Combinations Of Growth Regulators Research Articles

Establishment and application of highly efficient regeneration, genetic transformation and genome editing system for cucurbitacins biosynthesis in Hemsleya chinensis

Background Hemsleya Chinensis is a perennial plant in the Cucurbitaceae family containing antibacterial and anti-inflammatory compounds. The lack of genetic transformation systems makes it difficult to verify the functions of genes controlling important traits and conduct molecular breeding in H. chinensis.ResultsHighly efficient calli were induced on MS medium added 1.5 mg·L− 1 6-benzylaminopurine (6-BA) and 0.02 mg·L− 1 1-naphthylacetic acid (NAA) with high efficiency (> 95%). The frequency of shoot induction was increased to 90% with a plant growth regulator combination of 1.5 mg·L− 1 6-BA and 0.1 mg·L− 1 NAA. Our results also showed that 100% of shoot regeneration was achieved in a shoot regeneration medium. Additionally, more than 92% of kanamycin-resistant plants were confirmed. Furthermore, we achieved 42% genome editing efficiency by applying this transformation method to HcOSC6, a gene that catalyzes the formation of cucurbitadienol. HPLC analysis showed OE-HcOSC6 lines exhibited significantly higher cucurbitadienol levels than the genome-edited lines. Transcriptomic analysis revealed that some downstream genes related to cucurbitadienol biosynthesis, such as HcCYP87D20, HcCYP81Q58, and HcSDR34, were up-regulated in OE lines and down-regulated in mutants.ConclusionsHere, we established a process for regeneration, transformation, and genome editing of H. chinensis using stem segments. This provides valuable insight into the underlying molecular mechanisms of medicinal compound production. By combining high-efficiency tissue culture, transformation, and genome editing systems, we provide a powerful platform that supports functional research on molecular mechanisms of secondary metabolism.

Development of In Vitro Regeneration Protocol for Sweet Pepper (Capsicum annuum L.) using Cotyledon as Explant

Plant tissue culture techniques offer a promising avenue for the efficient propagation of sweet pepper (Capsicum annuum L.), a valuable crop with significant agricultural importance. An effective and repeatable in vitro plant regeneration protocol was developed for the CKN-1 (Lemon yellow) and CKN-8 (Red) genotypes of sweet pepper. Utilizing cotyledonary leaf and cotyledonary node explants, various concentrations and combinations of growth regulators were tested for shoot rejuvenation. For both explants, MS medium supplemented with 8 mg/L BAP, 0.02 mg/L NAA, and 0.5 mg/L IAA was found to be most effective for achieving high rates of response and shoot induction. Notably, cotyledonary node explants outperformed cotyledonary leaf explants, displaying a remarkable 80% response within 11-12 days. Additionally, the supplementation of 1 mg/L IBA with MS medium significantly improved root initiation, with cotyledonary node explants exhibiting the highest responsiveness at 83.77%. Throughout all stages, the genotype CKN-8 (Red) consistently outperformed CKN-1 (Lemon yellow), including in the percentage of regenerated plant establishment. J Bangladesh Agril Univ 22(3): 310–316, 2024

Amorphophallus muelleri Blume Shoot Induction on Different Media Types and Plant Growth Regulator Combinations

Amorphophallus muelleri Blume Shoot Induction on Different Media Types and Plant Growth Regulator Combinations

In vitro Propagation of Leucas biflora (Vahl) Sm. - A Vulnerable Medicinal Herb

A rapidly well-developed in vitro regeneration system has been established for Leucas biflora using nodal segments as explants and MS medium supplemented with various concentrations and combinations of plant growth regulators (PGRS). MS medium containing 2.0 mg/l BAP and 0.5 mg/l NAA showed the highest response (95.99%) in induction of shoot buds. This media combination also produced the maximum number (11.20 ± 0.49) of multiple shoot bud per explant after 3 subcultures at an interval of 14-days. Microshoots showed the maximum elongation (4.70 ± 0.07 cm) in the same medium along with subcultures. These elongated shoots were further grown on rooting media, producing vigorous and healthy root systems. The best response (95.99 ± 1.63%) for rooting was obtained when half-strength MS medium was supplemented with 0.5 mg/l IAA + 0.5 mg/l IBA in 21 days of culture. In this combination, the average number and length of roots were 5.20 ± 0.33 and 4.80 ± 0.30 cm, respectively. The seedlings were removed from culture tubes and transplanted into the pots via several stages of acclimatization. Finally, 98% of the seedlings were found to survive under natural condition. Plant Tissue Cult. & Biotech. 34(1): 49-54, 2024 (June)

Effect of Plant Growth Regulators on Mass Propagation of Strawberry (Fragaria sp.)

Efficient in vitro regeneration protocols for strawberries are crucial for accelerating the propagation of superior cultivars and improving overall productivity through controlled manipulation of plant growth regulators. This study investigated the influence of different concentrations and combinations of plant growth regulators on in vitro regeneration process of strawberries. BARI Strawberry-1 runner tips were sterilized and placed on Murashige and Skoog medium (MS) supplemented with various combinations of cytokinins, including 6-benzylaminopurine (BAP) and kinetin (KIN), to initiate shoot growth. Notably, the combination of 1.5 mg/L BAP + 1.0 mg/L KIN led to the shortest duration for shoot initiation (9 days), resulting in the highest number of shoots per explants (6.33), longest shoot length (2.87 cm), and maximum number of leaves per shoot (4.00). Additionally, this combination also exhibited the highest rate of shoot regeneration (70.33%). Subsequently, the proliferated shoots were transferred to MS medium supplemented with various concentrations and combinations of auxins, specifically 3-indolebutyric acid (IBA) and 1-naphthaleneacetic acid (NAA), for root induction. The in vitro regenerated shoots cultured in MS medium supplemented with 1.5 mg/L IBA + 1.0 mg/L NAA showed the shortest duration for root formation (8 days), the highest number of roots per explants (4.00), and the longest root length (2.77 cm). Furthermore, this combination also resulted in the highest rate of root regeneration (40%). These findings contribute valuable insights for the mass production of healthy strawberry plantlets using in vitro culture techniques. J Bangladesh Agril Univ 22(2):178-184, 2024

Clonal Propagation of Turmeric (Curcuma longa) and Confirmation of Genetic Fidelity of the Micropropagated Shoots by RAPD Markers

Turmeric (Curcuma longa) is used as a spice and has excellent medicinal value. However, the field multiplication rate of turmeric plants is not satisfactory. In vitro methods could be a feasible alternative to vegetative propagation in the field. This study was aimed at developing a suitable protocol for the clonal propagation of turmeric confirming the genetic fidelity of the in vitro regenerated plantlets. The explants of shoot bud (full, half, and quarter), root, rhizome, and leaf were cultivated on MS medium with BAP, NAA, and Kinetin. Cultured shoot buds showed a higher regeneration frequency and shoots per explant than those obtained from other explants. The highest shoot regeneration was 72.49% from shoot buds, followed by rhizome (68.76%) and the lowest was 53.93 % from ¼ shoot buds. There were significant effects of growth regulator combinations and concentrations in shoot regeneration from shoot bud explant. The highest shoot regeneration per shoot bud explant was 4.42 with BAP at 20.0 mg/l and NAA at 0.5 mg/l at 24 weeks. The best rooting response with 9.75 roots per explant was reported with 1.0 mg/l BAP with 1.0 mg/l NAA. The rooted plants were successfully transferred to pots for their acclimatization. Both the micropropagated shoots and their mother plant produced monomorphic bands with the six RAPD markers that confirm the genetic fidelity of micropropagated clones. These results showed that in vitro raised plantlets of turmeric had no risk of somaclonal variations and were found to be true-to-type in nature over the culture period. The protocol developed through this investigation has the potential application for the rapid multiplication of turmeric plants. Plant Tissue Cult. & Biotech. 34(1): 55-69, 2024 (June)

Applications of combinations and concentrations of different phytohormones for the induction of callus, shoot, and root of cannabis sativaunder tissue culture technique

Cannabis sativa is a significant crop of the family (cannabaceae).Interest has been resurgent in the usage of the cannabis plant for medical purposes, industrial fibre, seed oil, food, and recreation.Auxin and cytokinins act antagonistically with each other in which cytokinins promote shoot branching and inhibit root branching, while auxin inhibits shoot branching and promotes root branching. Young cannabis leaves and petioles were used and inoculated in the MS medium with various combinations. Murashige and Skoog (MS) medium for explant were developed in additions of 25µl-450µl 2, 4 D, 25µl-450µl Kinetin, and 50µl-450µl BAP for callus induction. Shoot induction was developed on 2-3ml/l BAP-supplemented MS media from the callus. Root induction was developed from the shoot on MS5ml/2 litters (indole-3-acetic acid) IAA + 8ml/2 litters (indole-3-butyric acid)IBA-supplemented media.Seeds of cannabis were inoculated in the MS medium. After one week, seed germination was observed in the seeds of the DG Khan variety of Cannabis. MS media found better and more abundant Callus induction with 450µl 2, 4D +450µl Kinetin+450µlBAP from cannabis sativa leaves. 3ml/l BAP observed the high growth of the shoot. Better growth of roots was observed by 5ml/2 litters (indole-3-acetic acid) IAA + 8ml/2 litters (indole-3-butyric acid) IBA. In the present study, seed, callus, shoot, root, and meristem growth were observed using the combination and concentration of different growth regulators. By using different phytohormones, the growth rate and productivity of Cannabiswere significantly increased, which can be valuable for medical purposes.

Evaluation of In-vitro Response of Dennettia tripetala (Pepper Fruit) Bak. f. Seedlings as Shoot Explants to a Combination of Plant Growth Regulators

Evaluation of In-vitro Response of Dennettia tripetala (Pepper Fruit) Bak. f. Seedlings as Shoot Explants to a Combination of Plant Growth Regulators

Response of Shoots from Porang Leaf Bulbs to Cytokinins and IAA in Shoot Multiplication In Vitro

The growth regulators in media have the potential to significantly influence the process of in vitro plant regeneration. The response of explants in forming shoots also depends on the type and the combination of growth regulators. This research aimed to identify the most optimal type and concentration of cytokinin for the multiplication of in vitro porang shoots. Additionally, it determined whether the addition of cytokinins should be accompanied by auxin/IAA for porang shoot multiplication. To achieve these objectives, a completely randomized design with six replications was employed, followed by a 5% BNJ. The tested treatments consisted of various combinations of growth regulators. Several observations were recorded, including the speed of explants forming prospective shoots/shoots (days), the age of the explants forming prospective shoots/shoots, the number of prospective shoots and shoots produced per explant, and shoot height (cm). The results showed that 1) Tdz 2 and IAA 0.5 mg.l-1 formed the highest shoots candidates and shoots in porang multiplication in vitro, and 2) the addition of BAP and Tdz 2 mg.l-1 should be combined with IAA 0.5 mg.l-1, while BAP and Tdz 3 mg.l-1 did not require IAA.

Micropropagation and Shoot Tip Cryopreservation of 'Sunny Gold' Freesia.

Cryopreservation is a promising method for the long-term preservation of plant germplasm, especially for vegetatively propagated species like freesias. In this study, we investigate streamlining the cryopreservation process for 'Sunny Gold' Freesia, starting from effective in vitro initiation and proliferation using various plant growth regulator combinations. We also assess the impact of subculture on regrowth rates after cryopreservation. The shoot tips were successfully initiated in vitro after sterilization. The shoots were multiplied an average of three times in media containing N6-benzyladenine and kinetin. The regrowth rates of non-cryopreserved shoot tips excised from different subculture cycles did not differ significantly, with rates of 44% observed for plants from more than five subcultures and 47% for those from three subcultures. However, only the shoot tips excised from cultures subjected to three subculture cycles were able to recover after cryopreservation, with a regrowth rate of 31%. Our findings lay the groundwork for the development of an efficient cryopreservation protocol for freesias in the future.

Phytochemical Profiles and Cytotoxic Activity of Bursera fagaroides (Kunth) Engl. Leaves and Its Callus Culture.

Bursera fagaroides, popularly used in México, possesses bioactive lignans. These compounds are low in the bark, and its extraction endangers the life of the trees. The aim of the present investigation was to search for alternative sources of cytotoxic compounds in B. fagaroides prepared as leaves and in vitro callus cultures. The friable callus of B. fagaroides was established using a combination of plant growth regulators: 4 mgL-1 of 2,4-dichlorophenoxyacetic acid (2,4-D), 1 mgL-1 Naphthaleneacetic Acid (NAA) and 1 mgL-1 Zeatin. The maximum cell growth was at day 28 with a specific growth rate of μ = 0.059 days-1 and duplication time td = 11.8 days. HPLC quantification of the dichloromethane callus biomass extract showed that Scopoletin, with a concentration of 10.7 µg g-1 dry weight, was the main compound inducible as a phytoalexin by the addition of high concentrations of 2,4-D, as well as by the absence of nutrients in the culture medium. In this same extract, the compounds γ-sitosterol and stigmasterol were also identified by GC-MS analysis. Open column chromatography was used to separate and identify yatein, acetyl podophyllotoxin and 7',8'-dehydropodophyllotoxin in the leaves of the wild plant. Cytotoxic activity on four cancer cell lines was tested, with PC-3 prostate carcinoma (IC50 of 12.6 ± 4.6 µgmL-1) being the most sensitive to the wild-type plant extract and HeLa cervical carcinoma (IC50 of 72 ± 5 µgmL-1) being the most sensitive to the callus culture extract.

A highly efficient organogenesis system based on 6-benzylaminopurine and indole-6-butyric acid in Suaeda glauca-a medicinal halophyte under varying photoperiods

Suaeda glauca (Bunge) is a pivotal halophyte species, well-known for its rich nutrient content and its importance as a superior vegetable and oil crop, offering significant health benefits for both human and animal consumption. However, the absence of an efficient regeneration technique has severely hindered its genetic development. Consequently, there is an urgent need to establish an effective in vitro organogenesis initiation method for this valuable plant. In this study, our primary objective was to develop a comprehensive protocol for micropropagating S. glauca, primarily capitalizing on the synergistic effects of 6-benzylaminopurine (6-BAP) and indole-6-butyric acid (IBA). We explored the impact of different combinations of 6-benzylaminopurine (6-BAP), 2,4-dichloroacetic acid (2,4-D), and IBA on callus induction, organogenesis, and rooting using various explants (cotyledons, hypocotyls, and leaves) under two different photoperiod regimes during in vitro culture. Our results unveiled noteworthy variations in several variables related to organogenesis. The optimal combination of plant growth regulators (PGRs) for inducing embryonic callus from cotyledon (COT) was pinpointed at 4.44 μM 6-BAP and 2.71 μM 2,4-D. By employing 6-BAP concentrations ranging from 8.88 μM to 13.33 μM, alongside 0.49 μM IBA, 3% sucrose, and 0.3% phytagel in full MS media under light conditions, we achieved a remarkable 100% regeneration rate through the direct organogenesis pathway. Our findings underscore the pivotal role of 6-BAP in triggering multiple shoots and attaining a 3.6% callus differentiation rate, while 0.98 μM IBA resulted in a 65.28% root induction rate, facilitating the successful acclimatization of plants. Based on the synergistic effects of 6-BAP and IBA, we propose a robust protocol for S. glauca organogenesis, which holds great promise for streamlining micropropagation in S. glauca tissue culture. This standardized protocol has the potential to advance genetic improvement and promote the commercial cultivation of S. glauca as a valuable halophyte species.

Mass Scale Micropropagation Of Andrographis Paniculata (Burm. F.) Wall Ex Nees

A reliable and efficient protocol was developed for in vitro propagation of Andrographis paniculata (Burm. f.) Wall ex Nees with the use of nodal and shoot tip explants. Nodal segments and shoot apex of in vitro grown seedlings were cultured on agar solidified MS medium supplemented with different concentrations and combinations of plant growth regulators (PGRs). Both the explants underwent direct organogenesis and produced multiple shoot buds. In case of shoot apex, maximum number of multiple shoot buds were formed when cultured on MS medium containing 1.5 mg/l Kn + 1.0 mg/l IAA. But nodal segments produced highest number of shoot buds on MS medium fortified with 1.5 mg/l BAP + 1.0 mg/l IAA. Multiple shoot buds underwent rapid elongation on elongation media and maximum elongation took place on MS medium fortified with 2.0 mg/l Kn + 1.5 mg/l IAA. Elongated shoot buds, on further culture in rooting media, produced strong and stout root systems. Half strength MS medium fortified with 2.0 mg/l IAA was most effective for induction and proliferation of roots. The healthy complete plantlets were successfully acclimatized and established in outside pots through successive phases of acclimatization. The Chittagong Univ. J. B. Sci.,Vol. 7(1 &2):121- 130, 2012.

Biochemical properties and pigment contents of Satureja genotypes affected by plant growth regulators and temperature stress.

There is little data, to our knowledge, on the biochemical properties of different Satureja sp. genotypes affected by plant growth regulators (PGR) under temperature stress. A split plot research on the basis of a complete randomized block design with three replicates examining temperature stress (planting dates, 8th of April, May and June) (main factor), and the factorial combination of plant growth regulators (PGR, control (CO), gibberellic acid (GA), fertilization (MI), and amino acid (A)), and genotypes (Khuzestani, Mutika, and Bakhtiari) on plant biochemical properties, was conducted. Plant pigment contents (chlorophyll a, and b and carotenoids (car)), antioxidant activity (catalase (CAT), ascorbate peroxidase (APX) and guaiacol peroxidase (GP)), and leaf protein were determined. Treatments significantly and differently affected the genotypes performance. PD3 and PD1resulted in significantly higher activity of APX (0.059 U. mg-1) and GP (0.190 U. mg-1), respectively (P ≤ 0.05). Temperature stress significantly affected plant CAT activity (U. mg-1) at PD1 (0.084) and PD3 (0.820). Higher temperature significantly enhanced leaf Pro, MI increased plant APX (0.054) and CAT activities (0.111 U. mg-1) significantly, and GA resulted in the highest and significantly different GP activity (0.186 U. mL-1). Treatments T1 and T3 significantly enhanced Chla and Car content, and MI resulted in significantly higher Chlb content (0.085mgg-1 leaf fresh weight). Car and CAT are the two most sensitive biochemical traits under temperature stress and can more effectively regulate Satureja growth and activity. It is possible to alleviate temperature stress on Satureja biochemical properties by the tested PGR.

A Comprehensive Study on the Micropropagation of Costus igneus: Media Composition, Growth, and Development

Costus igneus, commonly known as insulin plant, is an ornamental plant valued for its spiral ginger- like foliage and medicinal properties. Micropropagation is an efficient method for rapid multiplication of Costus igneus. This study aims to determine the optimal media composition and culture conditions for in vitro propagation of C. igneus. Axillary bud explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators including auxins (indole-3-acetic acid, indole-3-butyric acid and naphthaleneacetic acid) and cytokinins (6- benzylaminopurine and kinetin). Microshoots cultured on MS + 1.5 mg/L BAP showed the highest shoot proliferation rate (98%) and maximum number of shoots per explant (12.6). Elongated microshoots were rooted on half-strength MS medium supplemented with different auxins. The highest rooting percentage (95%) and maximum number of roots per shoot (9.8) were observed on medium containing 2.5 mg/L IBA. The regenerated plantlets were acclimatized and successfully transferred to pots with 80% survival rate. Morphological and phytochemical analysis showed no significant differences between the in vitro propagated and mother plants. This study demonstrates the potential of micropropagation for large scale production of quality planting material of C. igneus. Further studies on genetic and epigenetic stability are recommended to validate this protocol for commercial applications.

In Vitro Shoot Multiplication and Rooting of 'Kashan' and 'Hervy Azerbaijan' Damask Rose (Rosa damascena Mill.) Genotypes for Cosmetic and Ornamental Applications.

The damask rose (Rosa damascena Mill.) is an ornamental-medicinal plant from the Rosaceae family, and its aromatic compounds and essential oils are applied globally in the food, cosmetic, and pharmaceutical industries. Due to its economic value, this research aimed to establish a protocol for an efficient, rapid, and cost-effective method for in vitro shoot multiplication and rooting of the R. damascena 'Kashan' and 'Hervy Azerbaijan' genotypes. Nodal segments (as primary explants) were cultured on the Murashige and Skoog (MS) medium with combinations of various plant growth regulators (PGRs) such as gibberellic acid (GA3), 6-benzylaminopurine (BAP), and indole-3-butyric acid (IBA), as well as a PGR-like substance, phloroglucinol (PG), vitamins such as ascorbic acid (AA), and activated carbon in the form of active charcoal (AC). For the establishment stage, 0.1 mg·L-1 PG, 0.2 mg·L-1 GA3, and 1 mg·L-1 BAP were added to the media. Secondary explants (nodal segments containing axillary buds produced from primary explants) were obtained after 30 days of in vitro culture and transferred to the proliferation media supplemented with different concentrations of BAP (0, 0.5, 1, 1.5, 2, and 2.5 mg·L-1) and GA3 (0, 0.1, 0.2, 0.4, 0.8, and 1 mg·L-1) together with 0.1 mg·L-1 PG and 20 mg·L-1 of AA. The rooting media were augmented with different concentrations of BAP and GA3 with 0.1 mg·L-1 of IBA, PG and 20 mg·L-1 of AA and AC. The results showed that the highest regeneration coefficient (4.29 and 4.28) and the largest number of leaves (23.33-24.33) were obtained in the explants grown on the medium supplemented with 2 mg·L-1 BAP and 0.4 mg·L-1 GA3 for the 'Kashan' and 'Hervy Azerbaijan' genotypes, respectively. Likewise, this PGR combination provided the shortest time until bud break (approximately 6.5 days) and root emergence (approximately 10 days) in both genotypes. The highest number of shoots (4.78 per explant) and roots (3.96) was achieved in this medium in the 'Kashan' rose. Stem and root lengths, as well as stem and root fresh and dry weights, were also analyzed. In most measured traits, the lowest values were found in the PGRs-free control medium. Rooted plantlets were transferred to pots filled with perlite and peat moss in a 2:1 proportion and were acclimatized to ambient greenhouse conditions with a mean 90.12% survival rate. This research contributes significantly to our understanding of Damask rose propagation and has practical implications for the cosmetic and ornamental plant industries. By offering insights into the manipulation of regeneration processes, our study opens up new possibilities for the effective production of high-quality plant material.

Effect of Different Concentrations of Auxin as Well as Cytokinin on Shoot Initiation, Formation and Multiplication of Pepino (Solanum muricatum Ait.) cv. Valentia with MS Semi-solid Medium

The pepino (Solanum muricatum Aiton) is conventionally propagated through vegetative means. But, this approach of propagation is not feasible for commercial production of plants and has at more time the chances of reduction yield. Therefore, the availability of quality planting material is of urgent need. Recently, there has been a promising approach for pepino multiplication at large scales through tissue culture because it generates a large number of contamination free plantlets in a minimum space and time. The combination of different growth regulators may be tried to maximize the initiation, growth and development of shoot in commercial propagation of pepino plants. The Minimum days taken for shoot initiation (5.21 days) was noted under BAP 3.00 mgl-1 + IBA 0.50 ppm, and minimum days taken for shoot development (40.82 days) was noted under of BAP 3.00 ppm + IBA 1.00 ppm. The highest length of shoot (9.52 cm) was produced in media supplemented with BAP 3.00 ppm + IBA 1.00 ppm. Maximum percent of developed shoots obtained from the established culture (67.79) was noted under the treatment of BAP 3.00 ppm + IBA 1.00 ppm; While maximum shoots per explants/ innoculums from the established culture (4.06) was noted under the treatment of BAP 3.00 ppm + IBA 1.00 ppm. Maximum number of leaves / plantlet from the established culture (10.85) was noted under the treatment of BAP 3.00 ppm + IBA 1.00 ppm and maximum fresh weight (mg) of shoot / plantlet obtained from the established culture (26.05 mg) was noted under the treatment of BAP 3.00 ppm + IBA 1.00 ppm.

Establishment of plant regeneration protocol through callus induction of sugarcane (Saccharum officinarum l.) var. China using leaf-sheath

Establishment of a plant regeneration system through direct organogenesis or via callus is also a prerequisite to further in vitro genetic manipulation of the cultivar through somaclonal variation, in vitro mutagenesis and genetic transformation. Production of sufficient numbers of plants of a unique genotype is possible using in vitro culture system. In this study, the effect of various concentrations and combinations of plant growth regulators for in vitro regeneration of sugarcane via callus was described using leaf-sheath as explants. Thus, a reproducible protocol for induction of callus and regeneration into plantlets were developed in sugarcane var. china using leaf-sheath as explants. Potential callus induction was observed on MS medium supplemented with 4.5 mg/l 2,4-D, in which 80% explants induced callus after culture initiation of 90 days. Shoot formation from the callus was optimum on MS + 1.0 mg/l GA3 + 0.5 mg/l Kin., in which 70% of the cultured calli formed shoots. The average number of shoot formed per culture callus was 27.0 ± 2.50 and the average shoot length of 6.5 ± 0.50 cm were achieved in this medium after culture of 60 days. Regenerated shoots rooted well when they were transferred into half strength of MS + 2.50 mg/l NAA, in which 90% shoots rooted within 30 days of culture. The average number of root produced per shoot was 18.0 ± 2.25 and the average root length of 7.50 ± 1.20 cm were observed in this medium. Ninety percent of the in vitro raised plantlets were survived in the natural environment.

Rapid Multiplication of Geographical Indication (GI) Tagged Mysore Mallige (Jasminum azoricum L.) Using Single Node through In vitro Culture

Aims: The present study was undertaken to rapid multiplication of (GI) tagged Mysore Mallige (Jasminum azoricum L.) using single node through in vitro culture. Study Design: The present study follow Completely Randomized Design (CRD). Place and Duration of Study: The study was conducted at the Plant Tissue Culture Laboratory, Department of Horticulture, University of Agricultural Sciences, Gandhi Krishi Vignana Kendra, Bengaluru, from 2020 to 21. Methodology: Single node cutting from the young shoots of field growing plants were used as explants to conduct the experiment. The explants were sterilized and placed on MS medium supplemented with different concentrations and combinations of growth regulators, namely BAP and Kinetin. Results: Among the different combinations of growth regulators, the combination of BAP 1.5 mg L-1 and Kin 1.5 mg L-1 produced maximum number of shoots from single node. Very low intensity of callus formation was observed from the cut ends of single node explants that were cultured on half strength MS medium consisting of 1.5 mg L-1 BAP + 0.0 mg L-1 Kin and 1.5 mg L-1 BAP + 1.0 mg L-1 Kin. Conclusion: The use of growth regulators such as BAP and KIN is reliable for shoot regeneration even when the field explants are used.

Exploring somatic embryogenesis in Ajuga multiflora Bunge: Profiling lipophilic metabolites via HPLC, GC-FID, and GCMS– analysis

The present study aims to assess the effect of different concentrations of auxins and combinations of plant growth regulators (PGRs), specifically auxin and cytokinin, on somatic embryogenesis in Ajuga multiflora. We also aim to compare the lipophilic metabolite profiles, such as carotenoids (CAs), fatty acids (FAs), phytosterols (PSs), and tocopherols (TPs), in globular somatic embryos (SEs), germinated SEs, and shoots of SEs derived plantlets. This assessment is carried out utilizing high performance liquid chromatography, gas chromatography-flame-ionization detection, and gas chromatography–mass spectrometry techniques. The results reveal that the application of 2,4-dichlorophenoxyacetic acid (2,4-D) had the most significant impact on somatic embryogenesis, inducing both SE initiation (34.4%) and the number of SEs (3.5) per explant. Among the explants used, leaf explants were found to be the most effective for SE induction (SEI). Additionally, the study explores the combined effect of 2,4-D and cytokinins (N6-benzyladenine, N6-BA, or 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea, TDZ) on somatic embryogenesis, highlighting optimal conditions for SEI percentages and the number of SEs per explant. The maximum SE induction rate (100%) and the number of SEs (45.9) per leaf explant were obtained on an SEI medium with 30 µM of 2,4-D and 2.3 µM of TDZ. The maturation and conversion of SEs were also examined, emphasizing the role of GA3 in achieving the highest conversion percentage (94.0 ± 2.7 at 0.8 µM GA3). Furthermore, the study details the biochemical composition during various stages of somatic embryogenesis, including CAs, TPs, PSs, and FAs. Globular SEs displayed comparatively lower concentrations of CAs than their germinated counterparts and shoots from SE-derived plantlets, while the total PSs content remained relatively consistent across all stages. Conversely, fatty acids tended to accumulate more in globular SEs, with the exception of α-Linolenic acid. These findings provide valuable insights into the optimization of somatic embryogenesis protocols in A. multiflora, with implications for plant tissue culture, regeneration and lipophilic metabolites production.