Comprehensive Medicinal Plant Database

Tissue Culture Literation

Plant latin namePerilla frutescens (L.) Britton var. acuta Kudo
Literature codePerilla_frutescens_var._acuta-Ref-1
ReferenceZhang T et al., BIOLOGIA PLANTARUM 49 (3): 423-426 (2005)
SummaryA rapid plantlet regeneration system for Perilla frutescens was established from cotyledon and hypocotyl explants. A maximum of 91.06 % cotyledon and 76.4 % hypocotyl explants could directly produce shoots (3.09 ± 0.18 shoots per explants) on Murashige and Skoog (MS) medium. The optimum hormone combinations were 4.44 μM 6-benzylaminopurine (BA) for cotyledon and 2.22 μM BA + 2.85 μM indole-3-acetic acid (IAA) for hypocotyls. Rooting was induced on half-strength hormone-free MS medium. After transplantation to soil, approximate 80 % of the regenerated plantlets could survive, flower and fruit. Moreover, some morphological abnormalities were found among the regenerated plants.
ObjectivesIn present study, we focused on establishing a high frequency in vitro regeneration system of Perilla frutescens, aiming to get useful somatic mutants and to pave a way for further genetic manipulation of this plant.
MaterialsThe seeds were provided by Gansu Academy of Agricultural Science
ExplantThe seeds were surface-sterilized in 70 % ethanol for 30 s, then in 0.1 % mercuric chloride for 10 min, and rinsed 4 times (5 min each) in sterile distilled water.
Initial cultureThe sterilized seeds were placed on MS basal medium without any growth regulator in a 200 cm3 conical flask to germinate at 25 ± 2 °C with 16-h photoperiod (irradiance of 65 μmol m-2 s-1, cool white fluorescent light).
Shoot multiplicationAbout 3 - 5 mm long hypocotyls segments of 5-d-old seedlings:solidified MS medium supplemented with 3 % sucrose, 0.7 % agar and +2.22μM BA+2.85μM IAA.About 0.2 cm2 cotyledon segments: solidified MS medium supplemented with 3 % sucrose, 0.7 % agar and 4.44μM BA.
RootingAfter 40 d, the regenerated shoots (2 - 5 cm) excised and transferred onto half-strength MS media without growth regulators rooted easily.
Acclimation10 d later, the resultant plantlets with well-developed roots were pulled up from media, washed with tap water and then transplanted to pots filled with autoclaved Vermi-compost. The pots were covered with polyethylene film and kept in the growth chamber. Two days later, the film was removed and the plantlets were irrigated with half-strength MS solution.
PlantingAfter two weeks of acclimation, the alive were transplanted to larger pots containing garden soil to allow a further growth for 10 d, and then moved to outdoors.
Cultivation conditions
Traints of regenerantsWe found two morphological abnormalities, one with initial stem showing inhibited apical growth and new branches growing up from axillary bud, and another having calyxes without epidermal hairs. The regenerated plantlets and wild types of Perilla frutescens generally having very dense and long hairs on intra- and extra-epidermis of their calyxes. However, the possibility that the abnormalities were induced by environmental factors, such as nutrient, hormones and illumination period cannot be excluded.
Ingredients analyzed
Analitical methods