In vitro Propagation of Vanda testacea (Lindl.) Reichb. F, a Medicinally Important Threatened Orchid

The present study was planned to enable in vitro conservation of Vanda testacea, a highly medicinal orchid species through in vitro asymbiotic seed germination technique in Mitra orchid medium supplemented with cytokinins (Kn - 4.65 μM, BAP - 4.44 μM), and auxin (NAA- 5.37 μM). The germination frequency and initiation of germination was higher in NAA augmented medium and seedlings developed in 12.50 ± 0.50 weeks. Coconut water (20%) proved optimum for the multiplication of protocorm like bodies. Activated charcoal successfully checked the release of brownish exudates in the cultures. Plant Tissue Cult. & Biotech. 31(2): 153-160, 2021 (December)

In vitro propagation from nodal segments of Lippia origanoides (chemotype A)

ABSTRACT: This research described an efficient micropropagation protocol for Lippia origanoides (Verbenaceae). Sterile seeds were used to obtain germinated seedlings in Murashige and Skoog medium (MS) supplemented with sucrose and agar. The nodal segments obtained from seedlings were grown on MS medium supplemented with different concentrations of gibberellic acid (GA), benzylaminopurine (BAP) and 1-naphthalenacetic acid (NAA) with BAP. The callus induction, shoots length, shoots number and root length, were analyzed. The treatments showed high percentage of callus formation at 0.5 to 1.5 mg L-1 of BAP alone or in combination with NAA (0.1 mg L-1). The highest value of shoot number per nodal segments was obtained at 1.5 mg L-1 of BAP (4.3 ± 0.8). The obtained plantlets were better rooted in vitro in the absence of plant growth regulators (PGRs) and they showed acclimatization rate of 90%. We reported a protocol for in vitro propagation and acclimatization of L. origanoides for A chemotypes from Colombia.

In vitro propagation of select tomatillo (Physalis ixocarpa Brot. ex Horm.) plant families

The objective was to propagate select Physalis ixocarpa plants in vitro, acclimatize them and describe their phenological cycle. The in vitro response of stem apices was evaluated in families from the Tecozautla 04, Manzano Tepetlixpa and Morado San Miguel varieties. The apices were cultured in a medium containing Murashige and Skoog inorganic salts (100 %), supplemented with 0.4 mg·L-1 thiamine, 60 mg·L-1 L-cysteine, 100 mg·L-1 myo-inositol, 0.5 mg·L-1 nicotinic acid, 0.5 mg·L-1 pantothenic acid, 3 % sucrose and 7 g·L-1 agar, without growth regulators and the pH adjusted to 5.7 ± 0.1. in vitro rooting was done for 30 days, with 16 h of light at 3,000 μmol∙m-2∙s-1. The variables evaluated in vitro were seedling height, vigor, callus presence, root length, and number of leaves, roots, stems and buds. Plants produced in vitro were acclimatized and transplanted in greenhouses to follow their phenological cycle. The variables evaluated in acclimatization and phenological cycle were plant height and number of leaves, buds, flowers and set fruits. A completely randomized design was used for the in vitro evaluation, and randomized complete blocks for the greenhouse. The families with the best morphogenic responses in vitro were Tecozautla 04 and Manzano, and in phenological development they presented greater plant height. In acclimatization, survival was 100 % in all clones. in vitro responses, acclimatization and phenology depended on the variety and families.

In Vitro Propagation of XXY Undifferentiated Mouse Spermatogonia: Model for Fertility Preservation in Klinefelter Syndrome Patients

Klinefelter syndrome (KS) is characterized by a masculine phenotype, supernumerary sex chromosomes (usually XXY), and spermatogonial stem cell (SSC) loss in their early life. Affecting 1 out of every 650 males born, KS is the most common genetic cause of male infertility, and new fertility preservation strategies are critically important for these patients. In this study, testes from 41, XXY prepubertal (3-day-old) mice were frozen-thawed. Isolated testicular cells were cultured and characterized by qPCR, digital PCR, and flow cytometry analyses. We demonstrated that SSCs survived and were able to be propagated with testicular somatic cells in culture for up to 120 days. DNA fluorescent in situ hybridization (FISH) showed the presence of XXY spermatogonia at the beginning of the culture and a variety of propagated XY, XX, and XXY spermatogonia at the end of the culture. These data provide the first evidence that an extra sex chromosome was lost during innate SSC culture, a crucial finding in treating KS patients for preserving and propagating SSCs for future sperm production, either in vitro or in vivo. This in vitro propagation system can be translated to clinical fertility preservation for KS patients.