Plant Regeneration and In Vitro Growth Performance of Male-Sterile Somatic Plantlets of Sugi (Japanese Cedar, Cryptomeria japonica) Derived from Different Embryogenic Cell Lines

With the spread of pollinosis caused by sugi (Japanese cedar, Cryptomeria japonica) pollen, the use of pollen-free somatic seedlings of sugi is expected in Japan. However, there is a lack of knowledge on the relationship between the abilities during somatic embryogenesis, initial in vitro growth traits, and subsequent growth of somatic seedlings. In the present study, we provide the first basic information on somatic embryo maturation efficiency, somatic embryo germination, and plantlet conversion frequencies, as well as on in vitro growth performance of pollen-free somatic plantlets derived from different embryogenic cell lines (ECLs). Somatic embryo maturation efficiency varied from 34 to 514 cotyledonary embryos per plate and the average for the 19 ECLs tested was 244 embryos per plate. Subsequently, the overall average rates of somatic embryo germination and conversion among ECLs were 87.8% and 85.3%, respectively. The results of in vitro growth performance of pollen-free somatic plantlets showed significant differences in growth rate among ECLs.

Differential Influence of Growth Regulators during Somatic Embryogenesis of Gynodioecious Papaya Varieties‘CO.7’ and ‘Red Lady

The study involved two auxins viz., 2,4-D (2,4-Diclorophenoxyacetic acid) and picloram at three different concentrations (1,2, 3 mg/L) in full strength MS media to study their comparative influence on induction of somatic embryogenesis from immature zygotic embryos of two gynodioecious varieties of papaya ‘CO.7’ and ‘Red Lady’. In papaya cultivar ‘CO.7’, 2,4-D at 2 mg/L gave the highest callus induction frequency of 90.93%, whereas comparatively higher concentration of 3 mg/L 2,4-D was found suitable for ‘Red Lady’ (87.26%). Although there was profuse callus formation, 2 mg/L 2,4-D recorded comparatively higher frequency of embryogenic calli in ‘Red Lady’ (51.67%) when compared to ‘CO.7’ (30.00%). Somatic embryo maturation was achieved upon transfer of embryogenic calli exhibiting globular stage embryos on to maturation medium (MS medium + ABA (Abscisic acid) and BAP (Benzyl amino purine) in different concentrations + glutamine 400 mg/L). In the maturation medium, the combination of 1.5 mg/L ABA and 0.4 mg/L BAP registered better conversion of the globular embryo to cotyledonary embryos than other levels. The frequency of somatic embryo germination was higher in ‘Red Lady’ (50.00%) as compared to ‘CO.7’ (31.67%) on half-strength MS medium devoid of growth regulators.

Pengaruh Komposisi Media dan Sumber Eksplan Terhadap Induksi Kalus, Perkecambahan, dan Pertumbuhan Tunas Embrio Somatik Jarak Pagar

<p>Jarak pagar (Jatropha curcas L.) merupakan tanaman penghasil minyak nabati sebagai bahan baku bio-diesel. Selama ini, kebutuhan bahan tanam diperoleh dari benih dan setek. Teknik mikropropagasi khususnya melalui embriogenesis somatik merupakan alternatif untuk penyediaan bahan tanam dalam jumlah besar dengan waktu relatif lebih singkat. Jenis eksplan, genotipe, dan kondisi fisiologis tanaman donor serta jenis dan kondisi fisik mediummerupakan faktor utama yang mempengaruhi keberhasilan embriogenesis somatik. Penelitian ini dilakukan untuk mengetahui eksplan dan komposisi media yang tepat untuk induksi kalus embriogenesis somatik, perkecambahan embrio somatik dan pertumbuhan tunas hasil embriogenesis somatik. Penelitian dilaksanakan di laboratorium Kultur Jaringan, Balai Penelitian Tanaman Pemanis dan Serat mulai bulan April sampai dengan November 2011, meliputi tiga tahap, yaitu 1) menguji komposisi media untuk induksi kalus embriogenesis somatik antara lain M1=MS+0,5 mg/l BAP+0,5 mg 2,4 D; M2= MS+1 mg/l BAP +0,5 mg/l 2,4 D; M3= MS+0,5 mg/l BAP+0,2 mg/l TDZ, dan M4= MS+1 mg/l BAP+0,2 mg/l TDZ; 2) menguji komposisi media untuk induksi perkecambahan embrio somatik antara lain MK1= MS+0,5 mg/l BAP+0,1 mg/l NAA dan MK2= MS+0,5 mg/l BAP+0,4 mg/l IBA; dan 3) menguji komposisi media untuk pertumbuhan tunas embrio somatik antara lain MP1= MS+0,5 mg/l BAP+0,1 mg/l IBA dan MP2= MS+0,5 mg/l BAP+0,1 mg/l IAA. Bahan tanam yang digunakan adalah genotipe IP-3A dan IP-3M dengan sumber eksplan kotiledon dan daun. Hasil penelitian menunjukkan kombinasi MS+0,5 mg/l BAP+0,2 mg/l TDZ dengan sumber eksplan kotiledon paling sesuai untuk induksi kalus embriogenesis somatik. Genotipe IP-3M memiliki respon yang lebih baik disbanding IP-3A dan stabil dari tahap induksi kalus embriogenis somatik, induksi perkecambahan embrio somatik, dan pertumbuhan tunas embrio somatik.</p><p> </p><p>Jatropha (Jatropha curcas L.) is an oil producing plants as source of bio-diesel. Planting materials usually are obtained from seeds and stem-cuttings. Micro-propagation techniques especially through somatic embryo-genesis is an alternative to provide a large number of planting material in a relatively short time. Explant sources, genotype and physicological conditions of donor plants, also composition and physical condition of medium are the main factors affecting the successful of somatic embryogenesis. The study was conducted to determine the most suitable combination of explant and media composition for embryogenic calli induc-tion, somatic embryo germination, and shoots growth derived from somatic embryogenesis. The experiment was conducted in the Tissue Culture Laboratory, of Indonesian Sweetener and Fiber Crops Research Insti-tute from April to November 2011 covering three phases: 1) testing media composition to induce somatic embryogenic calli i.e. M1=MS+0.5 mg/l BAP+0.5 mg 2.4 D; M2 = MS+1 mg/l BAP+0.5 mg/l 2.4 D; M3 = MS+0.5 mg/l BAP+0.2 mg/l TDZ and M4 = MS+1 mg/l BAP+ 0.2 mg/l TDZ; 2) testing media composition to induce somatic embryo germination i.e. MK1 = MS+0.5 mg/l BAP+0.1 mg/l NAA and MK2 = MS+0.5 mg/l BAP+0.4 mg/l IBA; and 3) testing media composition to induce somatic embryo shoot growth i.e. MP1 = MS+0.5 mg/l BAP+0.1 mg/l IBA and MP2= MS+0.5 mg/l BAP+0.1 mg/l IAA. Plant material used are genotype IP-3A and IP-3M with cotyledone and leaf as explant sources. The results showed that combination of MS+0.5 mg/l BAP+0.2 mg/l TDZ and cotyledons as explants source is the most suitable for somatic embryogenic calli. IP-3M genotype showed a better response to IP-3A and stable from induction of somatic embryogenic calli, somatic embryo germination, and somatic embryo shoots growth.</p>