Concentrations Of Growth Regulators Research Articles

Organogenesis of corn plants (Zea mays L.) at various concentrations of auxin and cytokinin plant growth regulators in vitro

This study aimed to determine the effect of auxin and cytokinin concentrations as plant growth regulators (PGR) on the organogenesis of corn plants in vitro. This research was conducted from June to September 2021 in the plant tissue culture laboratory of Cokroaminoto University, Palopo City, South Sulawesi, Indonesia. The research method was a completely randomized design (CRD) consisting of 15 experimental units, i.e., P0 (control), P1 (2 ml NAA (naphthaleneacetic acid) + 1 ml BAP (benzylaminopurine)), P2 (2 ml NAA + 1.5 ml BAP), P3 (2 ml NAA + 2 ml BAP), and P4 (2 ml NAA + 2.5 ml BAP). The results revealed that the administration of auxin and cytokinin plant growth regulators had a significant effect on the parameters of germination age and plantlet weight, but it had no significant effect on the parameters of plant height, number of roots, root length, and number of leaves. The effective concentration of auxin and cytokinin growth regulators on the organogenesis of corn plants was P2 (2 ml NAA + 1.5 ml BAP) on the parameters of 11 cm plant height, 7 root strands, 1 leaf, and 1 gram plantlet weight. This was due to the influence of the given concentration of PGR, which plays multiple important roles in plant development.

PENGARUH KONSENTRASI ZAT PENGATUR TUMBUH (ZPT) ROOT UP TERHADAP PERTUMBUHAN STEK BATANG JARAK PAGAR (Jatropha curcas)

Jatropha curcas (Jatropha curcas) is a woody shrub that is widely found in tropical areas. This plant is known to be very drought tolerant and easily propagated by cuttings. The oil that comes from the fence is used as fuel (biodiesel). The object of research was to find out the influence of Root Up Growth Regulating Substance (ZPT) concentration on the growth of Jatropha stem cuttings. The method of research used is an experimental method using a Group Randomized Design (RAK) with the treatment of Root Up Growth Regulators (ZPT) concentrations of 0 mg, 20 mg, 25 mg, 30 mg, and 40 mg repeated 10 times. The observed variables are, number of roots, average length of roots, number of shoots, and average length of shoots. The results obtained from giving ZPT Root up with concentrations of 0 mg, 20 mg, 25 mg, 30 mg, and 40 mg had no significant effect on the growth of the number of roots, average root length, number of shoots, and average shoot length. No significant influence of concentration was found, but seen from DMRT there was a tendency for the P2 treatment with a concentration of 25 mg to show the best average shoot length.

An assessment for in vitro propagation and genetic stability of Phoebe goalparensis Hutchinson, an endemic valuable timber tree of North East India

BackgroundPhoebe goalparensis is an endemic forest species of North East India that belongs to Lauraceae family. P. goalparensis is used as timbers yielding plants for commercial importance in the local furniture markets of North East India. A rapid in vitro micropropagation protocol was established by using apical and axillary shoot tips on Murashige and Skoog medium with varied concentrations of plant growth regulators. ResultsIn this study, 5.0 mg/l BAP augmented medium was chosen as the best for shoot multiplication of the plant. However, IBA (2.0 mg/l) was the most responsive for root induction. Moreover, 70% of root induction was recorded during rooting experiment and 80–85% survivability was observed during the acclimatization of this species. Clonal fidelity of P. goalparensis was determined with ISSR marker and it was observed that in vitro raised plantlets were polymonomorphic. ConclusionHence, an efficient protocol with high proliferation and rooting was established for P. Goalparensis that could aid in massive propagation in future.

Somatic embryogenesis induction of Syzygium cumini

Somatic embryogenesis serves as an effective alternative system for in vitro cultivation of endangered plants (Syzygium cumini), as it allows for the propagation of plants under a controlled environment. So produce hundreds of embryos that can be used as artificial seeds. Somatic embryos of Syzygium cumini, family Myrtaceae, were induced from the calli of a sterile leaf explant in Murashige and Skoog's medium with 6 ppm 2,4-D. After the transfer of the developed calli into liquid media supplemented with the same concentration of growth regulator, all three embryonic stages (globular, heart and torpedo) were observed after 6 weeks. Liquid media with growth regulators appeared to enhance the development to torpedo-stage embryos, especially at six weeks of age cultures. 12 weeks after the transfer of the callus into liquid media, flasks containing predominantly one microscopic stage were pooled and plated on fresh solid media lacking a growth regulator, where the embryogenic calli germinated showing shoots and aerial parts. Embryogenesis production protocol is considered a good tool to save plants from extinction, especially in Syzygium cumini which is a very important anti-diabetic drug.

In Vitro Propagation of Three Date Palm (Phoenix dactylifera L.) Varieties Using Immature Female Inflorescences.

Immature female inflorescences are promising materials for use as explants for the tissue culture of date palm. Four types of MS media were used in this study during the four micropropagation stages-starting media (SM), maturation media (MM), multiplication media (PM) and rooting media (RM)-to micropropagate three elite date palm varieties, Amri, Magdoul and Barhy using the immature female inflorescences as explant. The highest percentage of callus induction in all the varieties studied was obtained on the SM1 (9 µM 2,4-D + 5.7 µM IAA + 10 µM NAA). Culturing on the MM1 (4.5 µM 2,4-D + 9.8 µM 2-iP + 1.5 AC) allowed us to obtain the best value in terms of callus weight. After culturing on the PM1 (4.4 µM BA + 9.8 µM 2-iP) produced the highest numbers of somatic embryos and shoots. The explants on RM2 (0.5 µM NAA + 1.25 µM IBA + 3 g AC) showed the highest root numbers and root lengths, while the highest shoot length was achieved on RM3 (0.5 µM NAA + 0.5 µM IBA + 3 g AC). The Amri variety presented the best response among the three varieties in all parameters, followed by the Magdoul and Barhy varieties. In all the stages of micropropagation, the analysis of variance revealed highly significant variations among varieties and culture media, and a significant difference in the number of roots during the rooting stage. The results also showed non-significant differences in the interaction between varieties and culture media, except for shoot length in the rooting stage. The results also reveal the broad sense heritability ranging from low to high for the measured parameters. It can be concluded that the immature female inflorescences can be used as a productive explant source for successful date palm micropropagation using the SM1, MM1, PM1 and RM2 culture media. It can also be concluded that the success of date palm micropropagation not only depends on the concentrations of growth regulators, but also on their types.

Assessment of Genetic Stability on In Vitro Propagation of Ardisia crenata var. bicolor Using ISSR Markers

Ardisia crenata var. bicolor is a multi-purpose plant and has important ornamental and medicinal properties. Conventional methods of propagating the species from seeds and cuttings have low efficiency because of the recalcitrant properties of seeds and low survival rate of high-quality cuttings. This work aims to study the in vitro regeneration protocol for direct organogenesis from nodal segments of A. crenata var. bicolor on Murashige and Skoog (MS) medium, with different combinations and concentrations of plant growth regulators (PGRs). The treatments used for the establishment and proliferation of shoots included MS medium supplemented with different concentrations of Benzyl-aminopurine (BAP) and indole-3-butyric acid (IBA). For rooting, IBA was used in combination with naphthaleneacetic acid (NAA) in full- and half-strength MS media. Maximum shoot establishment (76.67%) and the highest shoot length (6.6 cm) were observed on MS medium with 1.0 mg·L−1 BAP with 0.5 mg·L−1 IBA, while BAP at 1.0 mg·L−1 with 0.25 mg·L−1 IBA obtained the highest shoot proliferation (4.5 ± 1.53). The best rooting response (83.33%) was achieved on half-strength MS including 1.0 mg·L−1 IBA with 0.25 mg·L−1 NAA, and the maximum survival rate of 84.4% was observed after acclimatization under 75% shading. To define their genetic stability, using eleven primers of ISSR markers to assess the genetic stability of the unstable leaf color samples compared with their mother plant, the ISSR markers demonstrated a level of genetic polymorphism in plantlets, but without other morphological variations. This indicates the genetic resemblance to the mother plant and the reliability of this protocol for the efficient micropropagation of A. crenata var. bicolor.

A protocol for Agrobacterium-mediated genetic transformation of Lens culinaris Medik (lentil)

A reliable protocol for Agrobacterium-mediated genetic transformation of Lens culinaris Medik (lentil) was developed. Using cultivar Laird, the protocol yielded rooted shoots from an average of 6.8 independent events per hundred seeds. The protocol utilized longitudinal slices of embryo axes from imbibed mature seed as a starting explant and a plasmid containing a β-glucuronidase:neomycin phosphotransferase (gus:nptII) fusion gene in Agrobacterium strain EHA105. A series of four media, each with appropriate levels of kanamycin selection were identified and other factors tested included the optical density of the Agrobacterium suspension, and type and concentration of plant growth regulators. The expression of the gus reporter gene was visualized through histochemical staining, and further molecular analysis through PCR, qPCR, ddPCR and Southern hybridization confirmed transformation and provided copy number. The inserted genes were inherited into the T1 generation and chimaeras were not identified. The time from co-cultivation to the planting of rooted shoots ranged from 4 to 7 months, as transgenic clusters continue to produce additional clonal shoots.

Role of Phytohormones in Biomass and Polyphenol Accumulation in Salvia bulleyana In Vitro Culture.

Salvia bulleyana is a plant native to the Chinese Yunnan Province. This species has been used in traditional Chinese medicine as a substitute for Danshen (the roots of Salvia miltiorrhiza). The aim of our study was to establish an effective system for propagating S. bulleyana shoots to obtain large amounts of material rich in bioactive compounds. Phytohormones were used to regulate shoot growth and regeneration potential and influence plant secondary metabolism. The shoot tips were incubated on a Murashige and Skoog agar medium supplemented with 0.1 or 0.5 mg/L IAA (indole-3-acetic acid) and the cytokinins benzylaminopurine (BAP), meta-topoline (M-T), 6-benzylaminopurine riboside (RBAP), N-benzyl-9-(2-tetrahydropyranyl)-adenine (BPA) or kinetin, (K) at concentrations of 0.5, 1 or 2 mg/L. It was observed that the type and concentration of growth regulator significantly influenced the regeneration potential of S. bulleyana shoots. The highest multiplication rate was obtained when 0.1 mg/L IAA and 2 mg/L BPA were used. Under these conditions, 100% of shoot tips formed buds and almost seven buds/shoot per explant were obtained after five weeks. Meanwhile, the highest biomass was found for shoots growing on a medium supplemented with 0.1 mg/L IAA and 1 mg/L M-T: 1.2 g of fresh weight and 0.17 g of dry weight. However, a medium with 0.1 mg/L IAA and 2 mg/L RBAP was most favorable for bioactive phenolic acid content, with a total polyphenol level (37.7 mg/g dw) 4.5 times higher than in shoots grown on medium without growth regulators (8.23 mg/g dw). Finally, optimal conditions were selected by TOPSIS (technique for order of preference by similarity to the ideal solution); the culture of S. bulleyana grown on an MS medium containing 0.1 mg/L IAA and 1 mg/L M-T was found to be the most efficient for polyphenol accumulation and can be used for the production of medicinally relevant compounds.

Variation in Acacia senegal Seed size and Germination Collected from Different Regions of Rajasthan

Acacia senegal is common in Rajasthan and its seeds are used in Panchkutta vegetable. It is good for fencing and also on bunds fencing. Seeds from different locations were collected and their seed size and other parameters were studied. Its germination in laboratory with different treatments as well as with different concentrations of Growth regulators was studied. Results shows that seed size varies with different locations even in same districts also. However there is no correlation with germination behavior. Phenological study and reproductive biology of seeds need to be studied in details.

Germination and Seed Viability of <i>Helichrysum maracandicum</i> Popov Ex Kirp. Sterilized under <i>in Vitro</i> Conditions

Most medicinal plants are on the verge of extinction. In this regard, biotechnology is facing the challenge of developing alternative ways to produce biomass with the desired Biological active substance. The maximum yield of high growth performance of the cell culture mainly depends on the selection of optimal ratios and concentrations of growth regulators. This problem, namely the search for the optimal composition of the nutrient medium has become one of the main tasks in the cultivation of H. maracandicum Popov ex Kirp plant cells. Methods and results of seed sterilization of H. maracandicum are discussed in the article. This endemic, rare species of medicinal plant from the flora of Uzbekistan family Asteraceae Dumotr has a unique composition of secondary metabolites. For example, from the biomass of immortelle were isolated flavonoids, coumarins, lipids, phenols, purines, steroids, triterpenoids, glycosides, coumarins, cerines, bitter tannins, essential oils, etc. Used in folk medicine for cholecystitis and diseases of the liver, bladder, and gastrointestinal tract.

Physiological and anatomical aspects of the rooting of Brunfelsia pauciflora cuttings

• Cuttings represent a viable alternative for the vegetative propagation of Brunfelsia . • The limited rooting of the cuttings was not associated with an anatomical barrier. • Greater quantities of visibly detected starch in B. uniflora favored its rooting. Brunfelsia is an important genus of the Solanaceae family due to its ornamental aspects and pharmacological properties. Brunfelsia pauciflora stem cuttings represent an alternative to seedling production for its propagation. There is no scientific data currently available concerning B. pauciflora cuttings, although previous studies with Brunfelsia uniflora indicated that it may be difficult to root. Anatomical barriers have been reported as limiting factors for some species that are difficult to root. We investigated the effects of the plant growth regulator indolebutyric acid (IBA) and the influence of the seasons of the year on the rooting of B. pauciflora stem cuttings and compared its rooting to B. uniflora . We also investigated the possible influence of anatomical barriers and starch contents on their rooting. Cuttings of B. pauciflora (7 ± 1 cm long) were made in the autumn of 2019, spring/2019, and summer/2020, and treated with IBA at concentrations of 0, 2000, 4000, and 8000 mg L −1 . For comparison, similar cuttings of B. uniflora were prepared during spring/2019 and treated with IBA concentrations of 0, 4000, and 8000 mg L −1 . The rooting variables of both species were analyzed after 120 days of growth in a greenhouse. For the anatomical analyses, segments of the base of the cuttings were collected on the day of their collection, and again after the 120 day growth period. The samples were embedded in historesin, sectioned, and stained with toluidine blue and Lugol's iodine solution. No combined influence of plant growth regulator concentrations and the season of the year was identified, indicating that those factors act separately on the adventitious rooting of B. pauciflora . The IBA plant growth regulator did not significantly affect any of the variables analyzed. The greatest rooting of B. pauciflora (38.8%), the highest leaf maintenance (82.0%), and the lowest mortality rate (11.2%) were observed in the summer. The species can be classified as difficult-to-root. B. uniflora , on the other hand, evidenced a greater rooting capacity (52.2%) in the spring than B. pauciflora (12.2%). The use of IBA (4000 mg L −1 ) resulted in an increased rooting rate of B. uniflora . Anatomically, it was possible to identify the origin of the root primordia as the vascular cambium region, with root formation being defined as direct. A ring of perivascular fibers, often indicated in the literature as an anatomical barrier, was not observed as such in B. pauciflora , as rooting not only overcame the fibers, but also the callus mass. The two species of Brunfelsia differed in terms of their starch contents, which was greater in B. uniflora and appeared to favor its rooting.

Growth Performance of Black Pepper (P. nigrum) Cuttings in Different Rooting Media and Growth Regulators

Black pepper (Piper nigrum) is recognized as "Black Gold" and "King of Spices" because of its widespread use in culinary preparation, food flavoring, and condiment seasoning. It has worldwide fragrance and medical use. The purpose of the conducted research was to investigate growth performance of P. nigrum cuttings in various rooting media and growth regulators concentrations in a low-cost non-mist propagation chamber at the Agroforestry Field Laboratory, Department of Agroforestry and Environmental Science, Sylhet Agricultural University, Sylhet during the period starting from August 2019 to June' 2020. The experiment consisted of two factors: different rooting media and different concentrations of growth regulator, IBA (Indole 3-Butyric Acid). Under the first factor, there were four treatments, namely M1= biochar, M2= vermicompost, M3= well-decomposed cow dung, and M4= field soil as control. Under the second factor, there were four treatments, namely R1= 0.2%, R2= 0.4% and R3= 0.8%, and R4= 0.0% of IBA. The whole experiment was designed in a factorial RCBD (Randomized Complete Block Design) with three replications. Among the different media, M1 treatment (biochar) exhibited superior performance concerning vine length and the number of nodes per vine. On the other hand, R2 treatment (0.4%) concentration of IBA showed better performance regarding the number of roots and vine length. The research found that the growth performance of P. nigrum cuttings are accelerated through integrating rooting media and growth regulators. Consequently, the demand for planting materials could be met.

MICORPROPAGATION OF VIBURNUM OPULUS (ROSEUM) BY USING SINGLE NODES

The present study was carried out to investigate the micropropagation of viburnum opulus (Roseum) by using single nodes with axillary buds excised from soft cuttings using full strength MS salts, 30g.l-1 sucrose, 7g.l-1 Agar and different concentrations of plant growth regulators in culture medium. Results of the experiment at initiation stage revealed that the culture of single nodes of viburnum on a medium containing 4 mg.l-1 BA gave the highest number of shoots (2.21 shoots/explant). Concerning the interaction, a nutrient medium containing (2 mg.l-1 BA+ 0.4 mg.l-1 IAA) gave the highest values of mean number of shoots, leaves and length of new shoots (1.61 shoots/explant, 2.01 leaves/explant, 1.61 cm respectively). At shoot multiplication stage, the addition of 2.0 mg.l-1 from both BA and kinetin gave the highest number of shoots per explant estimated at 2.65 and 3.31 shoots/explant respectively at root formation stage; the interaction treatment of full MS salt strength with the use of 0 .5 mg.l-1 NAA gave the highest rooting percentage reaching 85.5 %. The highest number of roots (18 roots/ explant) was recorded as well from the combined treatment of full MS salt strength with the use of 0.5 mg.l-1 NAA. While the longest roots (6.65 cm) were recorded from the interaction treatment of half salt strength and 0.50 mg.l-1 IBA..

KOMBINASI JENIS BIBIT DAN KONSENTRASI ZAT PENGATUR TUMBUH GIBERELIN TERHADAP PERTUMBUHAN TANAMAN PORANG (Amorphophallus oncophillus)

Porang is a tuber plant that has the opportunity to be developed in Indonesia. Efforts have been made to increase the production potential of porang, including through nurseries. The purpose of this study was to determine the effect of tuber origin and concentration of gibberellin growth regulators on porang growth. The research was conducted in Wonosobo Village, Srono District, Banyuwangi Regency from August to November 2020. This study used a Randomized Block Design (RBD) with two factors, namely the origin of the seeds and the concentration of gibberellin growth regulators with three replications. Factors of seed origin consisted of tubers (B1) and air bulbs (B2) while ZPT gibberellin concentration factors were 1 ml/l (A1), 2 ml/l (A2), 3 ml/l (A3), 4 ml/l ( A4) and 5 ml/l (A5), resulting in 10 treatment combinations. Observational variables include shoot height and number of leaves. On the seedling origin factor, the B2 treatment was the highest in terms of the observed parameters of shoot height and number of leaves. In the ZPT gibberellin concentration factor, it was shown that factor A1 was the highest in the observed variable of shoot height 20 dap (days after planting), while in the variable number of leaves 50 dap, factors A4 and A5 had the highest number of leaves. The interaction of seed origin and gibberellin concentrations showed that factor B1A4 had the highest shoot height at 20 dap observations, while B1A5 had the highest number of leaves at 40 and 50 dap observations.

Phenotypic variation and epigenetic insight into tissue culture berry crops.

Berry crops, a nutrient powerhouse for antioxidant properties, have long been enjoyed as a health-promoting delicious food. Significant progress has been achieved for the propagation of berry crops using tissue culture techniques. Although bioreactor micropropagation has been developed as a cost-effective propagation technology for berry crops, genetic stability can be a problem for commercial micropropagation that can be monitored at morphological, biochemical, and molecular levels. Somaclonal variations, both genetic and epigenetic, in tissue culture regenerants are influenced by different factors, such as donor genotype, explant type and origin, chimeral tissues, culture media type, concentration and combination of plant growth regulators, and culture conditions and period. Tissue culture regenerants in berry crops show increased vegetative growth, rhizome production, and berry yield, containing higher antioxidant activity in fruits and leaves that might be due to epigenetic variation. The present review provides an in-depth study on various aspects of phenotypic variation in micropropagated berry plants and the epigenetic effects on these variations along with the role of DNA methylation, to fill the existing gap in literature.

THE GROWTH OF PIMPINELLA ALPINA HOST CALLUS AT VARIOUS TREATMENTS OF PLANT GROWTH REGULATOR CONCENTRATIONS OF NAA. 2,4 D AND ITS COMBINATION WITH BAP

Purwaceng (Pimpinella alpina) is an herbal plant that has aphrodisiac, diuretic, and tonic properties. The imbalance between the growth and the exploitation of this plant makes it included in Appendix I (endangered), so its conservation will be crucial. One method of the conservation of this plant is tissue culture. This method is considered one way to induct the callus. The research objective is to find the effect of various treatments of plant growth regulator concentrations of NAA. 2,4 D and its combination with BAP on the growth of purwoceng callus. Concentration of plant growth regulator NAA (0.5 mg L-1, 1 mg L-1, 2 mg L-1), 2,4-D (0.4 mg L-1, 0.8 mg L-1, 1.2 mg L-1 ), a combination of NAA with BAP (0.5 mg L-1 NAA: 1 mg L-1 BAP, 1 mg L-1 NAA: 2 mg L-1 BAP, mg L-1 NAA: 3 mg L-1 BAP) and a combination of 2, 4- D with BAP (1 mg LL 2,4-D : 0.5 mg L BAP, 1 mg L-1 2,4-D : 1 mg L-1 BAP, 2 mg L-1 2,4-D : 1 mg L-1 BAP) into Murashige Skoog medium. Then purwaceng leaf explants were planted on the medium until the growth was seen. This research shows that the plant growth regulator concentration has a different influence on the growth of callus that is recognized by the day of callus induction, colour and callus texture. MS media with growth regulator 1.2 mg L-1 2,4-D can grow callus faster (17 days) with friable texture.

Effect of Plant Growth Regulators in African Marigold: A Review

African marigold, being tall enough and producing large flowers suitable for preparing garlands, is the most cultivated cultivar of marigold in Nepal. Besides, pinching and other improved cultural techniques, a number of chemicals, hormones and phyto-hormones are being tested and found to impart a varying response in African marigold. This review has its importance for standardizing the type and concentration of plant growth regulators in African marigold. Online journals, scientific papers, proceedings, books, thesis and lecture notes are the key reference site for this review. Gibberellins or gibberellic acids (GA3), ethrel, paclobutrazol, salicylic acid, Indole 3 Acetic Acid, Naphthalene Acetic Acid (NAA), Chlormequat Chloride and sea weed extract are the most widely used and proven biochemical with pronounced effect on growth, development and differentiation in marigold. Among these, 300 ppm, 350 ppm and 450 ppm of GA3 results higher plant height, minimum days for flower initiation and higher basal diameter respectively. Similarly, 450 ppm for higher number of branches and leaves per plant and 400 ppm of GA3 or 600 ppm of CCC has been found to impart higher number and weight of flowers per plants. Besides, 1000 ppm of CCC has been found more effective to impart higher productivity (yield/ ha) in African marigold. Hence, plant growth regulators have been a valuable agent for optimizing yield and yield potential in African marigold.

Aplikasi Zat Pengatur Tumbuh Asam Giberelat (Ga3) dan Pengaruhnya terhadap Pertumbuhan dan Produksi Bawang Merah (Allium ascalonicum L.) di Desa Datar Kecamatan Sumbang pada Musim Hujan

The purpose of this research was to determine the effect and the best concentration of GA3 growth regulator on the growth and yield of shallots. The research was carried out in the rice fields of Datar Village, Sumbang District, Banyumas Regency, from February to April 2022. This study was a non-factorial field experiment using a randomized block design with 5 treatments and 5 replications. The treatment was the concentration of GA3 with 5 levells, namely 0 ppm, 250 ppm, 500 ppm, 750 ppm and 1000 ppm. The conclusion of this study is the application of GA3growth regulator affected increasing the number of tillers leaf area, bulb number/clump, fresh bulb weight/clump, dry bulb with dry leaves weight/clump, absolute dry bulb weight/clump, yield/plots and yield/hectare. Application of 500 ppm GA3growth regulator showed the best growth and yield of shallot. It was indicated by the number of tillers, leaf area and yield/hectare of 11.54 plants, 1162.18 cm2 and 16.54 ton, respectively

Encapsulation of Brunfelsia pauciflora microprogated shoots as a suitable conservation protocol

The percent work aimed to establish an efficient in vitro propagation and conservation protocol for Brunfelsia pauciflora plant using encapsulation techniques that allow the genetic stability of conserved material. The effect of plant growth regulators types (BAP, Kin, 2ip, and NAA) and concentrations at 0.0, 1.0, and 2.0 mg/l were stud-ied during the establishment phase in comparison with a hormonal-free medium. In vitro proliferated shootlets were encapsulated using various levels of sodium alginate at 1, 2, and 3% (W/V) and calcium chloride at 2.5,5 and 10 %. The regrowth ability of encapsulated explants was evaluated after conservation for 3, 5, and 7 weeks and compared with non-encapsulated buds. The findings of the current study proved that during the establish-ment phase, applying BAP at 2.0 mg/l gave the best results during the three successive subcultures. The best treatment 2ip (2.0mg/l) + BAP (2.0mg/l) + NAA (2.0mg/l) that was considered ideal propagation protocol for shooting and rooting of Brunfelsia. Successful plant recovery from encapsulated explants using low concentra-tions of sodium alginate (1%) and calcium chloride at 2.5% proved that the explants could to survive at the high-est percent (100%) during conservation periods 3, 5, and 7weeks. Increasing calcium chloride to 5% with the same concentration of sodium alginate (1%) caused a gradually reduced percent of survival to 53.33% with an increasing conservations period (from 3 to 7weeks). It is the first paper on the application of the syndetic seed technique on Brunfelsia for the shortest period of conservation and successive renovation of shootlets whose genetic stabilization was evaluated by analysis of RAPD that proved that the genetic similarity ranged from 56.1 to 80%. These results were confirmed by anatomical sections made in Brunfelsia leaf blade.

Modeling and optimizing in vitro percentage and speed callus induction of carrot via Multilayer Perceptron-Single point discrete GA and radial basis function

BackgroundCallus induction is the first step in optimizing plant regeneration. Fit embryogenesis and shooting rely on callus induction. In addition, using artificial intelligence models in combination with an algorithm can be helpful in the optimization of in vitro culture. The present study aimed to evaluate the percentage and speed of callus induction optimization in carrot with a Multilayer Perceptron-Single point discrete genetic algorithm (GA).Materials and methodsIn this study, the outputs included callus induction percentage and speed, while inputs were different types and concentrations of plant growth regulator (0. 5, 0.2 mg/l 2,4-D, 0.3, 0.2, 0.5 mg/l BAP, 1, 0.2 mg/l Kin, and 2 mg/l NAA), different explants (shoot, root, leaf, and nodal), a different concentration compound of MS medium (1 × MS, 4× MS, and 8× MS) and time of sampling. The data were obtained in the laboratory, and multilayer perceptron (MLP) and radial basis function (RBF), two well-known ANNs, were employed to model. Then, GA was used for optimization, and sensitivity analysis was performed to indicate the inputs’ importance.ResultsThe results showed that MLP had better prediction efficiency than RBF. Based on the results, R2 in training and testing data was 95 and 95% for the percentage of callus induction, while it was 94 and 95% for the speed of callus induction, respectively. In addition, a concentration compound of MS had high sensitivity, while times of sampling had low sensitivity. Based on the MLP-Single point discrete GA, the best results were obtained for shoot explants, 1× MS media, and 0.5 mg/l 2, 4-D + 0.5 mg/l BAP. Further, a non-significant difference was observed between the test result and predicted MLP.ConclusionsGenerally, MLP-Single point discrete GA is considered a potent tool for predicting treatment and fit model results used in plant tissue culture and selecting the best medium for callus induction.