Lipid production from tapioca wastewater by culture of Scenedesmus sp. with simultaneous BOD, COD and nitrogen removal

The use of microalgae to produce biodiesel or possibly remove nutrients from industrial wastewater has gained important attention during recent years due to their photosynthetic rate and its versatile nature to grow in various wastewater systems. In this study, a microalgae, Scenedesmus sp., was cultured to enhance the lipid production and nutrients removal from tapioca wastewater sample. To assess lipid production, Scenedesmus sp. was cultured in different concentration of tapioca wastewater sample (from 0 to 100 %), and nutrient removal including BOD, COD, NH4, NO2, NO3 level by Scenedesmus sp. was assessed in 100% of tapioca wastewater culture. After 8 days of culture, it was found out that 50% of tapioca wastewater sample resulted in highest concentration of lipid content than that of the other concentrations. The level of environment indicator as nutrient removal such as BOD, COD, NH4, NO2, NO3 were also decreased up to 74%, 72%, 95%, 91%, and 91%, respectively. The pH condition changed from initial condition acidic (pH: 4) to neutral or basic condition (pH: 7-8) as recommended in wastewater treatment system. This research provided a novel approach and achieved efficient simultaneous lipid production and nutrients removal from tapioca wastewater sample by Scenedesmus's culture system

Dampak penambahan bahan organik terhadap pertumbuhan dan patogenisitas jamur Rhizoctonia solani Kühn penyebab rebah kecambah pada tanaman kapas

ABSTRAK Kapas merupakan salah satu bahan baku sandang di Indonesia. Kurang lebih 78 % bahan baku sandang berasal dari serat kapas alam, namun kapas di Indonesia berkembang tidak sesuai dengan harapan. Salah satu kendala budidaya tanaman kapas adalah adanya serangan patogen. Rhizoctonia solani merupakan patogen utama pada tanaman kapas. Jamur ini mampu membentuk sklerosia dan mampu bertahan hidup dalam waktu yang lama di dalam tanah. Kondisi tanah yang lembab menyebabkan sklerosia berkecambah dan menginfeksi organ tanaman, sehingga menimbulkan penyakit. Tujuan Penelitian ini adalah untuk mengetahui dampak penambahan bahan organik dan waktu inokulasi terhadap pertumbuhan dan patogenisitas jamur R. solani. Penelitian terdiri dari 2 macam; pertama dampak penambahan ekstrak bahan organik terhadap pertumbuhan R. solani secara in vitro dan kedua penambahan bahan organik dan waktu inokulasi terhadap patogenisitas R. solani. Kedua penelitian tersebut merupakan penelitian eksperimen. Pada penelitian dampak penambahan ekstrak bahan organik terhadap pertumbuhan R. solani secara in vitro rancangan yang digunakan adalah rancangan acak lengkap (RAL). Ekstrak bahan organik yang digunakan adalah : ekstrak pupuk kandang ayam (PKA) dan sapi (PKS), kulit rajungan, biji mimba, bunga cengkeh dan umbi bawang putih dengan perbandingan 1 : 1 (100 gram bahan organik : 100 ml pelarut/alkohol). Analisis data yang digunakan adalah analisis varian satu arah. Pada penelitian penambahan bahan organik dan waktu inokulasi terhadap patogenisitas R. solani digunakan rancangan acak lengkap (RAL) faktorial yang terdiri dari dua faktor, yaitu penambahan bahan organik dan waktu inokulasi. Penambahan bahan organik terdiri dari pemberian campuran pupuk kandang ayam (PKA) dengan serbuk biji mimba (SBM), pemberian campuran pupuk kandang sapi (PKS) dengan SBM, dan pemberian serbuk kulit rajungan (SKR) dengan SBM dan kontrol (tanah pasir tanpa campuran apapun) masing-masing dengan perbandingan 1: 3. Waktu inokulasi terdiri dari 2 taraf, yaitu waktu inokulasi R. solani bersamaan dengan pencampuran bahan organik dan waktu inokulasi R. solani dilakukan setiap bulan setelah pencampuran bahan organik. Masing-masing pengamatan terdiri dari 4 jenis perlakuan dan 3 kali ulangan. Analisis yang digunakan adalah analisis varian dua arah. Hasil penelitian menunjukkan bahwa penambahan ekstrak bahan organik dengan konsentrasi 1% hanya mampu memperlambat pertumbuhan R. solani, sedangkan pada konsentrasi 4% mampu menekan pertumbuhan R. solani hingga 100%. Ekstrak PKS, kulit rajungan dan bunga cengkeh merupakan ekstrak yang paling efektif menekan pertumbuhan R. solani secara in vitro. Penambahan bahan organik berupa campuran PKA dengan SBM dan PKS dengan SBM mampu menurunkan persentase kecambah tanaman kapas terserang R. solani, sedangkan pada penambahan SKR dengan SBM belum mampu menurunkan persentase kecambah tanaman kapas terserang R. solani, bahkan bersifat racun bagi kecambah tanaman kapas. Masa inkubasi bahan organik berupa campuran PKA dengan SBM dan PKS dengan SBM yang paling efektif menekan patogenisitas R. solani adalah 3 bulan. Pemberian waktu inokulasi R. solani yang berbeda dan kombinasi antara waktu inokulasi dan penambahan bahan organik tidak memberikan pengaruh nyata terhadap patogenisitas R. solani

Solid Waste from Winery as a Value-Added Food

[[abstract]]Grape seeds and peels contain high level total phenolic that can reduce oxidative stress and may prevent chronic diseases, such as cancer and cardiovascular disease. Tyrosinase also known as polyphenol oxidase (PPO), is responsible not only for melanization in animals but also browning in plants. The inhibitors of this enzyme are important in cosmetics for skin-whitening and should be clinically useful for the treatment of some dermatological disorder. The objective of our research is to identify, analyze polyphenolic content in grape seeds and peels of solid grape waste and their inhibitory effect on activity of mushroom tyrosinase. We propose that a valuable food and cosmetic ingredient could be obtained after drying and grounding to powder without large losses of phytochemicals. Total phenolic, flavonoid and anthocyanin content were measured by Folin-Ciocalteu method, colorimetric method and spectrophotometric method, respectively. The antioxidant capacity was determined as were expressed as Trolox equivalents antioxidant capacity (TEAC). Diphenolase activity of mushroom tyrosinase was assayed using 1 mg per ml of L-3,4-dihydroxyphenylalanine (L-DOPA) as substrate and 10 μl of mushroom tyrosinase has been used. Our current results revealed that post-fermentation solid grape waste contain higher level total phenolic, flavonoid and anthocyanin content than the pre-fermentation ones, significantly. Based on DMRT analysis, addition of 0.5 % ascorbic acid in the prefermentation peels could preserve total phenolic compounds, significantly. Likewise in the mixture of seeds and peels of post-fermentation, addition of 0.5 % ascorbic acid also could increase total phenolic compounds. The total phenolic compounds increase in post-fermentation seeds under treatment of 1 s steaming. The only treatment that can increase total phenolic compounds in the post-fermentation peels was addition of 0.5 % citric acid in the solution. The optimal processing condition for the post-fermentation seeds to preserve total phenolic content was 1 s steaming, then oven-dried at 60°C of 37.591±0.610 mg gallic acid equivalent per g of grape seeds powder. Whereas for the peels of post-fermentations, the best processing condition to preserve total phenolic content were dipped in 0.5 % citric acid then oven-dried at 80°C of9.062±0.682 mg gallic acid equivalent per g grape peels powder. For the mixture of seeds and peels of post-fermentations, total phenolic content could be retained by dipping in 0.5 % ascorbic acid solution then oven-dried at 60°C and 80°C of 25.729±0.113 and 22.229±0.225 mg gallic acid equivalent per g of grape seeds and peels powder, respectively. Addition of 0.5 % ascorbic acid then oven-dried at 80°C of 5.763±0.009 mg gallic acid equivalent per g of grape peels powder, were the best processing condition to increase total phenolic content in the prefermentation peels. Post-fermentation seeds steaming for 1 s also had higher antioxidant capacity of 1705.79 ± 4.99 μM trolox equivalents per g of solid grape waste powder, than peels or mixture seeds and peels. In this research, the inhibitory effects of grape solid waste on the activity of mushroom tyrosinase had been studied. Our result indicated that in 1 mg/ml of seeds of post-fermentation with 1 s steaming oven-dried at 60°C and air-dried had higher inhibitory effect of 88.03%±0.72 and 79.24 %±0.72 on the activity of mushroom tyrosinase. Based on these findings, grape seeds and peels powder could be utilized to obtain compounds with rich total phenolic compounds, which can be used as a valuable and attractive addition to healthy food products and cosmetic products

Transgenic resistance against tospoviruses conferred by a single-chain antibody against the common epitope of the gene silencing suppressor

利用轉基因植物表現抗體使植物避免病毒感染是一種抗病毒之可行策略。本研究室先前研發出西瓜銀斑病毒 (Watermelon silver mottle virus, WSMoV) 基因沉寂抑制子 (gene silencing suppressor, NSs protein) 之單株抗體 (monoclonal antibodies, MAbs)，此抗體可以辨認WSMoV 血清群 NSs 之高保留區抗原決定基 (common epitope, WNScon)。本研究將此單株抗體融合瘤細胞株239F1B9之單鏈抗體變異區 (single-chain variable fragment, scfv) 構築於載體中，並以農桿菌為媒介進行菸草轉殖。首先利用 scFv 專一性引子確認30個擬轉基因菸草中可增幅出0.74 kb的專一性片段。經 WSMoV 接種後，結果有一個品系不發病被歸類為免疫抗性，四個為品系高度抗性，兩個為中度抗性及一個為微度抗性。以 WSMoV 血清群病毒進行廣泛抗病毒能力分析，結果顯示轉基因菸草對彩色海芋黃化斑點病毒 (Calla lily chlorotic spot virus)、番椒黃化病毒 (Capsicum chlorosis virus)、花生頂芽壞疽病毒 (Peanut bud necrosis virus)、西瓜頂芽壞疽病毒 (Watermelon bud necrosis virus) 具有延遲性抗性，但對同一血清群關係較疏遠的甜瓜黃斑病毒 (Melon yallow spot virus) 則不具抗性。北方及西方轉漬法結果顯示一個免疫品系、三個高抗及一個中抗品系具有較高量的scFv 轉錄體及蛋白累積，此表示 scFv 轉基因植物之抗性與其轉錄體及蛋白表現呈正相關性。利用純化的NSs-HA蛋白當作探針，以蛋白-蛋白套疊法分析顯示 scFv 與 NSs 具結合能力，因此scFv轉基因植物對WSMoV具有抗性應該是藉由scFv 與 NSs之結合產生的蛋白主導的抗病性狀。Expression of antibodies in transgenic plants is a possible strategy to prevent plant viral infection. The monoclonal antibodies (MAbs) against a common epitope WNSscon (amino acid 98-120) of the gene silencing suppressors (NSs proteins) of Watermelon silver mottle virus (WSMoV) serogroup tospoviruses were previously produced from our laboratory. In this study, a clone encoding a single-chain variable fragment (scFv) against a common epitope WNSscon was generated from a hybridoma cell line 239F1B9 by polymerase chain reaction (PCR) and used for Agrobacterium-mediated transformation of Nicotiana benthamiana plants. Using scFv-specific primers, a DNA fragment of 0.74 kb was amplified from 30 putative transgenic tobacco lines by PCR. The R0 scFv transgenic lines challenged inoculated with WSMoV showed various degrees of resistance: one line immune, four lines resistant, two lines moderately resistant, and one line weakly resistant to WSMoV. Evaluation of their broad-spectrum resistance against other WSMoV serogroup tospoviruses showed delayed-type resistance to Calla lily chlorotic spot virus (CCSV), Capsicum chlorosis virus (CaCV), Peanut bud necrosis virus (PBNV), and Watermelon bud necrosis virus (WBNV), but not obvious resistance to Melon yellow spot virus (MYSV). Northern hybridization revealed that one immune line, three resistant lines and one moderately resistant line accumulated detectable levels of scFv transcript. Moreover, immunoblot analysis demonstrated that the one immune line, three resistant lines and one moderately resistant line expressed high levels of scFv protein. Our results indicated that the expression levels of scFv transcript and protein are correlated to the levels of transgenic resistance. The protein-protein overlay assay using purified NSs-HA protein as a probe indicated the binding activity of expressed scFv and NSs protein.We conclude that the resistance to the WSMoV infection is resulted from the binding activity of the scFv and NSs protein.Abstract...................................................i Table of Content.........................................iii Introduction...............................................1 Materials and Methods......................................6 Virus sources and propagation..............................6 Construction of the scFv from WNSs MAb gene................6 Agroinfiltration and transformation of scFv-KDEL transgenic N. benthamiana plants................................. 7 DNA extraction and polymerase chain reaction analysis......9 Northern hybridization analysis............................9 Construction of pET32a-scFv and protein expression........10 Purification of bacteria-expressed scFv...................11 Production of mouse scFv antiserum........................12 Western blotting analysis.................................13 Resistance evaluation of scFv transgenic plant to WSMoV under greenhouse conditions...............................14 Indirect enzyme-linked immunosorbent assay (ELISA)........14 Resistance evaluation of scFv transgenic lines to WSMoV serogroup viruses under greenhouse conditions.............15 Construction of pBCO-scFvKDEL-myc and pBCO-scFvKDEL-HA....15 Construction of pMAL-NSs-HA and protein purification......16 Protein-Protein overlay assay.............................17 Results...................................................19 Construction of transgene encoding scFv of MAb to WNSscon.19 PCR analysis for the presence of the transgene in putative transgenic tobacco lines..................................19 Resistance evaluation of scFv transgenic lines to WSMoV under greenhouse conditions...............................20 Resistance evaluation of scFv transgenic lines to WSMoV serogroup viruses under greenhouse conditions.............21 Northern hybridization analysis of scFv transcript in transgenic plant.....................................................22 The expression of the scFvKDEL reading frame in pET32a system....................................................22 Purification of scFv fusion protein.......................23 The production of scFvKDEL antiserum......................23 Western blotting analysis of the expression of scFv protein in transgenic plants......................................24 Binding activity of expressed scFv-KDEL and WSMoV NSs protein...................................................24 Discussion................................................26 References................................................3