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• Multiple shoots were successfully regenerated from shoot tip explants of Tecoma stans L., under the influence of meta-topolin (mT). • in vitro raised leaf tissues revealed higher phytochemical diversity and significant increase in antidiabetic alkaloid. • No genetic variation was observed among micropropagated plants with the explant donor plant (mother plant) via RAPD marker assessment. Tecoma stans (Bignoneaceae) is a valuable medicinal shrub having commercial and industrial importance for its bioactive phyto-constituents. In the current study, multiple shoots were successfully regenerated from shoot tip explants of Tecoma stans L., under the influence of meta -topolin (m T). On evaluation of the morphogenic effect of its various doses either singly or in combination with different auxins, MS medium fortified with 7.50 μM m T and 0.50 μM NAA yielded maximum shoots (11.8 ± 0.37) per explant with an average length (5. 08 ± 0.28 cm), after 8 weeks. For studying the effect of subculturing passages, the cultures were grown on to optimized medium and an enhanced shoot multiplication upto 35 shoots per explant were observed at 8th subculture passage. For ex vitro rooting, healthy microshoots were excised and pulse treated for 3 h with higher doses (100 - 500 μM) of auxins IAA, IBA or NAA separately before their transplantation to Soilrite. Of the tested treatments, best rooting was obtained in 300 μM IBA treated microshoots, after 4 wk of plantation in Soilrite. Further, on studying the effects of secondary metabolite production, in vitro raised leaf tissues revealed higher phytochemical diversity and significant increase in antidiabetic alkaloid (Tecomine) from 0.36% control plant to 1.68% in in vitro raised plants. Inspite of different phytochemical profiles, no genetic variation was observed among micropropagated plants with the explant donor plant (mother plant) via RAPD marker assessment. The established micropropagation system appears to be practicable for large scale production of T. stans progenies with higher number of biochemical compounds. [ABSTRACT FROM AUTHOR]

Establishing vegetative propagation techniques to promote the rejuvenation/reinvigoration of genotypes is essential for the rescue of adult trees of the Eucalyptus genus used in species and provenance tests. The aim of this study was to evaluate the induction of epicormic buds and shoots in pruned branches and the in vitro establishment, multiplication, genetic fidelity, and elongation of three 44-year-old Eucalyptus pilularis selected plants. M1, M2 and M3 represent selected plants. M1 produced the highest number of epicormic buds, number of shoots, and with the lowest rate of tissue oxidation and non-reactive explants. In the establishment stage, M3 exhibited the lowest contamination percentage, number of shoots, and shoot length. The emission of shoots occurred only for M3, and it was the unique genotype subjected to the other stages (i.e. multiplication, elongation, and rooting). In the multiplication stage, the highest vigour and shoot length values of were found in the 15th subculture. Phenolic oxidation had its highest value in the 12th subculture, decreasing from the 13th. The highest value for number of shoots was found in the 11th subculture. No polymorphism was observed in the selected plant (M3) and the clonal plants obtained in the 15th subculture. For shoots elongation, the use of culture medium containing 0.10 mg L-1 BAP and 1.00 mg L-1 NAA provided the lowest means for oxidation, and the highest for vigour, number, and length of shoots. The emission of adventitious roots was observed, demonstrating that, through micropropagation, it was possible to induce the competence to root the material, even at an advanced ontogenetic age. [ABSTRACT FROM AUTHOR]

Purple flesh sweet potato var. 'Bhu Krishna' is a rich source of vitamins, minerals, and several important secondary metabolites, including anthocyanin. The tuber of this variety contains a high level of anthocyanin (85-90 mg/100 g). This plant is propagated through stem cutting, which is time-consuming and diseases are transmitted to the next generation. Therefore, there is an urgent need to develop a micropropagation system for the production of huge number of healthy plants to fulfil the demand of farmers as well as industrial utilization. The present study reports a reproducible micropropagation system for purple flesh sweet potato var. 'Bhu Krishna' using mature nodal explants on Murashige and Skoog's (1962) medium fortified with various plant growth regulators (PGRs). Maximum shoots regeneration was observed on MS medium fortified with 2.0 mg/l meta-Topolin. In vitro nodal segments were excised from established shoot cultures and inoculated on the above said optimum medium to produce more shoots (ca. 1300 shoots). In vitro regenerated shoots were successfully rooted on half-strength MS medium augmented with 0.5 mg/l Indole-3-butyric acid. The micropropagated plants were acclimatized on soil, coco peat, and sand (1:1:1) and about 95% of plants survived. Finally, these plants were transferred to the field. The genetic unifomity and phytochemical analysis of micropropagated plants vis-à-vis mother plant were demonstrated using Inter Simple Sequence Repeat markers and spectrophotometric method, respectively. The therapeutic activity in terms of anticancer activity of tubers of micropropagated plants with the mother plant was also tested. For the first time we are reporting a detailed micropropagation system for purple flesh sweet potato var. 'Bhu Krishna' for its mass propagation to fulfil the demand of farmers as well as industries. [Display omitted] • Micropropagation protocol was developed for anthocyanin-rich Sweet potato first time. • Multiple shoot proliferation from nodal segment on MS + 2.0 mg L−1 mT. • Rooting of in vitro regenerated shoot on ½ MS + IBA 0.5 mg L−1. • Genetic fidelity was analyzed by molecular marker (ISSR). • Phytochemical and anticancer activity were evaluated both the mother vis-à-vis micropropagated plant tuber. [ABSTRACT FROM AUTHOR]

The prized Red banana, selected for superior qualities, demands strong genetic uniformity for successful clonal propagation and preservation. Ensuring this uniformity early in the growth of in vitro Red banana plants is essential, as gene mutations and chromosome rearrangements during tissue culture can jeopardize both cloning and germplasm conservation. In this situation, molecular markers play a pivotal role in confirming genetic stability. Thus the study aims to discover a marker that identifies tissue-cultured Red bananas from their virescent variants during initial sub-culturing. A marker linked to anthocyanin has been identified which effectively differentiated Red bananas from virescent variants and it was further validated in various banana cultivars, ornamental Musa species and their interspecific hybrids. The PCR-based marker showed remarkable specificity, discerning Red bananas from virescent variants during tissue culture. It also distinguished green and red offspring, cutting time and resource costs, and shortening the banana breeding cycle. [ABSTRACT FROM AUTHOR]

Solena amplexicaulis (Lam.) Gandhi is a creeping cucurbit with high medicinal values in terms of its therapeutic uses. Traditionally, this plant species is used as a folklore medicine for the treatment of various disorders like spermatorrhea, urinary retention, jaundice, and cardiovascular disorders. Owing to high medicinal importance, it is overexploited, which has resulted in decrease in its population size in the wild. Furthermore, its natural propagation through root tubers and seeds is limited. In view of this constraint, the present study of in vitro micropropagation was chosen as an alternative for its propagation and conservation. Murashige and Skoog (MS) media amended with plant growth regulators like 6-benzylaminopurine (BAP) and thidiazuron (TDZ) independently and with a combination of indole-3-acetic acid (IAA) were tested for in vitro shoot induction by using nodal explants. The highest number of shoots (32.84±0.02) per explant was achieved on Murashige and Skoog (MS) medium with 1.0 mg L-1 TDZ and 1.5 mg L-1 IAA. The induced shoots were elongated with a maximum shoot length of 10.25±0.01 cm on MS medium augmented with 2.0 mg L-1 gibberellin (GA3) and 1.5 mg L-1 TDZ. Rooting of in vitro raised shoots was attained on half-strength MS medium fortified with 1.0 mg L-1 indole-3-butyric acid (IBA) and 2.0% sucrose. The hardening of resultant complete plantlets was carried out in plastic cups by using a 2:1 ratio of sterile black soil and sand; initially, they were kept in greenhouse conditions and subsequently shifted to natural field conditions with 78% of survival rate. The genetic fidelity of tissue-cultured plants was confirmed by ISSR (inter simple sequence repeats) and SCoT (starting codon targeted) primer-based analysis. [ABSTRACT FROM AUTHOR]

Artemisia vulgaris is a medicinally important essential oil–yielding plant belonging to Asteraceae, used as a traditional remedy against chronic diseases. The essential oil of this plant has been used as an insect repellent and antimicrobial agent. The growing concern of antibiotic resistance has forced to find an alternative way to control multi-drug-resistant urinary tract–infecting bacterial pathogens by using the essential oil and leaf extract of A. vulgaris for the first time. The antibacterial activity showed that the essential oil (in situ, ex vitro) and leaf crude extract (ex vitro) are highly effective on all human urinary tract–infecting pathogens. In contrast, ex vitro plant-mediated essential oil is most significant with an 18.07 ± 0.17-mm inhibition zone against Klebsiella pneumoniae. The ex vitro plants are also superior to the in situ plants for obtaining greater extract and essential oil content (950 µl), over a time period of 12 wk. To obtain the ex vitro plants, meta-topolin is used for in vitro regeneration of A. vulgaris for the first time, showing the highest mean shoot number (98.11 ± 0.31) regeneration after 42 d, which is more significant in comparison with other studies conducted till date. The cytogenetic stability of the regenerated plantlets has been checked using start codon targeted polymorphism and cytological studies, conferring the homogeneity among regenerants along with in situ plant. Furthermore, the essential oil of the ex vitro plants was analysed through gas-chromatography mass spectroscopy, which detected a few compounds with bio-significance from A. vulgaris, including 2-carene, 2-(4-nitrophenyl) acetamide, β-guaiene, α-acorenol, and 10,12-Tricosadiyonic. [ABSTRACT FROM AUTHOR]

• Biotechnological strategies and phytochemical profiling of medicinally important Saussurea spp. are discussed. • Medium-term and long-term conservation strategies are reviewed. • Plant cell and hairy root culture proved a non-destructive way of metabolites production. • Genetic transformation serves as an effective approach for engineering metabolic pathways. Saussurea DC. is a well-recognized genus for its commercially important secondary metabolites with its maximum diversity in the Himalayas and East Asia. The rising demand for versatile metabolites (costunolide, syringin, hispidulin, jaceosidin, rutin) poses a threat to the natural habitat of Saussurea spp. As a result, plant cell and tissue culture technology have attained cutting-edge progress for its conservation and secondary metabolites production. To date, various protocols of in vitro micropropagation, cell culture, somatic embryogenesis, and cryopreservation have been studied extensively for the conservation and specialized metabolite production in the genus Saussurea. Furthermore, to enhance in vitro biosynthesis of these metabolites, the application of elicitors has also played a significant role. The evolution of protocols that have been achieved during the past years for the augmentation of secondary metabolites is envisaged to design species-specific elicitation strategies in this genus. Also, molecular cues unpinning the genes involved in secondary metabolite biosynthesis have been studied in this genus. Therefore, the present review summarizes the up-to-date information on biotechnological prospects such as plant, cell, tissue, and organ culture, cryopreservation, and secondary metabolites production in Saussurea spp. This comprehensive review provides a better understanding of biotechnological tools implemented for conservation and specialized metabolite production to support its sustainability. [ABSTRACT FROM AUTHOR]

Rubber tree is economically important rubber producing plant of Thailand. At present, a rubber tree plantation is susceptible to white root disease. Therefore, the use of rootstock from early introduce clone that proved to be resistant to white root disease could help sustain growing of rubber tree. Thus, the objectives of this research were to study the effects of plant growth regulators and different explants on somatic embryo (SE) induction of this rubber clone and assessment genetic stability. The results revealed that mix flower explant cultured on MS medium supplemented with 2.0 mg/L 6-benzyladenine (BA) and 1.5 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) provided callus subsequent somatic embryo (SE) formation at the highest frequency of 39.84 % and number of cotyledonary embryos (CEs) at 3.25 embryos/callus after 3 passages of subculture in the same culture medium (4 weeks/passage). SEs germinated into embryonic axis at 50 % and further development into shoot at 25 % after transfer to 0.25 mg/L GA3 containing MS medium with the best concentrations of BA and 2,4-D for 4 weeks. The assessment gene stability by RAPD and SSR markers showed no variation between mother plant and in vitro plantlets. In this work, a novel explant source - the floral section of the rubber tree - was used for the first time in order to design an effective technique for in vitro somatic embryogenesis. [ABSTRACT FROM AUTHOR]

Teucrium (Teucrium polium L.) is one of the valuable medicinal plants, but its poor seed germination is one of the principal problems of its extensive cultivation. Somatic embryogenesis is crucial for medicinal plant genetic improvement, germplasm preservation, and in vitro mass propagation. Synthetic seed production is a suitable alternative for natural seed production in biotechnology and plant development. The objectives of the present study were to optimize somatic embryogenesis, produce synthetic seeds, and evaluate the genetic fidelity of the regenerated T. polium. Results suggested that MS medium supplemented with 1.0 mg L−1 kinetin (Kin) alone and without auxin demonstrated the maximum embryogenesis and regeneration rates in leaf and petiole explants. The simultaneous occurrence of several somatic embryonic developmental stages, such as the globular, heart-shaped, torpedo-shaped, and cotyledonary stages, on an embryogenic callus clearly suggested that T. polium embryogenesis accurately captures an asynchronous process. The petiole and leaf explants cultured on MS medium with 1.0 mg L−1 Kin showed the highest rates of plant regeneration with spontaneously root development. Compared to somatic embryos, synthetic seeds with meristem sources lacked callogenesis. At 25 °C for 4 wk and at 4 °C for 3 wk, the meristem source caused the highest percentage of synthetic seeds to germinate (96.66%), while − 18 °C had a negative impact on the germination of the synthetic seeds. The results suggested that there was no genetic variation identified in the ISSR analysis on in vitro–generated T. polium plants. [ABSTRACT FROM AUTHOR]

• First report on application of meta -Topolin for augmented in vitro mass propagation of stevia. • meta -Topolin outperformed the other cytokinins during large-scale shoot proliferation. • Growth pattern assessed via micromorphological and leaf gas exchange measurement. • 100% genetic fidelity during flow cytometry and CDDP, DAMD, ISSR and SCoT markers analyses. • 2.4-folds higher rebaudioside-A production from acclimatized plantlets compared to mother plant. Present report ascertains the efficacy of meta -Topolin (m T) for improved in vitro mass propagation of stevia (Stevia rebaudiana Bert.) with high-frequency shoot initiation, multiplication, and proliferation over other cytokinins. Shoot tips were inoculated on Murashige and Skoog (MS) medium fortified with 6-benzyladenine, kinetin, m T, thidiazuron, and zeatin individually, at different concentrations ranged from 0.5—1.5 mg L−1. Among various combinations, MS medium + 1.5 mg L−1 m T developed the maximum number (15.33) and length (57.33 mm) of shoots with a greater number of leaves (38.67) after 4 weeks of culture. Elongated shoots were then transferred for root induction on MS medium fortified with a indole-3-butyric acid (IBA), indole-3-acetic acid, and α-naphthalene acetic acid individually, at concentrations ranged from 0.5—1.5 mg L−1. MS medium + 1 mg L−1 IBA was found as the superior rooting medium with the maximum number (10.67) and length (19.00 mm) of roots after next 4 weeks of culture. Rooted plantlets were acclimatized with 95% survival rate in sand followed by sand-soil (1:1; w w −1) mixture in next 8 weeks. Proper growth and development of acclimatized plantlets without any abnormalities was confirmed by micromorphological assessment (at cellular level), plant pigment analyses as well as leaf gas exchange measurement. Stability of ploidy level of the acclimatized plantlets was confirmed by flow cytometry. Furthermore, genetic fidelity of the acclimatized plantlets with their mother plant was confirmed by various molecular markers such as primers from directed amplification of minisatellite-region DNA, conserved DNA-derived polymorphism, inter simple sequence repeats, and start codon targeted polymorphism markers. High-performance liquid chromatography analysis indicated that acclimatized plantlets produced significantly higher (2.4-folds) rebaudioside-A (96.50 mg g−1 dry weight) in comparison to mother plant (40.20 mg g−1 dry weight). The current report, for first time, accounts the m T-mediated large-scale in vitro propagation of stevia ensuring genetic fidelity and simultaneously ameliorating the rebaudioside-A production. [ABSTRACT FROM AUTHOR]

Indian tea germplasm is known for its diversity, which not only assists the development of superior planting material for tea breeding but also functions as a genetic resource for other tea-producing countries. In this study, the aim was to develop an improved cryo-protocol for long-term conservation of tea germplasm. Shoot tips (approximately 2 mm) of Camellia sinensis were cryopreserved using vitrification and V cryo-plate techniques. The recovery rate of cryopreserved shoot tips obtained using vitrification was 33.2% at 90 min of Plant Vitrification Solution 2 (PVS2) exposure. To improve recovery rates and tolerance to cryo-injury, shoot tip donor plants were preconditioned with cold hardening, high sucrose, abscisic acid (ABA), and proline prior to cryopreservation. The vitrification technique resulted in an increase of regrowth of shoot tips to 60.5% after preconditioning of shoot tip donor plants for 30 d at 25 °C with 10 mg L−1 ABA and 3 mg L−1 proline. Regrowth further increased to 75.6% with the V cryo-plate technique using shoot tip donor plants preconditioned with the above concentrations of ABA and proline. After cryopreservation, shoot tips were recovered on regrowth medium that developed into shoot cultures in 5 wk. Random amplified polymorphic DNA (RAPD) analysis was performed to test genetic fidelity after cryopreservation and results showed 100% monomorphism among all the regenerants and 10-mo-old in vitro mother cultures. Results showed that the preconditioning strategies along with V cryo-plate technique influenced and improved the cryo-recovery rate of shoot tips of tea and provided a robust protocol for tea cryopreservation. [ABSTRACT FROM AUTHOR]

• In vitro multiplication of sweet potato using meta-topolin is reported for the first time. • An improved and reproducible micropropagation protocol for sweet potato was developed. • The multiplication rate and rooting efficiency of sweet potato has been significantly enhanced. • In vitro propagated plants showed 100% genetic similarity with mother plants. An improved in vitro propagation protocol by use of meta-Topolin (m T) hormone was developed in sweet potato (Ipomoea batatas (L.) Lam) using nodal explants derived from 12 weeks old in vitro plantlets of different sweet potato accessions. This is the first study that analyzes the effect of meta-Topolin (m T) hormone on sweet potato micropropagation. Enhanced in vitro multiplication was achieved on Murashige and Skoog (MS) medium supplemented with 0.54 µM 1-Naphthaleneacetic acid (NAA), 0.09 µM Gibberellic acid (GA 3) and varying concentrations of m T. A hundred percent shoot induction was obtained on m T concentrations ranging from 3.11 – 16.58 µM (except 4.15 µM). The best multiplication response with maximum number of shoots (7.26 ± 0.17) and axillary nodes (65.25 ± 1.81) was achieved on MS medium containing 8.29 µM m T with an average shoot length of 2.79 ± 0.27 cm. Shoots multiplied on m T containing medium gave 100% rooting with the best response (9.75 ± 1.21 roots per explant) recorded on MS medium supplemented with 0.49 µM Indole-3-butyric acid (IBA). Profuse rooting with multiple secondary roots was observed on this medium. Rooted plantlets were successfully acclimatized and hardened with a survival percentage of 90.48 ± 0.38%. The effect of different concentrations of m T (low, best and high) on the genetic integrity of regenerated plantlets was tested using 48 ISSR primers. All the successful primers (32) yield monomorphic bands in mother as well as tested plants, thereby confirming the uniformity and genetic integrity of regenerated sweet potato plants. The optimized protocol may be used for large-scale production of true-to-type quality planting material of sweet potato. [ABSTRACT FROM AUTHOR]

The species belonging to the Rhododendron genus are well-known for their colorful corolla. Molecular marker systems have the potential to elucidate genetic diversity as well as to assess genetic fidelity in rhododendrons. In the present study, the reverse transcription domains of long terminal repeat retrotransposons were cloned from rhododendrons and used to develop an inter-retrotransposon amplified polymorphism (IRAP) marker system. Subsequently, 198 polymorphic loci were generated from the IRAP and inter-simple sequence repeat (ISSR) markers, of which 119 were derived from the IRAP markers. It was shown that in rhododendrons, IRAP markers were superior to the ISSRs in some polymorphic parameters, such as the average number of polymorphic loci (14.88 versus 13.17). The combination of the IRAP and ISSR systems was more discriminative for detecting 46 rhododendron accessions than each of the systems on their own. Furthermore, IRAP markers demonstrated more efficiency in genetic fidelity detection of in-vitro-grown R. bailiense Y.P.Ma, C.Q.Zhang and D.F.Chamb, an endangered species recently recorded in Guizhzhou Province, China. The available evidence revealed the distinct properties of IRAP and ISSR markers in the rhododendron-associated applications, and highlighted the availability of highly informative ISSR and IRAP markers in the evaluation of genetic diversity and genetic fidelity of rhododendrons, which may facilitate preservation and genetic breeding of rhododendron plants. [ABSTRACT FROM AUTHOR]

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-1NAA, 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. [ABSTRACT FROM AUTHOR]

An efficient in vitro protocol for high-frequency polyploidization for the first time in gerbera hybrid (BGC-2019-01) was developed in the present study. Two-week-old in vitro-developed shoots (tips) were treated individually with 0.1%, 0.25% and 0.5% (w/v) colchicine solutions for 4, 6, 8, and 12 h. The colchicine-treated shoot tips were then inoculated on Murashige and Skoog (MS) medium fortified with 1.5 mg/l meta-Topolin for multiple shoot proliferation and later transferred into 1.5 mg/l indole-3-acetic acid-fortified MS medium for rooting of shoots. The ploidy levels of the colchicine-treated and regenerated plantlets along with the non-treated ones were confirmed via flow cytometry analysis and metaphasic chromosome count. The highest frequency of tetraploid plantlets (50%) were obtained when shoot tips were treated with 0.1% colchicine for 4 h. Morphological observations revealed that induced tetraploid plantlets exhibited delayed fresh shoot initiation, fewer but longer shoots, as well as fewer but broader leaves. Likewise, the study of stomata revealed that in comparison to their diploid counterparts, the tetraploid plantlets exhibited less frequent yet significantly larger stomata, and higher number of chloroplasts. The tetraploids were recorded with significantly higher chlorophyll, carotenoid, and anthocyanin content during the photosynthetic pigment analyses. During ex vitro acclimatization and field growth, the tetraploid plants exhibited delayed proliferation but with higher vigor and thickened broad leaves. The genetic uniformity among the diploid and the tetraploid plants was confirmed using conserved DNA-derived polymorphism (CDDP), directed amplification of minisatellite-region DNA (DAMD), inter simple sequence repeats (ISSR), and start codon targeted (SCoT) polymorphism marker systems. The tetraploids developed in the present study would be of immense importance for the genetic improvement of gerbera as far as its ornamental values are concerned. [ABSTRACT FROM AUTHOR]

Background: Codonopsis pilosula (Franch.) Nannf. is a medicinal plant traditionally used in China, Korea, and Japan to treat many diseases including poor gastrointestinal function, low immunity, gastric ulcers, and chronic gastritis. The increasing therapeutic and preventive use of C. pilosula has subsequently led to depletion of the natural populations of this species thus necessitating propagation of this important medicinal plant. Here, we developed an efficient and effective in vitro propagation protocol for C. pilosula using apical shoot segments. We tested various plant tissue culture media for the growth of C. pilosula and evaluated the effects of plant growth regulators on the shoot proliferation and rooting of regenerated C. pilosula plants. Furthermore, the tissues (roots and shoots) of maternal and in vitro-regenerated C. pilosula plants were subjected to Fourier-transform near-infrared (FT-NIR) spectrometry, Gas chromatography-mass spectrometry (GC–MS), and their total flavonoids, phenolics, and antioxidant capacity were determined and compared. Results: Full-strength Murashige and Skoog (MS) medium augmented with vitamins and benzylaminopurine (1.5 mg·L−1) regenerated the highest shoot number (12 ± 0.46) per explant. MS medium augmented with indole-3-acetic acid (1.0 mg·L−1) produced the highest root number (9 ± 0.89) and maximum root length (20.88 ± 1.48 mm) from regenerated C. pilosula shoots. The survival rate of in vitro-regenerated C. pilosula plants was 94.00% after acclimatization. The maternal and in vitro-regenerated C. pilosula plant tissues showed similar FT-NIR spectra, total phenolics, total flavonoids, phytochemical composition, and antioxidant activity. Randomly amplified polymorphic DNA (RAPD) test confirmed the genetic fidelity of regenerated C. pilosula plants. Conclusions: The proposed in vitro propagation protocol may be useful for the rapid mass multiplication and production of high quality C. pilosula as well as for germplasm preservation to ensure sustainable supply amidst the ever-increasing demand. [ABSTRACT FROM AUTHOR]

The present study was focused towards determining the factors governing direct organogenesis in kiwifruit cultivar 'Allison'. Using in vivo buds or nodal segments as explants, the maximum 75.5% uncontaminated explants were obtained upon consecutive individual disinfection treatment with 1% carbendazim (3 min), 0.1% HgCl2 (1 min), and 2% NaOCl (2 min), respectively. The maximum shoot regeneration response of 84.81% was recorded on Murashige and Skoog (MS) medium fortified with 0.45 μM BA, 0.1 μM IBA, and 0.13 μM GA3. The months of May displayed the highest in vitro explant regeneration response for two consecutive years. The highest shoot proliferation rate was 1:6 with a 5.34 mean number of shoots and a maximum mean shoot length of 6.32 cm on MS medium containing 0.22 μM BA, 0.1 μM IBA, and0.69 μM GA3. A two-step or sequential approach for in vitro root induction showed good quality rooting response of 91.03% producing a higher number of long (2.23 cm) roots per shoot, and afterwards, the plantlets were hardened successfully with a higher survival rate (95%). Also, ISSR markers confirmed the genetic fidelity of the in vitro-raised regenerants. [ABSTRACT FROM AUTHOR]

Uraria picta root is used in the polyherbal product 'Dashmula'. Its exploitation for formulation preparation has depleted its availability, leading to medicine adulteration. Direct shoot regeneration from the cotyledonary node holds promise as a source of raw material. This study aimed to develop a regeneration protocol for U. picta and validate its genetic and metabolic fidelity. The seeds of U. picta showed low germination rates, prolonged dormancy, and poor viability. Root exploitation in the wild poses a threat to its availability in nature. Seedling's derived cotyledonary nodes cultured on B5 medium supplemented with BAP (0.5–3 mg L−1), Kinetin (0.5–3 mg L−1), Thidiazuron (0.01–1 mg L−1), and meta -Topolin (0.1–4 mg L1). To address hyperhydricity in regenerated shoots, cotyledonary nodes were cultured on high-agar concentration media. Microshoots were exposed to IBA solution (50–800 mg L−1) pulse treatment for rooting. Tissue-cultured plants genetic fidelity was assessed using ISSR and SCoT markers, while metabolic fidelity was studied with HRMS. The chlorophyll content, antioxidant, and antidiabetic activity of micropropagated plants were evaluated. The highest shoot regeneration frequency, with a maximum of 6.57±0.278 shoots per explant, was achieved using 2 mg L−1 meta- Topolin. The shoots were elongated, had expanded leaves, and were hyperhydrated. BAP (2 mg L−1) induced a maximum of 9.83±0.333 shoot buds per explant. BAP caused explant browning, profuse callus formation, dwarfing, and hyperhydric shoots. Hyperhydricity was alleviated with a higher agar concentration (1 %). IBA (400 mg L−1) induced a maximum of 2.18±0.090 roots per shoot and a root length of 9.23±0.033 cm. Tissue-cultured and mother plants exhibited clonal fidelity, similar metabolite and chlorophyll content, strong antioxidant activity, and equal efficacy for inhibiting α -amylase and α -glucosidase. This method can propagate elite clones of U. picta and offer its improvement via genetic transformation. [Display omitted] • B5 medium containing 2 mg L−1 meta -Topolin, supported direct shoot regeneration from cotyledonary node, with a maximum of 6.57±0.278 shoots per explant. • Indole butyric acid (400 mg L−1) pulse treatment of microshoots for 5 minutes supported well roots development. • Acclimatized plants showed 90 % survival after transfer into the field. • Genetic fidelity of micropropagated plants with mother was confirmed using SCoT and ISSR markers. • HRMS analysis also confirmed metabolic fidelity of both plants. Regenerated plants possess good antioxidant and antidiabetic activity. [ABSTRACT FROM AUTHOR]

Zingiberaceae family, also known as the ginger family, comprises 53 genera and 1300 species. The members of this family are widely known for their ethnobotanical properties. Owing to their anti-inflammatory, antimicrobial, antidiabetic, and cardioprotective properties, the rhizomes have been used in traditional medicine to treat a wide range of illnesses like cold, inflammation, gastritis, arthritis, diabetes, and respiratory problems. The medicinal properties of the plants are attributed to the wide array of secondary metabolites present in them. They also find their use in food, spices, ornamentals, perfumes, and cosmetics. The current review focuses on biotechnological techniques like direct regeneration, indirect regeneration via callus induction and somatic embryogenesis, microrhizome induction, and in vitro secondary metabolite production that have been established to meet the increasing commercial and pharmaceutical demands. This review also identifies unrevealed possibilities, knowledge gaps, and considerable challenges that should be taken into account for developing novel concepts pertaining to the biotechnology of Zingiberaceae in the near future. [ABSTRACT FROM AUTHOR]

• Developed a high-frequency regeneration protocol via Polyethylene glycol (PEG)–induced somatic embryogenesis. • 3-day old cotyledonary explants used for the development of early-stage- somatic embryos. • Incorporation of ABA and proline was effective in the maturation of somatic embryos. • Regenerated plants are phenotypically identical to the mother plants. • Genetic fidelity analysis of in vitro regenerated Vigna radiata plants was reported newly. The results show that the in vitro regenerated plants are true-to-type with no somaclonal variation. • This protocol could be useful for genetic transformation studies as well as large-scale production strategies of true-to-type plants without any somoclonal variation. An efficient plantlet regeneration protocol of green gram [ Vigna radiata (L.) Wilczek] via somatic embryogenesis (SE) using Polyethylene glycol (PEG) is reported in this study. Embryogenic calli were induced using 3-day-old cotyledonary explants on MS medium supplemented with picloram (5 µM). Early-stage somatic embryos were generated by treating these 3-day-old cotyledonary explants or calli derived from those explants with osmolytes such as, sorbitol, mannitol and PEG, among which PEG was found to be the ideal. Early-stage embryoids were subsequently developed into torpedo and cotyledonary stage embryoids in liquid cultures supplemented with abscisic acid (5 μM), proline (130 μM). Furthermore, plantlet conversion was achieved in MS medium with 6-Benzylaminopurine (BAP) 1 μM + gibberellic acid (GA 3) 12 μM with a ca. 35% regeneration frequency. Plantlets were established under field conditions after the confirmation of their survival. The survival rate of plantlets post-transplantation was ca. 99%. The field-grown plants were confirmed to be phenotypically identical to the mother plant. According to RAPD profiles, in vitro regenerated plants were monomorphic and similar to the control/mother plants. The regeneration protocol developed in this study could be efficiently utilized for genetic transformation studies and large-scale production strategies of true-to-type plants without any somaclonal variation. [ABSTRACT FROM AUTHOR]

The improved sugarcane varieties grown in and around the world are majorly evolved through interspecific hybridization of Saccharum species. Only few promising hybrids have been developed from allied genera such as Erianthus and Sclerostachya. Several other genera, including Phragmites, Vetivera, Narenga and Neyurida, deserve further research. Successful exploitation of these allied genera resources require careful characterization, evaluation and conservation using different methods including tissue culture techniques. Among them, Phragmites australis is used to remediate waste water and remove heavy metals. In this study, meristem tip culture was standardized for five clones of Phragmites to evaluate their response and growth. Among the five clones of Phragmites studied, two clones (IND 89-711 and IND04-1326) showed better regeneration and growth. The in vitro derived plants were compared with the mother plants using four SRAP markers to check their genetic fidelity. Monomorphic bands were obtained in both the in-vitro and mother plants. No polymorphism was detected, thus proving the genetic uniformity of the micropropgated plants. [ABSTRACT FROM AUTHOR]

Eulophia dabia (D. Don) Hochr is an endemic, critically endangered, terrestrial orchid species having profound importance in traditional herbal drug preparations and pharmacopoeia worldwide. In the present investigation, we have developed an efficient and genetically stable micropropagation protocol of E. dabia using axenic rhizome as explant. Seeds inoculated on half-strength Murashige and Skoog (MS) medium showed 5% germination efficiency. Scanning electron microscopy analysis revealed differential effects of various sterilizing agents (NaOCl2 and ethanol) on the seed coat integrity leading to a significant improvement in seed germination rate. The rhizome-like bodies (RLBs) developed after 6 mo of seed germination showed best growth of rhizome (size: 3.3 × 1.5 cm and 5.0 nodes per rhizome) in MS medium containing casein hydrolysate and activated charcoal. One-year-old mature axenic rhizomes were further sub-cultured for multiple shoot induction, and maximum 96.1% response was observed in MS medium fortified with 4.4 μM BAP and activated charcoal with 4.3 mean shoot number and 13.4 cm shoot length. After 3 to 4 sub-cultures using the same medium, bipolar structures (plantlets with shoot and root) were developed. The properly developed plantlets were successfully acclimatized to open environment with 98% survival. Genetic stability of the discussed protocol was evaluated by random amplified polymorphic DNA (RAPD) markers, which proved true to typesets of the in vitro raised plants. In the present study we optimized in vitro propagation system for the E. dabia which can be successfully applied further for the genetic improvement studies and commercial scale propagation. [ABSTRACT FROM AUTHOR]

A highly effective method has been developed to examine the impact of different combination of meta-topolin and various cytokinins on Chlorophytum borivilianum Sant. et Fernand. using apical bud explants. The effects of benzyl adenine purine (BAP), kinetin (Kin), meta-topolin (mT), and thidiazuron (TDZ) were explored separately and in different concentrations (ranging from 0.5 to 3.0 mg L−1) on shoot proliferation, as well as the influence of auxins like Indole -3- acetic acid (IAA), α-Naphthalene acetic acid (NAA) and Indole-3-butyric acid (IBA) on rooting. When grown on MS medium supplemented with mT, highest growth rates were observed followed by BAP. Within the permutations tested, 2.0 mg L−1 of mT and 1.0 mg L−1 of BAP produced the best results with 96.3% success rate and average of 19.2 ± 1.92 shoots measuring 6.72 ± 0.49 cm in length after 8 weeks of culture. Among the various auxins used for in vitro root formation, 95.5% of multiple shoots effectively established roots on MS medium supplemented with 1.0 mg L−1 of IBA resulting in an average of 14.0 ± 1.39 root numbers and 3.36 ± 0.27 cm in length after 4 weeks of culture. Furthmore, 94% of the acclimated plants not only survived but also thrived when grown in a mixture of loam, sand and fym (in 1:2:1 ratio) with an average weight of 16 g. High-performance thin layer chromatography (HPTLC) analysis revealed the presence of diosgenin and stigmasterol in both in vitro regenerants and mother plant, although there were variations in their content. To ensure genetic fidelity among regenerants, ISSR and SCOT markers were used, but no somatic differences were detected. Therefore, the established procedure seems to be easy and consistent for large scale development of genetically identical clones with a greater content of bioactive compounds. [Display omitted] • First report on the use of meta-topolin on Chlorophytum borivilianum. • The in vitro regenerants were similar to the donor plant revealed by ISSR markers. • HPTLC analysis confirmed presence of diosgenin and stigmasterol in both in vitro regenerants and donor plant. • The protocol could be useful for commercial and conservation purposes. [ABSTRACT FROM AUTHOR]

In vitro plant propagation techniques play an important role in modern agriculture and horticulture, offering efficient methods for the accelerated multiplication of high-value plant species. Thin cell layer (TCL) culture technique has emerged as a promising approach for large-scale in vitro plantlets production. This study explores the potential of TCL culture as a cutting-edge strategy to enhance the efficiency of multiple shoots formation in Kaempferia parviflora. An attempt was also made to look into the synergistic effects of cytokinin and auxin [6-benzyladenine (BA) - indole-3-acetic acid (IAA)] during micropropagation. Shoot tip explants (from rhizome sprouts) were cultured onto Murashige and Skoog (MS) medium enriched with BA (2.0–8.0 mg/L) and IAA (0.2–0.8 mg/L) in combination where the highest shoot number (7.3 shoots/explant) and length (5.9 cm) was obtained in 8.0 mg/L BA and 0.8 mg/L IAA. Subsequently, the transverse thin cell layer (tTCL) culture system was successfully utilized whereby the shoot tip explants were individually sliced from tips at the meristem and cultured onto MS medium supplemented with BA (2.0–8.0 mg/L) and IAA (0.2–0.8 mg/L) in combination. The regenerated tTCLs exhibited the highest shoot multiplication (9.3 shoots/tTCL; 28 shoots/shoot tip) and length (5.3 cm) in 4.0 mg/L BA + 0.8 mg/L IAA, thus significantly increased number of plantlets were obtained from tTCL culture (953.0 plantlets) compared to shoot tip culture (527.7 plantlets) after six sub-culture cycles. The tTCL regenerants were successfully acclimatized (with 90% survival) and displayed active growth. Genetic fidelity assessment via inter-simple sequence repeats, conserved DNA-derived polymorphism, directed amplification of minisatellite-region DNA, and start codon targeted polymorphism marker systems suggested that a very low level (5.8%; 148 out of 2572 scorable bands) of polymorphism was detected whereas the ploidy level remained unchanged, confirmed by flow cytometry analysis. The implementation of tTCL culture, coupled with a controlled sub-culture regimen, holds significant potential for the efficient commercial-scale production of K. parviflora. • First report on utilization of transverse thin cell layer (tTCL) culture technique in K. parviflora. • Cytokinin-auxin (BA-IAA) synergistic effects on micropropagation was proven. • Affirmed ploidy and genetic stability via flow cytometry, CDDP, DAMD, ISSR and SCoT markers. • Bulk production of plantlets in subsequent sub-cultures via tTCL culture technique. [ABSTRACT FROM AUTHOR]

• Polyethylene glycol improved in vitro morphogenesis in Spathoglottis plicata. • Exogenous application of PEG exhibited superior anatomical traits compared to control. • Mild osmotic stress increased the dimensions of leaf mesophyll and vascular tissues. • PEG-derived plantlets performed better during in vitro rooting and ex vitro hardening. • Genetic uniformity of the regenerates was confirmed using SCoT markers. In vitro propagation is extremely important for the rapid multiplication of a wide range of orchids. This study aimed to investigate the effects of polyethylene glycol (PEG) on morpho-anatomical improvements and ex vitro development of Spathoglottis plicata , a terrestrial orchid. Various concentrations of PEG (100-500 mg L−1) were incorporated with the optimized medium composition [Murashige and Skoog's medium with 2.0 mg L−1 6-benzylaminopurine (BAP) and 0.25 mg L−1 indole-3-acetic acid (IAA)] to study the morpho-anatomical developments of cultures. Results showed that PEG at 300 mg L−1 in the nutrient medium had a significant effect on the development of plantlets with superior morphometric and anatomical characteristics. It improved the rate of shoot multiplication (83 shoots), shoot length (10 cm), fresh weight (4.0 g), and dry weight (1.5 g) of shoots per culture vessel than the control experiment (devoid of PEG). Light microscopic analysis revealed that PEG-derived leaves possessed improved leaf thickness, differentiation in dermal, ground, and vascular tissue systems than the leaves developed without PEG in the medium. Such shoots were responded significantly better during rooting (11.0 roots with 6.2 cm length) and acclimatization stages. Start Codon Targeted (SCoT) markers analysis confirmed the genetic uniformity of the regenerates with the mother plant. The improvements in morpho-anatomical structures of in vitro raised plantlets due to PEG-induced mild drought stress in ground orchid S. plicata could be a useful tool for its large-scale production and survival success in the field. [ABSTRACT FROM AUTHOR]

Micropropagation technique has been recommended for plant tissue rejuvenation/reinvigoration and the improvement of clonal plants production commercially. Concerning the limiting factors on in vitro cultivation of Eucalyptus grandis × E. urophylla hybrid clone, the effects of light intensity on in vitro introduction and multiplication phases were assessed. The experimental tissue was collected from ministumps cultivated in a semi-hydroponic system. The results of light intensity on in vitro introduction and multiplication were evaluated with four treatments (fluorescent lamp/40 µmol m−2 s−1, red/blue LEDs 20 µmol m−2 s−1, red/blue LEDs 40 µmol m−2 s−1, and red/blue LEDs 80 µmol m−2 s−1). The explants' morphological, anatomical, and genetic characteristics were evaluated at 30 d in the in vitro introduction, and 12 subcultures for in vitro multiplication. Based on the results, the treatments F/L 40 µmol m−2 s−1 and R/B 80 µmol m−2 s−1 are the most suitable during in vitro introduction and multiplication of Eucalyptus grandis × E. urophylla. It provides a higher percentage of responsive explants, shoots per explant, vigor, photosynthetic pigment content, adaxial epidermis thickness, abaxial epidermis, mesophyll, palisade parenchyma, spongy parenchyma, and vascular tissues. Furthermore, it was found that the micropropagated plants are identical clones of the Eucalyptus grandis × E. urophylla selected tree; that is, there was no somaclonal variation during the twelve subcultures on in vitro multiplication. [ABSTRACT FROM AUTHOR]

A highly efficient and reproducible in vitro plant regeneration method has been developed from shoot bud, half-shoot, and shoot slice explants of four genotypes of turmeric (Curcuma longa L.). The shoot slice explant produced the highest number of shoots (22.4 shoots explant−1) than half-shoot (18.6 shoots explant−1) and shoot bud explant (14.2 shoots explant−1) of Salem genotype on medium amended with 13.32 µM 6-benzylaminopurine (BAP) and 5.37 µM 1-naphthaleneacetic acid (NAA). Salem genotype was more efficient among the four genotypes tested, followed by Duggirala Red, Prathibha, and PCT-13 genotypes. Approximately 80% of shoots were induced spontaneous rooting on shoot induction media fortified with NAA. The complete plantlets were obtained after rooting the shoots on a medium augmented with 4.90 µM Indole-3-butyric acid (IBA). IBA was an efficient auxin to induce the maximum number of roots per shoot than Indole-3-acetic acid (IAA) and NAA. The plantlets were acclimatized under greenhouse conditions, and the survival rate was recorded as 96%. The plantlets obtained from all explants were morphologically similar to mother plants. Two genome-based molecular markers, namely inter simple sequence repeats (ISSR) and start codon targeted (SCoT) molecular markers, were employed to determine the genetic stability of plants obtained from the tissue culture of the Salem genotype. The DNA markers validated the genetic uniformity of the plants obtained from all three explants was similar to their mother plant. This plant regeneration procedure could be a helpful method for the genetic improvement of turmeric. [ABSTRACT FROM AUTHOR]

[Display omitted] • An effective and efficient in vitro protocol for Dioscorea deltoidea was established. • The in vitro regenerants were identical to the mother plant according to ISSR markers. • HPTLC analysis confirmed presence of diosgenin in micropropagated plants. • The protocol could be useful for commercial and conservation purposes. Dioscorea deltoidea (Family: Dioscoreaceae) is a high value endangered medicinal plant and the source of pharmaceutically important bioactive compound diosgenin, which is used in the synthesis of steroidal drugs. Establishing an efficient reproducible regeneration system is necessary for its sustainable utilization and conservation. The purpose of this research was to establish an effective and efficient in vitro regeneration protocol for D. deltoidea and assess its genetic and biochemical fidelity. In this report, Murashige and Skoog (MS) medium fortified with different types and concentration of Plant Growth Regulators (PGRs) were studied. Maximum shoot regeneration frequency was achieved in MS medium supplemented with 2.0 mg L −1 BA whereas higher shoot multiplication was observed in 2.0 mg L −1 BA + 1.0 mg L −1 IBA. Highest rooting was recovered on MS medium supplemented with 2.0 mg L −1 NAA. In vitro rooted plantlets were acclimatized with a survival rate of 96%. The genetic fidelity of the mother plant (MP), acclimatized plant (AP) and in vitro regenerants were found to be clonally similar as examined through ISSR fingerprinting. Of 7 ISSR primers tested, 5 produced scorable amplified products. The High Performance Thin Layer Chromatography (HPTLC) based chemical assessment showed varied chemical profile. The developed protocol would be beneficial in providing a germplasm conservation system, isolation of secondary metabolites and production of high value clonal regenerants for commercial production. [ABSTRACT FROM AUTHOR]

Lilium davidii var. unicolour Salisb is an important edible and medicinal plant. In vitro plantlet regeneration in L. davidii var. unicolour Salisb and identification of genetic stability are the premises of large-scale cultivation, and also a way to meet the increasing global demand. The purpose of this study is to establish an efficient, rapid plant propagation process for L. davidii var. unicolour Salisb, and to define the genetic stability of its regenerants in vitro under optimised conditions. The effects of different concentrations of 6-benzyladenine (6-BA) and thidiazuron (TDZ) on shoot multiplication and growth of L. davidii var. unicolour Salisb were investigated individually, or in combination with α-naphthalene acetic acid (NAA), using Murashige and Skoog (MS) solid or liquid medium. Inter-sample sequence repeat markers were used to assess genetic fidelity among plantlets. The results were that we achieved efficient and reliable regeneration, using scales as explants for shoot induction. The highest mean number (16.8) of shoots was differentiated de novo from the lower portion of the outer scales. After 4 weeks of incubation, superior multiplication rates (9.8%), mean shoot number (3.38), and length (5.46 cm) were achieved using MS liquid medium supplemented with TDZ. Auxins, such as indole butyric acid (IBA) and NAA, were employed for in vitro root induction, and the maximum rooting rate (100%) was observed when samples were cultured on half-strength MS medium supplemented with NAA. In vitro rooted plantlets of L. davidii var. unicolour Salisb were acclimatised in a greenhouse and plants grew well with a 90% survival rate. Inter-sample sequence repeat markers were used to assess genetic fidelity among plantlets, the best ISSR primer UBC 808 had the highest number of reproducible bands and produced 15 distinct bands and all micropropagated plants and the mother plant could be grouped in a single cluster with a 94% similarity level, indicating a low level of genetic polymorphism in micropropagated plants. The developed protocol described herein is simple and reliable for large-scale production of L. davidii var. unicolour Salisb, which provides a good technical support for large-scale cultivation of L. davidii var. unicolour Salisb. [ABSTRACT FROM AUTHOR]

Background: Somaclonal variation is considered as a source of genetic variation for crop improvement. It has been investigated using cytological, biochemical and molecular techniques. Metohds: Genetic stability in the embryo-derived axillary shoots of 4 chickpea genotypes was assessed using eight inter-simple sequence repeat (ISSR) and 19 random amplified polymorphic DNA (RAPD) primers. Result: RAPD primers produced 94 and ISSR primers produced 38 distinct and scorable alleles, with an average of 4.9 alleles for RAPD and 4.75 for ISSR primers. The polymorphic information content (PIC) ranged from 0.36 to 0.90 for RAPD and from 0.50 to 0.87 for ISSR. ISSR recognized a 90%, but RAPD recognized 82% similarity value. No absolute similarity value was between the mother plant and the regenerated shoots for the overall genotypes. At a 90% similarity value, 15 out of the 20 regenerated shoots from 'Giza 88' group with their mother plant using ISSR markers; however, 11 regenerated shoots grouped with their mother plant in one central cluster for 'Giza 4' using RAPD markers. The observed variations in the total number of polymorphic RAPD and ISSR bands and the number of bands specific to the mother and regenerated shoots, detected intra-clonal variation and genetic instability seem to be genotype-dependent. [ABSTRACT FROM AUTHOR]

Scaevola taccada (Gaertn.) Roxb. is a hemi-sclerophyllous littoral shrub of the family Goodeniaceae. It is a salt-tolerant plant and used in soil reclamation and coastal landscaping programs to control beach erosion. The present article reports a reproducible in vitro regeneration system using meta-topolin (mT) through liquid medium employing nodal segment cultures. The highest number of axillary shoots (5.8 ± 0.29) was differentiated on Murashige and Skoog's (MS) medium containing meta-topolin (1.5 mg L−1). The amplification of shoots (49.8 ± 0.37) and fresh growth was achieved by recurrent subcultures on liquid MS medium supplemented with 0.25 mg L−1mT and 0.15 mg L−1 indole-3-acetic acid (IAA) in the presence of additives (50 mg L−1 ascorbic acid + 25 mg L−1 citric acid, adenine sulphate and l-arginine). Rooting and acclimatization was simultaneously achieved using concurrent ex vitro rooting and acclimatization (CEVRA) technique by pulse treating the shoot base with 300 mg L−1 IBA for 5 min. The in vitro developed shoots represented a maximum percentage (100%) of rooting in autoclaved soilrite in a greenhouse. After 2 mo, the plantlets were established in the field with cent percentage survival success. The genetic fidelity analysis using inter simple sequence repeats (ISSR) DNA molecular markers developed maximum of 55 monomorphic bands and revealed the genetic homogeneity of in vitro regenerated plantlets with the mother plant. [ABSTRACT FROM AUTHOR]

Muntingia calabura L. is a potent medicinal plant. Because of that several researchers reported phytochemical and pharmacological studies, but only minimal information is obtainable on tissue culture and genetic stability evaluation studies. Therefore, in the present investigation we attempted to establish a reliable direct and indirect regeneration study through leaf and node explants on MS medium containing cytokinins BAP and KN (0.5-3.0 mg l-1) in combination with auxins IAA and NAA (0.5 mg l-1). The breakage of bud and shoot initiation were noticed with the initiation of callus at lower concentrations of auxin alone at 0.5 mg l-1. 2,4-D was found to be good in callus induction from leaf and nodal explants. A problem of browning callus was prevented by regular subculturing of callus cultures. Leaf explants exhibited maximum number of shoots (32±0.88a) with shoot length (7.6±0.17a) and nodal explants obtained optimal number of shoots (26±0.88a) with shoot length (9.6±0.30a) on MS medium supplemented with BAP (2 mg l-1) and IAA (0.5 mg l-1). Half strength MS medium fortified with IBA (2.0 mg l-1) was effective and achieved 70% of rooting. These welldeveloped plantlets were shifted to pots containing soil and vermicompost in 1:1 ratio for acclimatization. The acclimatized plants were field transferred with survival rate of 85%. The ISSR and RAPD markers analysis revealed the genetic stability of in vitro regenerated plants with the mother plant. [ABSTRACT FROM AUTHOR]

An in vitro regeneration system for propagation of a valuable medicinal tree, Pterocarpus marsupium, using shoot tip (ST) explants derived from 7-d-old axenic seedlings has been successfully developed. Murashige and Skoog (MS) medium containing cytokinins (BA, mT, or Kn) and auxins (NAA, IAA) in different concentrations and combinations showed to have a marked stimulatory effect on the regeneration output. Of cytokinins, meta-topolin (mT 7.0 μM) proved optimum dose for multiple shoot induction yielding 6.50 ± 0.49 shoots per explant with mean shoot length (4.00 ± 0.16 cm) in 70% cultures after 6 wk. Whereas, supplementation of low concentration of auxin (1.0 μM NAA) with optimal cytokinin (7.0 μM mT) favored enhanced shoot induction with increased percent response. At this level, a maximum of 13.54 ± 0.34 shoots per explant with (5.30 ± 0.10 cm) mean shoot length were recorded in 80% cultures after 12 wk. Thereafter, microshoots were isolated and their basal end treated with high doses of indole-3-butyric acid (250 μM) for 5 d followed to transplant onto various potting substrates for ex vitro root formation. A maximum of 3.63 ± 0.0.08 roots per microshoot and mean root length of 3.59 ± 0.07 cm with highest rooting frequency (67.7%) was observed after 4 wk of transplantation. During ex vitro rooting-cum-acclimatization, various physiological parameters were found to be fluctuating initially for 35 d; thereafter, an increasing trend was observed in net photosynthetic rate, intercellular CO2 concentration, and chlorophyll a/b estimation. Overall, the successfully acclimatized regenerants when transferred in natural condition showed about 96.7% survival rate after 3 mo. Assessment of genetic integrity of tissue culture raised plantlets was accomplished by DNA–based ISSR primers. A total of 35 bands with an average of 4.38 bands per primer were scored in monomorphic banding pattern. Thus, it is apparent that the mode of in vitro regeneration protocol was suitable for obtaining true-to-type and disease-free plants. [ABSTRACT FROM AUTHOR]

The present study describes the first successful report on an efficient in vitro protocol for direct and indirect organogenesis of an important medicinal and anticancerous plant, Solanum erianthum D. Don. An optimal response using leaf explants was achieved in half-strength Murashige and Skoog (MS)-medium supplemented with 5 mg L−1 naphthalene acetic acid for friable callus formation and 2 mg L−1 naphthaleneacetic acid and 0.5 mg L−1 6-benzylaminopurine amino purine for induction of compact callus. The maximum number of shoot buds (20.33 ± 11.55) was obtained from compact callus using indirect shoot organogenesis in MS medium supplemented with 0.5 mg L−1 thidiazuron among the four cytokinins tested, which included 6-benzyl amino purine, kinetin, 6-γ,γ-dimethylallylamino purine, and thidiazuron. Supplements of 3 mg L−1 6-benzyl amino purine played a crucial role in direct shoot bud formation from a leaf. The buds appeared as small protuberances, and finally developed into leafy shoots as the culture period progressed. Regenerated shoot buds were transferred to MS media that contained four cytokinins for shoot tip culture, in which 8 mg L−1 6-γ,γ-dimethylallylamino purine exhibited the best response (15 ± 2.52). Although efficient plant regeneration was achieved in the presence of all four tested cytokinins in the three organogenetic pathways, distinct differences exist in terms of shoot regeneration potential. Half-strength MS basal medium is recommended for root induction. An approximate 70% survival rate of regenerated plantlets was recorded following sequential transfer from culture conditions to the field. The genetic fidelity of the regenerated plants was evaluated using random amplified polymorphic DNA molecular markers, which showed a high number of uniform monomorphic bands. [ABSTRACT FROM AUTHOR]

An in vitro micropropagation protocol for Nyctanthes arbor-tristis was developed through the culture of seedling-derived nodal explants. The highest direct shoot induction rate, the mean number of shoots per explant and average shoot length was obtained on MS medium supplemented with 5 mg/l 6-benzylaminopurine (BAP). Full-strength MS medium was found better than other concentrations for organogenesis. Their lower or higher concentration from optimum level was inhibitory for shoot regeneration. Among different carbon sources tested for organogenesis, MS medium supplemented with 87.64 mM sucrose was superior to other carbon sources for in vitro morphogenesis. Best rooting of microshoots was achieved on one-quarter strength MS medium supplemented with 2 mg/l indole-3-butyric acid (IBA). Under this treatment, maximum mean number of roots/shoot and mean root length were 9.40 ± 0.70 and 2.47 ± 0.25 cm, respectively. The genetic fidelity among regenerates and donor plant has been confirmed by ISSR markers. [ABSTRACT FROM AUTHOR]

A simple and efficient protocol developed for plantlet regeneration through in vitro somatic embryogenesis in three ornamental bananas. In vitro propagation, especially somatic embryogenesis has been a method of choice to obtain homogenous planting material in ornamental bananas. In the present investigation, true-to-type plants via somatic embryogenesis were achieved in three ornamental bananas viz. Musa laterita, Musa beccarii and Musa velutina using immature male flower bud as explants. The evaluation was carried out at two stages of somatic embryogenesis, induction, and maturation of somatic embryos. Embryogenic calli desiccated up to 2 h at 25 ± 1 °C resulted in higher frequencies of embryo induction and maturation compared to non-desiccated embryos. The plantlets generated were hardened, and the genetic fidelity of the plantlets was confirmed using SRAP markers. It was observed that banana plants derived via somatic embryogenesis displayed normal phenotype as that of plants grown from a single elite mother plant. In the present study, the successful establishment of plants from male flower bud was achieved in 6 to 7 months which is a relatively short duration compared to other reports involving different species of banana. The protocol developed could be useful for large-scale micropropagation studies in these commercially important ornamental bananas. [ABSTRACT FROM AUTHOR]

The objective of this study was to develop DNA fingerprints of potato progenies by SSR markers for genetic fidelity purpose. Interspecific potato somatic hybrid progenies, developed earlier by crossing somatic hybrids with Indian potato varieties, were used in this study. A total of 81 genotypes (progenies and parents) were analysed by two wellcharacterized potato SSR markers (STU6SNRN and STIIKA), and developed a data matrix for genetic fidelity purpose and other applications. High polymorphism was observed in STIIKA (PIC: 0.98) than STU6SNRN (PIC: 0.95), and also higher number of alleles was observed in STIIKA (22) than STU6SNRN (7). In STU6SNRN, alleles size 179, 182, 190 and 200 bp were predominant; whereas in STIKA, alleles size 195, 201, 221, 235, 242 and 245 were observed frequently in more than 50 per cent of the genotypes. Diversity analysis showed a clear distinction among the genotypes based on the Jaccard dissimilarity coefficient by the Neighbour-joining tree method using the DARwin software. Our study suggests that SSR fingerprints would be of practical utility to strengthen genetic fidelity of these somatic hybrids. Further, SSR fingerprints would also be valuable resources in resolving issues related to identification of true-to-type clone during field evaluation in multiple environments. [ABSTRACT FROM AUTHOR]

The present investigation was carried out to establish an efficient and reproducible micropropagation protocol for the production of morphologically, genetically and chemically uniform plants of Curcuma zedoaria. Axillary bud explants of C. zedoaria were inoculated into MS basal medium supplemented with various combinations and concentrations of 6-benzyladenine (2.2–22.2 µM, BA), kinetin (2.3–23.2 µM, Kin), indole-3-acetic acid (2.9–11.4 µM, IAA), α-naphthalene acetic acid (2.7–10.2 µM, NAA) and adenine sulphate (33.9–203.6 µM, Ads). Almost 95% of rhizome buds sprouted on MS medium supplemented with 13.3 μM BA, 5.7 μM IAA and 63.9 μM Ads giving rise to an average of 12.89 ± 0.02 shoots within 6 weeks. However, the maximum number of roots (25.8 ± 0.07 roots per explant) was obtained on half strength MS medium supplemented with 7.4 µM of IBA after 4 weeks of inoculation. Morphological characteristics were similar in both conventionally propagated and micropropagated plants. Additionally, genetic homogeneity of in vitro plants was further confirmed through ISSR and flow cytometry analysis. A total of 27 ISSR primers were screened, out of which 13 ISSR primers generated 58 monomorphic and reproducible bands thereby confirming the genetic uniformity of obtained plants. The mean 2C DNA content of the mother plant (2.96 pg) was similar to that of in vitro derived plants (3.07 pg). Gas chromatography-mass spectrometry (GC–MS) analysis showed similarity in the qualitative profile of chemical constituents of essential oil and high-performance liquid chromatography analysis revealed no significant differences in curcumin content in the tissue culture regenerants and mother plants of C. zedoaria. Therefore, the present micropropagation protocol could be effectively employed to generate true to type plantlets of C. zedoaria. [ABSTRACT FROM AUTHOR]

To enhance the multiplication rate in Musa acuminata Colla (banana; 'Grand Nain') organogenesis, higher amounts of thiamine along with different sugar types and concentrations were evaluated at the proliferation phase. Thiamine at 1, 10, 50, 100, and 200 mg L−1 was compared with 0.1 mg L−1 thiamine found in conventional Murashige and Skoog (MS) medium. Maximum proliferation of banana was induced with 100 mg L−1 thiamine. Additionally, 15, 30, and 45 g L−1 sucrose, glucose, fructose, and sorbitol combined with regular and optimal levels of thiamine were tested. Glucose at 30 g L−1 most improved shoot proliferation alone and enhanced shoot proliferation further, when combined with 100 mg L−1 thiamine, followed by sucrose and fructose, whereas sorbitol completely inhibited growth and caused tissue browning. All evaluated vegetative traits were significantly affected by sugar type and concentration, and thiamine levels, unlike the photosynthetic pigments. Moreover, genetic stability of the plants recovered from the enhanced protocol was confirmed by inter-simple sequence repeats (ISSR) and randomly amplified polymorphic DNA (RAPD) analysis. A total of 230 bands generated by both marker types were monomorphic for the randomly selected regenerated plants, compared with their mother plant. Thus, the proliferation medium supplemented with 30 g L−1 glucose and 100 mg L−1 thiamine could be recommended for banana organogenesis. Results herein are of great importance and helpful in enhancing the commercial in vitro propagation protocols of banana, without the need of increasing the number of subcultures, which can cause somaclonal variation. [ABSTRACT FROM AUTHOR]

Neolamarckia cadamba is one of the fast-growing tree species selected for industrial tree plantations (ITPs). As demand and supply of true-to-type planting material are increasing, micropropagation of elite N. cadamba planting material is required for the sustainable development of ITPs. However, somaclonal variation among tissue culture-derived plantlets is a bottleneck in micropropagation. The present study described the genetic fidelity of in vitro regenerated N. cadamba plantlets from first, second, third and sixth subcultures. These plantlets were obtained from axillary shoot multiplication and repeated subcultures of microshoots with three weeks interval period, which initially developed from nodal explants. By using five random amplified length polymorphism (RAPD) and two inter-simple sequence repeats (ISSR) primers, a total of 9,334 bands and 2,760 bands were amplified respectively, from a total number of 164 tested plantlets. The banding profiles for each primer was highly uniform, and the DNA bands were monomorphic across all tissue culture-derived plantlets from every four subcultures compared to the stock plants. A target gene-specific marker was also employed to detect single nucleotide polymorphism (SNP) within the targeted genomic sequence of Susy gene. There was no SNP detected from all the analysed plantlets. The current findings ascertained the efficiency and reliability of the N. cadamba micropropagation protocol at least up to six subculture cycles for mass production of true-to-type plantlets. [ABSTRACT FROM AUTHOR]

Korean arbor vitae (KAV; Thuja koraiensis Nakai) is a critically endangered coniferous tree in Korea. Here, we report the somatic embryogenesis (SE) and cryopreservation system that can be used for micropropagation of KAV and long-term storage of KAV cultures. To induce SE in KAV, the influence of the developmental stage of zygotic embryos and the effect of basal medium on embryogenesis induction were examined. The developmental stage of zygotic embryos had a significant effect on the embryogenesis induction (P < 0.0001). The highest frequency of embryogenesis induction occurred in megagametophytes with zygotic embryos at precotyledonary (P) and late embryogeny (L1) stage (36%). The highest frequency of embryogenesis induction was obtained on initiation medium containing IM basal salts with 2.2 μM 6-benzylaminopurine and 4.5 μM 2,4-dichlorophenoxyacetic acid (35%). The effect of abscisic acid (ABA) on production of somatic embryos was tested. The highest number of somatic embryos per 50 mg of embryogenic tissue was achieved on maturation medium with levels of 100 μM ABA (24.0 ± 2.4). The effect of cryopreservation treatment to embryogenic tissues on the maturation capacity of somatic embryos was also tested. No significant differences between noncryopreservation and cryopreservation treatment were observed (P = 0.1896), and the highest mean number of somatic embryo per 50 mg of embryogenic tissues was obtained in noncryopreserved cell line (28.17 ± 5.66). Finally, the genetic identities of the plantlets regenerated from non- and cryopreserved embryogenic cell lines were verified and there was no genetic variation in the regenerated plantlets from cryostored embryogenic cell lines. This study is the first report on SE and the successful cryopreservation of embryogenic culture of the genus Thuja. [ABSTRACT FROM AUTHOR]

Begonias are ornamental indoor plants and also cultivated for landscaping. Begonia malabarica Lam., found naturally in the hilly regions of South India and Sri Lanka called by the tribals as Narayana Sanjeevi, i.e. possess miraculous therapeutic properties. The present study targets in vitro culture, field establishment and evaluation of its genetic uniformity using anatomical, molecular markers followed by callus induction and establishment of cell suspension. Explants such as shoot tip and leaves from in vitro germinated seeds were cultured on Murashige-Skoog medium. Maximum callus induction was obtained in Murashige-Skoog medium with 0.3 mg L-1 2, 4-D + 0.5 mg L-1 BAP, i.e. 90.8 ± 2.8% with 2.5 ± 0.1 g callus growth. In vitro suspension of cells and their growth rate were also analysed. RAPD and ISSR experiments were carried to confirm the genetic fidelity among in vitro and in vivo plants. Twelve RAPD and seven ISSR primers raised 119 amplicons. Genomic DNA amplification showed similarity between the in vivo and the in vitro generated plantlets. The overall results revealed the uniformity of the in vitro raised B. malabarica plantlets in terms of histoanatomical genetical features and substantiated the assumption that in vitro micropropagation is the secure mode for mass propagation of true to type plants without any somaclonal variations. [ABSTRACT FROM AUTHOR]

Highlights • The results obtained in the present study are a step forward to develop improved genotype through hybridization in wild pomegranate. • Phenotypic study helps in the selection of best performing individuals of wild pomegranate for domestication. • Attack of fruit borer on fruits showed that controlled pollinated fruits are more prone as compare to fruits open pollinated sources. • The important hybrids obtained were tested for their paternity using 21 RAPD markers of which thirteen primers revealed the clue for hybridity. Abstract The study was conducted in locations Tatool (L 1) district Solan, Narag (L 2) and Daron (L 3) district Sirmour, H.P. The half sibs from Narag and Daron location performed best for seedling height, minimal fruit borer incidence and oval shape. The highest titratable acidity was recorded from genotype L 3 T 3 (Daron location) with elliptical shape which is beneficial for Anardana processing. The tree genotypes bearing ovate and elliptical shaped fruits can be preferred for better fruit size selection. High heritability followed by genetic advance was observed for percent germination and seedling at six month age. In cross L 2 T 2 X L 3 T 3 , reputed fresh weight of seed without aril and germination was better whereas L 1 T 1 X L 2 T 1 cross showed better fruit quality biochemical estimates. The fruit crosses viz. L 2 T 4 X L 2 T 1 and L 2 T 2 X L 2 T 1 were found to have lower incidences of the borer attack. Maximum value in fruit traits were obtained from open pollination as compared to controlled cross combination for all the characters except fruit weight and fresh seed weight with aril. The hybrids were tested for their paternity using 21 RAPD markers of which thirteen primers revealed that 7 hybrids out of 15 hybrids have similar pattern as their parent. [ABSTRACT FROM AUTHOR]

Crinum malabaricum , Lekhak & S.R. Yadav (Amaryllidaceae) is a critically endangered bulbous plant endemic to India. Establishing an efficient plant regeneration protocol is necessary for its conservation and large-scale propagation. In the present study, regeneration was achieved through somatic embryogenesis from callus produced on MS media supplemented with various different concentrations of 2,4-D alone and in combination with N6-benzyl-adenine (BA). Different advanced stages of embryo development (globular, torpedo and cotyledonary) and maturation were obtained on MS medium, with combinations of picloram and thidiazuron (TDZ). A high rate of somatic embryos (55.89 ± 0.60) was obtained after eight weeks. SEM examinations showed the occurrence of cell clusters (embryogenic) which converted to somatic embryos. Well-developed cotyledon embryos were successfully germinated on PGRs free full and half strength MS medium, however, 94.03% of somatic embryos germinated on half-strength MS medium supplemented with 1.0 mg L−1 gibberellic acid. The true-to-type genetic conformity of regenerated plants was examined by SCoT, ISSR and RAPD primers based molecular analysis. The amplification bands produced by SCoT, ISSR and RAPD primers applied were monomorphic across all the regenerated plants. This confirmed their genetic homogeneity compared to the mother plant and also demonstrated the reliability of our somatic embryogenesis system for C. malabaricum. A rapid method for phytochemical analysis based on LC-ESI/MS exhibited better separation and analysis of galanthamine and lycorine from methanolic extracts of in vitro raised plants derived from somatic embryogenesis. The protocol developed should be helpful in reintroduction, restoration and ex situ conservation of this valuable plant in the natural condition as well as in the pharmaceutical sectors. [Display omitted] • An efficient somatic embryogenesis system was developed in Crinum malabaricum. • Different concentrations and combinations of plant growth regulators influenced the development of somatic embryos. • The clonal fidelity of somatic embryo-derived plants was confirmed by molecular markers. • Galanthamine and lycorine were efficiently identified by using LC-ESI-MS/MS analysis. • This study will be helpful for large scale clonal propagation and production of secondary metabolites. [ABSTRACT FROM AUTHOR]

An effective and rapid method for propagating genetically true plantlets of a pharmaceutically important plant Bacopa monnieri L. was developed from thidiazuron (TDZ) pulse treated nodal segment explants. Explants treated for 24 h with different concentration of 0.0, 5.0, 10, 20 or 40 μM thidiazuron (TDZ) prepared in liquid Murashige and Skoog (MS) medium induces multiple shoot on MS basal medium without growth regulators. The highest frequency 95.3 ± 2.3% of regeneration and maximum number 43.7 ± 2.0 of shoots per explants with an average shoot height of 5.4 ± 0.29 cm were achieved after 08 weeks of culture from the explants treated with 20 μM TDZ. Simultaneously, the regenerated microshoots were rooted on the same growth regulator free media. Elongated shoots with roots were successfully acclimated to ex vitro condition with a 97% survival rate. Genomic stability of in vitro developed plants of B. monnieri was first time determined using flow cytometry and single primer amplification reaction (SPAR) methods viz., random amplified polymorphic DNA (RAPD), intersimple sequence repeat polymorphic DNA (ISSR) and directed amplification of minisatellite DNA (DAMD) and compared with donor plant. Nuclear DNA (nDNA) content of in vitro propagated B. monnieri plants has been estimated as 2.08 pg/2C based on the flow cytometric investigation of nuclei isolated from leaf tissues. No significant variations in nDNA content, plant genome size and ploidy level were observed between micropropagated plants and field grown donor plants. Likewise, the SPAR markers also revealed monomorphic banding profiles in micropropagated plantlets of B. monnieri and similar with to that of the donor plants verifying their genomic stability. [ABSTRACT FROM AUTHOR]

In the present study, a simple, eco-friendly, biogenic protocol for synthesis of silver nanoparticles (AgNPs) using leaf extract of in vitro banana plantlets (Musa acuminata Colla cv. Grand Nain) was established. The synthesized AgNPs were characterized by X-ray diffraction, transmission electron microscopy, scanning electron microscopy, and fourier transform-infrared spectroscopy. Shoot tips of stevia (Stevia rebaudiana Bert.) were cultured on three forms of regeneration medium (Murashige and Skoog basal medium + 0.5 mg. L−1 6-benzyladenine) i.e. 'semi-solid', 'liquid', and 'semi-solid followed by liquid' supplemented with 100–400 µM of AgNPs to study the effect of different concentrations of AgNPs along with different forms of medium on plant growth and rebaudioside-A content in stevia under in vitro conditions. Among different concentrations, 400 µM AgNP showed the best growth response irrespective of different forms of medium. Nonetheless, the overall best growth response was recorded in 'semi-solid followed by liquid' medium. High performance liquid chromatography analysis of regenerants showed gradual incline in rebaudioside-A content with the increase in AgNP concentrations. Regenerants from medium containing 400 µM AgNPs produced the highest rebaudioside-A of 334.8 mg. g−1 dry weight (>2-fold) while control ones produced 154.3 mg. g−1 dry weight. Genetic fidelity of AgNP-treated and control regenerants was confirmed using 20 inter simple sequence repeats and start codon targeted primers. The present study can be useful for commercial-scale propagation of stevia ensuring higher fresh and dry biomass as well as ameliorated rebaudioside-A production. • First report on green synthesis and characterization of silver nanoparticles (AgNPs) from in vitro grown banana leaf. • Among three forms, 'semi-solid followed by liquid' medium showed superior growth of stevia. • Ameliorated biomass and rebaudioside-A production with rising AgNP concentrations in stevia. • AgNP-treated regenerants via ISSR and SCoT markers confirmed 100% genetic fidelity. [ABSTRACT FROM AUTHOR]

In vitro clonal propagation of small fruit species has been one of the most broadly exploited approaches in biotechnology. However, the most critical aspect of this propagation technology has been the maintenance of genetic fidelity of tissue culture-raised plants. In the present study, three techniques were deployed in the assessment of genetic fidelity of micropropagated plants (TC plants) of blackberry cultivar ‘Čačanska Bestrna’ and raspberry cultivar ‘Meeker’ in the second year after planting in the open field, in comparison to plants originating from the standard planting material (ST plants). Flow cytometry analysis deployed to estimate the DNA ploidy level and relative nuclear DNA content in TC and ST plants revealed no significant differences in nuclear DNA content between plants of different origin. Chromosome counting in root/shoot tip meristems also showed a normal tetraploid chromosome number (2 n = 4 x = 28) in TC blackberry plants and a normal diploid chromosome number (2 n = 2 x = 14) in TC raspberry plants. Polyacrylamide gel electrophoresis of peroxidase (POX) extracted from the leaf tissue revealed no differences in POX profiles between TC and ST plants either. The results obtained in this study verify the suitability of micropropagation by axillary branching for commercial exploitation in blackberry and raspberry. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop an efficient protocol for the in vitro regeneration of chayote ( Sechium edule ), an important Mexican crop and the evaluation of the genetic fidelity. This species produces recalcitrant seeds, which complicates germplasm storage and makes in vitro conservation a desirable alternative. Proliferation of axillary shoots was induced from axenic nodal segments obtained from in vitro germinated seedlings. Nodal segments were cultured on MS medium supplemented with 0.1 mg/L 6-benzylaminopurine (BA), producing 8.0 ± 0.4 shoots per explant with a regeneration frequency of 92%. Plant regeneration by organogenesis was optimally achieved when leaf and petiole explants were cultured on MS medium supplemented with 0.1 mg/L BA and 0.05 mg/L gibberellic acid (GA3), obtaining an average of 5.3 ± 1.9 shoots per explant. All of the regenerated shoots originated directly from petioles without previous formation of callus. Adventitious shoots were successfully rooted in MS medium without growth regulators after 2 weeks. Since somaclonal variation is a potential hindrance to the in vitro propagation of plants, genetic stability was evaluated through RAMP markers using 9 combinations of RAPD and ISSR primers. The amplification products obtained from the regenerated plants were electrophoresed and showed banding patterns similar to that of the mother plant, demonstrating the homogeneity of the individuals obtained through the regeneration protocols reported here. [ABSTRACT FROM AUTHOR]

The aim of this study was to produce sugarcane plantlets from cell suspension culture and study its genetic fidelity using molecular markers. The study was carried out using sugarcane varieties Co 86032 and Q117. Callus cultures of both the varieties were optimized using six different callus induction media. After screening the growth response of callus on six different callus inductionmedia, itwas observed that medium no. VI supplemented with 500 mg l-1 of each PVP, Casein hydrolysate and MES buffer showed high amounts of callus in Co 86032 (79.66 ± 0.44%) and Q117 (82.83 ± 1.69%). Addition of PEG 8000 at 2.5% to this medium had a profound impact on inducing somatic embryogenesis in Co 86032 (54.66 ± 1.76%) and Q117 (66.66 ± 2.60%) as compare to control (24.33 ± 1.76%) and (27.33 ± 2.73%), respectively. Cell suspension cultures were established by culturing embryogenic calli in liquid medium showed well established suspension cultures with fever cell aggregates. There was negligible cell division during initial 2 days of incubation and cell count increased rapidly between 2 and 8 days. Further incubation beyond 8 days resulted in a decrease in cell viability. Enhanced callus proliferation in Q117 while enhanced shoot regeneration in Co 86032 was observed from cell suspension culture. The clonal fidelity of in vitro regenerated plants was assessed by using RAPD and ISSR markers. Analysis of the ten RAPD markers indicated that 90.48 and 86.95%true-to-type regenerated plantlets in Co 86032 and Q117, respectively. However, in the ISSRmarkers, Co 86032 did not show any polymorphism and in the Q117, 92.18% true-to-type plantlets were found. These results confirmed that somaclonal variation occurs during the process of indirect organogenesis and RAPD and ISSR marker based molecular analysis is a suitable method for an early detection of variation in sugarcane. [ABSTRACT FROM AUTHOR]

The aim of this study was to produce sugarcane plantlets from cell suspension culture and study its genetic fidelity using molecular markers. The study was carried out using sugarcane varieties Co 86032 and Q117. Callus cultures of both the varieties were optimized using six different callus induction media. After screening the growth response of callus on six different callus induction media, it was observed that medium no. VI supplemented with 500 mg l of each PVP, Casein hydrolysate and MES buffer showed high amounts of callus in Co 86032 (79.66 ± 0.44%) and Q117 (82.83 ± 1.69%). Addition of PEG 8000 at 2.5% to this medium had a profound impact on inducing somatic embryogenesis in Co 86032 (54.66 ± 1.76%) and Q117 (66.66 ± 2.60%) as compare to control (24.33 ± 1.76%) and (27.33 ± 2.73%), respectively. Cell suspension cultures were established by culturing embryogenic calli in liquid medium showed well established suspension cultures with fever cell aggregates. There was negligible cell division during initial 2 days of incubation and cell count increased rapidly between 2 and 8 days. Further incubation beyond 8 days resulted in a decrease in cell viability. Enhanced callus proliferation in Q117 while enhanced shoot regeneration in Co 86032 was observed from cell suspension culture. The clonal fidelity of in vitro regenerated plants was assessed by using RAPD and ISSR markers. Analysis of the ten RAPD markers indicated that 90.48 and 86.95% true-to-type regenerated plantlets in Co 86032 and Q117, respectively. However, in the ISSR markers, Co 86032 did not show any polymorphism and in the Q117, 92.18% true-to-type plantlets were found. These results confirmed that somaclonal variation occurs during the process of indirect organogenesis and RAPD and ISSR marker based molecular analysis is a suitable method for an early detection of variation in sugarcane. [ABSTRACT FROM AUTHOR]