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Title:	Analisis filogeni dan pengoptimunan pengkulturan miselium cendawan susu harimau (Lignosus rhinocerotis) yang dikutip dari negeri Pahang, Malaysia
Authors:	Lai Wei Hong (P58664)
Supervisor:	Fauzi Daud, Prof. Madya Dr.
Keywords:	Cendawan susu harimau Lignosus rhinocerotis Vitrifikasi Dissertations, Academic -- Malaysia Universiti Kebangsaan Malaysia -- Dissertations
Issue Date:	11-Jan-2014
Description:	Cendawan susu harimau merupakan cendawan liar berubi keras yang digunakan oleh Orang Asli untuk merawat pelbagai jenis penyakit termasuk kanser. Dalam kajian ini, analisis filogenetik molekul menggunakan gen subunit kedua terbesar RNA polimerase II (RPB2) telah mengenal pasti cendawan susu harimau liar yang dipencil dari negeri Pahang, Malaysia adalah Lignosus rhinocerotis (Cooke) Ryvarden dari order Polyporales dengan 99% homologi. Kultur tulen (miselium) sampel cendawan ini berjaya diawetkan menggunakan kaedah vitrifikasi bagi tujuan penyimpanan yang lama. Analisis proksimat berdasarkan 100 g berat basah mendapati tudung cendawan mengandungi 56.30 ± 0.26 g karbohidrat, 6.60 ± 1.38 g protein, 0.21 ± 0.09 g lemak, 1.77 ± 0.28 g/100 g abu dan 32.22 ± 5.54 g kelembapan; batang cendawan mengandungi kandungan 38.47 ± 0.49 g karbohidrat, 46.31 ± 14.00 g kelembapan, 2.76 ± 0.60 g protein, 0.27 ± 0.07 g lemak dan 1.76 ± 0.93 g abu; manakala sklerotium cendawan mengandungi 51.30 ± 0.10 g karbohidrat, 39.82 ± 2.22 g kelembapan, 3.04 ± 0.59 g protein, 0.46 ± 0.32 g lemak dan 4.38 ± 2.71 g abu. Dalam menentukan medium pertumbuhan, didapati miselium cendawan ini hidup dengan optimum di atas medium agar glukosa-pepton dan medium agar ekstrak yis pepton dekstrosa pada suhu 30 o C di antara pH 6 dan 7. Bagi kultur tenggelam pula, miselium hidup dengan optimum dalam medium yang mengandungi 80 g/L glukosa, 4 g/L kalium nitrat, 0.4 g/L FeSO4.7H2O dan 0.1 g/L CaCl2 pada suhu 25 o C dan pH 6; manakala penghasilan eksopolisakarida (EPS) tertinggi dalam medium yg mengandungi 80 g/L glukosa, 4 g/L kalium nitrat, 1.4 g/L FeSO4.7H2O dan 1.1 g/L CaCl2 pada suhu 25 o C dan pH 6. Dalam mengenal pasti protein yang terlibat dalam pembentukkan primordium, analisis pengekspresan protein secara elektroforesis gel dua dimensi natrium sulfat dodesil poliakrilamida (2D-SDS-PAGE) dengan Progenesis Samespot (Versi 4.0) menghasilkan lebih kurang 1000 titik protein dalam proteom sel vegetatif miselium manakala sel primordium mengandungi lebih kurang 100 titik protein. Perbandingan proteom sel primordium dengan sel vegetatif miselium menunjukkan peningkatan pengekspresan lima protein sel primordium yang dilabel sebagai P1, P2, P3, P4 dan P5. Pencapjarian Jisim Peptida telah mengenal pasti P1 dan P2 sebagai protein DnaJ domain, P3 dan P5 sebagai protein hipotetikal manakala P4 sebagai faktor transkripsi AP-2rep. Di samping itu, keadaan dan nutrisi pengkulturan yang optimum untuk pertumbuhan miselium dan penghasilan EPS L. rhinocerotis telah berjaya dikenal pasti. Pemprofilan pengekspresan protein sel primordiuml pula mencadangkan bahawa mekanisme tindak balas kepayahan suhu terlibat dalam pembuahan cendawan ini. Di samping itu, sklerotium L. rhinocerotis berjaya dihasilkan dengan menanam substrat yang terdiri daripada 80% habuk kayu getah, 18% dedak padi, 1% glukosa, 1% CaCO3 dalam tanah.,Tiger's milk mushroom is a wild tuberious mushroom that is being used by Aborigines to treat a variety of diseases including cancer. Molecular phylogenetic analysis utilizing second largest subunit RNA polymerase II gene (RPB2), identified the wild Tiger's Milk mushrooms collected from the state of Pahang, Malaysia for this study as Lignosus rhinocerotis (Cooke) Ryvarden in the order Polyporales with 99% homology. Pure culture (mycelia) of this indigenous sample was successfully cryopreserved using vitrification method. Proximate analysis of 100 g wet weight of this mushroom yielded the pileus contains 56.30 ± 0.26 g carbohydrate, 32.22 ± 5.54 g moisture, 6.60 ± 1.38 g protein, 0.21 ± 0.09 g fat and 1.77 ± 0.28 g ash; stipe of this mushroom contains 38.47 ± 0.49 g carbohydrate, 46.31 ± 14.00 g moisture, 2.76 ± 0.60 g protein, 0.27 ± 0.07 g fat and 1.76 ± 0.93 g ash; while tuber of this mushroom contains 51.30 ± 0.10 g carbohydrate, 39.82 ± 2.22 g mositure, 3.04 ± 0.59 g protein, 0.46 ± 0.32 g fat and 4.38 ± 2.71 g ash. In determining the solid growth mediun, it was observed that the mycelial growth of this mushroom is optimal in glucosepeptone mediun and yeast extract peptone dextrose mediun at 30 o C between pH 6 and 7. While for submerged culture, the optimal compositions for mycelial growth were 80 g/L of glucose, 4 g/L of potassium nitrate, 0.4 g/L of FeSO4.7H2O and 0.1 g/L of CaCI at 25 o C and pH6; consequently, the optimal compositions for exopolysaccharides (EPS) production were 80 g/L of glucose, 4 g/L of potassium nitrate, 1.4 g/L of FeSO4.7H2O and 1.1 g/L of CaCI2 at 25 o C and pH6. In identifying the proteins involved in the formation of primordium, protein expression analysis utilising two dimensional-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D-SDS-PAGE) with Progenesis Samespot (Version 4.0) yielded approximately 1000 distinct protein spots in the vegetative mycelial cells' proteome, while proteome of primordium cells contained nearly 100 spots. Further comparison between proteome of promordia cells to proteome of vegetative mycelial cells yielded up-regulation of 5 proteins from primordium cells that were labeled as P1, P2, P3, P4.,Tiger's milk mushroom is a wild tuberious mushroom that is being used by Aborigines to treat a variety of diseases including cancer. Molecular phylogenetic analysis utilizing second largest subunit RNA polymerase II gene (RPB2), identified the wild Tiger's Milk mushrooms collected from the state of Pahang, Malaysia for this study as Lignosus rhinocerotis (Cooke) Ryvarden in the order Polyporales with 99% homology. Pure culture (mycelia) of this indigenous sample was successfully cryopreserved using vitrification method. Proximate analysis of 100 g wet weight of this mushroom yielded the pileus contains 56.30 ± 0.26 g carbohydrate, 32.22 ± 5.54 g moisture, 6.60 ± 1.38 g protein, 0.21 ± 0.09 g fat and 1.77 ± 0.28 g ash; stipe of this mushroom contains 38.47 ± 0.49 g carbohydrate, 46.31 ± 14.00 g moisture, 2.76 ± 0.60 g protein, 0.27 ± 0.07 g fat and 1.76 ± 0.93 g ash; while tuber of this mushroom contains 51.30 ± 0.10 g carbohydrate, 39.82 ± 2.22 g mositure, 3.04 ± 0.59 g protein, 0.46 ± 0.32 g fat and 4.38 ± 2.71 g ash. In determining the solid growth mediun, it was observed that the mycelial growth of this mushroom is optimal in glucosepeptone mediun and yeast extract peptone dextrose mediun at 30 o C between pH 6 and 7. While for submerged culture, the optimal compositions for mycelial growth were 80 g/L of glucose, 4 g/L of potassium nitrate, 0.4 g/L of FeSO4.7H2O and 0.1 g/L of CaCI at 25 o C and pH6; consequently, the optimal compositions for exopolysaccharides (EPS) production were 80 g/L of glucose, 4 g/L of potassium nitrate, 1.4 g/L of FeSO4.7H2O and 1.1 g/L of CaCI2 at 25 o C and pH6. In identifying the proteins involved in the formation of primordium, protein expression analysis utilising two dimensional-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D-SDS-PAGE) with Progenesis Samespot (Version 4.0) yielded approximately 1000 distinct protein spots in the vegetative mycelial cells' proteome, while proteome of primordium cells contained nearly 100 spots. Further comparison between proteome of promordia cells to proteome of vegetative mycelial cells yielded up-regulation of 5 proteins from primordium cells that were labeled as P1, P2, P3, P4
Pages:	146
Call Number:	QK989.L338 2014 tesis
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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