Different Concentrations Of NAA Research Articles

Clonal propagated 'Ek Pothi Lehsun' as a potential antifungal agent against Candida sp.

'Ek Pothi Lehsun', also known as snow mountain garlic, is a type of garlic grown in the high mountainous region of Jammu and Kashmir state of India. The present study aimed to develop a protocol for propagating snow mountain garlic in-vitro using corm seed as an explant. The study also assessed the antifungal potential of in vitro-grown bulbils against different Candida species. Four different concentrations of NAA and 2,4-D were tested for their effectiveness in promoting root formation, and eighteen different combinations of BAP (µM), KN (µM) and TDZ (µM) were investigated for effective proliferation of shoots with varied lengths. Shoot with maximum length (5.03±1.40) was obtained in MS medium containing 1.0 µM TDZ after 24 days of inoculation, whereas MS basal media was found effective for rooting plantlets. Rooted micro shoots were acclimatized successfully in hardening treys with a percent survival of nearly 80%. Seven different concentrations of Sucrose, i.e. 5%, 7%, 10%, 15%, 17%, 20%, and 25% were investigated for effective bulbil formation. Bulbil with a maximum diameter of 0.86 cm was obtained in 20% sucrose-containing MS media in 5 days. Further, the antifungal potential of aqueous extract (TC-SMG) of in vitro grown bulbils was investigated against three Candida sp. A zone of inhibition of 22.30±0.33 mm, 17.3±0.33 mm and 19.3±0.33 mm was observed against C. albicans, C. tropicalis and C. glabrata respectively, by using 200 mg/mL extract after 24 hrs depicting the remarkable potential of TC-SMG as an antifungal agent. In vitro culture of snow mountain garlic has demonstrated promising antifungal properties against Candida species.

The Effect of Some Plant Growth Regulators on Callus Culture of Different Pistachio Varieties

Pistachio (Pistacia vera L) is one of the oldest cultivated plants in the world. Its fruits are rich in protein, minerals, carbohydrates, fiber, and vitamins. In addition, the demand for these plants is increasing due to the fact that they are very tasty and nutritious. On the other hand, pistachio cultivation is quite difficult. In addition, many problems are encountered in germination with seeds or reproduction with cuttings. These situations necessitate the development of different in vitro tissue culture protocols. In this study, callus culture optimization protocol was developed by using seeds of three different pistachio cultivars. Murashige and Skoog (MS) medium was supplemented with different concentrations of NAA, IAA, 2,4 D and BAP. When callus size (1,776 cm), callus weight (0.908 g) and embryogenic callus regenerations (27.94%) were considered, it was found that the best variety was Tekin. Again, in the evaluation made according to these factors, it was determined that the best improvement was in the MS medium containing 3 mg/L BAP and 1 mg/L 2,4D. The contamination rate detected throughout the studies ranged from 7.65% to 12.91%.

Response of Foliar Feeding of NAA, Urea, Nano-urea and Biofertisol on Fruit Quality of Mango (Mangifera indica L.) cv. Langra

A study was conducted during the season of 2022-23 at the fruit research station in Imalia, which is affiliated with the Department of Horticulture at the JNKVV in Jabalpur. The objective of the study was to evaluate the positive effect of foliar sprays containing NAA, Urea, Nano-urea, and Biofertisol on the fruit quality of mango (Mangifera indica L.) cv. Langra. The experiment was laid out in Randomized Block design in three replications and all the treatments were replicated thrice by using the single tree as a unit. Each treatment was carried out with one tree for each replication. At the full bloom and pea stage, the spraying of three different concentrations of NAA (20 ppm, 30 ppm, and 40 ppm), Urea (2%, 3%, and 4%), Nano-urea (0.2%, 0.3%, and 0.4%) and Biofertisol (5 ml/l, 10 ml/l, and 15 ml/l) was compared with the control, which consisted of no spraying. According to the findings of the experiment, applying a foliar spray containing 40 ppm of NAA resulted in a significant improvement in the quality parameter, as measured by a maximum TSS value of 22.22 oBrix, a maximum TSS: acid ratio of 117.13, maximum ascorbic acid content of 26.70 mg/100g, a minimum acidity content of 0.19%, and reducing sugars of 6.74% and total sugars of 17. During the evaluation, it performed at the same level as T3 (NAA 30 ppm), T2 (NAA 20 ppm), and T7 (Urea 4%).

Micropropagation of strawberry (Fragaria ananassa “pajaro”)by meristem culture method

Experiments were carried out to examine the effects of different combinations of plant growth regulators in vitro micropropagation of strawberry plants. Specimens were sterilised by a different concentration of sodium hypochlorite - NaOCl (1, 2, and 3%) for 20 mins. A meristem was separated and cultured on Murashige and Skoog (MS) containing 0.3 mg/l BAP for 8 weeks. Initial shoots were transferred to MS with various concentrations of BAP (0, 0.1, 0.3, and 0.5 mg/l) and kinetin (0, 0.1, 0.2, and 0.3 mg/l) in either single or combination. After 6 weeks of culture, the regenerated shoots were transferred on 1/2 MS with different concentrations of NAA (0, 0.1, 0.3, 0.5, 0.7, and 1.0 mg/l) and BAP (0, 0.1, 0.3, and 0.5 mg/l) in either single or combination. The results showed that the lowest percentage of contamination (10%) and the highest percentage of regeneration (80%) response were achieved on MS media by sterilisation with 3% NaOCl for 20 mins. The highest shoot proliferation per explant (16.2 shoots/explant) was reached on MS with 0.3 mg/l BAP plus 0.1 mg/l kinetin. 1/2 MS media with 0.1 mg/l NAA plus 0.1 mg/l BAP induced 14.4 roots per shoot and produced the longest roots (9.1 cm). However, the higher concentration of NAA (0.3-1.0 mg/l) or in combination with BAP resulted in callus formation. The plantlets, thus developed, were hardened and successfully established in soil containing organic fertiliser: Trichoderma: coconut peat (1:1:1 v/v/v).

Bidirectional Regulation of Sodium Acetate on Macrophage Activity and Its Role in Lipid Metabolism of Hepatocytes

Short-chain fatty acids (SCFAs) are important metabolites of the intestinal flora that are closely related to the development of non-alcoholic fatty liver disease (NAFLD). Moreover, studies have shown that macrophages have an important role in the progression of NAFLD and that a dose effect of sodium acetate (NaA) on the regulation of macrophage activity alleviates NAFLD; however, the exact mechanism of action remains unclear. This study aimed to assess the effect and mechanism of NaA on regulating the activity of macrophages. RAW264.7 and Kupffer cells cell lines were treated with LPS and different concentrations of NaA (0.01, 0.05, 0.1, 0.5, 1, 1.5, 2, and 5 mM). Low doses of NaA (0.1 mM, NaA-L) significantly increased the expression of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin 1 beta (IL-1β); it also increased the phosphorylation of inflammatory proteins nuclear factor-κB p65 (NF-κB p65) and c-Jun (p < 0.05), and the M1 polarization ratio of RAW264.7 or Kupffer cells. Contrary, a high concentration of NaA (2 mM, NaA-H) reduced the inflammatory responses of macrophages. Mechanistically, high doses of NaA increased intracellular acetate concentration in macrophages, while a low dose had the opposite effect, consisting of the trend of changes in regulated macrophage activity. Besides, GPR43 and/or HDACs were not involved in the regulation of macrophage activity by NaA. NaA significantly increased total intracellular cholesterol (TC), triglycerides (TG), and lipid synthesis gene expression levels in macrophages and hepatocytes at either high or low concentrations. Furthermore, NaA regulated the intracellular AMP/ATP ratio and AMPK activity, achieving a bidirectional regulation of macrophage activity, in which the PPARγ/UCP2/AMPK/iNOS/IκBα/NF-κB signaling pathway has an important role. In addition, NaA can regulate lipid accumulation in hepatocytes by NaA-driven macrophage factors through the above-mentioned mechanism. The results revealed that the mode of NaA bi-directionally regulating the macrophages further affects hepatocyte lipid accumulation.

Construction of sterile system of Buchloe dactyloides (Nutt.) Engelm.

The stems of male and female strains of Bison grass (Buchloe dactyloides (Nutt.) Engelm.) were taken as explants, and 1 / 2MS and MS medium were used, and different concentrations of NAA, KT and 6-BA were added to study the rapid propagation technology of ferison grass. Results indicated that 75% alcohol was treated for 30s with another 0.1 g ꞏL-1The disinfection effect of mercury solution for 12 min; The MS + 1.5 mg ꞏL-1 NAA + 0.4 mg ꞏL-1 KT female strain showed the best induction effect, The MS + 1.5 mg ꞏL-1 NAA + 0.6 mg ꞏL-1 KT male strain had the best induction effect; 1 / 2 MS + 2 mg + L-1 NAA + 1.5 mg ꞏL-1 6-BA the female and male strains had the best proliferation effect; The highest number of roots and root root rate of 1 / 2 MS + 1.5 mg ꞏL-1 female and male NAA strains were combined.

Changes in Photosynthetic Pigment of Wheat(Triticum Aestivum L. Var. Bari Gom-25) As Affected By Naa and N-Fertilizer

A field experiment was conducted to investigate the changes inphotosynthetic pigment of BARI Gom-25 as affected by foliar spray of NAA andN-fertilizers. The experiment consisted of seven different concentrations of NAA(NAA0 = 0, NAA1 = 10, NAA2 = 20, NAA3 = 30, NAA4 = 40, NAA5 = 50 and NAA6 =60 ppm) and five N-levels (F0=0, F1=25, F2=50, F3=75 and F4=100% of therecommended dose) and their 35 combination treatments. Results revealed thatfoliar application of 40 ppm NAA (NAA4) produced higher chlorophyll a contentat all stages (tillering, flowering and grain filling) and were significantly higherthan control at tilling and flowering stages. Chlorophyll b and carotenoidcontents did not show any general trend to NAA at any stage except at tillingstage. Application of 50% of the recommended N-fertilizer (F2) producedsignificantly maximum photosynthetic pigment contents at tilling stage whereas,at flowering stage the same results were obtained from 100% of therecommended N-fertilizers (F4) but with few exceptions. At grain filling stage,photosynthetic pigment did not show any general trend to N-fertilizerapplication. Findings on combined effects indicated that 20 and 30 ppm NAA incombination with 50% N-fertilizer (NAA2F2 and NAA3F2) had producedsignificantly maximum chlorophyll a content at tilling stage. However, atflowering stage the highest chlorophyll a content was noted from NAA2F4treatment (20 ppm NAA in combination with 100% N-fertilizer). Findings alsorevealed that combined application of NAA and N-fertilizer had not resulted anygeneral trend on chlorophyll b and carotenoid contents in most of the caseswhere the maximum value in photosynthetic pigment was found depending onthe concentrations of NAA, fertilizer dose and plant growth stages. Dhaka Univ. J. Biol. Sci. 31(1): 169-176, 2022 (January)

Efficacy of Different Concentrations of NAA on Selected Ornamental Woody Shrubs Cuttings

Ornamental woody shrubs are used in landscape design worldwide. Their propagation can be made generatively and vegetatively. Vegetative propagation methods are mostly used by nurseries, as such methods are quick and the newly propagated plants inherit the genetics of the mother plant. However, rooting in some woody plants is slow and, unfortunately, sometimes produces only a small number of rooted cuttings. In this study, shoot cuttings from six selected ornamental woody shrubs were subjected to different concentrations of rooting stimulators (0.5 (NAA5) and 0.8 (NAA8) % concentrations of 1-Naphthylacetic acid; cuttings without treatment were considered as control) and propagated in two different periods (spring and summer). Our results show that significant changes were obtained in the plants under the different treatments. Most of the plants showed a positive response to both treatments, expect for Cotinus coggygria ‘Royal Purple’, which, compared to control, registered decreases in all the tested parameters under NAA5 treatment. Ilex aquifolium was the species that showed increments in all the parameters when NAA treatments were applied. In conclusion, our research suggests that NAA increases rooting in ornamental woody shrubs, although in some cases rooting could be a species-dependent process.

Promotion of flower opening in cut rose cultivars by 1-naphthaleneacetic acid treatment

ABSTRACT Improving the quality and rate of opening of cut flowers is important to meet consumer demand. Thus, it is important to develop methods to control the rate of flower opening and senescence in ornamental plants. In this study, we investigated the effects of 1-naphthaleneacetic acid (NAA) in flower opening in rose (Rosa sp.) cultivars Princess Meg, Red Star and Madrid. Cut roses were maintained under different concentrations of NAA. Shoot bases were immersed in water solution containing 0, 100, and 1,000 μM NAA, in addition to 2% w/v sucrose with 0.02% w/v 8-hydroxyquinoline monohydrate. Subsequently, their vase life, flower opening, flower diameter and petal weight were measured. Flower opening in all three cultivars was clearly promoted by the 1,000 μM NAA treatment, resulting in higher petal fresh weight and flower diameter at 2 days following treatment. 100 μM NAA treatment also promoted flower opening and petal wilting in three cultivars, although the decrease in relative fresh weight of cut rose became slower and vase-life became longer than 1,000 μM NAA treatment in “Madrid”. This indicates that NAA promotes flower opening and petal growth in three cut rose cultivars. However, NAA treatment also promoted petal wilting, resulting in shorter vase-life. Although rose cultivars differed in their sensitivity to the NAA treatment, we conclude that NAA shows high potential as a chemical agent for controlling flower opening in cut rose cultivars.

Direct plant regeneration through micropropagation using selected explants of sugarcane

The experiment was conducted at the Laboratory of Biotechnology Division, Bangladesh Sugarcrop Research Institute (BSRI), Ishurdi, Pabna during the period from January 2009 to December 2009 to regenerate plants through micropropagation technique using selected explants of sugarcane. In this experiment shoot tip, leaf segment and leaf roll section explants of three sugarcane varieties (Isd 16, Isd 35 and Isd 36) were cultured on shoot inducing MS media with four combinations of NAA and Kn (NAA2.5Kn0.5, NAA5.0Kn0.5, NAA7.5Kn0.5 and NAA10.0Kn0.5 mg/l) to regenerate plants. The proliferated shoots were multiplied on liquid MS media with five combinations of BAP and Kn (BAP0.25Kn0.25, BAP0.5Kn0.25, BAP0.5Kn0.5, BAP1.0Kn0.5 and BAP1.0Kn1.0 mg/l) and further transferred to root inducing MS media fortified with six different concentrations of NAA (1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 mg/l) for adventitious root formation to raise full-fledged plantlets. The leaf roll section explant of variety Isd 35 cultured on MS medium containing hormonal combination of 7.5 mg/l NAA + 0.5 mg/l Kn produced the highest percentage (83.33%) of shoot regeneration. The number of shoots per explant was also found highest (10.23) in the same explant of same variety with the same hormonal combination. The highest multiplication rate (4.64) was obtained from the liquid MS medium containing hormonal combination of 1.0 mg/l BAP + 0.5 mg/l Kn in the variety Isd 35. The MS medium containing 5.0 mg/l NAA showed better performance for adventitious root induction to raise full-fledged plantlets in all the varieties within nine days of inoculation. Â

Propagation Studies in Passion Fruit (Passiflora edulis Sims.) Using Cuttings

Passion fruit is a high value export oriented crop, which is gaining popularity among farmers. One of the constraints in expansion of this crop is availability of quality planting material. Passion fruit is propagated through seed, cutting and grafting. Among these methods, cuttings encourage the production and multiplication of true to type planting material. Therefore, a study was conducted at the Fruits Crops Research Station, Kerala Agricultural University, Vellanikkara, Thrissur, during August 2019 - November 2019 to evaluate the effect of different concentrations of NAA and number of nodes on the rooting of cuttings and survival percentage. The experiment was laid out in completely randomized design with twenty treatments and two replications. Experiment consisted of two factors, viz. NAA concentration (200 ppm, 400 ppm, 600 ppm, 800 ppm and control) and number of nodes (one node, two node, three node and four node), following quick dip method. The study revealed that there was significant variation among the treatments. When the effect of number of nodes alone was considered, four noded cutting had the maximum survival percentage, and higher root and shoot parameters. Improved survival percentage, and enhanced root and shoot characteristics were noticed with the increasing NAA concentration. From the present study, it was concluded that combined effect of more number of nodes and higher NAA concentration had profound effect on survival percentage, shoot length, root length, number of leaves and leaf area and four noded cuttings dipped in 800 ppm NAA, for 3-5 seconds was found to be the best successful propagule for passion fruit.

Effect of Growth Regulators on Stevia rebaudiana Bertoni Callus Genesis and Influence of Auxin and Proline to Steviol Glycosides, Phenols, Flavonoids Accumulation, and Antioxidant Activity In Vitro.

Stevia is a plant containing many active compounds, but usually propagated by stem cuttings because of low seed-yield-germination ability. The aim of this study was to investigate the impact of plant-growth regulators on stevia callus induction and growth from somatic tissue, as well as to determine the effect α-naphthalene acetic acid (NAA) and proline (PRO) on the amount of stevioside, rebaudioside A, phenols, flavonoids, and antioxidant activity. Stem and leaf segments were inoculated on a Murashige and Skoog (MS) medium supplemented with different concentrations of NAA and 6-benzylaminopurine (BAP) for callus genesis. The amount of steviol glycosides (SGs) was evaluated using high-performance liquid chromatography (HPLC), and the amounts of total phenols, flavonoids, and antioxidant activity by spectrophotometric methods. The highest callus-induction frequency and callus-mass increase were obtained from the leaf explants in MS medium supplemented with 2.0 μM NAA. The highest amount of SGs, phenols, and flavonoids, and stronger antioxidant activity were determined in the cellular compounds of callus from leaf explant. PRO reduced the amount of SGs and flavonoids. The significantly highest amount of total phenolic compounds was obtained in the callus from leaf explants in the medium supplemented with 2.0 µM NAA and 2.0 µM PRO.

Somatic embryogenesis of hypocotyl derived calli from an eggplant cultivar

Optimization of tissue culture and regeneration conditions of eggplant is necessary for achieving different goals such as gene transformation and the development of somaclonal variations. In this study, hypocotyl explants ware used to produce callus in a medium containing different concentrations of NAA and BAP. Moreover, the concentration of the elements Ca, Mn, Mg, Fe and K were measured and analysed between embryogenic and non-embryogenic calli. For shoot elongation, embryogenic calli were transferred to a new culture medium containing 3.5, 4 and 4.5 mg l-1 BAP plus 2 mg l-1 GA3. Finally, produced shoots were rooted in a culture medium containing 1, 1.5 and 2 mg l-1 NAA. Results showed that the best treatment for the embryogenic callus induction was MS medium containing 0.5 mg l-1 BAP plus 0.25 mg l-1 NAA. Two elements, Fe and K, had the highest amount in non-embryogenic calli compare to the embryogenic one. For plant regeneration, MS medium containing 4.5 mg l-1 BAP plus 2 mg l-1 GA3 and 2 mg l-1 NAA were the best treatments for shooting and rooting, respectively. In this study, the best treatments for plant regeneration produced 35 shoots from an explant with 92 % shooting. This regeneration protocol could be useful for gene transformation and micro-propagation studies.

English

Plant growth and development is significantly influenced by phytohormones – endogenous molecules present naturally in plants. The best known plant hormones are auxins and cytokinins. This study examined the possible effect of externally applied plant hormone analogues (growth regulators): 1-naphthyl acetic acid (NAA) and 6-benzyl aminopurine (BAP) on industrial fibre hemp (Cannabis sativa L., variety Bialobrzeskie). Plants were treated with three different concentrations of NAA (5, 10 and 20 mg/l) and three different concentrations of BAP (10, 25 and 50 mg/l). Morphological and physiological characteristics, such as apical dominance, shoot branching, fibre properties, and flavonoid content were evaluated. The chosen variety of hemp had a significant response to exogenous application of growth regulators, as has been observed with other plant species. Most notably, completely understood and controlled synthetic auxin tretament has a potential to increase the bark fibre yield of hemp.

Clonal propagation of Curcuma aeruginosa Roxb.: A versatile medicinal plant

Curcuma aeruginosa Roxb. (blackturmeric) belonging to family Zingiberaceae is a versatile medicinal plant. This study aimed to evaluate the effects of MS-salts (Murashige and Skoog) and sucrose concentrations on the in vitro multiplication and rooting of C. aeruginosa. The MS medium supplemented with different concentrations of BAP alone or in combination with different concentrations of NAA produced a varying degree of multiple shoots. Supplementation of 3 mg/L BAP and 1 mg/L NAA gave the best result. The in vitro-established C. aeruginosa, plants were inoculated in a medium supplemented with 10, 20, 30, 40 and 50 g L−1 sucrose. Optimum sucrose concentrations 30g L−1 induced a greater number of shoot and root numbers. During the establishment of initial culture; explants of 1 to 2 cm took longer days for the first bud appearance and scored the highest percentage of survival rate after 45 days. Explants of 2cm produced 10±0.53 number of microshoots. The interactive effect between BAP (3 mg/L) and NAA (1 mg/L) on the cultures produced the highest number of shoots 24±0.79. The plantlets were then allowed to acclimatize and successfully established under field conditions

In Vitro Propagation of Wetland Medicinal Plant Lythrum salicaria L.

The aim of this experiment is to develop a protocol to establish the in vitro propagation potential of Lythrum salicaria L (purple loosestrife). Seeds of L. salicaria were germinated and grown in Murashige Skoog (MS) medium for 30 days. At the end of 30 days, shoot tips of L. salicaria were cultured on MS medium supplemented with different combinations and concentrations of 6-benzylaminopurine (BAP; 1, 2 and 3 mg/L) and 1-naphthaleneacetic acid (NAA; 0, 0.1 and 0.5 mg/L). Regenerated shoots were transferred to MS supplemented with varying concentrations of different auxins: NAA (0.5 and 1 mg/L) and indole-3-butyric acid (IBA; 0.5 and 1.0 mg/L) for rooting. Among the cytokinins investigated, BAP indicated the best response to initiate shoot elongation and multiple shoot formation alone. The highest number of shoot per explant (37±3.38) was obtained in MS medium with 1 mg/L BAP. Although, the highest shoot length (3.1 cm ± 0.48) was found on medium with 1 mg/L BAP+0.5 mg/L NAA, there were no statistically differences among all types of medium. Percentage of rooting, shoot length and root length were altered significantly with different concentrations of NAA and IBA. Thus, shoot length (9.5 cm±0.46), root length (4.1 cm±0.13) and the percentage of rooting (100%) increased significantly with a concentration of 1 mg/L NAA. It was concluded that MS medium with 1 mg/L NAA has been superior for root development compared to other hormone concentrations.

Efficient Adventitious Morphogenesis from In vitro Cultures of the Medicinal Plant Cymbopogon schoenanthus

Cymbopogon schoenanthus subsp. proximus is a medicinal grass, the source of sesquiterpene proximadiol and is well known in folk medicine as renal antispasmodic. The effect of growth regulators, different sugar types, methyl jasmonate and vitamins on de novo organogenesis from seed cultures was studied. Among different concentrations of NAA and BA, 4 mg/l NAA in combination with 0.5 mg/l BA produced 100% callus induction. Both 1.0 and 4.0 mg/l NAA in combination with 0.5 mg/l BA yielded the highest number of shoots. High concentration of sucrose 6% was efficient in root induction from organogenic shoots. Sugar types at a constant concentration of 3% had a significant effect on raising shoot numbers. Different concentrations of methyl jasmonate, biotin and Ca-pantothenate showed a decline in de novo shoot induction, but root formation frequency and numbers were significantly improved with all concentrations used.&#x0D; Plant Tissue Cult. &amp; Biotech. 28(2): 147-160, 2018 (December)

Effect of Paclobutrazol and NAA on Sex determination and Seed Yield of Medicinal Pumpkin (Cucurbıta pepo L.)

In order to evaluate the effect of plant growth regulators on sex determination and yield, an experiment was conducted by spraying different hormonal combinations on medicinal pumpkin. Experimental treatments were included different concentrations of NAA (25, 75, 100, 125 mg/l), paclobutrazol (50, 100, 200 and 300 mg/l) and combination of growth regulators (NAA 25 mg/l + PBZ 50 mg/l), (NAA 75 mg/l + PBZ 100 mg/l), (NAA 100 mg/l + PBZ 200 mg/l), (NAA 125 mg/l + PBZ 300 mg/l), and control (no application of growth regulators). Growth regulators were applied on the plants by spraying at the –two-leaf and four-leaf stages. Results showed that spraying paclobutrazol and NAA caused increase in number of female flowers, fruits per plant, seeds per fruits, 1000 seed weight and seed yield per hectare, but the number of male flowers and ratio of male to female flowers were decreased compared to their values in control plants. The highest fresh and dried seed yield of pumpkin seed was obtained with 50 mg/l paclobutrazol.

Somatic embryogenesis of Byrsonima intermedia A. Juss.: induction and maturation via indirect approach

Byrsonima, especially the species Byrsonima intermedia, is an endangered Brazilian plant that has been widely used as food and for its therapeutic characteristics. However, this species faces challenges with sexual propagation, and somatic embryogenesis has emerged as a viable alternative option for propagation. Therefore, this study aimed to establish a protocol for inducing somatic embryogenesis in B. intermedia. For the induction of callus from in vitro seedling leaves, different subcultures (three subcultures, 60 days each) and concentrations of different cytokinins (BAP, TDZ, Kin and ZEA) combined with varying NAA solutions were tested. Different concentrations of NAA were also analyzed in the induction of pro-embryogenic calli. For the induction of embryogenic calli and somatic embryos, the pro-embryogenic calli were subcultured on MS medium without adding growth regulators. The somatic embryos that originated were inoculated on a maturation medium containing different concentrations of gibberellic acid (GA3). The formation of secondary embryos was also analyzed using different concentrations (0, 2.88, and 8.66 µM) of GA3 and different types of lids (Conventional lid, Biossama® commercial lid and conventional lid with membranes). The results show that for the induction of somatic embryos, the use of kinetin with NAA presented the formation of somatic embryos in the second (4.76 µM CIN + 0.54 µM NAA) and third (5.17 µM CIN + 10.54 µM NAA) subcultures. The use of 28.87 µM GA3 favored the formation of seedlings. The Biossama lid and 2.88 µM GA3 showed higher formation of secondary embryos.

Establishment of in Direct Propagation of Mandarin (Citrus reticulata L.) Using Tissue Culture

THE PRESENT investigation deals with development of an efficient micropropagation protocol for mandarin (Citrus reticulata L.) via in direct propagation using cotyledons and juice vesicles as explant. The establishment of in vitro callus induction from leaf, stem segments and cotyledon system were excised from seedlings has been done. Best results for induction of callus were observed from cotyledons and stem segments 100% on MS medium supplemented 1 or 2mg/l 2,4-D. MS medium supplemented with 300mg/l glutamine or 1.5mg/l casein hydrolysate proved to be the most efficient additive in promoting callus formation from juice vesical with 4.22, 4.23g/callus fresh weight, 100% response and was the most favorable medium. Maximum callus fresh weight (7.96 g) and 100% response were obtained from juice vesicles when cultured on MS medium supplemented with 2mg/l 2,4-D and 10% coconut water. The highest callus regeneration and weight of callus observed from juice vesicles by adding 2mg/l 2,4-D, 300 or 400 mg/l malt extract. Callus raised from juice vesicles showed maximum shoots (96.67%) with 0.5mg/l Kin, 400mg/l malt extract and 2mg/l NAA. Regenerated shoots raised from juice vesicles showed better shoot multiplication and highest length of shoots on MS medium with 2mg/l BA and 0.4mg/l NAA. Regenerated shoots were rooted on MS medium supplemented with different concentrations of NAA and best response (98%) was observed with NAA (2 mg/l) and gave maximum, roots number, roots length and leaves number. Rooted plantlets were successfully acclimatized with survival rate reaching almost 87%. These plants grew normally without showing any morphological variation. Developed protocol can be useful for application of somatic embryogenesis from juice vesicles to improve mandarin.