Recent Articles

Below are the titles and abstracts from the most recent articles published in In Vitro Cellular and Developmental Biology – Plant. Click on the title to view the full article at Springerlink.


  • Overexpression of a Malus xiaojinensis WRKY transcription factor gene ( MxWRKY55 ) increased iron and high salinity stress tolerance in Arabidopsis thaliana
    on October 15, 2020 at 12:00 am

    Abstract Apple, as one of the most important economic forest tree species, is widely grown in the world. Abiotic stresses, such as high salinity and iron deficiency, affect apple growth and development. WRKY transcription factors (TFs) are widely involved in the responses of plants to adverse stresses. In the present study, a new WRKY gene was isolated from Malus xiaojinensis and designated as MxWRKY55. Subcellular localization showed that MxWRKY55 was localized to the nucleus. The expression level of MxWRKY55 was highly affected by salt, low-Fe, and high-Fe stresses in M. xiaojinensis seedlings. When MxWRKY55 was introduced into Arabidopsis thaliana, it greatly enhanced the salt and Fe tolerance. When dealt with high- and low-Fe stresses, the overexpression of MxWRKY55 in transgenic A. thaliana resulted in higher levels of root length and fresh weight as well as contents of chlorophyll and Fe than wild type (WT). Increased expression of MxWRKY55 in transformed A. […]

  • Physiological, biochemical, and metabolic responses of a Taxus baccata L. callus culture under drought stress
    on October 13, 2020 at 12:00 am

    Abstract The metabolic stimulation induced by abiotic stress is an efficient strategy for the production of secondary metabolites in sterile and controlled plant cell, tissue, and organ cultures. Paclitaxel (taxol), one of the most widely used therapeutic compounds for the treatment of various cancers, is mainly produced through cell culture of the European yew (Taxus baccata L.). In this work, a T. baccata callus culture was subjected to drought stress induced in vitro by mannitol and sorbitol (1, 2, 3, and 4%) and sucrose (6 and 8%) to evaluate its impact on physiological and biochemical traits as well as paclitaxel and 10-deacetyl baccatin III (10-DBA) production. It was observed that drought stress caused a significant increase (P < 0.05) in dry weight, proline, soluble sugars, hydrogen peroxide, lipid peroxidation, total phenolics, flavonoids, and flavonols, and a decrease in relative water content, fresh weight, relative growth rate, and cell viability. […]

  • Management of microbial contaminants in the In Vitro Gene Bank: a case study of taro [ Colocasia esculenta (L.) Schott]
    on October 7, 2020 at 12:00 am

    Abstract Ex situ conservation of vegetatively propagated crops by tissue culture is sometimes hampered by covert endophytic bacteria which become visible after repeated subculture over prolonged periods. In the present study, we identified bacterial contaminants and devised a strategy for their elimination from 20 in vitro conserved accessions of taro [Colocasia esculenta (L.) Schott] held in the In Vitro Gene Bank (IVGB) at ICAR-National Bureau of Plant Genetic Resources (NBPGR). Visually, the cultures were exhibiting white to dark brown exudation in tissue culture growth medium and undergoing slow degeneration as recorded by wilting and necrosis of leaves and shoots. On culturing the macerated pieces of shoots, corms, and leaf petioles from the contaminated taro cultures on bacterial indexing medium, six distinct bacterial colonies were observed. The identity of these bacteria was established through 16S rDNA sequencing as gram-negative strains (2) of Ralstonia […]

  • A hydroponic-based efficient hardening protocol for in vitro raised commercial kiwifruit ( Actinidia deliciosa )
    on October 7, 2020 at 12:00 am

    Abstract Actinidia deliciosa is a commercially important plant receiving recognition because of its high nutritive value. This study presents an efficient protocol for the hardening of in vitro raised Actinidia deliciosa plants using a hydroponic method. Leaf explants inoculated on Murashige and Skoog (MS) medium supplemented with 5.00 μM 6-benzylaminopurine (BAP) and 1.00 μM α-naphthaleneacetic acid (NAA) resulted in 5.28 ± 0.14 shoots per explants and 8.33 ± 1.80 cm average shoot length. Rooting was achieved through half-strength MS medium supplemented with 1.5 μM indolebutyric acid (IBA) and 0.6 μM BAP. Maximum root number (12.76 ± 1.08) and 6.78 ± 0.25 cm average root length were recorded from plantlets using half-strength MS medium supplemented with 1.5 μM indolebutyric acid (IBA) and 0.6 μM BAP. In hydroponic system, an average root length of 22.40 ± 0.59 cm, average root number of 21.50 ± 1.24, average leaf number […]

  • Effect of light quality and tissue origin on phenolic compound accumulation and antioxidant activity in Camellia japonica calli
    on October 6, 2020 at 12:00 am

    Abstract In Camellia japonica callus culture, the effects of light quality on the accumulation of phenolic compounds were examined. To this end, the calli were cultured under three monochrome [white (W), red (R), and blue (B)] and two mixed [red/blue (RB) and red/green/blue (RGB)] light sources. After 4 weeks of culture, fresh and dry callus weights were determined, and phenolic content was analyzed using high-performance liquid chromatography. We also examined the effects of RGB light on calli derived from four origins (leaf: CL; root: CR; petal: CP; and ovary: CO). Total phenolic and flavonoid content and DPPH radical scavenging activity were highest in calli exposed to RB and RGB. Catechin was detected only in calli exposed to mixed light (RB and RGB). Interestingly, red pigmentation development in CL and CR calli after 2 weeks of culture and organogenic calli in CR and CP samples was observed. Furthermore, expression of the phenylalanine ammonia lyase and […]

  • Morphological and physiological responses of tara ( Caesalpinia spinosa (Mol.) O. Kuntz) microshoots to ventilation and sucrose treatments
    on September 28, 2020 at 12:00 am

    Abstract One of the main problems of in vitro plant tissue culture is the poor ventilation in the culture vessels. This leads to morphological and physiological anomalies in regenerated in vitro plants, a crucial issue in woody species, such as Caesalpinia spinosa (Mol.) O. Kuntz, which is a legominous tree or thorny shrub of the Andes, adequate photoautotrophic and photomixotrophic in vitro rooting systems could eliminate these problems. Therefore, the purpose of the present study was to evaluate photoautotrophic and photomixotrophic treatments to optimize the in vitro rooting of C. spinosa microshoots. Micropropagated shoots were cultured in vessels sealed with a polypropylene lid with 0, 1, or 3 ventilation holes covered with a 0.45-μm adhesive polypropylene microfilter. The vessels contained MS salts with or without 30 g L−1 sucrose or, alternatively, a porous substrate (PRO-MIX®), which facilitated rooting and produced greater fresh and dry weights, a […]

  • Establishment of regeneration system of callus pathway for Iris sanguinea Donn ex Horn
    on September 28, 2020 at 12:00 am

    Abstract Iris is a perennial flowering plant, usually cultivated through seeds or bulbs. However, due to the limitations of traditional reproduction, the establishment of a callus regeneration system is particularly important, and callus induction and plant regeneration are the keys to transgenic technology. The callus regeneration system was studied using the I. sanguinea flower stem as explants. The experiment investigated the impact of different disinfectant solution concentrations, disinfection exposure times, and hormone concentrations on the explant growth status at various culture stages. The research analyzed the impact of the experimental variables on explant regeneration to determine the optimum callus regeneration system for I. sanguinea. In this research, it was determined that when I. sanguinea flower stems, that were used for explants, were treated with 75% alcohol for 30 s and 2% NaClO for 8 min for disinfection, there was a 54.48% callus induction […]

  • Molecular analysis of ERF subfamily genes during coffee somatic embryogenesis
    on September 25, 2020 at 12:00 am

    Abstract With ample cultivation and economic potential, coffee is one of the main commodities in the world. Despite its vast cultivation, there remain problems related to its productivity as a consequence of pests and diseases, questions regarding its physiology, and difficulty with its propagation. Due to such challenges, the application of biotechnological tools, such as tissue culture and molecular markers, has become essential in the search for improvements to coffee cultivation. Among tissue culture techniques, somatic embryogenesis is considered the best form of micropropagation in vitro since it is possible to produce plants that are identical to the parent plant. It is therefore essential to identify the genes involved in the process of somatic embryogenesis, as well as to understand their functions. Different studies have shown the expression of ethylene response factor (ERF) genes in different tissues and under different conditions, mainly in response to […]

  • Trevor Alleyne Thorpe: His academic life and scientific legacy
    on September 21, 2020 at 12:00 am

    Abstract This special review is written to commemorate the life and contributions of Dr. Trevor Alleyne Thorpe, who passed away on May 18, 2020, at the age of 83 yr ( As part of his academic legacy, Trevor had trained an impressive number of laboratory members—15 PhD students, 14 Master’s students, 25 postdoctoral fellows, and many visiting scientists—who collectively identify themselves as proud members of “Team Thorpe.” This executive summary version of his major scientific contributions was prepared by the team.

  • Embryo rescue response and genetic analyses in interspecific crosses of oilseed Brassica species
    on September 12, 2020 at 12:00 am

    Abstract Embryo rescue technique is widely utilized to save developing embryo from abortion in interspecific hybridization of plants. In Brassica species, success of an embryo rescue depends on the age of embryo, developmental stage of embryo, and media composition. In this study, ten crosses were conducted between diploid oilseed species B. rapa × tetraploid B. napus and B. rapa × tetraploid B. juncea along with their reciprocal crosses. The immature embryos from parents and crosses were cultured at 14 d after pollination in MS and ½ MS medium without any growth regulators for direct embryogenesis. Embryos were found at different stages of development during isolation, from globular to cotyledonary, indicating that rate of embryo development depends on genotype. Embryos isolated at the cotyledonary stage showed the highest regeneration percentage (av. 83.29%) and required the lowest time to regenerate (5.8 d) compared with another type of embryo, torpedo stage, […]

  • Vitrification and proteomic analysis of embryogenic callus of Panax ginseng C. A. Meyer
    on September 11, 2020 at 12:00 am

    Abstract Panax ginseng C. A. Meyer is a desirable medicinal plant due to its health benefits. With the growing interest in ginseng products, the safe preservation of genetically engineered material with unique attributes is increasingly important. In the present study, an effective protocol for cryopreservation of the ginseng embryogenic callus by vitrification is reported. After 2-wk subculture, the calluses were cold-acclimated at 4°C for 3 wk, then precultured for 2 d with a preculture medium containing 0.3 M sucrose. The precultured embryonic callus was then treated with a loading solution, followed by exposure to plant vitrification solution 2 at 0°C for 90 min. The dehydrated embryogenic calluses were then transferred into fresh 1.5 mL plant vitrification solution 2 and directly immersed in liquid nitrogen. With this protocol, an average of 68.3% regrowth rate for cryopreserved material was obtained, and the number of somatic embryos differentiated from […]

  • Optimization of efficient direct organogenesis protocol for Punica granatum L. cv. Kandhari Kabuli from mature leaf explants
    on September 11, 2020 at 12:00 am

    Abstract Punica granatum L. is an important horticultural fruit crop with high medicinal and economic value. Its rising commercial demand necessitates the production of high-quality planting material. Here, we describe an efficient protocol for direct organogenesis in Punica granatum L. cv. Kandhari Kabuli from mature leaf explant. The optimized sterilization procedure for explant includes sequential treatment with 70% ethanol (0.75 min), 0.2% Bavistin (15 min), and 0.5% sodium hypochlorite (2 min), which resulted in 83% axenic cultures. The accumulation of phenolics was effectively controlled by subculturing of leaf explants three to four times at a regular interval of 24 h. The organogenic capability of leaf segments was investigated on full-strength Murashige and Skoog (MS) medium supplemented with different plant growth regulators (PGRs), including the cytokinins 6-benzylaminopurine (BAP) and thidiazuron (TDZ) alone or in combination with α-naphthaleneacetic […]

  • Organogenesis and high-frequency plant regeneration in Caryopteris terniflora Maxim. using thidiazuron
    on September 11, 2020 at 12:00 am

    Abstract In vitro culture is employed as a means of producing plant materials from medicinal plants to meet the increasing demands of the market. Herein, we focused on an efficient protocol for organogenesis through two pathways in six explant types from Caryopteris terniflora Maxim (C. terniflora), a medicinal and ornamental plant. For direct organogenesis, the maximum number of shoots per internode explant (62.5) was achieved on Murashige and Skoog (MS) medium containing 3.0 mg L−1 thidiazuron (TDZ) and the maximum number of shoot tips per explant (45.5) was obtained by 1.5 mg L−1 TDZ, with 100% response. A protocol was also developed for indirect organogenesis from leaf, leaf-petiole, petiole, and root explants on MS basal medium containing a range of TDZ concentrations. Leaf-petiole explants induced more adventitious shoots (57.6 per explant) and leaves (31.4 per explant) than petioles (37.5 per explant) or roots (25.0 per explant), and the […]

  • In vitro propagation of three Iranian apricot cultivars
    on September 9, 2020 at 12:00 am

    Abstract In vitro propagation is a useful method for clonal propagation of apricot; however, it is a highly genotype-dependent procedure. Eight different experiments were conducted to optimize and establish an efficient in vitro propagation protocol for three Iranian apricot cultivars, including Ordubad, Shams, and Qaysi. Sterilization, in vitro establishment, proliferation, root induction, and acclimatization steps were assessed. The fungal and bacterial infections were significantly decreased when nanosilver (2.5%) was applied in sterilization of initial explants. The highest number of active buds was obtained from summer-season collected lateral bud explants. The establishment responses of apricot cultivars to investigated basal culture media were different and the best results for Ordubad, Shams, and Qaysi cultivars were obtained from Driver and Kuniyuki (DKW), Woody Plant Medium (WPM), and modified Quoirin and Lepoivre (QL) basal media, respectively. The highest […]

  • Application of cryobanking for Platycodon grandiflorum in vitro axillary buds using cryo-plate methods
    on September 8, 2020 at 12:00 am

    Abstract The application of the V cryo-plate method using either PVS2 or PVS3 and the D cryo-plate method to Platycodon grandiflorum in vitro axillary buds was investigated. Precultured buds (1 mm) were attached to cryo-plates using calcium alginate gel. Then, osmoprotection treatment (2 M glycerol and 1 M sucrose) was performed by immersing the cryo-plates for 30 min at 25°C. The procedures for each cryogenic method were as follows: in the V cryo-plate method, the buds on cryo-plates were exposed to PVS2 for 40 min (highest regrowth 71.1%) or PVS3 for 50 min (highest regrowth 82.2%). In the D cryo-plate method, the buds on the cryo-plates were dehydrated under the air current of a laminar air flow cabinet for 60 min (highest regrowth 80.0%) at 25°C. Then, the cryo-plates were plunged directly into liquid nitrogen. After cryopreservation, buds on the cryo-plates were rewarmed and unloaded by immersion in 1 M sucrose solution for 15 min at 25°C for subsequent plant […]

  • Direct shoot organogenesis from shoot tip explants of a highly medicinal valued tree Pterocarpus marsupium Roxb.
    on September 2, 2020 at 12:00 am

    Abstract 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 […]

  • Somatic embryogenesis and plant regeneration in Piper aduncum L
    on August 28, 2020 at 12:00 am

    Abstract An efficient protocol is reported for in vitro plant regeneration through somatic embryogenesis in Piper aduncum, a Brazilian Amazon species with high economic potential. The species is important due to a variety of components found in its essential oil, with emphasis on dillapiole. Leaf explants from five accessions identified for high oil yield and levels of dillapiole were evaluated for their embryogenic potential. To induce embryogenic calli, the explants were cultivated in MS medium supplemented with 5 mg L−1 of 1-naphthaleneacetic acid (NAA) and 2.5 mg L−1 of N6-benzylaminopurine (BAP) for 80 d. For somatic embryogenesis, the embryogenic calli were transferred to MS medium with 10 mg L−1 of NAA and 2.5 mg L1 of BAP and incubated for 45 d. The obtained somatic embryos were germinated in MS medium without regulators by 45 d and the obtained plantlets were subjected to acclimatization. Somatic embryos and calli from this process were […]

  • Overexpression of annexin gene in rice ( Oryza sativa L.) for salinity and water stress
    on August 28, 2020 at 12:00 am

    Abstract The aim of the study is to develop transgenic rice with annexin genes (AnnBj2) to determine its salinity and water stress tolerance through Agrobacterium-mediated genetic transformation. The transgenic somatic embryos were developed on Murashige and Skoog medium fortified with 0.5 mg L−1 kinetin and 3.0 mg L−1 2,4-D. About 80% of the transformed somatic embryos germinated and was successfully established in the transgenic greenhouse. The transformed plantlets (T0 and T1 generation) were identified through PCR using NPT-II and AnnBj2 gene–specific primers as well as positive uidA reporter gene expression. Compared with non-transformed plants, the transgenic rice plants overexpressing AnnBj2 gene exhibited salt tolerance at the seedling stage. Seeds generated at T0 and T1 generations were further studied to determine their salinity tolerance through nutrient culture and pot culture experiments for water stress. When compared with the non-transgenic […]

  • Development of acetosyringone-inducible Gateway® and Golden Gate expression vectors for heterologous gene expression in Agrobacterium tumefaciens
    on August 20, 2020 at 12:00 am

    Abstract As a plant genetic engineer, Agrobacterium tumefaciens utilizes phenolic compounds, such as acetosyringone, to activate its virulence genes during the infection and transformation process. In this study, two novel broad-host-range (BHR) acetosyringone-inducible expression vectors were constructed. Unlike Agrobacterium binary vectors which are developed for plant transformation, these vectors were designed for heterologous gene expression in A. tumefaciens. To this end, an acetosyringone-inducible virB promoter (PvirB) was placed upstream of a Gateway® cassette for plasmid vector pASE1 and Type IIS recognition sites, BpiI for plasmid vector pASE2, thus allowing the PvirB to drive the expression of the insert, which may be cloned into pASE1 and pASE2 via Gateway® LR Cloning and Golden Gate Cloning approaches, respectively. Here, we tested the functionality and inducibility of pASE1 and pASE2 vectors by expressing the reporter gene superfolder green […]

  • Improving of rooting and ex vitro acclimatization phase of Agave tequilana by temporary immersion system (BioMINT™)
    on August 17, 2020 at 12:00 am

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