Rajasegar, Arulvilee
(2015)
Cryopreservation Of Ascocenda Wangsa Gold Orchid Using Pvs2 Vitrification Method.
Masters thesis, Universiti Sains Malaysia.
Abstract
Satu kaedah pengkrioawetan larutan vitrifikasi tumbuhan 2 (PVS2) yang effisien telah dibangunkan untuk jasad seperti protokom (JSP) Ascocenda Wangsa Gold orkid. Kesan bahan tambahan organik seperti air kelapa, homogenat ubi kentang dan pisang ke atas pertumbuhan JSP Ascocenda Wangsa Gold telah dikaji. Bilangan JSP diaruh dari setiap eksplan telah dikira selepas empat minggu rawatan. Penambahan 2% homogenat ubi kentang, 2% pisang dan 15% air kelapa ke dalam media Vacin dan Went didapati adalah kultur medium terbaik untuk menginduksi JSP yang menghasilkan sebanyak 94.7%. Parameter yang dinilai termasuk size protokom, kepekatan sukrosa, kesan tempoh pendedahan kepada PVS2, tempoh pencairan, suhu, dan keadaan kultur berdasarkan bacaan penyerapan TTC dan melalui pemerhatian penjanaan semula protokom. Kadar pemulihan sebanyak 33.3% telah diperolehi selepas 2 bulan apabila JSP telah didedahkan pada PVS2 selama 30 min. Kadar pemulihan telah meningkat kepada 47% apabila dicairkan pada 45°C selama 85s. Kadar pertumbuhan semula yang paling tinggi (53.3%) telah diperolehi apabila JSP didedahkan kepada rawatan gelap 7 hari sebelum dipindahkan ke fotokala 16/8 jam cahaya/gelap.
An efficient plant vitrification solution 2 (PVS2) cryopreservation method was developed for protocorm-like bodies (PLBs) of Ascocenda Wangsa Gold orchid. The effects of organic additives such as coconut water, potato and banana homogenates on PLBs proliferation of Ascocenda Wangsa Gold in different media culture were studied. The numbers of PLBs induced from each explant were counted after four weeks of treatment. Addition of 15% of coconut water, 2% of potato and banana homogenate into Vacin and Went medium was found to be the best media culture for PLBs induction at 94.7%. Parameters assessed included the preculture sucrose concentration effect of PVS2 exposure periods, thawing duration, temperature, and culture conditions based on 2, 3, 5-triphenyltetrazolium chloride absorbance readings and regrowth rates. A regrowth rate of 33.3% was obtained after 2 months when the PLBs were dehydrated in PVS2 for 30 min. The growth rate was improved to 47% when thawed at 45°C for 85 s. The highest growth rate (53.3%) was obtained when the PLBs were subjected to a 7-day dark treatment before being transferred to a 16-h/8-h light/dark photoperiod.
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