet in ex vitro environment, and acclimatized plantlets showed 95% survival rate in outdoor condition which proved the effectiveness of using biotechnology to improve plant’s growth rate and mass production.

Induction of Embryogenic Callus in Cashew (Anacardium occidentale L.) by Cotyledon, Nucellus and Testa with Antioxidants Effect

Aims: This work aims to evaluate the influence of different growth regulators combined with antioxidants on the induction of callus with different explants in cashew.
 Materials and Methods: Nucellar tissues, cotyledons and testa from elite tree nuts are cultured on media that differ in their concentration of 6-Benzylaminopurine (BAP: 0 mg/l; 0.25 mg/l; 0.5 mg/l) and Acid 2,4 dichlorophenoxyacetic (2,4-D: 0 mg/l; 0.8 mg/l). Therefore, for the control of the browning of the explants, the effect of activated charcoal and polyvinylpyrrolidone was tested. The amount of callus formed is evaluated after 28 days and after 90 days by simple observation according to a given scale. 
 Results: Analysis of variance of callus formation 28 days after the culture of explants shows that the interaction between growth regulators and antioxidants significantly influences (p < 0.05) the induction of callus. The combination BAP 0.25mg/l and 2,4-D 0.8mg/l produces on average more callus (0.47 ± 0.00). There is a very significant difference (P < 0.05 to p<0.001) in the effect of growth regulators and antioxidants on obtaining callus after 90 days of cultivation. The 0.25 mg/l combination of BAP and 0.8 mg/l of 2,4-D still appears to be the best combination of growth regulators for callus with 58% of callus formed. Cotyledons in the presence of PVP respond better than activated charcoal. The nucellus are the explants that respond better in the presence of activated charcoal. The test for Least Significant Difference reveals that PVP significantly promotes (p <0.05) the production of a large number of calli (77%) unlike activated charcoal (9%) after 90 days of culture.
 Conclusion: Summary, for obtaining viable and embryogenic callus in cashew, the combination of BAP at 0.25 mg/L and 0.8 mg/L of 2,4-D is the best and this in the presence of PVP with cotyledons.

Effect of plant growth regulators on in vitro induction and maintenance of callus from leaf and root explants of Atropa acuminata Royle ex Lindl

Atropa acuminata, an important medicinal plant belonging to family Solanaceae is under tremendous threat of extinction in its natural habitat due to the overexploitation by pharmaceutical industries. Present study is an attempt of establishing callus cultures of this important medicinal plant as callus has considerable potential as an alternative for production of secondary metabolites for industrial use, hence reducing pressure on natural populations.Callus cultures were established from leaf and root explants of Atropa acuminate. Murashige and Skoog (MS) media containing different concentration and combinations of 6-Benzyl Amino Purine (BAP), Naphthalene Acetic Acid (NAA), Kinetin (Kn) and 2,4- Dichloropheoxyacetic acid (2,4- D) were used for callus induction. Different phytohormonal combinations resulted in different types and degrees of callus. The combination of BAP and NAA on MS media supplemented with 0.5 mg/l BAP in combination with 1.0 mg/l NAA, was found to be the most efficient for in vitro callus development from root explants and from leaf explants most effective combination and concentration was 1 mg/l of both BAP and NAA. The maximum mean fresh weight of callus formed using root explants was 33.13 mg per explant and maximum fresh weight obtained from leaf explants was 22.14 mg per explants.

Callus inducement of Toona sinensis: Potential agents against SARS-Corona virus replication

Bioprospecting of anti-SARS-corona virus phytochemical agents is an important issue today as an alternative to chemical drugs whose effectiveness has not been fully successful with no light effect. Quercetin, a component of Toona sinensis young leaves extract, was reported to have anti-viral activity against SARS-corona virus replication in vitro. Currently, the low efficiency of secondary metabolite production is an obstacle. Production of secondary metabolites from callus induction in vitro is considered more efficient and in a short time for commercial applications. The composition of hormones in callus media affects the secondary metabolites formed. The study was conducted to determine the effect of synthetic hormones auxin (BAP) and cytokinin (NAA) on callus percentage, fresh and dry callus cell weight and the average time of callus formation. The call percentage (100%) was obtained in different combinations of BAP and NAA in MS medium. The highest fresh weight of callus (165.50 gL−1) and dry cell weight (28.47 gL−1) were observed in MS medium (1.5 mgL−1 BAP and 1 mgL−1 NAA) within 7 days initial formation time of callus. The results showed that all the indices measured were positively correlated with callus induction in T. sinensis.

Response of Different Explants for Callus Induction in Cucumber

In vitro regeneration of cucumber is relatively difficult for genetic improvement. In this regard, different concentrations of growth regulators and three types of explants (cotyledon, hypocotyl and leaf disc) were investigated for their efficiency on callus induction potential. Among different explants explored for callus induction with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), leaf disc responded earlier (4.67 days) and showed higher percentage of callus induction (91.50%) with 2 mg/l 2,4-D supplemented Murashige and Skoog (MS) media. The same concentration of 2,4-D resulted in the maximum callus fresh (0.56 g) and dry weight (0.39 g) from leaf disc explant. Then the callus was transferred to untreated, 2.0 mg/l BAP + 0.2 mg/l NAA + 1.0 mg/l Kn, 2.0 mg/l BAP + 1.0 mg/l NAA + 1.0 mg/l Kn and 2.0 mg/l BAP + 1.5 mg/l NAA + 1.0 mg/l Kn fortified MS medium. After transferring the callus of different explants to shoot regeneration media containing different concentrations of 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA) and Kinetin (Kn), only cotyledon callus started to regenerate shoot. The combination of BAP (2 mg/l) + NAA (0.2 mg/l) + Kn (1 mg/l) showed highest shoot regeneration percentage (67.77%) and the maximum number of shoots (5.12) per explant were recorded in the treatment combination of 2 mg/l BAP + 0.2 mg/l NAA + 1 mg/l Kn. These results provided a basis for the optimization of the callus induction protocol of cucumber for genetic transformation.

Use of micropropagation in the vegetative rescue of adult trees of Cedrela odorata L.

The objective of this work was the vegetative rescue of Cedrela odorata L. adult trees from forest areas by micropropagation using shoot regeneration from axillary buds in vitro. Nodal segments (0.5 and 1.0 cm) of shoots formed from cuttings taken from canopy sprouts were used. The explants were successfully established in vitro on MS medium supplemented with 2.22 ?M BAP, 0.49 ?M AIB, 0.28 ?M GA3 and PPM (0 and 8.6 ?M). Shoot multiplication on MS with combinations of GA3 (0.28 and 1.12 ?M) and BAP (2.2, 3.3, 4.4, 6.6, and 8.8 ?M) and shoot rooting on medium with 4.9 ?M AIB, 5.7 ?M AIA or 5.37 ?M ANA were evaluated. The MS medium supplemented with 0.28 ?M GA3 and 8.8 ?M BAP generated 4.16 shoots per explant. The best rooting induction was observed on medium containing 4.9 ?M IBA, resulting in 60% of shoot rooting. The plantlets rooted were acclimatized and showed normal development with 97% survival rate. The use of canopy sprouts as explants is feasible in the rescue of C. odorata and the combination of BAP and GA3 favors in vitro multiplication.