Nurcahyani, Endang and Putri Najla, Azahra and Octaviani, Ratna and Santa Maria, Meilyana (2022) ASSEMBLY OF SUPERIOR VARIETY OF VANILLA (Vanilla planifolia Andrews.) RESISTANT TO FUSARIUM WILT DISEASE BASED ON MOLECULAR TECHNIQUES AND INDUCED RESISTANCE WITH FUSARIC ACID. In: ICON SMART 2022 -The 3rd International Conference On Natural Sciences, Mathematics, Applications, Research, and Technology, 3-4 June, 2022, Bali, Indonesia. (Unpublished)
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Abstract
Vanilla (Vanilla planifolia Andrews) is one of the plantation commodities with a fairly high economic value and has a fairly good name in the international market. In the international market, Indonesian vanilla is known as Java Vanilla Beans because it has the best quality with a vanillin content of 2.75%. The United Nations Development Program (UNDP), recommends that Indonesian vanilla be no different from "Bourbon vanilla" which has a very good image of the commodity in the international community. One of the production constraints in vanilla cultivation is stem rot disease which can cause very large losses with the result that the plant will die 50-100%, or even cannot produce and the quality of fruit from diseased plants is very low. This disease is caused by the fungus Fusarium oxysporum f. sp. vanillae (Fov) which until now still cannot be overcome effectively even though several studies have been done. The use of high-yielding varieties of vanilla that are resistant to stem rot is expected to be an important alternative for disease control. Resistance to disease can be obtained by means of Induced Resistance, namely treatment before pathogen infection with chemical compounds or by inoculation of non-pathogenic microorganisms. Induced resistance is resistance that is expressed after a plant is attacked by pathogens and can be used as an important tool for controlling plant diseases. The use of fusaric acid as a selection agent can produce variants that are insensitive to fusaric acid, so that after being regenerated into plants, the plants are resistant to Fusarium oxysporum infection. Vanilla plantlets resistant to Fov infection were initiated and selected in vitro in Murashige and Skoog medium with the addition of fusaric acid at different concentrations, and an indication of the concentration of fusaric acid tolerant for the selection of resistant vanilla plantlets was found. Inoculation of Fov fungus isolates on resistant vanilla has also been carried out, followed by analysis of DNA patterns, protein profiles, sequencing analysis, and specific expression characters on Fov resistant vanilla plantlets.This study aims to determine and determine: 1) the right combination of 2,4-D and NAA concentrations for callus initiation and shoot development from stem shoot explants and the appropriate BAP concentration for shoot initiation from stem node explants; 2) the concentration range of tolerant fusaric acid for selection of vanilla plantlets with optimum growth; 3) effective concentration of fusaric acid to suppress Fov fungus in vitro; 4) protein profile and DNA pattern of Fov resistant vanilla plantlets; and 5) sequencing analysis of ITS r-DNA regions; 6) The specific expression characters in Fov-resistant vanilla plantlets included total phenol content, peroxidase enzyme activity, lignin thickness, total chlorophyll, chlorophyll a, and chlorophyll b levels, as well as root and stem anatomical structures. The results showed that: 1) 2,4-D 2.0 mg/L + NAA 10 mg/L was the best concentration combination for callus and shoot initiation from shoot and shoot explants and BA 1.0 mg/L was the best concentration for shoot initiation from stem node explants; 2) the concentration range of tolerant fusaric acid for the selection of vanilla plantlets with optimum growth is between 90 ppm-110 ppm; 3) in vitro suppression of Fov fungi using fusaric acid concentration of 110 ppm was more effective than concentrations of 90 and 100 ppm; 4) the appearance of protein bands (±18 kD) indicated the formation of PR-protein (peroxidase) in vanilla plantlets that were resistant to Fov. The presence of new (specific) DNA bands in Fov-resistant vanilla plantlets with sizes of 930 bp (OPB_14), 430 bp (OPB_20), and 230 bp and 270 bp (OPD_19), can be predicted as candidate RAPD markers for resistance of vanilla plantlets to Fov; 5) the results of the sequencing analysis showed that the amplification of the ITS r-DNA region was detected in the 600-700 bp size band. Based on the comparison of the nucleotide base arrangement, vanilla treated with fusaric acid concentrations of 90 ppm, 100 ppm, and 110 ppm experienced mutations in the 18S; ITS 1; 5,8S; ITS2; and 26S rDNA regions; 6) the higher the concentration of fusaric acid, the higher the total phenol content, peroxidase enzyme activity, lignin thickness, and total chlorophyll, chlorophyll a, and chlorophyll b content in Fov-resistant vanilla plantlets. There were differences between the anatomical structure of the roots and stems of control, resistant and Fov-infected vanilla plantlets, namely the formation of tilosis in the trachea of roots and stems infected with Fov.
| Item Type: | Conference or Workshop Item (Speech) |
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| Subjects: | Q Science > Q Science (General) |
| Divisions: | Fakultas Matematika dan Ilmu Pengetahuan Alam (FMIPA) > Prodi Biologi |
| Depositing User: | ENDANG NURCAHYANI |
| Date Deposited: | 13 Jun 2022 01:17 |
| Last Modified: | 13 Jun 2022 01:17 |
| URI: | http://repository.lppm.unila.ac.id/id/eprint/42367 |
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