Judul: Density of Fusarium oxysporum and Phytonematodes of Banana Plantation at PTPN VIII Parakansalak
Abstrak: Banana is the most consumed of horticulture commodity plants. The government and society also cultivate some banana varieties such as Cavendish, Raja Buluh, and Barangan. Considering the production demand is also increased, the government also implemented the fruit self-sufficiency program in the Minister of Plantation Regulation Number 86/Permentan/OT.140/8/2013. Therefore, BUMN began to cultivate banana plants in several plantation sectors owned by the company. During the banana plants’ cultivation process, many problems are faced, one of which is the banana plant attacked by a pathogen. The primary pathogen that attacks the banana plant is Fusarium oxysporum causing the fusarium wilt. F. oxysporum is categorized as the soil-borne pathogen that infected banana plant through the wound of plant tissue. The wound found in the plants’ root could be identified as the natural wound result of plant growth process or caused by several vectors such as nematode. Many researchers reported that nematode has a significant role in the process of fusarium wilt disease, which is caused by F. oxysporum. This research aims to observe the abundance of F. oxysporum and phytonematode founded in the banana plant and the relation of both pathogens in causing fusarium wilt. The hypothesis of this research that phytonematode founded a positive correlation toward the severity of fusarium wilt disease. The method used in this research was observing the severity of fusarium wilt of banana plant Emas Kirana varieties that’s most know as Mas Kirana. Based on the predetermined score rating. Soil and root samples was collected which was conducted by purposive method. From every block, samples were taken from 5 different clumps with different disease severities. Samples were collected from 5 different blocks as replication. Sample from 5 different blocks with the same disease severity were then mix for future research. The abundance of F. oxysporum and phytonematodes were calculated by applying conventional and molecular methods. Isolation of phytonematodes from soil were conducted by extracting the phytonematodes from soil samples by using a floatation centrifugation method. Identification of phytonematodes to the genus level were carried out by observing the morphology of their anterior to posterior. The abundance of phytonematodes then calculated under microscope by placed 1 ml of phytonematode suspension on cyrus dish, and repeated 3 times. Furthermore, the number of phytonematodes was calculated using the formula of absolute population calculation. Calculation of F. oxysporum was conducted by two ways, conventional (growing on synthetic media) and molecular methods. The conventional method was performed by serial dilution of soil samples, followed by cultured on Martin Agar, and incubated for 3-4 days before counting the number of F. oxysporum colonies grow on medium. To prove that F. oxysporum growing on Martin Agar is Foc, molecular identification was carried out using specific primer Foc1/F (5’-CAGGGGATGTATGAGGAGGCT-3’) and Foc2/R (3’-GTGACAGCGTCGTCTAGTTCC-5’). PCR result then visualized on agarose gels. Real-time PCR technique was also conducted to count the abundance of F. oxysporum from soil samples. The standard curve construction is started by isolating the competent bacteria cell of DH5 alpha that has been added the synthesis gene of F. oxysporum. The cell transformation is becoming plasmid that
Keyword: Panama disease, real-time PCR, tropical race 4