The Effect of Different Seed Cutting Treatments and Concentrations of BAP for the Successful In Vitro Micrografting of Mangosteen (Garcinia mangostana L.)

The efforts of the rooting of regenerants resulting from gamma-ray irradiation require plant tissue culture, which known as micrografting. This technique can help irradiated regenerants to develop a well root system, by combining non-rooting shoots with rooted in vitro cultured shoots of plant rootstock. The purpose of this study was to examine the effect of seed explants cutting and the application of different BAP concentrations for the successful micro-grafting of mangosteen in vitro. This experiment employed Complete Randomized Design (CRD) Factorial with two factors and ten replications. The first factor was the cutting treatments of mangosteen seeds explants for rootstock shoots, consisting of 2 types of seeds: uncut and cut seeds. The second factor was BAP concentrations: BAP 0 mg/l and BAP 2 mg/l. The results showed that the division of the seeds had an influence on the results of micro-grafting mangosteen in vitro. Micrografted mangosteen, which rootstock applied from undivided seeds, possessed faster growth, longer shoots, and produce more leaves compared to rootstock shoots from the divided seeds. BAP concentrations also contributed to the results. The application of BAP 2 mg/ demonstrated better effect on all variables observed. There were no interactions between seed divisions and BAP concentrations in all observed variables.

Teknik sambung mikro in vitro kina Cinchona succirubra dengan C. ledgeriana In vitro micrografting technique of Chincona succirubra and C. ledgeriana

Summary In vitro micrografting is a technique for grafting scions to rootstocks of plantlets from tissue culture. In vitro micrografting of Cinchona plant has never been carried out. The objective of this research was to obtain the best method of in vitro micrografting, medium for micrografted plantlets, and acclimatization for Cinchona plantlets from micrografting. The research consisted of (i) optimization of micrografting method, (ii) optimization of medium for growing plantlets, and (iii) acclimatization of micrografted plantlet. Plantlets of four-month-old of C. ledgeriana QRC clone were used as scions, while of C. succirubra as rootstocks. Each of experiments was arranged according to Completely Randomized Design, consisted of combination of scion and rootstock and type of micro-grafting with 10 replicates. Parameters measured were the percentage of survived plantlet, leaf number, and callus productions on union area, and percentage of survived plantlet. The results show that V type of micrografting was the best for Cinchona micrografting. MS medium with the addition of 3 mg/L IBA was the best medium for growing of micrografted plantlet. Husk charcoal mixed with top soil (1 : 1) was the best medium for acclimatization. Acclimatization consisted of two steps: preaclimatization in a culture room with 12- hour photoperiod at temperature 25 – 27oC for two weeks, followed by aclimatization in a plastic house with 70% reduced light intensity for one month. Using this method, 90% of the seedlings were survived. It is concluded that in vitro micrografting can be used as a technique for clonal propagation of Cinchona sp.Ringkasan Teknik sambung mikro (mikrografting) in vitro adalah teknik penyambungan potongan batang atas pada batang bawah dalam kultur jaringan. Pada tanaman kina teknik sambung mikro in vitro belum pernah dilakukan. Tujuan penelitian ini adalah menetapkan tipe sambung mikro, medium terbaik untuk planlet hasil sambung mikro, dan perbanyakan tanaman kina dengan sambung mikro. Pelaksanaan percobaan meliputi (i) optimasi tipe sambung, (ii) optimasi medium, dan (iii) aklimatisasi planlet hasil sambung mikro. Bahan tanaman yang digunakan sebagai batang atas adalah planlet Cinchona ledgeriana klon QRC, sedangkan sebagai batang bawah digunakan planlet C. succirubra, berumur empat bulan. Masing- masing percobaan disusun dengan Rancangan Acak Lengkap terdiri dari dua taraf yaitu kombinasi batang bawah dengan batang atas bentuk sambung tipe V dan L dilakukan dengan 10 ulangan. Peubah yang diukur meliputi persentase planlet yang bertahan hidup, jumlah daun, berkalus atau tidak berkalus pada daerah pertautan, dan persentase planlet yang bertahan hidup. Hasil yang diperoleh menunjukkan bahwa tipe V merupakan cara sambung mikro yang terbaik. Medium MS dengan penambahan 3 mg/L IBA adalah medium terbaik untuk pertumbuhan dan perakaran planlet hasil sambung mikro. Aklimatisasi planlet dilakukan dengan medium tumbuh arang sekam : top soil (1 : 1) yang disterilkan. Tahapan aklimatisasi adalah pre-aklimatisasi dalam ruang kultur suhu 25 - 27 oCdengan pencahayaan 12 jam per hari dan diikuti dengan aklimatisasi di rumah plastik bernaungan 70% paranet. Dengan metode aklimatisasi ini 90% dari bibit mampu bertahan hidup. Kesimpulan dari penelitian ini menunjukkan bahwa teknik sambung mikro dapat digunakan untuk perbanyakan klonal Cinchona sp..

Teknik sambung mikro in vitro kina Cinchona succirubra dengan C. ledgeriana In vitro micrografting technique of Chincona succirubra and C. ledgeriana

Summary In vitro micrografting is a technique for grafting scions to rootstocks of plantlets from tissue culture. In vitro micrografting of Cinchona plant has never been carried out. The objective of this research was to obtain the best method of in vitro micrografting, medium for micrografted plantlets, and acclimatization for Cinchona plantlets from micrografting. The research consisted of (i) optimization of micrografting method, (ii) optimization of medium for growing plantlets, and (iii) acclimatization of micrografted plantlet. Plantlets of four-month-old of C. ledgeriana QRC clone were used as scions, while of C. succirubra as rootstocks. Each of experiments was arranged according to Completely Randomized Design, consisted of combination of scion and rootstock and type of micro-grafting with 10 replicates. Parameters measured were the percentage of survived plantlet, leaf number, and callus productions on union area, and percentage of survived plantlet. The results show that V type of micrografting was the best for Cinchona micrografting. MS medium with the addition of 3 mg/L IBA was the best medium for growing of micrografted plantlet. Husk charcoal mixed with top soil (1 : 1) was the best medium for acclimatization. Acclimatization consisted of two steps: preaclimatization in a culture room with 12- hour photoperiod at temperature 25 – 27oC for two weeks, followed by aclimatization in a plastic house with 70% reduced light intensity for one month. Using this method, 90% of the seedlings were survived. It is concluded that in vitro micrografting can be used as a technique for clonal propagation of Cinchona sp.Ringkasan Teknik sambung mikro (mikrografting) in vitro adalah teknik penyambungan potongan batang atas pada batang bawah dalam kultur jaringan. Pada tanaman kina teknik sambung mikro in vitro belum pernah dilakukan. Tujuan penelitian ini adalah menetapkan tipe sambung mikro, medium terbaik untuk planlet hasil sambung mikro, dan perbanyakan tanaman kina dengan sambung mikro. Pelaksanaan percobaan meliputi (i) optimasi tipe sambung, (ii) optimasi medium, dan (iii) aklimatisasi planlet hasil sambung mikro. Bahan tanaman yang digunakan sebagai batang atas adalah planlet Cinchona ledgeriana klon QRC, sedangkan sebagai batang bawah digunakan planlet C. succirubra, berumur empat bulan. Masing- masing percobaan disusun dengan Rancangan Acak Lengkap terdiri dari dua taraf yaitu kombinasi batang bawah dengan batang atas bentuk sambung tipe V dan L dilakukan dengan 10 ulangan. Peubah yang diukur meliputi persentase planlet yang bertahan hidup, jumlah daun, berkalus atau tidak berkalus pada daerah pertautan, dan persentase planlet yang bertahan hidup. Hasil yang diperoleh menunjukkan bahwa tipe V merupakan cara sambung mikro yang terbaik. Medium MS dengan penambahan 3 mg/L IBA adalah medium terbaik untuk pertumbuhan dan perakaran planlet hasil sambung mikro. Aklimatisasi planlet dilakukan dengan medium tumbuh arang sekam : top soil (1 : 1) yang disterilkan. Tahapan aklimatisasi adalah pre-aklimatisasi dalam ruang kultur suhu 25 - 27 oCdengan pencahayaan 12 jam per hari dan diikuti dengan aklimatisasi di rumah plastik bernaungan 70% paranet. Dengan metode aklimatisasi ini 90% dari bibit mampu bertahan hidup. Kesimpulan dari penelitian ini menunjukkan bahwa teknik sambung mikro dapat digunakan untuk perbanyakan klonal Cinchona sp..