13551

10.30848/PJB2026-5(13)

An efficient method of rooting from in vitro culture of zygotic embryos of Pinus roxburghii Sarg.

Aroosha Ashraf, Muhammad Akram and Faheem Aftab

This study was conducted to explore the possible rooting efficacy of the mature zygotic embryo of Pinus roxburghii. Megagametophytes with zygotic embryos were aseptically removed from surface-sterilized mature cones. After aseptic extraction, mature zygotic embryos (MZE) and megagametophytes (Mgt) were individually cultured on modified LP 505 media supplemented with 0.45 mg/L BAP, 2 mg/L NAA, and 0.43 mg/L Kinetin for initiation and extrusion of cultures in the dark for 20 days. The cultures were then changed to a 16-h photoperiod for an additional 20 days. In light (40-day-old cultures), embryos extruded 75% and Mgt 19.04%. The shoot length was 2 cm after 30 days of cultivation, while Mgt produced 3 cm shoots. Developing shoots (40 days old) were moved to DCR or LP semi-solid media enriched with 2, 4, 6, or 8 mg/L of indole-3-butyric acid (IBA) + α-naphthalene acetic acid (NAA) in a 16 h photoperiod. Rooting was unsuccessful in these media; however, shoots developed up to 4 cm in length on LP + 6 mg/L IBA + NAA after 20 days (using 2-month-old cultures). Both MS and DCR cultures exhibited 50% rooting with 2 cm long roots in half DCR, LP, or whole MS media. After a month, rooted shoots were transferred to 8 × 15 cm plastic pots filled with a 3:1 peat moss and sand potting media. Pots were wrapped in plastic bags to constrain moisture and kept under the culture room conditions. For one month, irrigation with ¼ DCR was done every 10 days. Plants from MS lasted 35 days, whereas DCR plants stayed green in the potting mix for 15 days and successfully grew under field conditions.

Published

2026 58 5

1088

1095