In Vitro Cellular & Developmental Biology - Plant最新文献

The objective of this study was to assess the effect of sodium nitroprusside (SNP) on the morphology, physiology, and biochemistry of Citrus indica Yu. Tanaka, which is considered a native and endangered species in northeast India. Moreover, C. indica seeds do not germinate well in their natural habitat, and the species does not have a good commercial standing in terms of valuable metabolite synthesis. Therefore, an efficient quick-growth method with improved phytochemical synthesis has been accomplished. For mass production employing micropropagation with embryos as explants, Murashige and Skoog (MS) medium was supplemented with 0.2 µM, 0.4 µM, and 0.6 µM of SNP, 3.0 µM of 6-benzylaminopurines (BAP), and 4.0 µM of thidiazuron (TDZ). The results showed that 0.4 µM of SNP alone or in conjunction with 3.0 µM of BAP was sufficient to improve morphological characteristics. In addition, established clones were examined for membrane stability by measuring electrolyte leakage (EL) and malondialdehyde (MDA), as well as photosynthetic pigment and carotenoids. In almost all the cases, high values were recorded when 0.4 µM of SNP was used, either singly or in conjunction with 3.0 µM of BAP in the MS medium. Clonally produced plantlets were also subjected to phytochemical screening and antioxidant activities, such as hydrogen peroxide (H₂O₂), superoxide anion, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, wherein these tests also revealed incorporation of SNP exhibited enhanced phytochemical concentrations and antioxidant activities. This research can thus be used to mass produce in vitro plantlets with considerably higher quantities of secondary metabolites, which may make them more resistant to environmental challenges than their natural counterparts without treatment.

Abstract

Pepper (Capsicum annuum L.) is one of the most widely cultivated species and is highly valued for its pungency. The pungency of pepper is primarily attributed to a group of chemical compounds known as capsaicinoids. These compounds are synthesized in the placental tissue of pepper fruits through the activation of specific genes and enzymes, thereby contributing to their pungency. Protoplast-based gene expression systems have been considered an efficient method for gene function studies, protein-protein interactions, promoter analysis, and subcellular localization. Here, we optimized an efficient protocol for isolating protoplasts from pepper placental tissues and the polyethylene glycol (PEG)-mediated transient expression of green fluorescent proteins (GFP). Several factors affecting GFP expression in intact protoplasts were evaluated and optimized in this study. Protoplast isolation was carried out using 2.0% cellulase “onozuka” R-10 and 0.3% macerozyme R-10 solution. Different amounts of plasmid DNA and various incubation times for transfection with 40% PEG resulted in different transfection efficiencies (78–85%). The highest GFP transformation efficiency was observed when 120 µL protoplast suspension (3 × 10⁷ to 5 × 10⁷/mL) was mixed with 15 µg plasmid DNA and incubated for 25 min with an equal volume of 40% PEG. The improved protocol described in this study can be helpful for the isolation and transfection of pepper placenta-originated protoplasts and for the rapid investigation of pepper gene functions.

The primary goal of this study was to compare the multiplication rates of yam varieties propagated through organogenesis and somatic embryogenesis (SE). Callus was induced from axillary bud explants of three genotypes of Dioscorea rotundata (Asiedu, Ekiti2a, and Kpamyo) and two genotypes of Dioscorea alata (Swaswa and TDa2014) cultured in Murashige and Skoog (MS) medium containing 9.1 µM 2,4-dichlorophenoxylacetic acid and 5.4 µM naphthaleneacetic acid. Plantlets were regenerated in MS containing 4.4 µM benzylaminopurine and 34 µM uniconazole-P through SE. Single-node cuttings of the five genotypes were grown in MS for 8 wk via organogenesis. The SE and organogenesis regenerants were acclimatized and potted in a 2 (propagation techniques (PTs)) × 5 (genotypes) factorial arranged in a completely randomized design (r = 10). The multiplication ratios (MR), number of tubers (NoT) of the SE, and organogenesis regenerants were collected and analyzed using ANOVA, and means were separated using DMRT (P ≤ 0.05). The SE and organogenesis MR ranged from 1:2 (TDa2014) to 1:8 (Asiedu) and 1:4 (Asiedu) to 1:5 (Ekiti2a and TDa2014), respectively. The NoT differed among genotypes, ranging from 1.15 ± 0.49 (Swaswa) to 2.45 ± 1.39 (Asiedu), and between PTs, ranging from 1.42 ± 0.70 (SE) to 1.86 ± 1.11 (organogenesis). The optimum propagation pathway was genotype-specific.

Lepidium ostleri S.L. Welsh & Goodrich (Ostler’s peppergrass) is an endemic plant species restricted to Ordovician limestone outcrops associated with the San Francisco Mountain Range in western Utah. Due to restricted population distribution and proximity to modern mining operations, L. ostleri is a species of conservation interest. This study focused on the development of a micropropagation protocol for propagating mature plants using plant tissue culture methods. Indirect shoot organogenesis was obtained from L. ostleri explants on Murashige and Skoog (MS) medium augmented with various concentrations of BAP (6-Benzylaminopurine), kinetin (N6-furfuryladenine), and IAA (indole-3-acetic acid). Plantlets supporting shoots grown in vitro were pulse treated with differing strengths of indole-3-butyric acid (IBA) and transferred to sterile soil. Following root induction, plantlets were acclimated to ambient conditions. The successful development of a micropropagation protocol supports management activities for L. ostleri and also contributes to in vitro propagation knowledge at the species, genus, and family levels.