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.


  • Micropropagation of Catasetum integerrimum Hook (Orchidaceae) through seed germination and direct shoot regeneration from pseudobulbs and roots
    on January 22, 2022 at 12:00 am

    Abstract Catasetum integerrimum is an orchid used in Mexican traditional medicine, which is overexploited due to its ornamental and medicinal properties. In this scenario, conservation strategies are mandatory to protect wild populations from this valuable orchid. The aim of the present study was to establish a micropropagation protocol by direct organogenesis, as a tool for Catasetum integerrimum production and conservation. Seeds from a mature capsule were cultivated in a Murashige and Skoog basal medium. Then, 6-mo-old vitroplants were transplanted into treatments fortified with 6-bencilaminopurin (BAP) or indole-3-acetic acid (IAA) to induce plant development. Treatment fortified with 2.5 mg L−1 BAP in combination with 5.0 mg L−1 IAA developed 5.73 ± 0.45 shoots per explant and 5.84 ± 0.48 leaves per shoot. For the rooting process, the best treatment was 2.5 mg L−1 IAA, which developed 11.20 ± 0.28 roots with a 13.20 ± 0.28-cm root […]

  • Development of a high-frequency adventitious shoot regeneration using cotyledon explants of an important oilseed crop Sesamum indicum L.
    on January 18, 2022 at 12:00 am

    Abstract Sesame (Sesamum indicum L.; 2n = 26) is an ancient oil-yielding crop known for its valuable nutrients such as essential fatty acids, proteins, and medicinally important lignans. Recalcitrant nature of sesame limits the use of modern biotechnology for its genetic improvement. For the development of an efficient in vitro regeneration protocol, the cotyledon (de-embryonated) explants were used for the development of adventitious shoots. In the present study, Murashige and Skoog (MS) medium with 25 μM BAP (6-benzylaminopurine) and 2 μM IAA (indole-3-acetic acid) showed high regeneration frequency in both GT-3 (33.3 ± 1.1%) and GT-4 (30 ± 0.0%) varieties within 6 wk of culture. Addition of 20 μM thidiazuron (TDZ) showed further improvement in regeneration frequency in both GT-3 (63.3 ± 1.1%) and GT-4 (26.7 ± 1.1%) varieties. In another experiment, 0-d-old cotyledons inoculated on MS medium supplemented with 4 μM abscisic acid (ABA) […]

  • An efficient micropropagation protocol, chemical components, and hypoglycemic activity for Cremastra appendiculata (D. Don) Makino pseudobulbs
    on January 17, 2022 at 12:00 am

    Abstract Cremastra appendiculata (D. Don) Makino is a valuable species in the ornamental field and pharmaceutical industry, while inefficient propagation limits the utilization of C. appendiculata. Here, we reported an improved and efficient method for large-scale micropropagation including seed germination, initiation, and proliferation of protocorm-like bodies (PLBs), “shoot cluster” induction, rooting, and acclimatization. Factually, the highest shoot multiplication coefficient 3.16 was achieved on 1/4 MS medium supplemented with 1.5 mg L−1 thidiazuron (TDZ), along with 0.4 mg L−1 indole-3-acetic acid (IAA), 3% sucrose, and additives. Likewise, the best medium for protocorm proliferation was 1.0 mg L−1 indole-3-butyric acid (IBA), 1.0 mg L−1 6-benzyladenine (6-BA), and 0.2 mg L−1 adenylate cyclase (AC) with proliferation of 170.07%, followed by root induction on 1/4MS with 0.5 mg L−1 α naphthaleneacetic acid (NAA), 0.5 mg L−1 AC, and 3% […]

  • In vitro culture in combination with aeroponics is an efficient means of mass propagation of Sarcostemma acidum: a rare medicinal plant of Indian arid zone
    on January 14, 2022 at 12:00 am

    Abstract In vitro culture in combination with aeroponics is observed to be an efficient means for mass propagation of Sarcostemma acidum in the present investigation. S. acidum is a rare leafless xerophytic shrub of the family Apocynaceae, commonly known as Som-lata or khir-kheep, a source of medicines and the religious drink “Somaras.” Aeroponic technique was used for the induction of adventitious roots from stem cuttings of field-grown plants for the development of quality plants in the greenhouse as a source of explants for micropropagation. Among various concentrations of IBA, NAA and NOA used for induction of adventitious rooting from stem cuttings 3.0 g L−1 NAA were found to be most suitable with average 90% induction response and 14 adventitious roots. Surface-sterilized nodal shoot segments of 6 to 7 cm length were inoculated on basic and modified Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% (w/v) agar, and various concentrations […]

  • Phytochemical and anti-oxidant evaluation of in vitro and in vivo propagated plants of Curculigo orchioides
    on January 11, 2022 at 12:00 am

    Abstract Curculigo orchioides (Kali Musli) is a medicinal plant commonly used in the Indian and Chinese traditional medicine system to treat jaundice and asthma. It is also used as an aphrodisiac in several countries. C. orchioides is treated as an essential resource in many pharmaceutical industries as a source of bioactive compounds. Indiscriminate use, depletion of its natural habitat, and its seasonal availability have resulted in its rarity. As per the IUCN status, Curculigo orchioides is now listed as an endangered plant. The present micropropagation study resolves its seasonal availability by ensuring a continuous supply of plants. In vitro somatic embryos were induced on the leaf explants of C. orchioides in one-fourth–strength Murashige and Skoog (MS) media. The embryos developed into whole plants in 8 wk. The in vitro plants produced were evaluated for their phytochemical evaluation and anti-oxidant activity along with their wild counterparts as controls. […]

  • In vivo and in vitro bulb multiplication of Lilium rhodopaeum Delip. and growth acceleration using hydroponic technologies
    on January 10, 2022 at 12:00 am

    Abstract Lilium rhodopaeum Delip. (Liliaceae) is a Balkan endemic, with limited distribution in Bulgaria and Greece, in the Rhodopes Mts. The species is included in the Red Data Book of Bulgaria as critically endangered, and protected by the national Biodiversity Act. The present study aimed to multiply L. rhodopaeum by in vivo or in vitro regeneration of bulblets and to accelerate their growth using soilless cultivation. An average of 7.2 ± 2.7 in vitro bulblets per half bulb scale was obtained for 4 mo on the best medium (MS supplemented with 0.5 mg/l α-naphthyl acetic acid (NAA) and 0.1 mg/l kinetin (Kin)). More than 800 bulblets were gradually ex vitro adapted in a phytotron, but only a few survived in the greenhouse. Growth of 72 in vitro bulblets evenly distributed in 3 variants was compared: on a flood and drain hydroponic system (with perlite as substrate); cutting board hydroponic system (deep water culture system, peat cubes as substrate); and […]

  • Correction to: Impacts of the regulatory environment for gene editing on delivering beneficial products
    on January 10, 2022 at 12:00 am
  • Cell-penetrating peptide-mediated transfection of proteins into tissues of Arabidopsis and Chinese cabbage
    on January 6, 2022 at 12:00 am

    Abstract Cell-penetrating peptides (CPPs) are a class of short peptides, which have the ability of intracellular delivery. CPPs can also be used as an ideal transfection tool whereby biomacromolecules can be transferred as cargo into recipient cells. Here, a synthesized cell-penetrating peptide (R9)-mediated DNA-free transfection is presented. DNA sequences of nine tandem arginine (R9), one cysteine (cys), reporter mCherry, and histidine label were sequentially constructed into pET-45b( +) expression vector and transformed into the Escherichia coli BL21(DE3) strain. The R9-cys-mCherry fusion protein was obtained by prokaryotic expression. The Arabidopsis root tips, Chinese cabbage microspores, and 3-wk-old microspore embryos (MDEs) were used as transfected recipients. With a working concentration of R9-cys-mCherry ranging from 10 to 100 μg mL−1 and overnight incubation, R9-cys-mCherry protein can be translocated into the recipient mentioned above. The […]

  • Better light spectral quality and thermal amplitude inside the greenhouse stimulate growth and improve acclimatization of in vitro–grown Cattleya warneri T. Moore
    on December 1, 2021 at 12:00 am

    Abstract Growth and photosynthetic capacity of Cattleya warneri T. Moore seedlings cultivated in vitro were evaluated in two environments: (1) growth room (GR) with constant light, humidity, and temperature; and (2) greenhouse (GH) with variable humidity, temperature, and light intensity and quality. In both environments, two different tissue culture vessel lids were used: transparent plastic lids (TCVplastic) and non-transparent metal lids (TCVmetal). After 11 months of in vitro cultivation, five seedlings from each tissue culture vessel were evaluated for growth and photosynthetic capacity, while the other five seedlings of each tissue culture vessel were transferred to a greenhouse for acclimatization. Increased biomass production in vitro was observed in GH and GR (GH>GR). However, the photosynthetic capacity was not altered by the GH environment, since the net photosynthetic rate at 300 μmol m−2 s−1 (NPR300) was low in the final of period in all […]

  • Shining light on anther culture, a poorly understood regeneration route in passion fruit (Passiflora gibertii N. E. Brown): histological, hormonal, and phytochemical aspects
    on December 1, 2021 at 12:00 am

    Abstract Plant regeneration from reproductive organs is a poorly understood route in passion fruit. The present study provides a morpho-hystological description of somatic embryogenesis from anther culture in Passiflora gibertii N. E. Br., as well as a phytochemical profile and genetic analysis of ploidy in the generated embryos. Flower buds were collected during the pre-anthesis period. Murashige and Skoog culture medium was supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Embryogenic calluses were collected at different ages for histological, biochemical, and genetic evaluation of their ploidy. Anatomical characterization revealed the cell layers of the epidermal tissue involved in morphogenic events. Specifically, regions of intense cell division led to embryo formation and its subsequent regeneration. Moreover, as shown by hormonal and secondary compounds’ profiles, these regions were characterized by the presence of reserve compounds and […]

  • Micropropagation of the endangered species Malus niedzwetzkyana for conservation biodiversity in Kazakhstan
    on December 1, 2021 at 12:00 am

    Abstract Biodiversity conservation requires advanced and effective ex situ plant propagation techniques. The present study was conducted to optimize the micropropagation of a rare and endangered species of Malus niedzwetzkyana. The objective was the ex situ conservation and reintroduction, via micropropagation, of this species. Further, we aimed to obtain healthy planting material on a large-scale for commercial implementation. To study shoot multiplication, we examined the effects of three cytokinins (6-benzylaminopurine, kinetin, and thidiazuron) supplemented to Quoirin–Lepoivre (QL) culture medium. To examine the rooting of the shoots, we tested the effects of the macronutrient concentration of the QL medium (at 1×, 0.5×, and 0.25× concentration), and of its sucrose content (0%, 10%, 20%, and 30%). The optimized micropropagation technology provided a high propagation rate (28.77 new shoots per explant) on QL medium supplemented with 0.5 mg L−1 […]

  • Establishment of adventitious root cultures and assessment of secoiridoid production in the Chinese medicinal plant Gentiana scabra
    on December 1, 2021 at 12:00 am

    Abstract Gentiana scabra is an important medicinal herb that is used to treat inflammation, tuberculosis, rheumatism, cholecystitis, and hepatitis. The primary bioactive components in G. scabra are secoiridoids derived from the roots and rhizomes of the plant. In this study, we sought to establish a means of reliably producing these secoiridoids through G. scabra in vitro adventitious root culture. To that end, we studied the effect of phytohormones, light condition, explant type, and medium composition on adventitious root induction and secoiridoid production. Maximal induction frequency (100%) was ultimately achieved when using leaf explants cultured on Murashige and Skoog (MS) medium containing 3.0 mg L−1 NAA and 0.25 mg L−1 TDZ with a 16/8-h photoperiod, yielding 53.20 ± 3.15 roots per explant. We explored root growth curves over an 8-wk period in MS liquid medium prepared as above, and observed maximal fresh weight (FW) and dry weight (DW) values of 26.94 […]

  • Cannabis sativa L. photoautotrophic micropropagation: a powerful tool for industrial scale in vitro propagation
    on December 1, 2021 at 12:00 am

    Abstract Global demands for an in vitro culture of cannabis have never been more sought after as countries shift their paradigm towards legalization. Cannabis conventional (photomixotrophic) micropropagation has not been suitable enough for large-scale propagation due to a high degree of plant hyperhydricity, low growth rate, poor rooting, and acclimation efficiency. In the present study, cannabis photoautotrophic micropropagation method is introduced with the purpose of overcoming the difficulties that conventional micropropagation entails when conducted at a large scale. The roles of rockwool medium pH and moisture content, cutting length, basal wounding methods, light intensity, and culture vessel gas exchange capacity were assessed with the intention of increasing productivity of micropropagation method. The results showed 300 mL per vessel of fertilizer solution containing 5-mM MES buffer stabilized medium pH and increased rooting success. Both 5- and 7-cm […]

  • In vitro response of Indica rice microspores subjected to cold stress: a cytological and histological perspective
    on December 1, 2021 at 12:00 am

    Abstract This study investigated developmental changes in cold stressed microspores of Indica rice variety At 303. After 3 d at 10°C, approximately 76% of microspores were in the late uni-nucleate stage. Even after 5 d, 49% of viable cells remained in the late uni-nucleate stage without advancing to the bi-nucleate stage. In comparison, microspores undergoing normal gametogenesis in planta progressed rapidly from uni-nucleate to bi-nucleate stage eventually forming tri-nucleate pollen during this period. Thus, cold stress prevented normal microspore development and retained cells in the late uni-nucleate stage which is the most favorable stage for in vitro induction of sporophytic development in rice. Uni-nucleate microspores subjected to cold stress showed a characteristic pattern consisting of several minute vacuoles surrounding a centrally positioned nucleus, which can be interpreted as an early indicator of sporophytic determination in Indica rice microspores. […]

  • The development of clonal propagation and determination of phenolic profiles of in vitro-raised and field-raised leaves of Astragalus brachypterus Fischer (milkvetch) by LC-ESI-MS/MS analysis
    on December 1, 2021 at 12:00 am

    Abstract Astragalus brachypterus Fischer (milkvetch) is a perennial medicinal plant in the family Fabaceae. Astragalus species have been utilized as antiviral, antibacterial, antifungal, antioxidant, anticancer, hepatoprotective, wound healing, anti-diabetic, and diuretic for centuries in folk medicine. Objectives of this study were to determine an in vitro regeneration system for A. brachypterus and then to compare phenolic constituents of in vitro-raised and naturally raised leaves. Various plant explants (leaf, petiole, stem, and node) were taken from sterile seedlings that were obtained from germinated seeds. The explants were transferred to Murashige and Skoog (MS) medium supplemented with different cytokinins (benzyladenine (BA), kinetin (KIN), and thidiazuron (TDZ)), auxins (indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), naphthaleneacetic acid (NAA), and 2,4-dichlorophenoxy acetic acid (2,4-D)), and gibberellic acid (GA3) at various concentrations. […]

  • Flower power: floral reversion as a viable alternative to nodal micropropagation in Cannabis sativa.
    on December 1, 2021 at 12:00 am

    Abstract The legalization of recreational and medicinal Cannabis sativa L. has been gaining global momentum, therefore increasing interest in Cannabis micropropagation as growers look for large-scale solutions to germplasm storage and clean plant propagation. Mother plants used in commercial propagation are susceptible to pests and disease and require considerable space. While micropropagation can produce disease-free starting material in less space, current published in vitro methods are not robust and few report high multiplication rates. Furthermore, these micropropagation methods rely on photoperiod-sensitive plants maintained in a perpetual vegetative state. Current methods are not adaptable to long-term tissue culture of day-neutral cultivars, which cannot be maintained in perpetual vegetative growth. In this study, we developed a micropropagation system which uses Cannabis inflorescences as starting materials. This study used two Cannabis cultivars, two […]

  • In vitro plant regeneration from mature embryos of amphidiploid spelt Triticum spelta L.
    on December 1, 2021 at 12:00 am

    Abstract The present study reports an efficient in vitro plant regeneration system for amphidiploid (2n = 42) spelt wheat (Triticum spelta L.). Plant regeneration from mature embryos of winter spelt variety “Europe” was carried out as a two-step process. The first step was callus induction on a medium supplemented with 2 mg L−1 dichlorophenoxyacetic acid and 10 mg L−1 silver nitrate resulting in 96% callus formation. The second step resulted in 30% plant regeneration frequency from calluses transferred to Murashige and Skoog medium supplemented with 2 mg L−1 6-benzylaminopurine, 0.5 mg L−1 1-naphthaleneacetic acid, and 10 mg L−1 silver nitrate. Regeneration medium supplemented with 1 mg L−1 6-benzylaminopurine, 0.2 mg L−1 picloram, and the hormone-free medium turned out to be less effective in our experiments. Regenerated plants formed roots after transfer to half-strength Murashige and Skoog medium supplemented with […]

  • Estimation of endogenous indole-3-acetic acid in leaf explants and a protocol for direct regeneration in Lilium cv. Pavia
    on December 1, 2021 at 12:00 am

    Abstract A method for regeneration via direct organogenesis in Lilium cv. Pavia using leaf primary explants has been established. Outer leaves from in vitro plantlets of 10 to 12 cm were used as the primary explant for direct organogenesis. Shoots were directly regenerated from leaf explants within 25 d of culture on Murashige and Skoog (MS) augmented with 4.5 μM 2, 4-dichlorophenoxyacetic acid (2, 4-D), 5.4 μM α-naphthaleneacetic acid (NAA), and 2.2 μM 6-benzylaminopurine (BAP). Different regions of the same leaf explant exhibited varied responses. The acropetal 3 to 4 cm of the leaf (L1) produced the maximum number of shoots (75%) indicating its high regeneration ability whereas the response of middle portion (L2) and basipetal portion of the leaf (L3) exhibited a significantly decreasing trend of shoot induction with 37.5 and 12.5%, respectively. HPLC analysis of endogenous indole-3-acetic acid (IAA) concentration established significantly higher […]

  • Morpho-physiological investigations in transgenic tomato (Solanum lycopersicum L.) over expressing OsRGLP1 gene
    on December 1, 2021 at 12:00 am

    Abstract The OsRGLP1 gene was overexpressed under the control of CaMV 35S promoter in tomato (Solanum lycopersicum L.) plants using Agrobacterium-mediated transformation. The transgene expression was validated using RT-qPCR (real-time quantitative polymerase chain reaction) in the transgenic plants. Noticeable morpho-physiological traits, higher SOD activity, and elevated histological H2O2 were manifested by transgenic tomato lines. An increase in total chlorophyll and relative water content (RWC) in the leaves of transgenic lines was noted. The electrolytic leakage/conductivity (EC) in leaves of transgenic lines was lower than the control. The transgenic tomato lines showed dwarf phenotypic features in vegetative and floral traits. Interestingly, the vascular cambium regions in cross sections of stem showed secondary growth in radial symmetry. The transgenic lines also produced higher density and diversity of trichomes. The differences observed in transgenic lines […]

  • Shoot proliferation, embryogenic callus induction, and plant regeneration in Lepturus repens (G. Forst.) R. Br.
    on December 1, 2021 at 12:00 am

    Abstract Lepturus repens (G. Forst.) R. Br. is a perennial grass that grows on coral reef outcrops and sandy beaches and has drought tolerance and salt resistance. In this study, the internodes of L. repens were used as explants and cultured on Murashige and Skoog (MS) medium supplemented with 1.13 mg L−1 6-benzyladenine (BA) to successfully induce axillary shoots. Additional experimental results showed that cytokinins were a key factor for the successful induction and proliferation of shoots. BA induced 7.2 axillary shoots per explant, significantly more than kinetin (KIN), which induced 3.5 axillary shoots per explant. Shoots were proliferated more successfully on medium supplemented with 1.13 mg L−1 BA and a low concentration of 0.09 mg L−1 1-naphthaleneacetic acid (NAA) than BA alone. Shoot bases were used as explants for embryogenic callus induction and subsequent plant regeneration, which was possible on MS medium supplemented with 0.11 mg L−1 2,4-D, […]

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