Mgl-1 Kinetin Research Articles

Enhancing Clitoria ternatea Propagation Through Indirect Somatic Embryogenesis: Optimal Plant Growth Regulators Type and Concentration for Efficient in Vitro Plant Production

The propagation of Clitoria ternatea poses several challenges, including low germination rates, vulnerability of young seedlings in natural environment, and a slow flowering rate. This study aims to identify the best Plant Growth Regulators and their optimal concentration to stimulate indirect somatic embryogenesis from C. Ternatea leaves explants. The samples were cultured on Murashige and Skoog (MS) medium supplemented with various concentration of thidiazuron (TDZ; 1, 2, 3, 4, and 5 mgL-1 ) and a combination of 2,4-dichloro phenoxy acetic acid (2,4-D; 1, 2, and 3 mgL-1 ) and 0.1 mgL-1 kinetin. The findings revealed that all treatments resulted in the formation of both embryogenic and non embryogenic calluses. The addition of TDZ to the MS medium led to a greater percentage of embryogenic callus formation compared to using the combination of kinetin and 2,4-D. The highest percentage of 84.37% was achieved when samples were cultured on MS medium with 3 mgL-1 TDZ. The most effective induction period of 21 days was observed with the medium supplemented with 3 mgL-1 TDZ, followed by 4 and 5 mgL-1 TDZ, which took 23 days. The research findings suggest that a concentration of 3 mgL-1 TDZ is the optimal PGR type and dosage for achieving rapid and efficient in vitro plant production compared to various concentrations of 2,4-D in combination with kinetin.

In vitro Propagation and Genetic Assessment by using Markers in Potato (Solanum tuberosum L.) cv Kufri Ganga

The purpose of this study was to determine the best types and concentrations of plant growth regulators to use during different stages of micropropagation, as well as to assess the genetic fidelity of tissue culture raised plants of a cv Kufri Ganga using reliable markers. According to the data on shoot bud initiation, a combination of 0.25 mgl-1 BAP and 0.5 mgl-1 Kinetin results in the greatest number of shoots per explant (3.4±0.11) in the shortest amount of time (3.2±0.09) days. The maximum number of in vitro shoots per shootlet (4.7±0.15) was recorded when auxins were combined with cytokinin (MS medium + 0.25 mgl-1 BAP + 0.01 mgl-1 IAA) for shoot proliferation. Root initiation was observed in (2.2±0.10) days on MS medium fortified with 1.5 mgl-1 NAA. The maximum number of in vitro roots per shoot (14.2±0.31) was observed when MS media was fortified with 2.0 mgl-1 IBA. Molecular markers were used to assess genetic fidelity in plants grown in vitro. Monomorphic DNA banding patterns were found in all tissue culture raised plants and mother plants, indicating true to type planting material. This protocol for tissue culture propagation, as well as testing its genetic fidelity, may be beneficial for the successful application of better conservation of germplasm.

Efficient Callus and Shoot Induction Protocol from Leaf and Node Explants of Javanese Ginseng (Talinum paniculatum (Jacq.) Gaertn.)

This present study was carried out to determine an efficient and reproducible callus and shoot induction protocol of Talinum paniculatum by using leaf and node explants. Explants were cultured in MS medium supplemented with a combination and concentration 2,4-D and kinetin (0, 1, 2, 3 mgL-1) in single or combination treatment for 14 days. The best response of callus and shoot induction (100%) and the fastest callus (6,0 ± 2,31 days) and shoot induction (5,0 ± 1,15 days) from leaf explants were obtained from MS medium supplemented with (1 mgL-1 2,4-D and 1 mgL-1 kinetin) and (3 mgL-1 2,4-D and 1 mgL-1 kinetin) respectively. In the case of node explants, none of calli were produced, whereas the best response of shoot induction (100%) and the fastest shoot induction (4,0 ± 0 days) were obtained from MS medium supplemented with 3 mgL-1 kinetin. Callus produced from leaf explant has a bright yellow color and friable texture. The optimized protocol of callus and shoot induction for Talinum paniculatum plant could be used effectively for mass propagation to fulfill the demand of the pharmaceutical industry as well as for its cultivation.

Optimization of Protocol for Callus Formation and Shoot Development in Sugarcane (Saccharum officinarum L.) Cultivar CPF-248

Sugarcane production face numerous difficulties, including infestations of insects and diseases, particularly red rot, a lack of appropriate seed supply, a lack of industrial support, etc. Using a process called plant tissue culture, huge amounts of authentically grown, disease-free plant material may be produced quickly. Plant tissue culture can also be used to quickly reproduce recently released varieties with crucial agronomic characteristics. For this purpose, sugarcane callus culture was collected from the inner soft leaf sheath to increase genetic diversity. Ten different concentrations viz., 1.5, 1.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5 and 6.0 mgL-1 of 2, 4-dichlorophenoxy acetic acid in MS medium were used for callus development, along with 0 mgL-1 control was used. Different combinations and concentrations of BAP+ Kin (1.0+1.0, 1.0+1.5, 1.0+0.5, 1.0+1.0, 1.5+1.0 and 1.0+1.5 mgL-1) were used for regeneration of shoots and auxin. IBA with 6 different concentrations (0. 0.3, 0.5, 0.7, 1.5 and 1.8 mgL-1) were used for rooting of the shoots. Among all the growth regulators, 2, 4-D at 3.0 mgL-1 demonstrated the greatest auxin for callus development with BAP. The 6-12 mm meristem was grown in MS medium with NAA. It was observed that at concentrations viz., 3.5mgL-1 and 4.0mgL-1 of 2, 4-D, maximum callus (79.0-84.5 %) was developed with 4-5 mm in size. Maximum root growth and length (3.7 mm) were reported at 1.0 mgL-1 IBA and 1.0 mgL-1 Kinetin and 1.5 mgL 1 BAP in MS medium. It was concluded that auxin concentration is best for all sugarcane in vitro callus culture, which will assist researchers in future work.

Establishment of an efficient in vitro propagation system for Holostemma annulare (Roxb.) K. Schum. using SH medium

Holostemma annulare (Roxb.) K. Schum. (Asclepiadaceae) is an important medicinal herb and its root tubers are an important ingredient of the drug ‘jivanti’. Here, a more reliable procedure of in vitro propagation of the plant is discussed using SH nutrient medium formulation that facilitates the in vitro multiplication and establishment without callusing from the basal region of shoots that was prominent in the earlier protocols. Surface sterilization of the nodal explants with 10% (v/v) NaOCl for 12 minutes followed by 0.1% (w/v) HgCl2 for 8 minutes exhibited 90% shooting response. The in vitro initiated shoots subcultured to SH medium fortified with 2.0 mgl-1 BA in combination with 0.5 mgl-1 Kinetin and 1.0 mgl-1 IBA produced maximum 7.16±0.15 shoots. Better root induction of the in vitro raised shoots was noticed in SH medium augmented with 0.01 mgl-1 NAA that induced a mean number of 5.65±0.23 thin, long, white coloured roots. The plantlets were transferred to polythene bags containing river sand:garden soil (1:1) for acclimatization, further transplanted in garden soil and river sand mixture (3:1) in earthern pots in the greenhouse and subsequently to the field got established at 80% efficiency. They produced tuberous roots after 4 months of field establishment. Thus, the study standardized an improved system of in vitro propagation of H. annulare using SH medium and this protocol can be extended for long-term conservation methods such as cryoconservation and also for the in vitro production of bioactive metabolites.

Callus induction of mountain papaya endosperm (Vasconcellea pubescens A. DC) with different combination of 2,4-Dichlorophenoxyacetic acid and Kinetin

Endosperm as a result of double fertilization in Angiospermae shows high level chromosomes and polyploidy. It is also considered as dead tissue that unable to be generated to form plantlet. The aim of this research is to determine the effect of kinetin and 2,4-Dichlorophenoxyacetic acid (2,4-D) in induction of callus formation of mountain papaya. This research used A factorial randomized block design with 18 groups, 1 fruit was used for 1 experimental group. Culture using Murashige and Skog (MS) media with combination of three level of kinetin (0, 2, 4 mgL-1) and six level of 2,4-D (0, 1, 2, 3, 4, 5 mgL-1). Maximum endosperm callus induction (0.88%) was achieved from endosperm explant cultured on MS medium fortified with 2.0 mgL-1 Kinetin and 4.0 mgL-1 2,4-D. The fastest day induction (24,66 day) was observed with 5.0 mg L-1 2,4-D. The maximum number of browning (0,10) was induced by 2.0 mgL-1 Kinetin and 5.0 mgL-1 2,4-D. Combination Kinetin and 2,4-D proved could induces callus formation from mountain papaya endosperm.

Shoots multiplication of vanilla (Vanilla planifolia) with benzyl amino purine and kinetin modification

Micro propagation techniques for vegetative propagation of vanilla have been developed to overcome limited planting material and the attack of Fusarium disease. The ability of vanilla explants to regenerate and differentiate to form multiplication of shoots is still limited. Therefore it is necessary to regulate the addition of Benzyl Amino Purine and Kinetin into the media for optimal shoot multiplication so that the quality of the vanilla culture seedlings can be improved. The study was conducted at Tissue Culture Laboratory of Politeknik Negeri Jember using a factorial Complete Randomized Design. Factor 1 is BAP concentration which is 0.5, 1.0, 1.5, 2.0, 2.5, 3.0 mg/liter and factor 2 is Kinetin concentration which is 0.0, 1.0, 2.0 mg/liter. Further data analysis used Duncan’s average difference test. The research result show that The addition of BAP affected the shoots growth rate of vanilla explants with the fastest mean rate of 9 days after inoculation with the addition of 2.5 mgL-1 and 67-100% sprouting ability 14 days after inoculation. The average number of shoots was highest in the addition of 0.5 mg L-1 BAP and 2.0 mgL-1 Kinetin at 56 days after inoculation with a mean of 6.00 shoots / explant.

In vitro Callus Induction and Regeneration of Popular Indica Rice Genotypes

Five rice cultivars viz., ASD 16, White Ponni, Pusa Basmati 1, Pusa Sugandh 4 and Pusa Sugandh 5 belonging to subspecies indica were compared for its ability in callus formation and regeneration. In this experiment, the different parameters viz., the effect of hormones (2,4-D and kinetin), organic supplement (coconut milk O1-CM 100 mll-1, O2-CM 75 mll-1, O3-CM 50 mll-1), explants (seed and immature embryo), media (MS and N6), carbon source (sucrose and maltose) using five genotypes on callus response was studied. The effect of hardening methods was also assessed. Results showed that for enhanced callus induction was with MS medium supplemented with 2.0 mgl-1 2, 4-D + 0.5 mgl-1 kinetin + 30 gl-1 maltose irrespective of explants used. Addition of 100 ml l-1 coconut milk was found have improvement in callus response. The performance of immature embryo was better than seed for callus induction, emrbyogenic callus formation, rhizogenic callus formation and regeneration. MS media provided superiority over N6. Among the genotypes Pusa Basmati 1 rendered outstanding performance in callus behavior. The treatment combination MS + 2.5 mgl-1 BAP + 0.5 mgl-1 NAA + 1.0 mgl-1 KN gave the highest organogenesis response and regeneration of plantlets. Hardening in mist chamber was recognized as the best method to give the highest per cent of regenerated plant lets.

Callus Production Protocol for Aegle marmelos (L.) Corr.: As a Tool for Extraction of Secondary Metabolites

Background: Aegle marmelos (L.) Corr. is an important medicinal and a fruit tree belongs to the family Rutaceae possessing numerous valuable secondary metabolites. The growing commercial importance for secondary metabolites has led to a great demand in the pharmaceutical industry in recent years. Therefore, an efficient callus production protocol was developed as a tool for extracting valuable secondary metabolites from Aegle marmelos.Methods: For seeds, callus induction was observed under three conditions as with seed coat, after removing seed coat and split into two halves after removing seed coat. For callus multiplication, 1cm2 pieces of initiated calli were used. These explants were established in MS medium supplemented with combinations of 2, 4 D either with BAP or Kinetin. All experiments were arranged according to the completely randomized design (CRD) with 20 replicates at the Tissue Culture Laboratory, Department of Crop Science, Faculty of Agriculture, University of Ruhuna, Sri Lanka, for a period of 1 year. Percentage of fungal and bacterial contaminations and percentage of bleached explants were observed to select the best explant/s. Percentage of responded explants were observed to select the best condition for callus induction and quality of callus. Growth of callus was observed visually by giving a score. Best hormonal combination for callus multiplication was observed as fresh weight and dry weight of callus produced under each treatment.Result: High quality callus with higher growth was observed in all combinations of BAP and 2, 4 D tested: ranging from 0.5 mgL-1 to 1.5 mgL-1 BAP and 1.0 mgL-1 to 2.0 mgL-1 2,4 D in Murashige and Skoog (MS) medium. Initiated calli were further multiplied in MS medium supplemented with 2,4 D combined with either BAP or Kinetin. Highest amount of callus biomass was recorded in the MS medium with 0.5 mgL-1 2, 4 D and 1.0 mgL-1 Kinetin (132.58 gL-1 fresh weight). The optimized protocol could be used to produce higher amount of callus in order to extract secondary metabolites from Aegle marmelos (L).

In vitro response and effect of gamma irradiation on four local indica rice varieties

In vitro response of four local Indica rice cultivars viz. Sadamota, Kachamota, Moulata and Dudhkalam was evaluated. The aim of this study is to develop an efficient protocol for callus induction, plant regeneration and to observe the effect of gamma radiation on plant regeneration for creating possible genetic variability. In Different concentration of 2,4-D and growth regulators were supplemented with MS medium (Murashige and Skoog’s) to observe their callus induction frequency using mature embryo as explant. Among the cutivars, the highest primary callus (92.55%) as well as embryogenic callus induction (56.26%) was showed in sadamota at 3.0 mgl-1 2,4-D and 10 mgl-1 kinetin under dark condition. Twenty one days old embryogenic calli were exposed to 0, 2, 4 and 6 Gy of gamma rays and transferred to regeneration medium. Both callus growth and regeneration capacity were found to be decreased with increasing level of exposure to gamma rays. The doses of 4 Gy of gamma radiation were found to be the 50% inhibition dose for callus growth and plant regeneration in sadamota and kachamota, repectively whereas the 50% inhibition dose for moulata and dudhkalam at 2 Gy. This results indicate that sentivity of gamma radiation on rice callus depends on genotype of a genus.

Organogenesis of Tomato (Lycopersicon esculentum Mill.) cv. Sandra and Rocky by Hypocotyl, Cotyledon and Leaf Explants

In vitro regeneration protocols were established for two Tomato (Lycopersicon esculentum Mill.) cultivars (Sandra and Rocky) grown in Kurdistan region using hypocotyl, cotyledonous leaves and leaf explants. The explants excised from 12-15 days old of in vitro plantlets cultured on MS medium supplemented with different concentrations and combinations of plant growth regulators. High organogenesis percentage (100%) was achieved from hypocotyl grown in MS medium augmented with 1.0mgl-1 BA+ 0.5mgl-1 kinetin, which record approximately 3.75 in Sandra and 4.09 in Rocky cultivar as average number of shoots produced from each explant. While the highest organogenesis percentage (100%) was formed from cotyledon leaves using 1.5 mgl-1 BA+0.5mgl-1 kinetin that gave around 3.5 in Sandra cultivar as average number of shoots / explant. In contrast, the highest organogenesis was found on MS medium supplemented with 2.0mgl-1 BA+1.0 mgl-1 kinetin and the average number of shoots produced from each explant were approximately 3.17 in Rocky cultivar. The results of combination between 5.0mgl-1 BA with (0.0, 0.2, 0.5 and 1.0)mgl-1 NAA, indicated that the highest organogenesis percentage was recorded from hypocotyl (91.67 % with 6.5 shoots/explant and 66.67% with 4.0 shoots/ explant) in Sandra and Rocky cultivars respectively grown on MS medium supplemented with 5.0 mgl-1 BA alone. While the highest organogenesis percentage (100%) was induced from cotyledonous leaves in both cultivars (3.67 shoots for Sandra and 3.08 for Rocky) on MS medium supplemented with 5.0mgl-1 BA +0.2mgl-1 NAA. Meanwhile, the highest organogenesis percentage was obtained from leaf explants for both cultivars on the combination of 1.0mgl-1 BA +0.5mgl-1 kinetin, (77.78% and 66.67%) in Sandra and Rocky cultivars respectively. The regenerated plants were successfully transplanted to the field for acclimatization

SOME FACTORS AFFECTING MICROPROPAGATION OF Haplophyllum tuberculatum L

This study was carried out at Plant Tissue Culture Laboratory, Faculty of EnvironmentalAgricultural Sciences, El-Arish University, North Sinai Governorate, Egypt during 2014 to 2017.Effect of three medium types, Murashige and Skoog (MS), Linsmaier and Skoog (LS) and Lloyd andMcCown (WPM), two explant types (shoot-tip and single node cutting), three cytokinin types(Benzyladenine, Kinetin and 2-isopenteny1 adenine), two additives; viz., s (thiamine and pyridoxinehydrochloride) and amino acids (asparagine and arginine) and two gelling agents (guar seed powderand locust bean seed powder) combined with agar on micropropagation of Haplophyllum tuberculatumwere investigated. For rooting, the effect of three auxins (Indole acetic acid, Indol butyric acid andNaphthalene acetic acid) were studied in combination with three medium strengths (full, half andquarter) combined with the best mixture (0.50 mgl-1 Kin + 6 gl-1 agar with 2 gl-1guar seed powder) ofgelling agents. Obtained results showed that Murashige and Skoog (MS) was the best medium withsingle node explant as starting material during establishment stage. Multiple shoots were obtained onMS medium supplemented with 1.0 mgl-1 Kinetin (Kin) combined with 0.1 mgl-1 α-naphthalene aceticacid (NAA). Thiamine and arginine were the best vitamins and amino acid at 0.1 mgl-1 and 0.50 mgl-1,respectively. Agar at 6 gl-1 + guar seed powder at 2 gl-1 was the best gelling agent treatment. Halfstrength MS medium with 0.50 mgl-1 Kin + 0.5 mgl-1 NAA and 6 gml-1 agar with 2 gml-1 guar seedpowder was the best combination for rooting were successfully (85% survival) acclimatized in mixtureof peatmoss, vermiculite, washed sterilized sand and perlite (1:1:1:1, V/V/V/V).

Effects of carbendazim on <em>In vitro</em> micropropagation and flowering of pepper elder herbal plant

The present study reports on application of carbendazim for micropropagation and in vitro flowering of Peperomia pellucida. In vitro multiplication of shoot tips and axillary nodal explants were used in 25-100 mgL-1 carbendazim in 70% alcohol treatment for 5 min in MS (Murashige and Skoog) medium and MS hormone medium The explants were cultured on MS medium supplemented with auxins, cytokinins and additives (Casin hydrolyte and glutamine) used to proliferate the shoots. The maximum number of shoots (21.5/shoot tip) were developed on MS medium supplemented with 2.0 mgL-1 KIN (Kinetin) 0.5 mgL-1 NAA (α- Naphthalene acetic acid), 100 mgL-1 casein hydrolysate and 100 mgL-1 glutamine additives. The highest number of roots per shoot (17.6) and highest rooting frequency (94.2%) were obtained when the adventitious shoots were inoculated on MS medium with 1.5 mgL-1 IAA (Indole-3-acetic acid). The highest number of shoot development (75.4 %) was achieved on MS medium supplemented with 2.0 mgL-1 KIN 2.0 + 0.5 mgL-1 ABA (Abscisic acid) with 5% sucrose. It was found that MS medium devoid of 1.5 mgL-1 IAA promote the root formation and maturation of in vitro flower in P. pellucida. Acclimatized plantlets were transferred to botanical garden and the characters compared with mother plant of P. pellucida.

Effect of benzyl Aminopurine and Kinetin hormones on vegetative and reproductive growth of two Sri Lankan traditional rice varieties (Oryza Sativa L.); suwadal and Kahata wee

The study was carried out as a pot experiment in the green house condition to observe the effect of BAP and Kinetin on the growth and yield performances of Sri Lankan traditional Rice varieties (Oryza sativa L) Suwadal and Kahata wee. At the time of applying fertilizer applications to the plants, each pot was treated by 200 ml of BAP and Kinetin in different concentrations (0, 0.1, 0.3, 0.5 and 0.8 mgL-1) separately. Each pot was treated by one hormone; BAP or Kinetin throughout the experiment was over. Plant height in vegetative stage, number of tillers plant-1, number of panicles plant-1, number of seeds panicle-1 and weight of thousand seeds were measured. The experiment was conducted in a Complete Randomized Design (CRD) with 3 replications. The best vegetative growth and yield performances were observed in 0.3 mgL-1 BAP and 0.8 mgL-1 Kinetin in traditional rice varieties Kahata wee and Suwadal. The effect of BAP and Kinetin for two rice varieties was simillar in plant growth and development. Number of tillers plant-1 and thousand grain weight significantly increased in both varieties with hormones BAP or Kinetin whereas number of tillers plant-1 increased by 10-17% compared to control. Therefore it is possible; BAP or Kinetin (cytokinin) extracting plants could be integrated into rice ecosystem for better productivity.

Effects of 6-Benzyl aminopurine and Kinetin on In Vitro Shoot Multiplication of Sugarcane (Saccharum officinarum L.) Varieties

In vitro shoot multiplication of two commercial sugarcane varieties grown in Ethiopian sugar estates initiated using shoot tip explants was carried out with the objective to assess the multiplication responses of the sugarcane varieties (B41-227 and N14) under five levels of 6-benzylaminopurine (0, 0.5, 1, 1.5 and 2 mgL-1) and kinetin (0, 0.1, 0.25, 0.5, and 1 mgL-1) in a completely randomized design with 5x5x2 factorial treatment combinations arrangements. Analysis of variance (ANOVA) showed that the interaction effects of 6-benzylaminopurine, kinetin and the sugarcane genotypes on number of shoots per explant, average shoot length and number of leaves per shoot was very highly significant (P < 0.0001). Murashige and Skoog (MS) medium fortified with 1 mgL-1 6- benzylaminopurine (BAP) and 0.5 mgL-1 kinetin for B41-227; and 2 mgL-1 6-benzylaminopurine (BAP) without kinetin for N14 were found to give optimum shoot proliferation results. On these medium, B41-227 gave 34 ± 1.54 shoots per explant with 6.95 ± 0.01cm average shoot length and 12 ± 0.17 leaves per shoot while N14 produced 21 ± 0.58 shoots per explant with 5.63 ± 0.01 cm average shoot length and 5.4 ± 00 leaves per shoot after 30 days of culture transfer to multiplication medium. Thus, the optimized protocol is useful for rejuvenation, rapid In vitro propagation and production of large quantity quality sugarcane planting materials and hence help minimizes the major limitations hampering sugarcane production in the Ethiopian sugar Estates.

In vitro Shoot Multiplication in Physalis minima var. Indica L

Apical shoot buds were taken from in vitro germinated seedlings of Physalis minima. var. indica L. These were placed aseptically on MS (Murashige and Skoog,1962),B5 (Gamborg,1968)basal medium and supplemented with auxins and cytokinins alone. Maximum healthy shoot buds(100 percent) were obtained on MS medium supplemented with 0.5 mgl-1BAP (6benzyl amino purine) but length of shoot buds was more on (1.0,2.0 mgl-1) Kinetin. Multiplied shoots were transferred for rooting on MS and B5 basal medium and supplemented with auxins (IAA or IBA). Rooting was achieved in 100 percent explants on MS medium supplemented with all the levels of IAA and IBA.Higher root length (11.5+0.50cm) was observed in shoots incubated on 0.5mgl-1 IAA.

In vitro propagation of Ethiopian ginger (Zingiber officinale Rosc.) cultivars: Evaluation of explant types and hormone combinations

Ginger ( Zingiber officinale Rosc.) is the second most widely cultivated spice in Ethiopia, next to chilies. Recently, there has been huge demand for clean planting material of improved ginger cultivars, though it is difficult to meet the demand of planting materials using the conventional propagation techniques due to production inefficiency and disease transmission. Therefore, the present study was carried out with the objective of assessing the potential of axillary buds and shoot tips as explant sources and determination of suitable growth regulators for in vitro propagation of two ginger cultivars. Murashige and Skoog (MS) medium with four levels of benzyl adenine (BA) and kinetin was used for shoot multiplication in combination with two explant sources. A highly significant difference (p<0.0001) was observed between explant sources and among growth regulators for shoot multiplication. From this study, it was found that shoot tip explants on 2 mgl-1 BA and 1 mgl-1 kinetin was found to be better than other explant-media combinations which gave an average of 7 shoots per explant within six weeks of culture. Consecutively, the plantlets developed an average of 8.75 roots within four weeks of culture period and performed well in acclimatization and subsequently in the field. Key words : Ginger, Zingiber officinale Rosc., growth regulators, explants, micropropagation.

Effect of Different Concentrations of Kinetin and NAA on Micropropagation of Gardenia Jasminoides

The goal of this study was to use plant tissue culture technique in vegetative propagation of&#x0D; Gardenia (Gardenia jasminoides) by using single nodes and shoot tips excised from soft cuttings and&#x0D; treated with different concentrations of growth regulators.The results revealed that the use of different&#x0D; disinfistants was highly effective in reducing cultures contamination. The use of mercuric chloride (0.1%,&#x0D; HgCl2) for 10 minutes was very effective in preventing contamination and gave the highest survival&#x0D; percentage (99%). During the culture of shoot tips at initiation stage, a nutrient medium contained (2&#x0D; mgl-1 kinetin+ 0.6 mgl-1 NAA) gave the highest values of average number of shoots and leaves and growth&#x0D; length (2.3 shoots/ explant, 2.4 leaves and 1.9 cm respectively).At vegetative multiplication stage, The&#x0D; culture of shoot tips on a nutrient medium containing (3 mgl-1 kinetin) gave the highest values of&#x0D; average number of shoots and leaves (2.9 shoots/ explant, 3.6 leaves respectively).As for growth length,&#x0D; the results revealed that the highest value was achieved by MS medium supplemented by 2 mgl-1 kinetin +&#x0D; 0.1 mgl-1 NAA (3.3 cm). Regarding rooting process, MS medium supplemented by 4 mgl-1 NAA gave the&#x0D; highest average number and length of roots (4.0 roots and 2.50 cm respectively).Also, the medium&#x0D; supplemented by 8 mgl-1 IAA gave the highest number and length of roots (3.40 roots and 3.50 cm&#x0D; respectively). Plantlets obtained were transferred to pots and acclimatized with 95 % success.

English

&nbsp; Transgenic&nbsp;Cucumis melo&nbsp;cv. Maazoun containing the neomycin phosphotransferase II (NPT II)&nbsp;chimeric&nbsp;gene conferring resistance to kanamycin were obtained from cotyledons explants inoculated with&nbsp;Agrobacterium tumefaciens&nbsp;(GV3101) that contained the binary vector plasmid pADI. Transformed shoots were obtained on Murashige and Skoog medium supplemented with 1.50 mgl-1&nbsp;IAA, 0.10mgl-1&nbsp;BAP, 0.01 mgl-1&nbsp;NAA and 6 mgl-1&nbsp;kinetin.&nbsp;Transformants&nbsp;were selected by using only 100 mgl-1&nbsp;of kanamycin and 4 days of pre-culture. Putative transformants were confirmed for&nbsp;transgene&nbsp;insertion through polymerase chain reaction (PCR) analysis. From the inoculated explants, 6.66% produced transgenic shoots.&nbsp;&nbsp;&nbsp; &nbsp; Key words:&nbsp;Cucumis&nbsp;melo, genetic transformation, kanamycin, culture medium, PCR analysis.

Long-term effects of growth regulators on growth and turnover of symplastic and apoplastic sugars in the suspension subculture of kidney bean

Suspension cells of kidney bean were grown for 42 d in MS medium supplemented with growth regulators (2.0 mgL-1 2,4-D and 0.5 mgL-1 kinetin) or without At the stationary growth phase (42 d), the sugars were fractionated into the symplastic (ethanol and starch) and apoplastic [low-molecular pectin (lm-pectin), high-molecular pectin (hm-pectin), hemicellulose, and cellulose] sugars. The neutral sugars (NS) of hm-pectin and hemicellulose fractions were analyzed by GLC. The growth of the suspension cells in the liquid MS media, in terms of settled cell volume (SCV), remained similar, to the end of the experiment, irrespective of the presence or absence of growth regulators, indicating the nonnecessity of the exogenous growth regulators for the subculture. Total sugar (TS) of the ethanol fraction and NS of the Im-pectin of the suspension cells grown in the medium with growth regulators were higher than in the medium without growth regulators. However, starch content in the starch fraction and uronic acid (UA) content of the Im-pectin fraction did not exhibit any differences. From these results, it was suggested that the growth regulators modulated the structure of the cell wall polysaccharide. Analysis of the NS composition of the hm-pectin fractions revealed that the Rha, Arb, and Gal contents in the presence of growth regulators were higher than in the absence, while the Xyl, Man, and Glc contents in the presence of growth regulators were higher than in the absence, indicating the turnovers of rhamnogalacturonan and/or arabinogalactan. On the other hand, analysis of NS composition of hemicellulose fractions revealed that the Ara and Glc contents in the presence of growth regulators was higher than in the absence, whereas Xyl and Glc contents were nearly consistent, indicating the turnovers of arabinaogalactan I or II. The cellulose contents remained similar, irrespective of the presence (19.1%) or absence (18.7%) of growth regulators.