Endophytic Bacteria Utilization to Increase Yield and Andrographolide Content on King of Bitter (Andrographis paniculata).

Abstract

King of bitter (Andrographis paniculata) is widely used in the pharmaceutical industry as antimalarial, antidiabetic, antimicrobial, antiviral, anti-inflammatory, and anticancer. The problems in its cultivation were low yield and andrographolide content since appropriate cultivation techniques have not been standardized and minimal research. One of the solutions was the application of endophytic bacteria and phosphate. Endophytic bacteria have roles as growth promoter and producer of secondary metabolites. They generate plant hormone such as auxin, gibberellins, and cytokinins, and also can supply nutrients such as Nitrogen fixation and Phophate solubilization. Phosphate acts as a constituent of macro molecules and compound storage; transferring energy; and regulating biochemical reactions. This research aimed to increase yield of king bitter plant both in quantity and quality. This study had general objective to find endophytic bacteria consortia and appropriate phosphate dosage to increase yield and andrographolide content of king of bitter. Specifically it aimed to: 1) obtain endophytic bacteria consortia at king of bitter indigenous from several agro-climatic conditions which can promote plant growth, 2) obtain potential endophytic bacteria consortia which can stimulate plant growth and increase yield and andrographolide content, 3) obtain the information of the association of endophytic bacteria consortia and phosphate on growth, yield and andrographolide content of king of bitter in the field, 4) obtain the information of association of endophytic bacteria consortia and phosphate in increasing the yield and content of andrographolide of king of bitter in nutrient solution media. This dissertation consisted of four experiments. The title of first experiment was “The exploration, isolation and selection of endophytic bacteria consortia from king bitter plant as plant growth promoter”. The activity in this study was to isolate endophytic bacteria from four locations which has different agro-climatic conditions viz. West Java (Bogor), Central Java (Blora), and East Java (Madiun and Pasuruan). The results showed there were 24 endophytic bacteria consortia from roots, stems, and leaves. Media used for isolation were 5% and 20% Trytone Soya Agar (TSA). The consortia isolated from root tissue (105-106 CFU ml-l) had higher population than from leaves and stems (103-105 CFU ml-l). All the 24 consortia were potential as plant growth promoter. However, only four consortia (5MD, 20BB, 20BD, and 20CD) had better effect on plant growth than others, so chosen and tested for further research. The title of the second experiment was “The potency of endophytic bacteria consortia to promote growth and andrographolide content on king bitter plant”. Four selected endophytic bacteria and one bacteria isolated from Graminae leaves, were tested in the greenhouse. The experiment was arranged in randomized block design (RBD), 6 treatments and 4 replications. Treatments consisted of: control, 5MD, 20BB, 20BD, 20CD consortia, and isolate 90AA. The results revealed that the consortia were able to enhance plant height, number of primary branches, shoot dry weight and andrographolide content at 14 weeks after planting (WAP). Shoot dry weight increased 25-82.81%, as well as N, P, and K uptake. The highest increasing of shoot dry weight and andrographolide yield was indicated by 20CD. Andrographolide content of 20CD, 20BB and 20BD treatments were significantly higher than control. The title of the third experiment was “The application of endophytic bacteria and P fertilizer on king of bitter in the field”. This research was designed in RBD factorial, 9 treatments, and 3 replications. The first factor was endophytic bacteria consortia treatments 1) no endophytic bacteria consortia, 2) 20CD, and 3) 20BB. The second factor was the rate of P fertilizer consisted of: 1) without P, 2) 27 kg ha-1 P, and 3) 54 kg ha-1 P. The results showed that both endophytic bacterial consortia were capable of enhancing growth, and increasing yield and andrographolide content at 14 WAP. The 20CD and 20BB treatments had similar effect in increasing dry matter production, content and yield of andrographolide. Andrographolide content of 20CD and 20BB treatments were 2.42 and 2.69% respectively. The application of P fertilizer significantly increased growth, dry matter yield than without P. The increased of dry matter yield were indicated by physiological characters of plant such as increase in leaf area, relative growth rate and net assimilation rate. The application of 27 and 54 kg ha-1 P gave similar result at dry matter yield and andrographolide yield. The application of 27 kg ha-1 P did not increase andropholide content. Endophytic bacteria consortia or P fertilizer application increased N, P, and K uptake. There were no interaction between endophytic bacteria consortia and P fertilizer treatments. The title of the fourth experiment was “The agronomic response of king of bitter to the endophytic bacteria consortia and phosphate in nutrient solution media”. This experiment consisted of two stages: a) the response of king of bitter to several P concentrations in nutrient solution media. The trial was designed in randomized block design, 6 treatments and 4 replications. P concentrations tested were 0, 0.01, 0.05, 0.1, 1.0, dan 2.0 mM KH2PO4, b) the response of king bitter plant to endophytic bacteria consortia and P concentration in nutrient solution media. The trial was arranged in RBD (randomized block design), 4 treatments and 6 replications. The treatments consisted of: 0.1 mM KH2PO4, 0.1 mM KH2PO4 + 20 CD, 1.0 mM KH2PO4, and 1.0 mM KH2PO4 + 20CD. The first stage experiment resulted standard concentrations of P were P deficient (0.1 mM KH2PO4) and P sufficient (1.0 mM KH2PO4). The results of the second stage revealed that the application of P sufficient produced the highest dry matter production and nutrient uptake, but produced low andrographolide content (2.33%). Phosphate deficient produced lower dry matter production, but produced highest andrographolide content (2.7%). The application of endophytic bacteria consortia at condition of P deficient or P sufficient on nutrient solution media had no significant effect on parameters observed. The conclusion from the four experiments were the acquisition of 24 endophytic bacteria consortia from king of bitter from several agro-climatic potential as plant growth promoter. In the nutrient solution media, at condition of P sufficient increased dry matter production, but at condition of P deficient increased Andrographolide content, and endophytic bacteria consortium no had effect on parameters observed. The 20CD and 20BB had the best effect to promote plant growth, yield, andrographolide content and N, P, and K uptake at 14 WAP. P fertilizer could enhance plant growth and increase dry matter yield and andrographolide content. Phosphate fertilizer did not increase andrographolide content. The application of endophytic bacteria consortia of 20CD and 20BB or 27 kg ha-1 P fertilizer were recommended for king of bitter cultivation at Latosols or soils that have the similar soil chemistry properties with experiment land.