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In Vitro Seed Germination and Shoot Growth of Nepenthes jamban Chi. C. Lee, Hernawati & Akhriadi, A Unique Pitcher Plant from Indonesia Prawestri, Apriliana Dyah; Rahayu, Resa Sri; Kurniajati, Wulan Septiningtyas; Sunardi, Sunardi; Mansur, Muhammad
Journal of Tropical Biodiversity and Biotechnology Vol 9, No 2 (2024): June
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.87674

Abstract

The study to optimize in vitro propagation of the Indonesian native and critically endangered species, Nepenthes jamban, in order to support the ex-situ conservation efforts has been done.  Using Murashige and Skoog (MS) as a basal media, disinfected seeds of N. jamban were germinated on five types of germination media, viz. ¼ MS, ½ MS, MS, ¼ MS+benzyl adenine (BA)+Biotin and MS+BA+Biotin. Afterwards, in vitro shoots with 6-7 leaves were inoculated on growing media, i.e., ¼ MS, ¼ MS 60 (3:1) (MS modification with a higher concentration of nitrogen), and ¼ MS+naphtalene acetic acid (NAA)+BA. The results showed that the germination of N. jamban seeds was slow, indicated by the percentage of germination being less than 20% after 6 months of being planted on germination media. The highest percentage of germination was after the 6th month and the greatest pitcher development at the 10th month were obtained on ¼ MS medium. Furthermore, shoot growth and pitchers development consistently increased for 12 months in ¼ MS 60 (3:1) medium while other media resulted in a decrease in pitcher formation. It seemed that low concentrations of nutrient in the medium proved to be more effective to induce in vitro seed germination and enhance shoot growth which was also supported by higher nitrogen (nitrate) concentration in the medium. This study provides information that supports ex situ conservation action of native and critically endangered Nepenthes species from Indonesia. 
Histological Analysis of Somatic Embryogenesis from Immature Zygotic Embryo of Wild Banana Musa acuminata ssp. malaccensis Handayani, Tri; Martanti, Diyah; Prawestri, Apriliana Dyah; Tihurua, Eka Fatmawati; Maharijaya, Awang; Wahyu, Yudiwanti; Sobir; Witjaksono
Journal of Tropical Crop Science Vol. 11 No. 03 (2024): Journal of Tropical Crop Science
Publisher : Department of Agronomy and Horticulture, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jtcs.11.03.307-319

Abstract

Somatic embryogenesis, a crucial plant regeneration method, has become indispensable for crop improvement, particularly for species reliant on somatic cell manipulation techniques. Optimization of this process necessitates an understanding of the developmental stages involved. This study investigates the histological aspects of somatic embryogenesis in Musa acuminata ssp. malaccensis derived from immature zygotic embryos. Through detailed histological analysis, we aimed to elucidate the morphological changes and cellular organization occurring during the various stages of somatic embryogenesis, from induction, culture proliferation, and somatic embryo development to plantlet conversion. The initial stages of embryogenesis, characterized by nodules, were primarily composed of meristematic cells with high cell division activity. These cells contained tetrad-like structures that could develop into distinct two- and four-celled proembryoids or proembryogenic aggregates. Our histo-anatomical analysis revealed that embryogenic cultures proliferated through multiple pathways simultaneously: somatic embryo budding, proembryo formation, and pro-embryonic mass formation from both internal and peripheral cells. At the stage of somatic embryo development, embryos with a well-defined protoderm layer, containing cells with prominent nuclei and dense cytoplasm, potentially regenerate into plantlets. Furthermore, histological examination revealed the presence of procambium within mature somatic embryos, which subsequently developed into the vascular system of the complete plantlet
An Efficient Somatic Embryogenesis and Plant Regeneration from Immature Embryo of Wild Banana Musa acuminata ssp. malaccensis Handayani, Tri; Martanti, Diyah; Prawestri, Apriliana Dyah; Maharijaya, Awang; Wahyu, Yudiwanti; Sobir; Witjaksono
Journal of Tropical Crop Science Vol. 12 No. 01 (2025): Journal of Tropical Crop Science
Publisher : Department of Agronomy and Horticulture, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jtcs.12.01.172-184

Abstract

Wild banana Musa acuminata ssp. malaccensis, an ancestor of cultivated bananas, possesses valuable genetic diversity, including resistance genes to fusarium wilt, and demonstrates high environmental adaptability. These traits are important for pre-breeding programs, whether by conventional breeding, which is a lengthy process, or by taking advantage of somatic cell manipulation techniques such as somatic hybridization, which requires an efficient plant regeneration system like somatic embryogenesis. We have established an efficient and comprehensive protocol for somatic embryogenesis of this wild Musa using immature zygotic embryo explants covering culture induction and proliferation, somatic embryo development, and subsequent plant conversion. Embryogenic culture was induced on a complex-modified MS medium supplemented with 1 mg.L-1 2,4-D auxin or 5 mg.L-1 picloram. The embryogenic cultures proliferated in the form of granular or nodular structures, which was best obtained by reducing the picloram concentration to 1 mg.L-1 and combining it with the same concentration of 2,4-D at a half-strength macro-nutrient of basal medium. Embryo development from embryogenic cultures and regeneration into shoots. Proembryos as granular structures spontaneously matured into early-stage somatic embryos upon withdrawal of the strong auxin inducer. Increasing the sucrose and gelling agent concentrations in the growth medium improved somatic embryo formation from embryogenic cultures. The frequency of shoot formation from developed somatic embryos was increased by incorporating 0.5 mg.L-1 BA and 0.5 mg.L-1 GA3 into the regeneration medium.