Kloning Dan Karakterisasi Edna Dari Gen Proteinase Inhibitor Klon Kakao (Theobroma Cacao L) Tahan Dan Tidak Tahan Terhadap Penggerek Buah Kakao Serta Uji Toksisitasnya

Isda, Novializa Mayta (2011) Kloning Dan Karakterisasi Edna Dari Gen Proteinase Inhibitor Klon Kakao (Theobroma Cacao L) Tahan Dan Tidak Tahan Terhadap Penggerek Buah Kakao Serta Uji Toksisitasnya. S3 thesis, Universitas Andalas.

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Abstract

Cacao {Theobroma cacao L.) is a commodity crop that it has economically significance for tropical countries in generally and Indonesia in especially. Indonesia is the third largest cocoa producer in the world after Ghana. More than 80% of the world's cocoa production are lossing due to fungi, viruses and insect pests. The main pests that attack cocoa production in Indonesia, including Southeast Asia are the cacao pod borer (CPB) which which caused by Conopomorpha cramerella (Lepidoptera). In Indonesia, yield loss due to attacks CPB interpreted approximately 60,000 tons per year. Based on the nature of the cacao pod borer larvae, these pests are very difficult to overcome by the chemical and biological control. Various efforts have been made for tackling the CPB attack, but there is no way of controlling the effective and efficient against a very rapid spread of these pests. One of solution is the center of attention today is the provision of seeds of plants that are resistant to fruit borer cocoa. To search for new alternatives in terms of molecular cocoa crop is attacked at CPB and socially acceptable, it is necessary to harness the potential of resistance genes contained in the cocoa itself. One of the genes for traits such as resistance in plants against insect pests is the gene coding for proteinase inhibitors (PIN)- gene is expected to help efforts of cocoa breeding and selection of resistant clones from several centers CPB cocoa in Indonesia. The research for tihs ne^' discovered gene derived from the pin cocoa beans that function only as storage protems. To eet the oin gene that functions as a defense gene against the cacao pod borer in this study sought genes that pin borer resistance derived from the frmt itself. In this research has isolated a gene derived from the bark of pin cocoa pods of the resistant clones and susceptible CPB by PCR and constructed the expression vector pBRf. The result showed that the PIN genes isolated that ±600 bp and has a high homology with the genes encoding PIN proteins 21 kDa in size. There is a difference between the PiV gene amino acids derived from resistant and susceptible to cocoa pod borer seen from the alignment that the existence of differences in some amino acids in area 5' on the amino acid sequence to 33 to 100. The differences in gene expression from various tissues of the pin outer pod wall, inner pod wall, pulp, seed and leaves. The expression of TcP/V gene was stronger in the cocoa pod wall that are resistant to a pod borer cocoa (KW5I4) more than the wall that a susceptible to pod borer cocoa (ICS60) with quantitative values in a row on the pod wall of resistant cocoa and susceptible is 47 771 and 5680 (for outer pod wall), 30 890 and 3631 (for inner pod wall). The based on the results of SDS-PAGE there are differences in trypsin activity contained in the cocoa pod wail of resistant or susceptible clones. The trypsin activity in vitro was higher in the cocoa pod wall resistant clones (KW514) more than those susceptible (ICS 60) to CPB.

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