Optimization of sterilization methods and PPM™ supplementation in culture media for in vitro culture of leaf and nodal explants of Cryptocarya massoy

Syifa Fajrisani; Zozy Aneloi Noli; Mansyurdin; Syamsuardi

doi:10.55779/nsb18212941

Authors

Syifa Fajrisani Universitas Andalas, Faculty of Mathematics and Natural Sciences, Doctoral Program in Biology, Padang 25163 (ID) https://orcid.org/0000-0002-9975-0957
Zozy Aneloi Noli Universitas Andalas, Faculty of Mathematics and Natural Sciences, Department of Biology, Padang 25163 (ID) https://orcid.org/0000-0003-1483-2652
Mansyurdin Universitas Andalas, Faculty of Mathematics and Natural Sciences, Department of Biology, Padang 25163 (ID) https://orcid.org/0000-0003-0705-0820
Syamsuardi Universitas Andalas, Faculty of Mathematics and Natural Sciences, Department of Biology, Padang 25163 (ID) https://orcid.org/0000-0001-8351-6528

DOI:

https://doi.org/10.55779/nsb18212941

Keywords:

antifungal, browning, contamination, Masoyi, PPM™, survival

Abstract

Masoyi (Cryptocarya massoy (Oken) Kosterm.) is a valuable aromatic plant with limited information on its in vitro propagation. This study aimed to evaluate sterilization methods and PPM™ supplementation in culture media for the in vitro culture of leaf and nodal explants of Cryptocarya massoy (Oken) Kosterm. This study employed a two-factor completely randomized design (CRD) consisting of 12 treatment combinations, 10 replications, and a total of 120 experimental units. The first factor was the soaking duration in PPM™, namely 1, 2, and 3 days, while the second factor was the concentration of PPM™ in the MS medium, namely 0, 0.2, 0.4, and 0.6%. Results showed a strong explant-dependent response. Leaf explants exhibited consistently poor performance, with 100% contamination across all treatments, no survival, and no shoot or callus formation. Contamination was predominantly fungal and occurred despite variations in sterilization intensity. In contrast, nodal explants showed significant variation among treatments, with contamination ranging from 10% to 100%, and delayed contamination under certain conditions. Nodal explants also demonstrated improved physiological responses, including higher survival rates, reduced browning in specific treatments, and limited morphogenic responses. The highest shoot formation (5.00%) and callus induction (4.00%) were observed in treatment A3B3, where compact callus formation was also recorded. Statistical analysis confirmed that treatments significantly affected contamination, time of browning, survival, and shoot formation (p ≤ 0.05). Meanwhile, callus formation was not significantly different among treatments (p > 0.05). Overall, nodal explants treated with 3 days of PPM™ soaking and 0.4% PPM™ in the culture medium showed the most promising response for establishing aseptic in vitro cultures of C. massoy, whereas leaf explants were unsuitable under the tested conditions.

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