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Sisunandar Sisunandar
Biology Education Department, University of Muhammadiyah Purwokerto Jl. Raya Dukuhwaluh, Kembaran, Purwokerto, 53182, Indonesia
Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Civil Engineering, Building, Construction & Architecture Electrical & Electronics Engineering Engineering Health Professions Materials Science & Nanotechnology Mechanical Engineering Medicine & Pharmacology Public Health Other

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Cryopreservation for Germplasm Conservation: Progress Report on Indonesian Elite Mutant Coconut “Kopyor” Sisunandar, Sisunandar
Proceeding International Conference on Global Resource Conservation Vol 4, No 1: Proceeding of 4th ICGRC 2013
Publisher : Proceeding International Conference on Global Resource Conservation

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Abstract

Among Indonesian elite materials, the Kopyor mutant is a paramount interest, as it is much sought after by customers for its soft and sweet flesh, and high prices with more than 10 times higher than the one of a normal fresh coconut. However, this mutant cannot be conserved by normal means such as field genebanks or seed genebank. An alternative conservation method would be one that involves a degree of dehydration and then ultra-cold storage (cryopreservation). However, as yet such protocols developed for Kopyor have been untested and no field-growing plants have been produced back from these techniques. The cryopreservation of zygotic embryos was achieved through the following steps viz.: a rapid tissue dehydration step prior to storage and a rapid warming step upon recovery followed by acclimatization to soil-supported growth. The best protocol was one based on an 8-hour rapid dehydration step followed by rapid cooling step, rapid warming step and an optimized in vitro culture technique. Following this protocol almost 20 % of cryopreserved embryo could be returned to normal seedlings growing in soil. Moreover, 23 % of recovered embryos were viable but unable to produce normal plantlets, mostly showed stunted shoot. These results indicate that it is possible to store coconut kopyor germplasm on a long-term basis using cryopreservation approach, even though the production of abnormal seedlings following cryopreservation as has been seen in many recalcitrant species before remains a major challenge for the cryopreservation protocol.Keywords: cryopreservation, Embryo culture, Germplasm conservation, Kopyor, Rapid dehydration
KEMAJUAN PENELITIAN INDUKSI DAN PEMANJANGAN TUNAS MERBAU (INTSIA BIJUGA [COLEBR.] O. KUNTZE) SECARA IN VITRO Ramadhani, Latifah Esti; Husin, Arief; Sisunandar, Sisunandar
Proceeding Biology Education Conference: Biology, Science, Enviromental, and Learning Vol 11, No 1 (2014): Prosiding Seminar Nasional XI Biologi
Publisher : Universitas Sebelas Maret

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Abstract - Merbau is one of the high-quality timber tree with the demanding of international market increase every year. However, the over-exploitation of the tree put merbau as a vulnerable species in the future years. Moreover, seedling production of merbau using traditional technique showed several limitations  due to the low number of seed production every year. One of the most promising technique for mass seedling production of merbau is using shoot-tip culture. However, the protocol of shoot-tip culture is remain under develop with low success rate of shoot induction (less then 15 %), without protocols on shoot multiplication, root induction and acclimatisation. Our research at the Laboratory of Botany and Genetics, University of Muhammadiyah Purwokerto showed that the axillary buds isolated from 1-month seedlings can be maintained to induce shoot formation and elongation on the DKW medium (Driver and Kuniyuki, 1984) supplemented with 10-8 M kinetin. Two new shoots started to elongate from each explant after 4 weeks culture (60 % success rate) with each shoot has the lenght of c.a. 4 cm and 4 internode. The inducted shoots were then cut them of into four new axilary buds and subcultured onto the same medium every two week for shoots multiplication. The next step of shoot-tip culture technique, root induction and acclimatization stages is remain developed. Keywords : kinetin, shoot-tip culture,multiplication,micropropagation.
PRODUKSI BIBIT KELAPA KOPYOR TRUE-TO-TYPE MELALUI TEKNIK KULTUR EMBRYO Sisunandar, Sisunandar
Proceeding Biology Education Conference: Biology, Science, Enviromental, and Learning Vol 11, No 1 (2014): Prosiding Seminar Nasional XI Biologi
Publisher : Universitas Sebelas Maret

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Abstract

Kopyor is one of the most valuable germplasm in Indonesia. However, the production of true-to-type seedlings with 100 % kopyor nuts is still limited because the nut would not germinated naturally. The only technique available to solve the problem is by using embryo culture technique. However, the technique is limited especially on the low of success rate after acclimatisation step (less than 40 %). Therefore, we report a new breakthough technique for producing kopyor seedling using embryo culture technique especially on acclimatisation step. Our results showed that the best acclimatisation step could be done by culturing 4-month old seedlings in a culture room with 14 hour light with intensity around 1400 lux and 10 hours in the dark. Seedlings were cultured for three months in mini growth chamber with 95 % relative humidity level and temperature around 26 to 28 0C. The survival rate after acclimatisation step on the seedlings with full root was more than 95 %, while for the seedlings without root was c.a 90 %. The acclimatized seedlings then underwent to screenhouse for 1 year befor being field planting. Up to now, more than 170 seedlings of Kopyor have been transfer to the field for developing seed garden of Kopyor in Purwokerto, Indonesia. In the future, creating a breeding program to produce hybrid of Kopyor will be useful for poverty reduction programmes in Indonesia. Keywords: acclimatization, growth chamber, hybrid of kopyor, seed genebank
Analisis Histologi Pelepah Daun Kelapa Kopyor Hibrida (Kopyor dalam Sinumpur (KDS) × Kopyor Genjah Kuning Sinumpur (KGKS) Latifah, Nisa Khoeri; Sisunandar, Sisunandar
Proceedings Series on Physical & Formal Sciences Vol. 7 (2024): Prosiding Seminar Nasional Fakultas Pertanian dan Perikanan
Publisher : UM Purwokerto Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.30595/pspfs.v7i.1201

Abstract

Perbaikan kualitas kelapa kopyor khususnya melalui persilangan belum banyak dilakukan. Di samping memerlukan waktu yang cukup lama, metode deteksi keberhasilan persilangan juga belum banyak dikembangkan. Saat ini metode morfologi sebagai alat deteksi keberhasilan persilangan memiliki keterbatasan sedangkan metode molekuler relatif mahal dan membutuhkan teknologi tinggi. Analisis keberhasilan persilangan secara histologi perlu dilakukan. Penelitian ini bertujuan untuk membandingkan histologi pelepah daun kelapa kopyor hibrida (KDS × KGKS) dengan tetua jantan (KDS) dan tetua betinanya (KGKS). Pelepah daun diisolasi dari dua sampel yaitu bibit kelapa in vitro berumur 9 - 12 bulan setelah tanam dan bibit kelapa ex vitro berumur 8 – 12 bulan pasca aklimatisasi. Preparat penampang melintang pelepah daun dipotong dengan tangan kemudian diwarnai dengan 2% safranin (30 detik) dilanjutkan 1% fastgreen (50 detik). Preparat diamati menggunakan mikroskop Olympus BX51 dan gambar diambil menggunakan kamera Olympus DP74. Gambar yang diperoleh dianalisis menggunakan software ImageJ untuk menghitung jumlah, luas total, dan luas rata- rata sklerenkim, serta jumlah, luas total, dan luas rata-rata ikatan pembuluh berserat (IPB). Data yang diperoleh dianalisis menggunakan ANOVA pada tingkat kepercayaan 95% dilanjutkan dengan uji DMRT. Hasil penelitian menunjukkan tidak ada perbedaan yang signifikan pada struktur histologi antara pelepah daun dari bibit in vitro dan bibit ex vitro. Pelepah daun bibit KDS memiliki jumlah maupun luas total sklerenkim yang lebih tinggi dibandingkan dengan pelepah daun bibit KGKS. Demikian pula bibit kelapa hibrida memiliki pelepah daun dengan jumlah dan luas total sklerenkim yang sama dengan pelepah daun bibit KDS dan lebih tinggi dibandingkan dengan pelepah daun bibit KGKS. Namun demikian, pelepah daun bibit kelapa hibrida tidak menunjukkan perbedaan yang signifikan dalam hal jumlah, luas total dan luas rata-rata IPB dibandingkan dengan bibit KGKS. Hasil tersebut dapat disimpulkan bahwa analisis histologi pelepah daun dapat dikembangkan lebih lanjut menjadi salah satu kunci identifikasi awal keberhasilan persilangan pada produksi bibit kelapa kopyor hibrida.
Co-Authors Arief Husin, Arief Latifah Esti Ramadhani Latifah, Nisa Khoeri