Regeneration potentials in Gerbera jamesonii Bolus ex. Hook f. from tissues culture system was studied using leaf, petiole and root explants. In vitro regeneration, callus induction and root formation were optimized by manipulation of growth regulators during organogenesis. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), alpha-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2,4-D), Indole-3-acetic acid (IAA), Indole-3-Butyric acid (IBA), N6-[2-Isopentenyl]adenine (2iP), Kinetin and Zeatin were used to initiate cultures. These plant growth regulators were added to Murashige and Skoog medium in different combinations and concentrations. Adventitious shoots were obtained from petiole explants cultured on Murashige and Skoog (MS) medium supplemented with 2.0 mg L(-1) BAP and 0.5 mg L(-1) NAA. Effectiveness of shoot regeneration medium, type of growth regulator used and duration of induction period were investigated. Leaf explants cultured on MS medium supplemented with 1.0 mg L(-1) BAP and 2.0 mg L(-1) 2, 4-D showed the best results for callus induction. Root explants were found to be non-regenerative in all experiments conducted. Petiole segment was identified as the best explant for regeneration of this species. Regenerated plants were rooted on Murashige and Skoog basal medium. Plantlets were then transferred to field with 75% survival rate.
Seven different hormone treatments, namely 6-benzylaminopurine (BAP) at 2, 3 mg L(-1) was applied singly and in combination with Indole Acetic Acid (IAA) at 0.18, 0.8 and 1.8 mg L(-l), BAP at 3.3 mg L(-l) in combination with IAA at 1.8 and 3.3 mg L(-l) and triple combination of BAP at 2.3, IAA at 1.8 and Gibberellic acid (GA3) at 1.0 mg L(-1) were tested, over four different incubation periods of 30, 45, 60 and 75 days, for their effect in the proliferation and growth of Smooth cayenne pineapple shoot-tip culture. Combined application of BAP at 3.3 and IAA at 1.8 mg L(-1) induced the highest proliferation of 19 shoots/explant and the highest total of 121 and 125 shoots over 4 cycles of multiplication. Raising the IAA to 3.3 mg L(-1) resulted in the lowest proliferation and stunted shoots. Incorporation of GA3 improved the shoot length but caused drastic reduction in proliferation. The other treatments showed an intermediate effect.
Plants tend to acclimatize to unfavourable environs by integrating growth and development to environmentally activated signals. Phytohormones strongly regulate convergent developmental and stress adaptive procedures and synchronize cellular reaction to the exogenous and endogenous conditions within the adaptive signaling networks. Gibberellins (GA), a group of tetracyclic diterpenoids, being vital regulators of plant growth, are accountable for regulating several aspects of growth and development of higher plants. If the element of reproduction is considered as an absolute requisite then for a majority of the higher plants GA signaling is simply indispensable. Latest reports have revealed unique conflicting roles of GA and other phytohormones in amalgamating growth and development in plants through environmental signaling. Numerous physiological researches have detailed substantial crosstalk between GA and other hormones like abscisic acid, auxin, cytokinin, and jasmonic acid. In this review, a number of explanations and clarifications for this discrepancy are explored based on the crosstalk among GA and other phytohormones.
Phytohormones comprise a variety of trace bioactive compounds that can stimulate cell growth and promote metabolic shifts. In the present work, a two-stage screening strategy was innovatively established to identify positive phytohormones for enhancement of astaxanthin and lipid coproduction in microplate-based cultures of mixotrophic Chromochloris zofingiensis. The results showed that auxins were the most efficient stimulators for astaxanthin accumulation. The maximum content of 13.1 mg/g and yield of 89.9 mg/L were obtained using indole propionic acid (10 mg/L) and indoleacetic acid (7.8 mg/L), representing the highest levels of astaxanthin in this microalga reported to date. Total lipids with the highest content (64.5% DW) and productivity (445.7 mg/L/d) were coproduced with astaxanthin using indoleacetic acid. Statistical analysis revealed close relations between phytohormones and astaxanthin and lipid biosynthesis. This study provides a novel original strategy for improving astaxanthin and lipid coproduction in C. zofingiensis using the selected phytohormones as positive stimulators.
Efficient plant regeneration of Saintpaulia ionantha (African violet) has been obtained in the present study. MS medium supplemented with 1.0 mg L(-1) IAA and 2.0 mg L(-1) Zeatin resulted in 100% shoot regeneration and induced the highest number of shoots (average 15.0 +/- 0.8 shoots per explant) after being cultured for 8 weeks. The above hormone combination was optimum for shoot regeneration. Most of Saintpaulia ionantha plantlets derived from tissue culture system could be hardened and transferred to the greenhouse conditions with 84.0 +/- 1.6% success rate. However, regenerated plantlets of Saintpaulia ionantha (even after 12-months-old) failed to flower. Morphological characters of regenerated plantlets of Saintpaulia ionantha were observed and compared with in vivo (intact) plants. Regenerated plantlets showed some differences in morphological characters, such as height and leaf size, texture and colour, but the plantlets showed no variation in leaf arrangement and leaf margin. However, the morphological characters of the regenerated plantlets were found to be unstable.
Differential effect of plant growth regulators and additives in proliferation of 18-month-old calli of Ananas comosus L. cv. Moris were assessed in vitro. The proliferation of callus relied on the growth regulators and additives. Of the different auxins supplemented in the Murashige and Skoog (MS) media, 32.22 microM alpha-naphthaleneacetic acid (NAA) gave the highest mean fresh weight of callus (46.817 g). Medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) was inferior to NAA, while b-naphthoxy acetic acid (BNOA) and p-chlorophenoxy acetic acid (4-CPA) were not effective in proliferating 18-months old callus. Addition of casein hydrolysate and coconut water to NAA supplemented medium showed better proliferation and production of callus. However, in terms of callus production, NAA at 32.22 microM was economically better.
This study represents the first paper of the effects of growth regulators on the physiochemical and phytochemical properties of the wax apple fruit, a widely cultivated fruit tree in southeast Asia. Net photosynthesis, sucrose phosphate synthase (SPS) activity, peel color, fruit firmness, juice content, pH value, total soluble solids (TSSs), and the sugar acid ratio were all significantly increased in growth regulators (PGRs) treated fruits. The application of gibberellin (GA(3)), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxy acetic acid (2,4-D) significantly reduced titratable acidity and increased total sugar and carbohydrate content compared to the control. The 50 mg/L GA₃, 10 mg/L NAA, and 5 mg/L 2,4-D treatments produced the greatest increases in phenol and flavonoid content; vitamin C content was also higher for these treatments. PGR treatment significantly affected chlorophyll, anthocyanin, and carotene content and produced higher phenylalanine ammonia lyase (PAL) and antioxidant activity levels. There was a positive correlation between peel color and TSS and antioxidant activity and both phenol and flavonoid content and PAL activity and anthocyanin formation. A taste panel assessment was also performed, and the highest scores were given to fruits that had been treated with GA₃ or auxin. The study showed that application of 50 mg/L GA₃, 10 mg/L NAA, and 5 mg/L 2,4-D once a week from bud development to fruit maturation increased the physiochemical and phytochemical properties of wax apple fruits.
The SCF complex is a widely studied multi-subunit ring E3 ubiquitin ligase that tags targeted proteins with ubiquitin for protein degradation by the ubiquitin 26S-proteasome system (UPS). The UPS is an important system that generally keeps cellular events tightly regulated by purging misfolded or damaged proteins and selectively degrading important regulatory proteins. The specificity of this post-translational regulation is controlled by F-box proteins (FBPs) via selective recognition of a protein-protein interaction motif at the C-terminal domain. Hence, FBPs are pivotal proteins in determining the plant response in multiple scenarios. It is not surprising that the FBP family is one of the largest protein families in the plant kingdom. In this review, the roles of FBPs, specifically in plants, are compiled to provide insights into their involvement in secondary metabolites, plant stresses, phytohormone signalling, plant developmental processes and miRNA biogenesis.
This review paper discussed about publications related to micropropagation of bamboo species. In recent years, the application of tissue culture technique like in vitro micropropagation has been used to meet the demands for bamboo planting materials. In the past 30 years, protocols for micropropagation of various bamboo species have been established by researchers from all over the world. The controlling factors for cultures such as the explants, culture medium, carbon sources, combination and concentration of plant growth regulators and other additional additives are varied. The controlling factors are crucial in developing successful regeneration protocols for various bamboo species. This paper attempts to review and summarize the available and up-to-date information regarding in vitro micropropagation of bamboos.
Plant tissue or cell culture keeps a significant role in micro-propagation in the plant production industry. Combination of 6-Benzylaminopurine (BAP) and other plant growth regulators like 1-Naphthaleneacetic acid (NAA) or Indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was used in the most of the research in tissue culture. The study was carried out to investigate the optimization of the concentration of IBA and BAP combination (0, 0.25, 0.50, 1.0, 1.50, 2.0, 2.5, 3.0 and 3.5 mg/l) for the root, callus and leaf proliferation from the leaf cutting slice. The highest number (6.75) of root proliferation was observed in the concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination. The callus initiation was found in the concentration of IBA 1.0-3.5 mg/l+BAP 1.0-2.0 mg/l. However, the highest callus weight was observed at the concentration of IBA 1.5 mg/l+BAP 1.0 mg/l combination than other combination of concentrations. Positively leaf initiation and formation was better in the concentration of IBA 1-3.5 mg/l+BAP 1.0-2.0 mg/l combination. In addition, the 2,2-diphenyl-2-picrylhydarzyl (DPPH) free radical scavenging potential was higher (70.1%) in leaves extract than in callus extracts (46.3%) at the concentration of 10 mg/ml though both extracts had lower DPPH free radical scavenging activity compared to the positive control, vitamin C and BHT. Theresults conclude that the optimum concentration was IBA 1.5 mg/l+BAP 1.0 mg/l combination to produce callus cell proliferation and concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination was the optimum for root proliferation of broccoli in vitro.
In this paper, a micropropagation protocol of sugar palm (Arenga pinnata Wurmb Merr) through callogenesis and somatic embryogenesis was examined. Callus induction frequency and somatic embryogenesis response were dependent on plant growth regulators (PGRs) and genotype. Semi-compact and compact embryogenic calluses were induced from excised immature zygotic embryo (IZE) cultured on semi-solid MS (Murashige & Skoog, 1962) medium supplemented with various concentration and combination of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyl aminopurine acid (BAP). MS medium supplemented with 0.4 mg/L 2,4-D and 0.5 mg/L BAP was found optimum to induce 100% rate of embryogenic calluses and maximum degree of callus formation after 8 and 12 weeks of culture. The incorporation of increased sucrose concentration (60.0 g/L) and 2.0 g/L casein hydrolysate (CH) to the culture medium with similar PGRs composition enhanced the induction of globular somatic embryos (SEs), while addition of silver nitrate (AgNO3) produced SEs of different stages. SEs maturated in MS medium containing 1.0 mg/L BAP and 1.0 mg/L naphthalene-acetic acid (NAA) formed cotyledon-stage embryos. Clonal roots regeneration was obtained on half-strength MS devoid of PGRs after 4 months of culture. Frequent subcultures increased embryogenesis rate favourably.
Callus was induced from mangosteen (Garcinia mangostana L.) young purple-red leaves on Murashige and Skoog basal medium with various combinations of plant growth regulators. Murashige and Skoog medium with 4.44 µM 6-benzylaminopurine and 4.52 µM 2,4-dichlorophenoxyacetic acid was the best for friable callus induction. This friable callus was used for the initiation of cell suspension culture. The effects of different combinations of 6-benzylaminopurine and 2,4-dichlorophenoxyacetic acid, carbon sources and inoculum sizes were tested. It was found that combination of 2.22 µM 6-benzylaminopurine + 2.26 µM 2,4-dichlorophenoxyacetic acid, glucose (30 g/l) and 1.5 g/50 ml inoculum size was the best for cell growth. Callus and cell suspension cultures were then treated either with 100 µM methyl jasmonate as an elicitor for 5 days, or 0.5 g/l casein hydrolysate as an organic supplement for 7 days. Metabolites were then extracted and profiled using liquid chromatography-time of flight mass spectrometry. Multivariate discriminant analyses revealed significant metabolite differences (P ≤ 0.05) for callus and suspension cells treated either with methyl jasmonate or casein hydrolysate. Based on MS/MS data, methyl jasmonate stimulated the production of an alkaloid (thalsimine) and fatty acid (phosphatidyl ethanolamine) in suspension cells while in callus, an alkaloid (thiacremonone) and glucosinolate (7-methylthioheptanaldoxime) was produced. Meanwhile casein hydrolysate stimulated the production of alkaloids such as 3ß,6ß-dihydroxynortropane and cis-hinokiresinol and triterpenoids such as schidigerasaponin and talinumoside in suspension cells. This study provides evidence on the potential of secondary metabolite production from in vitro culture of mangosteen.
An in vitro protocol has been established for clonal propagation of Nothapodytes nimmoniana which is an important source of Camptothecin (CPT). Elite source was identified based on the chemical potency to accumulate the optimum level of CPT. Different types and concentrations of plant growth regulators were used to study their effect on inducing multiple shoots from the explants regenerated from embryos of N. nimmoniana. Of these, a combination of N6-benzyladenine (0.2 mg L(-1)) and Indole-3-butyric acid (IBA) (0.1 mg L(-1)) proved optimum for differentiating multiple shoots in 90.6 % of the cultures with an average of 10.24 shoots per explant obtained within 8 weeks of inoculation. Nearly, 92 % of the excised in vitro shoots rooted on half strength Murashige and Skoog (MS) medium containing 0.05 % activated charcoal, supplemented with 1-naphthaleneacetic acid and IBA at 0.1 mg L(-1) each. The micropropagated plants were evaluated for their genetic fidelity by employing inter simple sequence repeats (ISSR) markers. Ten individuals, randomly chosen from a population of 145 regenerants, were compared with the donor plant. The regenerated plants were also evaluated for their chemical potency using high-performance liquid chromatography (HPLC) analysis of CPT content. The true-to-type nature of the micropropagated plants was confirmed based on their monomorphic banding profiles with that of the mother plants using ISSR markers. Besides, HPLC evaluation of the CPT content confirmed the existence of chemical uniformity among the regenerated plants and the elite mother plant.
Black pepper (Piper nigrum L.) is one of the most popular and oldest spices in the world with culinary uses and various pharmacological properties. In order to satisfy the growing worldwide demand for black pepper, improved productivity of pepper is highly desirable. A primary constraint in black pepper production is the non-synchronous nature of flower development and non-uniform fruit ripening within a spike. The uneven ripening of pepper berries results in a high labour requirement for selective harvesting contributes to low productivity and affects the quality of the pepper products. In Malaysia, there are a few recommended varieties for black pepper planting, each having some limitations in addition to the useful characteristics. Therefore, a comparative study of different black pepper varieties will provide a better understanding of the mechanisms regulates fruit development and ripening. Plant hormones are known to influence the fruit development process and their roles in black pepper flower and fruit development were inferred based on the probe-based gene expression analysis and the quantification of the multiple plant hormones using high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). In this study, jasmonic acid and salicylic acid were found to play roles in flowering and fruit setting, whereas auxin, gibberellin and cytokinins are important for fruit growth. Abscisic acid has positive role in fruit maturation and ripening in the development process. Distinct pattern of plant hormones related gene expression profiles with the hormones accumulation profiles suggested a complex network of regulation is involved in the signaling process and crosstalk between plant hormones was another layer of regulation in the black pepper fruit development mechanisms. The current study provides clues to help in elucidating the timing of the action of each specific plant hormone during fruit development and ripening which could be applied to enhance our ability to control the ripening process, leading to improving procedures for the production and post-harvest handling of pepper fruits.
A study was conducted to determine the effects of a plant growth regulator (paclobutrazol, PBZ) and commercial
fertilizer (Krista-K Plus) as a source of potassium nitrate (KNO3
) on the growth of Xanthostemon chrysantus. It was
also attempted to investigate the anatomical changes in the leaf and stem after the treatment. Nine treatments, i.e.
control (no PBZ and Krista-K Plus application), 0 PBZ gL-1 + 100 g Krista-K Plus, 0 PBZ gL-1 + 200 g Krista-K Plus,
0.125 PBZ gL-1 + 0 g Krista-K Plus, 0.125 PBZ gL-1 + 100 g Krista-K Plus, 0.125 PBZ gL-1 + 200 g Krista-K Plus, 0.25
PBZ gL-1 + 0 g Krista-K Plus, 0.25 PBZ gL-1 + 100 g Krista-K Plus and 0.25 PBZ gL-1 + 200 g Krista-K Plus, were
tested. PBZ was soil drenched at the commencement of the study while Krista-K Plus was applied at three-month
intervals. Plant growth performances such as tree height, diameter at breast height, canopy diameter and leaf area
were recorded monthly throughout the study period. Stem and leaf samples were collected before the application
of treatments and after six months of treatments for anatomical observation by using electron microscope. Plant
height, diameter at breast height, crown diameter and leaf area were significantly reduced with the application of
PBZ. Palisade parenchyma thickness was increased by 33.83% with 0.25 PBZ gL-1 + 200 g Krista-K Plus, while only
2.44% increment recorded in the control tree. Xylem thickness in the stem was reduced by 21.81% after treated with
the highest dosage of PBZ, while the control tree only had 1.78% increment. Spongy parenchyma thickness in the leaf
was unaffected. However, palisade parenchyma was found the thickest after combined treatment with 0.25 PBZ gL-1
+ 200 g Krista-K Plus. Micrograph images of the cross-section of leaf lamina and stem showed that the cells were
tightly arranged in response to the application of PBZ.
Xylem is an essential conductive tissue in vascular plants, and secondary cell wall polymers found in xylem vessel elements, such as cellulose, hemicellulose, and lignin, are promising sustainable bioresources. Thus, understanding the molecular mechanisms underlying xylem vessel element differentiation is an important step towards increasing woody biomass and crop yields. Establishing in vitro induction systems, in which vessel element differentiation is induced by phytohormonal stimuli or by overexpression of specific transcription factors, has been vital to this research. In this review, we present an overview of these in vitro induction systems, and describe two recently developed in vitro induction systems, VISUAL (Vascular cell Induction culture System Using Arabidopsis Leaves) and the KDB system. Furthermore, we discuss the potentials and limitations of each of these new in vitro induction systems for advancing our understanding of the molecular mechanisms driving xylem vessel element differentiation.
The production of nitrogenase enzyme and auxins by free living diazotrophs has the potential to influence the growth of host plants. In this study, diazotrophs were grown in the presence of various concentrations of nitogen (N) to determine the optimal concentration of N for microbial growth stimulation, promotion of gaseous N (N2) fixation, and phytohormone production. Therefore, we investigate whether different levels of N supplied to Herbaspirillum seropedicae (Z78) have significant effects on nitrogenase activity and auxin production. The highest nitrogenase activity and the lowest auxin production of H. seropedicae (Z78) were both recorded at 0 gL(-1) of NH4Cl. Higher levels of external N caused a significant decrease in the nitrogenase activity and an increased production of auxins. In a subsequent test, two different inoculum sizes of Z78 (10(6) and 10(12) cfu/ml) were used to study the effect of different percentages of acetylene on nitrogenase activity of the inoculum via the acetylene reduction assay (ARA). The results showed that the optimal amount of acetylene required for nitrogenase enzyme activity was 5% for the 10(6) cfu/ml inoculum, whereas the higher inoculum size (10(12) cfu/ml) required at least 10% of acetylene for optimal nitrogenase activity. These findings provide a clearer understanding of the effects of N levels on diazotrophic nitrogenase activity and auxin production, which are important factors influencing plant growth.
Auxin and cytokinin regulate different critical processes involved in plant growth and environmental feedbacks. These plant hormones act either synergistically or antagonistically to control the organisation, formation and maintenance of meristem. Meristem cells can be divided to generate new tissues and organs at the locations of plant postembryonic development. The aboveground plant organs are created by the shoot apical meristem (SAM). It has been proposed that the phytohormone, cytokinin, plays a positive role in the shoot meristem function, promotes cell expansion and promotes an increasing size of the meristem in Arabidopsis, whereas it has the reverse effects in the root apical meristem (RAM). Over the last few decades, it has been believed that the apically derived auxin suppresses the shoot branching by inactivating the axillary buds. However, it has recently become clear that the mechanism of action of auxinis indirect and multifaceted. In higher plants, the regulatory mechanisms of the SAM formation and organ separation are mostly unknown. This study reviews the effects and functions of cytokinin and auxin at the shoot apical meristem. This study also highlights the merger of the transcription factor activity with the actions of cytokinin/auxin and their complex interactions with the shoot meristem in rice.
A procedure was developed for in vitro propagation of Curculigo latifolia through shoot tip culture. Direct regeneration and indirect scalp induction of Curculigo latifolia were obtained from shoot tip grown on MS medium supplemented with different concentrations and combinations of thidiazuron and indole-3-butyric acid. Maximum response for direct regeneration in terms of percentage of explants producing shoot, shoot number, and shoot length was obtained on MS medium supplemented with combination of thidiazuron (0.5 mg L(-1)) and indole-3-butyric acid (0.25 mg L(-1)) after both 10 and 14 weeks of cultures. Indole-3-butyric acid in combination with thidiazuron exhibited a synergistic effect on shoot regeneration. The shoot tips were able to induce maximum scalp from basal end of explants on the medium with 2 mg L(-1) thidiazuron. Cultures showed that shoot number, shoot length, and scalp size increased significantly after 14 weeks of culture. Transferring of the shoots onto the MS medium devoid of growth regulators resulted in the highest percentage of root induction and longer roots, while medium supplemented with 0.25 mg L(-1) IBA produced more numbers of roots.
The use of in vitro culture has been accepted as an efficient technique for clonal propagation of many woody plants. In the present research, we report the results of a number of experiments aimed at optimizing micropropagation protocol for tea (Camellia sinensis (L.) O. Kuntze) (clone Iran 100) using nodal segments as the explant. The effect of different combinations and concentrations of plant growth regulators (PGR) (BAP, TDZ, GA₃) on shoot multiplication and elongation was assessed. The influence of exposure to IBA in liquid form prior to transfer to solid media on rooting of tea microshoots was investigated. The results of this study showed that the best treatment for nodal segment multiplication in terms of the number of shoot per explant and shoot elongation was obtained using 3 mg/L BAP in combination with 0.5 mg/L GA₃. TDZ was found to be inappropriate for multiplication of tea clone Iran 100 as it resulted in hyperhydricity especially at concentrations higher than 0.05 mg/L. Healthy shoots treated with 300 mg/L IBA for 30 min followed by transfer to 1/2 strength MS medium devoid of PGR resulted in 72.3% of shoots producing roots and upon transferring them to acclimatization chamber 65% survival was obtained prior to field transfer.