ISOLASI, IDENTIFIKASI DAN DETEKSI GEN ICE NUCLEATION ACTIVE BAKTERI PADA TUMBUHAN BERDAUN LEBAR DI JALUR PENDAKIAN CEMORO SEWU GUNUNG LAWU

KARNO, RIA (2014) ISOLASI, IDENTIFIKASI DAN DETEKSI GEN ICE NUCLEATION ACTIVE BAKTERI PADA TUMBUHAN BERDAUN LEBAR DI JALUR PENDAKIAN CEMORO SEWU GUNUNG LAWU. Masters thesis, Universitas Sebelas Maret.

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    Abstract

    Ice nucleation Active Bacteria (INA) is a filosfer bacterium that can cause frost injury on the surface of plant leaves, besides, this bacteria also has a role in the process of bioprecipitation. Research and information on INA bacteria in subtropical regions have been carried out a lot, however they are still very little to be done in the tropical regions. This research aims to obtain isolates, to determine species based on 16S rRNA gene and to find out the genetics relationship of INA bacteria which is isolated from broadleaf plants in the Tracking route of Cemoro Sewu, Gunung Lawu which consists of 6 species of Lawu broadleaf plant and also to find out the protein-coding genes of ice nucleating bacteria of INA. Isolation of INA bacteria was carried out by using spread plate method on the media of Nutrient agar + glycerol (NAG) and King's B media. The ice nucleation activity was determined by the method of tube nucleation test using circulating alcohol bath. The estimation of population were determined based on the MPN table. 16S rRNA gene amplification was done by Polymerase Chain Reaction (PCR) using 63 Forward primer and 1387 reverse primer. Genetic relationship was determined based on the phylogenetic tree in MEGA 5.1. program. Ina gene detection was done with PCR using forward and reverse primer of inaZ, inaA, inaW and inaX. DNA sequences were analyzed to find out its similarity with genes from other INA bacteria that exist in the GeneBank of NCBI online. The results of the research obtained 3 isolates of INA bacteria. A2N4 isolates which was detected from the plants of Dodonae viscose Jacq had 96% similarity with Pseudomonas syringae. C2N9 isolates from Vaccinium laurifolium plants had 97% similarity with Pantoeasp and F1N6 isolates from Polygonum chinense L plant identified had 98% similarity with Pseudomonas sp. The estimation of population of bacteria found were relatively low at <3 x 103 to 5.4 x 103/ g of leaf. The genetic relationship of INA positive bacteria from the Tracking route of Cemoro Sewu, Gunung Lawu and INA bacteria which was found previously had very close genetic relationship. The results ina gene sequence analysis showed that isolates A2N4 were 99% detected as ice nucleation protein coding genes in Pseudomonas syringae existing in ice4 gene, strain INA4. Keywords: Ice nucleation Active Bacteria (INA), isolation, identification, detection of genes, phylogenetic, broadleaf plants.

    Item Type: Thesis (Masters)
    Subjects: Q Science > Q Science (General)
    Divisions: Pasca Sarjana > Magister > Biosains
    Depositing User: Minardi Aris
    Date Deposited: 27 Sep 2015 14:41
    Last Modified: 27 Sep 2015 14:41
    URI: https://eprints.uns.ac.id/id/eprint/19614

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