Combinations Of Plant Growth Regulators Research Articles

Optimization of in vitro micropropagation of ethnomedicinally important underexplored plant Maytenus emarginata

In vitro callus induction and shoot emergence protocols for Maytenus emarginata (Willd) Ding Hou. have been established using young leaves and stem explants of in vivo growing, one and half year-old plants. Plants were growing in the month of August 2016 at department of Botany, Ch. Charan Singh University, Meerut. However, explants were collected in the month of August, 2019. MS (Murashige and Skoog) medium and WPM (Woody Plant Medium), supplemented with different concentrations and combinations of plant growth regulators (PGRs) such as 0.5-2.0 mg/l Naphthalene acetic acid (NAA), 0.5-1.0 mg/l Benzyl Amino Purine (BAP), 2, 4-D (1.0-2.0 mg/l) and 1.0-1.5 mg/l Gibberellic acid (GA3) and temperature regimes were used for callus induction and shoot emergence. The presence of GA3 was found necessary for callus induction and shoot emergence from both explants (leaf and stem). The best response of callus formation from leaf explants was obtained at 24°C on WPM supplemented with 0.5 mg/l NAA+1.0mg/l BAP+1.5mg/l GA3 and MS medium supplemented with 2.0 mg/l NAA+ 0.5 mg/l BAP+ 1.0mg/l GA3. Stem explants exhibited best degree of callus induction only on MS medium supplemented with 0.5mg/l NAA+ 1.0 mg/l BAP+1.5 mg/l GA3. Light and temperature play very crucial role in shoot emergence because exposure of callus or nodal section of stem, to 12 hours dark period followed by 12 hours PAR (photosynthetically active radiation) at 27±2 °C induced shoot emergence on MS medium supplemented with 0.5mg/l NAA+1.0mg/l BAP+1.5 mg/l GA3 after 42 days. This is the first optimization report on tissue culture and medicinal use of callus of Maytenus emarginata.

In vitro effect of phytosynthesised AgNPs to enhance plantlets and biomass production in Nyctanthes arbor-tristis L.

This work was aimed to establish an effective approach for direct regeneration of Nyctanthes arbor-tristis L. using nodal segment. Amongst the different combinations of plant growth regulators (PGR's), the maximum number of shoots/explant (32.94 ± 0.86) with 3.14 ± 0.08 cm shoot length was attained on Murashige and Skoog (MS) medium fortified with 3.5 mg /L 6-benzyladenine (BA) and 0.3 mg/L concentration of 1-phenyl-3-(1, 2, 3- thiadiazol-5-yl)- urea (TDZ). Besides, nodal segments of Nyctanthes grown on MS medium were used to analyze the effect of biogenic AgNPs (0,2,4,6,8 and 10 mg/L) on plant growth & multiplication in solid & suspension culture medium. After 35 days of in vitro shoot proliferation, the shoot number, shoot length, chlorophyll content, fresh & dry matter, guaiacol peroxidase (GPOX) and catalases enzyme (CAT) activities of plants were quantified. The administration of AgNPs (6 mg/L) along with the lower concentration of BA (1 mg/L) could enhance shoot multiplication from (1.29 ± 0.14) to (25.17 ± 0.45), chlorophyll content as well as the GPOX, and CAT enzyme activities in suspension culture medium. Optimized rooting response (12.15 ± 0.63) was noted on the half-strength MS medium supplemented with 0.5 mg/L concentration of Indole acetic acid (IAA), The maximum in vitro raised plantlets was hardened & acclimatized in a ratio of 6:2:2:1(garden soil: cocopeat: vermiculite: pearlite) potting mixture. The regenerated plantlets showed 80% survival rate under greenhouse conditions. Out of twenty PCR-based RAPD (Random Amplified Polymorphic DNA) molecular markers, 8 primers produced a total of 350 scorable bands which showed 90–99% similarity with mother plant.

In vitro propagation for conservation of the rare date palm (Phoenix dactylifera L.) ‘Amri’ using immature inflorescence

Immature female inflorescence plays a significant role in date palm micropropagation because inflorescences are available with no practical limit as the source of explants. Moreover, using floral buds for propagation helps in the conservation of date palm biodiversity and the enhancement of socioeconomically valuable landraces. With the goal of avoiding undesirable genetic variability, the optimal combinations and concentrations of plant growth regulators and other medium compositions were investigated to achieve direct organogenesis and multiplication from the immature female inflorescence of date palm (Phoenix dactylifera L.) cultivar Amri. For the initiation stage, the best response was achieved using Murashige and Skoog (MS) medium containing 1.0 mg L−1 zeatin and 1.0 mg L−1 thidiazuron (TDZ) after 16 wk of culturing. For the multiplication stage, the best culture medium contained 0.5 mg L−1 TDZ solidified with GelriteTM, without activated charcoal for four subcultures, and then supplementing 30 mg L−1 glutathione to this medium composition for two additional subcultures. Plantlets were multiplied and grown for 12 wk on elongation medium and then transferred to the rooting stage in two steps. Compared with other treatments, foliar spraying and watering with 30 g L−1 sorbitol and 40 g L−1 salicylic acid twice a week yielded the best results in terms of survival percentage (95%), leaf width (2.9 cm), and growth vigor (4.4 lateral branch). This was the best combination of plant growth regulators and other medium compositions for micropropagation of date palm (Phoenix dactylifera L.) cultivar Amri without the need for callus formation to avoid undesirable genetic variability.

Development of a Highly Efficient Shoot Organogenesis System for an Ornamental Aeschynanthus pulcher (Blume) G. Don Using Leaves as Explants.

Aeschynanthus pulcher (Blume) G. Don, the “lipstick plant” is a prized ornamental plant with distinctive flowers. Here, we introduce a novel in vitro regeneration method for A. pulcher using leaf explants and an optimized combination of phytohormone plant growth regulators (PGRs). The optimal conditions for shoot regeneration included 1 mg L−1 polyvinyl pyrrolidone (PVP) plus 3 mg L−1 thidiazuron (TDZ), inducing a response rate of 82.4% and a shoot/explant ratio of 38.6. When the Murashige and Skoog (MS) medium contained indole-3-butyric acid (IBA) alone, leaves first differentiated into adventitious roots and then adventitious shoots. Leaves cultured on MS medium containing 1 g L−1 PVP, 3 mg L−1 TDZ, 5 mg L−1 casein, and 0.1 mg L−1 α-naphthaleneacetic acid (NAA) for 30 d exhibited the highest embryogenic callus (EC) induction rate (95.6%). The optimal shoot proliferation coefficient (21.5) was obtained when shoots derived from EC were cultured on the same medium as that used for EC induction for 5 weeks. The most effective medium for rooting of elongated shoots was MS medium containing 1 g L−1 PVP, 5 mg L−1 casein, 3 mg L−1 6-benzyladenine (BA), and 0.1 mg L−1 NAA, and the number of roots reached 18.8. The regenerated plants grown in a greenhouse had 100% survival following one week of hardening. Overall, our effective and efficient propagation method should result in shortened culture periods and reduced production costs, allowing for the future selective breeding and genetic improvement of A. pulcher.

Conservation Horticulture: In Vitro Micropropagation and Acclimatization of Selected Florida Native Orchids

Florida is home to 106 native orchid species, of which 77 are listed as endangered or threatened by the State of Florida. The Institute for Regional Conservation (IRC) has classified 62 of these species as either critically imperiled, imperiled, or rare in South Florida. Because of lack of endosperm, orchid germination rates are very low in nature, as they depend on an obligate relationship with mycorrhizal fungi for nutrients. Most orchid seeds can be germinated in vitro without the need for specific mycorrhizal fungi. This study aims are to establish a fast and efficient protocol for in vitro seed germination using different nutrient media and plant growth regulator (PGR) combinations, and to optimize seedling acclimatization protocols using different greenhouse media. To determine germination preferences, three different in vitro seed germination media were tested supplemented with PGRs, including 1) Murashige & Skoog (MS) (control), 2) MS supplemented with 1.5 mg/L 6-benzylaminopurine (BAP), and 3) MS supplemented with 1 mg/L BAP and 0.5 mg/L 1-Naphthaleneacetic acid (NAA) on Cyrtopodium punctatum, a state-listed endangered, IRC critically imperiled epiphytic orchid. There was no significant difference amongst the treatments after 2 and 6 weeks of seed sowing culture. To understand post-culture survivorship, two epiphytic and one terrestrial native species (Trichocentrum undulatum, Encyclia tampensis, and Oncidium ensatum) were chosen for the acclimatization study with two commercially available potting substrates (coir, sphagnum). To measure seedling growth rates, phenotypic measurements [leaf number, leaf length, root length, plantlet height, light intensity, pH, and electrical conductivity (EC)] and Soil Plant Analysis Development (SPAD) and Normalized Difference Vegetation Index (NDVI) values were recorded monthly for five months. All media/PGR combinations resulted in an initial high percentage of stage I growth but inhibited Protocorm-like bodies (PLBs) development, suggesting more research is necessary to determine later improvements or detriments to MS basal media with BAP and NAA. Comparing acclimatization media with the three species of orchid chosen for this experiment, neither O. ensatum nor E. tampensis showed a marked preference for sphagnum moss or coir. However, T. undulatum did perform better with coir compared with sphagnum. This research will help botanical gardens and commercial plant tissue culture laboratories to have a better understanding on selection of PGR combinations for in vitro cell culture and acclimatization media on increasing the viability and plant health and decreasing the mortality of endangered plants.

Preliminary Study of <i>Pleurotus ostreatus</i> (Jacq.) P. Kumm. Spent Mushroom Compost as Nutrient Supplement on the Shoot Induction of Fig Tissue Culture

Spent mushroom compost is identified as waste substrate. As the mushroom agriculture industry is growing rapidly, the production also increases and contributes to large amounts of spent mushroom compost annually. The conversion of spent mushroom compost as waste to wealth is a great concept that can give value to agricultural industry and solid waste management of mushroom cultivation. The use of SMC as nurient supplement may reduce the production cost for in vitro culture of fig plant. This research is to study the use of Pleurotus ostreatus (Jacq.) P. Kumm. spent mushroom compost (SMC) as a nutrient supplement of the media in shoot induction of fig plant. Nodal segment of fig’s stem was cultured in MS media supplemented with different concentrations of SMC. With presence of SMC, 10% SMC have the highest shoot proliferation (2.167 ± 1.169) and length of shoot (0.154 ± 0.060 cm). The study revealed that in presence of 10% of SMC could produce new shoots at a frequency which was comparable to the control (p<0.05; n=3; Tukey’s multiple range test). Among the different media combinations of plant growth regulators with SMC, 15 μM Kinetin with 10% SMC is the most effective treatment which gave maximum number of shoot proliferation (3.00 ± 1.27). 30 μM IAA with 10% SMC media have the highest leaves regenerated which were 1.33 ± 0.84. In conclusion, the SMC added in media for plant tissue culture had positive effect on shoot induction thus highlighted the potential of waste substrate of SMC to act as nutrient supplement for plant tissue culture.

Influence of Plant Growth Regulators on Glandular Trichome Density and Steviol Glycosides Accumulation in Stevia rebaudiana.

With the advent of modern lifestyles, diabetes-related comorbidities attributed the importance of low-caloric natural sweetener plants such as Stevia rebaudiana. This plant is the viable source of steviol glycosides (SGs) and other economically important secondary metabolites. Glandular trichomes (GTs) play the role as a reservoir for all secondary products present in the plant species. Therefore, the present study was carried out to evaluate the influence of different plant growth regulators (PGRs) on GT density and its impact on the SG content. The direct shoot regeneration system was developed on Murashige and Skoog (MS) + benzyl aminopurine (BAP) (1.0 mg/L) + naphthaleneacetic acid (NAA) (0.5 mg/L), and MS + BAP (1.5 mg/L) + NAA (0.5 mg/L) from nodal and leaf explants, respectively. Among the combination of PGRs used, MS medium fortified with BAP (1.0 mg/L) and 2,4-dichlorophenoxyacetic acid (2,4-D) (0.5 mg/L) played a significant role in increasing the GT density on leaf and stem tissues of S. rebaudiana. Furthermore, high-performance thin-layer chromatography and gas chromatography–mass spectrophotometry data confirmed a notable rise in SGs and other valuable secondary metabolites. Thus, the protocol developed can be used for the propagation of stevia with an improved metabolic profile at a large scale.

GC-MS Analysis of the in vitro leaf derived Callus Extracts of Ichnocarpus frutescens (L)W.T.Aiton

Medicinal plants are the treasure house of secondary metabolites. Production of seconmdary metabolites through micro propagation technology has immense application in new drug discoveries. Ichnocarpus frutescens (L.) W.T. Aiton - is a latex secreting medicinal herb commonly known as black creeper or Krishna Sariva belonging to the family Apocyanaceae.The present study was aimed at induction of callus from the leaf explants of I. frutescens using Murashige and Skoog (MS) medium supplemented with various combinations of plant growth regulators. Maximum amount of green compact mass of callus was proliferated from leaf explants on MS medium fortified with 3mg/L NAA + 2mg/L BAP. The main objective of the present study was to investigate the secondary metabolites in different extracts of callus by GC-MS analysis. To our best knowledge, no literature is available and this is the first report on GC–MS analysis of leaf derived callus extracts, which showed the presence of different phytocompounds out of which, Stigmasterol, Hexadecanoic acid, Beta-Sitosterol, Stigmasta-3,5-dien-7-one and Heptacosane are the major compounds that found to possess a wide range of biological activities. This protocol could be used to identify the secondary metabolites from leaf derived callus of the I. frutescens and further studies have to be carried out for the isolation and characterization of novel compounds for the formulation of new pharmaceutical drugs and also for its commercial production which will be used to cure some important ailments. ,

Influence of Boron Levels and Plant Growth Regulators on Growth, Yield and Economics of Cowpea (Vigna unguiculata L.)

Background: Cowpeas thrive in poor dry conditions, growing well in soils up to 85% sand. This makes them a particularly important crop in arid, semidesert regions where not many other crops will grow. As well as an important source of food for humans in poor, arid regions, the crop can also be used as feed for livestock. Cowpea is primarily used in the form of dry seeds, fodder, green pod, green manure, and cover crops.
 Objectives: Effects of boron levels and plant growth regulators on growth, yield and economics of cowpea (Vigna unguiculata L.).
 Methods: The field experiment was conducted in Zaid season of 2021, at crop research farm of Department of Agronomy at Sam Higginbottom University of Agriculture, Technology and Sciences, Prayagraj in North Eastern plains of Eastern Uttar Pradesh, under Randomized block design comprising of 10 treatments with different combination of boron levels and plant growth regulators which are replicated thrice.
 Conclusion: Based on the above findings it can be concluded that the application of boron and plant growth regulators performs effective and improves the Plant height, Dry weight, No of Branches, No of pods per plant, No of seeds per pod, Test weight, Seed yield and stover yield of cowpea, the findings are based on research conducted during a single season, they may be repeated for further confirmation.

Studies on the induction of basal stem cluster buds and nodes propagation of Amomum villosum Lour.

Amomum villosum Lour. as Chinese herbal medicine and seasoning, which has a huge potential economic value. This plant is increasing shortage of resources by the slow sexual reproduction and the low artificial reproduction rate. The plants with strong stress resistance and high yield were selected as the female parent, on the basis of single factor and complete combination, the orthogonal test of L9 (34) and L16 (45) was further conducted. 6-BA induced basal stem cluster bud formation was obviously better than that of KT or ZT. 2, 4-D significantly induced callus occurrence and node enlargement, while NAA was more beneficial to adventitious root formation The combination of 6-BA and NAA was more beneficial to induce the formation of cluster buds from basal stem, but the proliferation effect was not ideal. Thus, KT or 2, 4-D was introduced for two orthogonal tests of L9 (34), and the highest proliferation coefficient was only 4.56. Then it was found that adding 0.5 mg·L-1 2, 4-D to the above combination, appeared a unique phenomenon of node propagation. Next, L16 (45) orthogonal test was conducted using 4 plant growth regulators combinations of 6-BA, 2, 4-D, KT and NAA. The optimal medium for proliferation culture was the MS medium with 7.5 mg·L-1 6-BA, 5.0 mg·L-1 NAA, 1.5 mg·L-1 KT, 0.5 mg·L-1 2, 4-D, and the proliferation coefficient reached above 10.00. The optimal rooting medium was the 1/2 MS medium with 2.0 mg·L-1 NAA. With the node propagation, a rapid propagation system of A. villosum was established which provided a possible solution for improve the efficiency of artificial planting, solve the market demand and quality problems.

Development of an efficient in vitro regeneration system in Costus speciosus - an important herbal insulin plant

Costus speciosus is being rapidly eliminated in its natural habitats in India and driven to a nearly threatened degree of extinction because of non-selective collection and over-exploitation. The commercial cultivation of C. speciosus is impeded owing to its poor seed viability, reduced germination rate, and weak rooting potential of vegetative cuttings. As costus could not be multiplied rapidly by conventional propagation methods, in-vitro regeneration technology was experimented with to standardize explants in addition to the combinations of plant growth regulators (PGRs) to fortify the growing media that helps to produce the enormous number of plants in a shorter duration. Several kinds of explants viz., leaves and nodes were cultured into the Murashige and Skoog (MS) medium reinforced with different combinations and doses of PGRs. A higher frequency (51.67 ± 1.7%) of the regenerated adventitious shoot was obtained from nodal segments cultured onto MS medium fortified with 0.5 mg/l Thidiazuron (TDZ) and 2.0 mg/l of 6-benzyl amino purine (BAP). TDZ in combination with BAP and Indole-3-acetic acid (IAA) exhibited a remarkable effect on shoot multiplication and elongation. Further, MS medium with 0.5 mg/l TDZ, 1.0 mg/l 2, 4-dichlorophenoxyacetic acid, and 0.5 mg/l IAA yielded a great range of callus initiation (86.66 ± 1.6%) from leaf explants. Leaf-derived callus results in the greatest shoot regeneration (56.66 ± 1.7%) response on TDZ (0.5 mg/l) and BAP (1.0 mg/l) reinforced MS medium. Elongated shoots from node and callus cultures when inoculated with the MS medium reinforced along with 1.0 mg/l of IAA and 0.5 mg/l of Indole-3-butyric acid (IBA) produced optimum rooting (88.33 ± 1.66%). The rooted young plantlets were effectively acclimatized to the environmental climate and their greenhouse survival rate was 85%.

The combination of plant growth regulators (GA3 and Gracilaria sp. extract) and several fertilisers in Salak Pondoh fruit production

Plant growth regulators (PGRs) and proper fertilisers are used to increase quality of the Salak Pondoh, such as in the plant production, fruit number and size. This study aims to determine the type and dosage of PGRs and the best type of fertiliser to increase the production bunch and fruit of Salak Pondoh. The method used was a Factorial Completely Randomised Design (FCRD) with 36 treatments (excluding the control) and 3 replications. Three treatments were conducted: first, as a control, no PGR treatment + fertilisation (no fertilisation, manure, and NPK). Second, a 3 × 3 factorial scheme. Treatment of PGR GA3 (50, 100, 150 ppm) + fertilisation (no fertilisation, manure, and NPK). Third, a 3 × 3 × 3 factorial scheme was used. Treatment of PGR Gracilaria sp. extract (50, 100, 150 ppm) with different solvents (n-Hexane, methylene-chloride, ethanol) + fertilisation (no fertilisation, manure, and NPK). The parameters observed were the number length, area of the bunch, length and area of the fruit. The results showed the highest number of bunches was found in PGR GA3 50 ppm. While, the highest bunch area was found in PGR Gracilaria sp. ethanol 150 ppm. The PGRs only increased the formation of the bunches and did not increase the length and width of the bunches and the fruit size. Fertilisation affected the Salak production in the form of the number, length and width of the bunches, and the fruit size. In addition, the fertiliser increased the number of the bunches, bunch area, and length and area of the fruit. The interaction of the PGR and fertilisation had no significant effect on increasing the Salak Pondoh production. The best result in increasing the Salak Pondoh production was PGR Gracilaria sp. ethanol 100 ppm with manure fertilisation.

Effect of Nipping and Plant Growth Regulator on Growth and Yield of Chick Pea (Cicer arietinum)

A field experiment was carried out at Crop Research Farm, Department of Agronomy, Naini Agricultural Institute, SHUATS ,Prayagraj(U.P.)in rabi 2021 to study the "Effect of nipping and plant growth regulator on growth and yield of Chick pea (Cicer arietinum)".It was consisting of three combinations of nipping and plant growth regulators. The experiment was laid out in Randomized Block Design with nine treatments each replicated thrice. The experiment results revealed that the growth parameters and yield parameters such as Plant height (23.93cm),Number of Nodules (22.00cm),Dry weight (11.41cm),Number of Branches (8.20cm), crop growth rate(16.96) Recorded highest At harvest, Significantly recorded in treatment T2 with application of Nipping and GA3 at 100ppm. Moreover, No.of pods/plant (56.40),No.of seeds/pod (1.53),Test weight(264.82),Seed Yield (3.43t/ha),Harvest index (41.34%),Gross return (154,350.00),Net return(107,252.50),and were also recorded significantly higher in the treatment T4 With application of Nipping and NAA at 50ppm among all treatments.

Callus via Shoot Organogenesis and Plant Regeneration of Stevia rebaudiana Bertoni

Stevia rebaudiana Bertoni is important medicinal plant which is native between southern Brazil and northern Paraguay. A reliable protocol for shoot organogenesis and regeneration was developed for an antidiabetic medicinal plant species. Regeneration via indirect shoot organogenesis was established from two types of explant (leaf and stem) on MS medium [1] with different concentrations and combinations of plant growth regulators (PGR) such as 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP) and gibberellic acid (GA3) for inducing organogenic callus of S. rebaudiana. In the present investigation the highest percentage of response was shown by 90% and 80% callogenesis from leaf and stem explants on MS medium supplemented with 2.0 mg/l 2,4-D + 0.5 mg/l BAP + 0.5 mg/l GA3 (MSC3) was observed. The high frequency of regenerants was observed from S. rebaudiana leaves at 0.5 mg/l 2,4-D + 2.0 mg/l BAP + 0.5 mg/l GA3 (MSR3). Optimal shoot multiplication and rooting were obtained on 2.0 mg/l BAP and 0.1 mg/l indole- 3-butyric acid (IBA), respectively, followed by successful acclimatization of regenerants (90% survival) to greenhouse condition. The purpose of this study was to establish a reliable S. rebaudiana regeneration protocol that can be used to facilitate the cultivation, propagation and planting of this plant species.

Plant growth regulators affecting corn (Zea mays L.) physiology and rab17 expression under drought conditions

Finding methods, which may improve corn ( Zea mays L.) growth and physiology under drought stress, is of significance. Accordingly, the effects of different plant growth regulators (PGRs) including benzyl adenine (BA6), gibberellins (GA 3+7 ), proline (Pr), and BA6 + GA 3+7 + Pr (sub plots), at different drought levels including control (S1), moderate (S2) and severe (S3) (main plots) on the physiology and genetic contents of corn (Single Cross 704) plants were investigated. A two-year field research, as a split plot on the basis of a randomized complete block design with four replications, was conducted in the Research Field of Islamic Azad University, Isfahan, Iran in 2015–2016. Corn physiological parameters including carotenoids (Car), total chlorophyll (Chlt), chlorophyll a (Chla), chlorophyll b (Chlb), plasma membrane resistance (PR), and rab17 expression were determined. Analysis of variance and comparison of means indicated the experimental treatments significantly (P ≤ 0.01) affected the measured parameters. The combination of PGRs and drought stress was not significant on PR, and just drought stress had a significant effect on rab17 expression at P ≤ 0.01. The year parameter significantly affected corn Car and PR. Although stress decreased plant Car, and Chl contents, it increased PR. The tested PGRs significantly improved corn physiology and rab17 expression, and interestingly their effectiveness enhanced with increasing the level of stress (significant interaction of PGRs and drought stress). The role of proline as an RNA binding protein affecting RNA activity and rab17 expression and subsequent corn tolerance under drought stress has been suggested. • Plant growth regulators (PGRs) affecting corn physiology and genetic under drought. • Proline and its combination with benzyl adenine and gibberellins the most effective. • Suggesting proline as and RNA binding protein in the alleviation of drought stress. • The possibility of alleviating drought stress in corn plants using the tested PGRs.

English

Sorghum is a tropical grass used for various purposes including food, animal feed, and biofuels source, but its in vitro propagation still found an obstacle due to tissue browning. This study aims to propagate and regenerate callus in the Indonesian sorghum hybrid, named Numbu cultivar from seed and meristematic leaf whorl explants. The combined effect of plant growth regulators (PGRs), namely 2,4-D and proline as well as NAA (naphthalene acetic acid) and BAP (6-benzyl amino purine) was investigated on embryogenic callus induction and regeneration, respectively. The results showed that callus induction was significantly faster in the seed explants compared to leaf whorl. The optimal medium used to obtain a large induction in the seed explants was MS salt containing 3 mg L-1 2,4-D and 5000 mg L-1 proline, which gave a 68% yield. Furthermore, the best medium used to obtain a high plant regeneration frequency was the MS medium containing 0.1 mg L-1 NAA and 3 mg L-1 BAP, which regenerated 78% of the callus into plantlet. The addition of 560 mg L-1 proline to the combination medium increased doubled the regeneration into the planlet. Based on the results, PGRs combinations are required to improve embryogenic callus induction and regeneration into planlet in sorghum. © 2022 Friends Science Publishers

Highly efficient Agrobacterium-mediated transformation and plant regeneration system for genome engineering in tomato

Tomato (Solanum lycopersicum L.) is an important vegetable and nutritious crop plant worldwide. They are rich sources of several indispensable compounds such as lycopene, minerals, vitamins, carotenoids, essential amino acids, and bioactive polyphenols. Plant regeneration and Agrobacterium-mediated genetic transformation system from different explants in various genotypes of tomato are necessary for genetic improvement. Among diverse plant growth regulator (PGR) combinations and concentrations tested, Zeatin (ZEA) at 2.0 mg l−1 in combination with 0.1 mg l−1 indole-3-acetic acid (IAA) generated the most shoots/explant from the cotyledon of Arka Vikas (36.48 shoots/explant) and PED (24.68 shoots/explant), respectively. The hypocotyl explant produced 28.76 shoots/explant in Arka Vikas and 19.44 shoots/explant in PED. In contrast, leaf explant induced 23.54 shoots/explant in Arka Vikas and 17.64 shoots/explant in PED. The obtained multiple shoot buds from three explant types were elongated on a medium fortified with Gibberellic acid (GA3) (1.0 mg l−1), IAA (0.5 mg l−1), and ZEA (0.5 mg l−1) in both the cultivars. The rooting was observed on a medium amended with 0.5 mg l−1 indole 3-butyric acid (IBA). The transformation efficiency was significantly improved by optimizing the pre-culture of explants, co-cultivation duration, bacterial density and infection time, and acetosyringone concentration. The presence of transgenes in the plant genome was validated using different methods like histochemical GUS assay, Polymerase Chain Reaction (PCR), and Southern blotting. The transformation efficiency was 42.8% in PED and 64.6% in Arka Vikas. A highly repeatable plant regeneration protocol was established by manipulating various plant growth regulators (PGRs) in two tomato cultivars (Arka Vikas and PED). The Agrobacterium-mediated transformation method was optimized using different explants like cotyledon, hypocotyl, and leaf of two tomato genotypes. The present study could be favourable to transferring desirable traits and precise genome editing techniques to develop superior tomato genotypes.

Thidiazuron Induced In Vitro Clonal Propagation of Lagerstroemia speciosa (L.) Pers.—An Important Avenue Tree

A high throughput regeneration protocol has been developed for Lagerstroemia speciosa through node explants under the regime of various plant growth regulators (PGRs). This protocol can provide an alternative mode to seed-grown plants and minimize the cost–time of regeneration, significantly. Murashige and Skoog (MS) medium containing various combinations of PGRs exhibited a marked stimulatory effect on morphogenesis. Of the various combinations tried, node explant pretreated with thidiazuron (TDZ; 5.0 µM) for 4 weeks and followed with transfer into MS medium containing 1.0 μM 6-benzyladenine (BA) and 0.25 μM α-naphthalene acetic acid (NAA) was reported to be the best treatment as it resulted in a maximum number of 24.5 shoots with an average shoot length of 7.1 cm per explant in 90% of cultures after 12 weeks of incubation. The in vitro-generated shoots rooted satisfactorily in the adopted ex vitro method of rooting, which saves time and cost. Among the different treatments, the greatest rooting percentage (85%) was observed in the 200 μM IBA-treated shoots, with the highest root number (8.7) and length (3.4 cm) occurring after 4 weeks. Four months after being transferred to ex vitro, some of the physiological attributes of the in vitro-propagated plants were examined and compared to the ex vitro plants. Further, analysis of the genetic integrity in tissue culture-raised plantlets along with the parental tree was accomplished through DNA-based RAPD technique. The monomorphic banding pattern obtained by the RAPD primers resulted in a high level of genetic uniformity in regenerated plants.

Comparison of Phytochemicals, Antioxidant, and Antimicrobial Activities of in Vitro Propagated and Wild Grown Potentilla Nepalensis, an Endemic Medicinal Plant from North Western Himalayas

ABSTRACT A reliable protocol for in vitro propagation of Potentilla nepalensis was developed and the phytochemical, antioxidant, and antimicrobial effects of in vitro propagated and wild grown P. nepalensis plants were compared. Leaves of P. nepalensis used as explants were cultured on conventional Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators. Optimum sterilization conditions were achieved by using 0.04% Bavistin for 10 min, 0.04% sodium hypochlorite for 1 min, and 0.04% mercuric chloride for 1 min and the optimum media composition for callus generation was MS minerals containing ammonium nitrate (165 mg L−1), calcium chloride (44 mg L−1), and growth regulators IBA (20 mg L−1), and BAP (4 mg L−1). The phenolic and flavonoid content were higher in wild grown plant (TPC-21.21 ± 0.54 mg g−1 GAE; TFC-4.24 ± 0.17 mg g−1RE) compared to in vitro propagated P. nepalensis (TPC-3.55889 ± 0.72 mg g−1 GAE; TFC-0.33 ± 0.07 mg g−1 RE). Although the antioxidant activity was high in wild grown plant (IC50-235 ± 92.120 µg mL−1) than in vitro propagated (IC50-682.989 ± 57.118 µg mL−1), the antimicrobial properties for bacterial and fungal pathogens were similar.

In vitro production of tropane alkaloids from Brugmansia suaveolens

For thousands of years, secondary metabolites have been utilized as medications, flavors, pesticides, and dyes. For the generation of valuable secondary metabolites, in vitro plant culture techniques have been viewed as beneficial alternatives to whole plants. Brugmansia suaveolens is an ornamental plant including anticholinergic agents which are employed in medicine. Callus induction was performed from leaf and internode explants cultured on Murashige and Skoog’s medium supplemented with different concentrations and combinations of plant growth regulators (PGRs) with 6 treatments. The highest callus induction response was obtained from the leaf-originated explants (73%) on the medium supplemented with 0.4 mg/L KIN and 0.2 mg/L NAA which produced friable callus in 4 weeks. The cell suspension culture of B. suaveolens was established in shake flasks using friable calli. The extraction protocol of tropane alkaloids was optimized, atropine and scopolamine were obtained efficiently. The data could provide technical support for the large-scale production of valuable alkaloids of B. suaveolens in vitro systems with improved strategies.