| Plant latin name | Perilla frutescens (L.) Britton var. acuta Kudo |
| Literature code | Perilla_frutescens_var._acuta-Ref-1 |
| Reference | Zhang T et al., BIOLOGIA PLANTARUM 49 (3): 423-426 (2005) |
| Summary | A 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. |
| Objectives | In 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. |
| Materials | The seeds were provided by Gansu Academy of Agricultural Science |
| Explant | The 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 culture | The 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 multiplication | About 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. |
| Rooting | After 40 d, the regenerated shoots (2 - 5 cm) excised and transferred onto half-strength MS media without growth regulators rooted easily. |
| Acclimation | 10 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. |
| Planting | After 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 regenerants | We 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. |
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