Evaluation of Somatic Embryogenesis Ability in Robusta Coffee (Coffea Canephora Pierre)

Embriogenesis somatik diharapkan sebagai metode perbanyakan tanaman yang sangat efektif pada kopi. Evaluasi dua jenis proses embriogenesis somatik, yaitu proses langsung dan tidak langsung akan bermanfaat untuk menggambarkan kemampuan proliferasi sel. Penelitian untuk mengevaluasi embriogenesis somatik kopi Robusta (Coffea canephora) yang mempunyai tingkat keragaman genetik tinggi telah dilakukan di Nestlé R&D Centre Tours, Perancis. Bahan tanam menggunakan kopi Robusta koleksi Nestle Perancis dan tiga klon koleksi Pusat Penelitian Kopi dan Kakao Indonesia (Puslitkoka). Tiga aspek, yaitu proses embriogenesis, keragaman embriogenesis dan kemantapan embriogenesis dievaluasi dalam penelitian ini. Hasil penelitian menunjukkan bahwa baik embriogenesis somatik langsung maupun tidak langsung dapat diamati. Penelitian ini menunjukkan bahwa kedua proses embriogenesis somatik tersebut merupakan dua mekanisme yang berbeda. Dalam penelitian ini ditunjukkan bahwa kemampuan embriognesis somatik tergantung pada genotipe, baik antar maupun di dalam kelompok genetik kopi Robusta, yaitu Congolese,Guinean dan Conillon. Lebih lanjut diketahui bahwa kedua proses embriogenesis somatik tersebut stabil terhadap indukan sebagai sumber eksplan. Kemampuan embriogenesis somatik tidak langsung ketiga klon Puslitkoka (BP409, BP961 dan Q121) sangat beragam, sehingga memberikan harapan adanya pola segregasi yang baik berdasarkan kemampuan embriogenesis somatik tidak langsung pada populasi yang dibuat dari silangan klon tersebut

Maintenance of embryogenic potential of calli derived from embryonic shoot of West Coast Tall cv. of coconut (Cocos nucifera L.)

Maintenance of embryogenic potential of calli is important as the totipotency is often lost in a short time in vitro. This caters to the need for year round availability of somatic embryos in a regenerable state. In the present study, 14 media combinations, with either 2,4-D or picloram as auxin source, were tested for maintaining embryogenic calli obtained from embryonic shoot explants of coconut. Irrespective of type and concentration of auxins, callusing was observed in all the media combinations. However, high dose of 2,4-D (above 74.6 μM) in the initial medium resulted in intense browning and lesser percentage of callusing. Embryogenic nature of calli could be maintained to a maximum of 21 weeks in medium supplemented with 2,4-D (74.6 μM) and subsequent culturing into higher concentration of 2,4-D (90.4 μM). Gene expression studies carried out using qRT-PCR revealed that genes such as ECP, GST, LEAFY and WUS were highly expressed in long term embryogenic calli (21 week old) and genes such as SERK, GLP, WRKY and PKL in initial embryogenic calli (21 days old). The study concludes that coconut plumular calli could be maintained for longer periods without compromising on the embryogenic potential of the calli

Morphological Characterization and Identification of Coffea Liberica Callus of Somatic Embryogenesis Propagation.

Compared with other types of coffee, Liberica coffee is more difficult to be propagates using clonal methods. Meanwhile, demand for planting materials and consumption of this type of coffee is increasing lately. The objective of this paper is to present results of the work on morpological characterization of Liberica coffee (Coffea liberica) callus produced by somatic embryogenesis propagation. This research used C. liberica Arruminensis clone. This clone was one of Liberica coffee clones which had superior taste. Main activitis carried out in this experiment were explant sterilization, explant induction and histological analysis on the callus produced. The result of this research showed that non embryogenic callus was higher (72%) than embryogenic callus (28%). The callus description can be used to identify type and characteristic of the callus. Therefore, it can be a parameter to choose type of callus for propagation material. This is important because choosing the right callus is determine of the succesfully process of Liberica somatic embryogenesis

A comparative study of three different methods of shoot meristem excision for induction of embryogenic calli in coconut

A protocol was standardized to maximize yields of embryogenic calli from shoot meristem culture of coconut. Three different shoot meristem excision methods were tested viz., excision of shoot meristem aseptically from in vitro germinated embryo after 10-12 days, excision of shoot meristem from in vitro germinated embryo subjected to GA3 treatment for five days and excision of shoot meristem from fresh embryo. The primary calli induction after 30 days of culture incubation for the three treatments were 21%, 27%  and 79% respectively.  Further, the primary calli formed from the shoot meristem excised from fresh embryo gave rise to 56% of embryogenic calli. The calli obtained from the shoot meristem which were excised from in vitro germinated embryo formed less percentage of embryogenic calli because of the presence of cotyledonary tissues which inhibited the multiplication of meristematic tissues. In the case of shoot meristem extracted from GA3-treated embryos, the percentage of non-embryogenic calli was more compared to the shoot meristem excised from fresh embryo. It was observed that the addition of GA3 in the initial stages of culture inhibited the formation of embryogenic calli and favored direct shoot development. Currently, the shoot meristem excised from fresh embryo is being employed for scaling up the planting material production from released varieties of coconut