Holy Wahyuni, 2019.Isolation and Characterization of Actinomycetes from
Several Sentra Plantation Onions AntagonistFusariumoxysporum f Cepaeand Test Capabilities on Plant Germination of Onion (Allium ascalonicum L.) varieties Tuktuk. Makassar State University (guided by Alimuddin Ali and Karim Hilda).
Shallots (Allium ascalonicum L.) including one multi-purpose tuber vegetable
which has a major commodity in the priority development of vegetable in Indonesia. As well as one source of income of farmers and the country's economy. However, the productivity of onion erratic, unusual increase or decrease. One cause of low production of onion is Fusarium wilt disease. This study aimed to isolate and characterize endophytic actinomycetes and rhizosphere of some onion plantation centers antagonist Fusarium oxysporum f Cepae and test capabilities on plant germination of onion (Allium ascalonicum L.) varieties tuktuk. Tests conducted with test methods antagonistic to pathogenic fungi F. oxysporum f.sp Cepae. Through the mechanism of infection, Actinomycetes isolates which have antifungal activity in plant root tissues infected red onion varieties tuktuk in vitro which then acts as a biological control in plant tissue in suppressing the growth of pathogenic fungi F. oxysporum f.sp Cepae. The next actinomycetes isolates were characterized to determine the characteristics of morphology, physiology and biochemistry. The results showed 25 isolates were isolated from onion plants, two isolates showed inhibitory activity against F. oxysporum isolates f.sp Cepae namely R2L5 and E5S5. Based on the test antagonist, it is known that isolates R2L5 have a clear zone at 47.41% and amounted to 33.48% E5S5. The characterization results of morphology, physiology, and biochemistry show that the isolated actinomycetes belonging to the genus Streptomyces. The test results measuring the levels of the pigment chlorophyll a, b, and c respectively show the difference of treatment. The treatment is not sterile in isolates of endophytic (ETS) has the highest chlorophyll content. Based on data analysis endophyte treatment just is not sterile and sterile rhizosphere effect on chlorophyll content.
oxysporum f.sp Cepae, rhizosphere, endophytic and chlorophyll. iv v
TABLE OF CONTENTS
TITLE PAGE ................................................................................................... i
ENDORSEMENT PAGE ................................................................................ ii ABSTRACT ...................................................................................................... iii TABLE OF CONTENTS .................................................................................. v LIST OF TABLES ............................................................................................ viii LIST OF FIGURES .......................................................................................... ix APPENDIX LIST .............................................................................................. x PART I PRELIMINARY A. Background ................................................................................................... 1 B. Formulation of the problem .......................................................................... 3 C. Research purposes ......................................................................................... 3 D. Benefits of research ....................................................................................... 4 CHAPTER II LITERATURE REVIEW AND FRAMEWORK FOR THOUGHT A. Literature review ........................................................................................... 5 1. General review Red onion (Allium ascalonicum L.)............................... 5 2. Overview Actinomycetes ........................................................................ 8 3. An overview of the rhizosphere and Endophytic .................................... 11 4. An overview of Fusarium oxysporum ................................................... 14 5. An overview of Onion varieties Tuktuk ................................................. 18 B. framework ..................................................................................................... 19 CHAPTER III RESEARCH METHODS A. Types of research .......................................................................................... 21 B. Time and Location Research ......................................................................... 21 C. Research execution procedures ..................................................................... 21 1. Preparation phase .................................................................................... 21 a. provision ........................................................................................... 21 b. provision of materials ....................................................................... 22 vi
c. Provision of study sample ................................................................ 22
2. Implementation phase ............................................................................. 22 a. Isolation of Fusarium oxysporum f.sp Cepae of rhizosphere Onion Sick Plants ........................................................................................ 22 b. isolation of Actinomycetes ............................................................... 23 c. Antagonist test isolates as antifungal producer ................................ 24 d. Phenotype characterization of isolates of Actinomycetes elected ... 26 1) Morphological characteristics and spore chain ornaments ........ 26 2) color mycelium ........................................................................... 26 3) Characterization of physiology and biochemistry...................... 27 a) The use of carbon and nitrogen sources ............................... 27 b) Optimization of pH growth .................................................. 27 c) Optimization of the growth temperature .............................. 28 e. Test biocontrol Fusarium Wilt Disease ............................................ 28 1) Preparation Seed ......................................................................... 28 2) Preparation Isolate Selected ....................................................... 28 3) Colonization Test Isolates .......................................................... 28 4) Measurement Photosynthesis Pigments ..................................... 29 CHAPTER IV RESULTS AND DISCUSSION A. Research result .............................................................................................. 30 1. Isolation of Fusarium oxysporum f.sp Cepae of rhizosphere Onion Sick Plants ....................................................................................................... 30 2. Insulationactinomycetes Originally rhizosphere and Plant Endophytic Shallots .................................................................................................... 31 3. Test antagonists actinomycetes as producers Antifungal Isolates .......... 36 4. Colonization Ability Test In Vitro .......................................................... 37 5. Morphological characterization and Ornaments Chain Spore Isolates Producing Antifungal Compounds .......................................................... 38 6. Characterization of Isolates Producing Mycelium Color Antifungal Compounds ............................................................................................. 39 7. Characterization of Physiology and Biochemistry .................................. 43 vii
B. Discussion ..................................................................................................... 49 1. Isolation of Fusarium oxysporum f.sp Cepae of rhizosphere Onion Sick Plants ....................................................................................................... 49 2. Insulationactinomycetes Originally rhizosphere and Plant Endophytic Shallots .................................................................................................... 51 3. Test antagonists actinomycetes as producers Antifungal Isolates .......... 52 4. Colonization Ability Test In Vitro .......................................................... 55 5. Morphological characterization and Ornaments Chain Spore Isolates Producing Antifungal Compounds .......................................................... 56 6. Characterization of Isolates Producing Mycelium Color Antifungal Compounds ............................................................................................. 58 7. Characterization of Physiology and Biochemistry .................................. 59 8. Measurement Photosynthesis Pigments .................................................. 61 CHAPTER V CONCLUSIONS AND RECOMMENDATIONS A. Conclusion .................................................................................................... 63 B. Suggestion ..................................................................................................... 64 BIBLIOGRAPHY ............................................................................................. 65 ATTACHMENT................................................................................................ 73 viii
LIST OF TABLES
Table Page
table 4.1Isolates and Images rhizosphere and endophytic actinomycetes isolated
from plant tissue of onion (Allium ascalonicum L.) .................. .. ............... ................. ............................................... 31
Table 4.2 Characterization of Color mycelium actinomycetes isolates producing
Figure 3.1.Actinomycetes isolates placement scheme and Fusarium oxysporum ............................................................................................................................. 25 Figure 4.1Morphology isolates of Fusarium oxysporum f. sp Cepae ................. 30 Figure 4.2 Test actinomycetes isolates antagonistic against Fusarium oxysporum ............................................................................................................................. 36 Figure 4.3 The observation of Fusarium oxysporum before and after the test antagonist ............................................................................................................ 37 Figure 4.4 The appearance of colonization In Vitro ........................................... 37 Figure 4.5 The appearance of morphology and spore isolates of actinomycetes chain ornaments .................................................................................................. 38 Figure 4.6 The results of chlorophyll .................................................................. 46 x
APPENDIX LIST
Table Page
Appendix 1.Karakterisasi mycelium color .......................................................... 73
Appendix 2 Biochemical characterization, ......................................................... 77 Appendix 3. Characterization Physiology........................................................... 85 Appendix 4. Table composition of the medium .................................................. 87 Appendix 5. Harmonica color ............................................................................. 89 Appendix 6. Data Analysis ................................................................................. 93