Growth and yield performance of four generations of high amylose transgenic Adira 4 cassava

Attempt to improve cassava starch quality have been conducted by producing the first high amylose transgenic Adira 4 Indonesian cassava. This study aimed to evaluate agronomic performances of transgenic plants, assessment of four generations in specific containment screenhouse and limited field trial were conducted with improved method. The survival and growth rate of cuttings was enhanced by applying growth regulator such as Atonik and improving planting procedures. The growth and yield performances were measured by comparing related variables across four generations of selected lines. Dipping in 1 mL.L-1 Atonik solution with shading in the early planting were only required when the plant growth was poor and not favorable soil and weather. The result showed that survival of third and fourth generations of Lines A55-5 and A55-15 could reach 100 % when it was treated with Atonik. According to 14 lines assessed, Line A55-5 was one of the promising ones due to its growth, multiplication performance, and almost double amylose content compared to wild type. Meanwhile, Line A55-12 showed the highest yield. The finding will benefit functional food industries when appropriate gene construct and approaches were applied for better growth and yield results.

CRYOPRESERVATION OF TRUE-SEED AND EMBRYO OF MAIZE AND SOYBEAN FOR LONG-TERM STORAGE

Study on cryopreservation of Indonesian local cultivars and improved  varieties of maize and soybean has never been done. This method may be used for long-term preservation of seeds of maize and soybean. In this study, the method was applied to maize and soybean, Arjuna and Wilis respectively, as a model for preserving germplasm of ortodox seeds. Whole seeds and excised embryos of both varieties were subjected to two methods of cryopreservation, i.e., two-stage cooling and rapid freezing with or without 15% dimethyl sulfoxide (DMSO) as cryoprotectant solution prior to immersion in liquid nitrogen (-196oC). Results indicated that there was no significant difference between the use of DMSO for both species in terms of viability, although pretreatment in DMSO was slightly reduced the percentage of viability of both species. Slow freezing to -30oC prior to immersion in the liquid nitrogen could give as high as 76.67% and 51.67% surviving whole seeds of maize and soybean, respectively. Preserving excised embryos of maize in the liquid nitrogen using either slow or rapid freezing significantly reduced the percentage of viability from 20-76.67% to 5-18.33% (four folds) depending on treatments applied. Results also showed that one day or 15 minutes of immersion of samples in the liquid nitrogen gave rise to similar values of viability of maize and soybean, i.e., 20-60% and 20-51.67%, respectively depending on  treatments applied. These results implied that for long-term storage of maize and soybean seeds as they could survive at the rate of 76.67% and 51.67% respectively, the seed can be treated by prefreezing to -30oCwithout the presence of DMSO prior to immersion in liquid nitrogen

Determination of the Optimum Gamma Ray Lethal Dosage for Mutation Breeding of Indonesian Cassava Genotype Mentega 2

This study aimed to determine the optimum lethal dosage (LD50) of gamma rays in cassava and to study the effect of gamma irradiation on the morphological characters. Gamma rays is one of the physical mutagens that has been widely used for mutation breeding to improve the genetic traits of several crops including cassava. A total of 72 plantlets of Indonesian cassava genotype (Mentega 2) were cultured on Murashige & Skoog (MS) medium containing 2% of sucrose and were exposed to 0, 5, 15, 30, 50, and 75 Gray (Gy) using a Cobalt-60 source at dose rate of 600 Gy/h. Morphological characters such as plant height, leaf number, leaf width and length, and root number and length were measured. The percentage of plant survival decreased with the increase of gamma rays dose. The lowest survival rate (17%) was shown by 75 Gy-treated samples. The morphological characters’ observation at 60 days of treatment showed the highest plant height was obtained from 15 Gy-treated samples (12.41±1.84 cm), while the lowest plant was from 75 Gy-treated samples (2.08±1.98 cm) compared to control (4.17+1.46 cm). The LD50 value was calculated to be 29.7 Gy which will be referred for further studies

Storage and the Use of Peroxydase Enzyme to Detect Germination Capability of Sandoricum koetjape Merr. Seeds- A Neglected Tropical Fruit Species

Sandoricum koetjape which belongs to the group of mahogany, possesses seeds with sticky white aril, is a neglected local fruit species that might extinct if conservation efforts are not made. Besides preserving the embryos and embryonic axis on different periods of storage (0, 3, 5, 7 days) on a vacuum glass container containing silica gel, the latter organs were also preserved in liquid nitrogen to study the possibility of long-term storage. The water content of the preserved organs was measured in relation to the length of storage and the germination rate. To determine the role of peroxidase in the germination rate of preserved zygotic embryos, the level of peroxidase was measured. Seeds of control and of 3-day storage were mostly germinated at day-6. The average rate of germination was reduced to 23.33% when the seeds were desiccated with silica gel for 7 days which resulted in 27.69% water content. This germination capability and the length of hypocotyls seem to correlate with peroxidase activity in the seeds. In general, the higher the percentage of germination, the longer the length of hypocotyls, the higher the peroxidase activity, except for seeds desiccated for 7 days. The range of peroxidase activity was 6.81-3856.20 A/2min/mg. When the seeds were desiccated for 7 days, they still could germinate at day-18 which indicated by a very high peroxidase activity. Peroxidase activity assay could detect the viability within 15 seconds while the TTZ requires 15 minutes. Although the highest percentage of survived embryonic axis after storage in liquid nitrogen was only 23.42%, the results showed that soaking in 10-20% DMSO for 20 minutes of prerequisite as without DMSO led to no survival. These results offer an alternative procedure to detect the germination ability of seeds at early stage and longer period of preservation which could contribute to future ex situ conservation

Molecular cloning of gene fragment encoding 4-coumarate: Coenzyme A ligase of Sengon (Paraserianthes falcataria)

4-coumarate:Coenzyme A ligase (4CL) plays an important role in lignin biosynthetic pathway thatcatalyzed the activation of coumaric acid, caffeic acid or ferulic acid to be a syringil monomer. Ligninbiosynthesis control through 4CL down regulating would support lower lignin wood production. Theobjective of this study was to clone conserved region cDNA of gene encoding 4CL. Gene fragment isolation wasconducted by means of reverse transcriptase polymerase chain reaction (RT-PCR) using degenerateheterologous primer. The RT-PCR products were purified, sequenced and analyzed to select the highlyhomologous fragment to 4CL. BLASTanalysis result showed that deduction of amino acid sequences from oneof two RT-PCR product nucleotide was highly homologous with the 4CL conserved region from Rubbus ideaus,Oryza sativa, Populus tomentosa, Populus balsamifera, Betulla platyphilla, Nicotiana tabacum, and Arabidopsisthaliana with identity ranging from 78-90%.Key words: 4-coumarate: Coenzyme A ligase, lignin, sengo

INDUKSI MUTASI DAN SELEKSI IN VITRO TANAMAN GANDUM (Triticum aestivum L.)

INDUCTION MUTATION AND SELECTION OF IN VITRO PLANT OF WHEAT (Triticum aestivum L.)The goal of this research was to produce wheat crop which is tolerant to lowland condition.Six varieties were used, Dewata, Selayar, Alibey, Oasis, Rabe and HP1744. This research consisted of 4 stages: production of the best callus on MS medium containing 3 g/L 2.4-D, induced mutation of embryogenic callus using EMS, in vitro selection of callus at temperature of 27–35°C, and callus regeneration. The best result for callus production was 76% for Dewata and 70% for Selayar varieties. Higher concentration of EMS and longer soaking time decreased the percentage of callus growth. LC50 for Dewata was 0.3% EMS at 30 minutes and that for Selayar was 0.1% EMS at 60 minutes. The higher the temperature was, the lower was the adaptation tolerant of the plants, and callus growth was inhibited. At the highest temperature (35°C) the callus did not grow at all.Keywords: Induced mutation, Triticum aestivum, EMS, in vitro selection, callusABSTRAKTujuan penelitian ini adalah untuk merakit tanaman gandum yang toleran pada dataran rendah. Varietas yang digunakan ada 6 yaitu Dewata, Selayar, Alibey, Oasis, Rabe dan HP-1744. Penelitian terdiri atas empat tahap yaitu induksi pembentukan kalus terbaik menggunakan media MS + 3 g/L 2,4-D (dipilih dua varietas yang terbaik), induksi mutasi kalus embriogenik menggunakan EMS, seleksi kalus in vitro pada suhu 27–35°C, dan regenerasi. Hasil induksi kalus terbaik terdapat pada varietas Dewata sebesar 76% dan Selayar sebesar 70%. Semakin tinggi konsentrasi EMS dan semakin lama waktu perendaman yang digunakan maka semakin menurun persentase pertumbuhan kalus. LC50 varietas Dewata adalah EMS 0,3% waktu 30 menit sedangkan LC50 varietas Selayar adalah EMS 0,1% waktu 60 menit.Semakin tinggi suhunya maka semakin berkurang toleran adaptasi tanaman tersebut, dan pertumbuhan kalus semakin sedikit. Bahkan pada suhu tertinggi yaitu suhu 35°C tidak ada pertumbuhan kalus sama sekali.Kata Kunci: Induksi mutasi, Triticum aestivum, EMS, seleksi in vitro, kalu

Evaluation of Adaptation and Production of Three Selected Cassava (Manihot esculenta Crantz) in Peat Land Area of Central Kalimantan

Along with the increasing the of the land-use change of the fertile agricultural land in Java, the better awareness has been subjected to the importance of function peat land for agricultural development. Cassava (Manihot esculenta Crantz) is a tuber plant that potentially developed as important carbohydrate source. The objective of this research was to identify superior cassava variety candidate of selected cassava developed by of Research Centre Biotechnology, LIPI which has high adaptation capability and yield in Central Kalimantan peat land area. This research was conducted from July 2010 through May 2011 in Central Kalimantan Province (Kalampangan dan Pulang Pisau in total of area 1.5 and 2.25 ha respectively). There were three genotypes of cassava tested, i.e. Iding (high amylose), Gebang (low amylose), and Menti (high starch) compared to Adira 4 (35 ton/ha) and Darul Hidayah (102.1 ton/ha). The variables observed were plant height (cm) and tuber weight (grams). Observations were conducted at the age of 10 months. Data was analyzed using SPSS version 16.0. The results showed that the plant height and yield were higher in Pulang Pisau than Kalampangan even though the difference is not significant (P ≤ 0,05). Darul Hidayah has highest plant height and yield, 229.8 cm and 2271 g respectively among other genotypes/varieties grown at Pulang Pisau even though the value was not significant, whereas in Kalampangan, Darul Hidayah showed the lowest yield (670 g). Based on the data obtained, the Darul Hidayah variety can be cultivated by farmers in Pulang Pisau peat land to fulfill the availability of superior cassava seedlings for food, feed and industry. Keywords: Peat, tuber yield, plant height, liquid organic fertilizer, yam wood

Regeneration Rate of Eggplant Somatic Embryogenic In Various Maturation Media

Ralstonia solanacearum is one of the most important pathogen that causes bacterial wilt disease in eggplant and inhibits eggplant production. Improvement of eggplant varieties resistant to bacterial wilt can be accomplished through genetic manipulation. Regeneration of in vitro plants isone of the important tools to supports plant improvementthrough biotechnology. This study was aimed to determine the rate of eggplant regeneration in various maturation media, and to find the best medium for eggplant regeneration based on maturation rate and the number of cotyledon produced. We used resistant eggplant (accession 032) as the material to produce somatic embryogenic.There were 7 types of regeneration media used in this research. MS medium was supplemented with a certain concentration of plant growt regulators , such as: 1 mg / L + BAP 1 mg / L, NAA 4mg / L, TDZ 0.005 mg / L, TDZ 0.001 mg / L, CuSO4 2mM + BAP 1 mg / L, CuSO4 2mM + BAP 2 mg / L and Kinetin 1 mg / L + CuSO4 2mM. Three clumps of callus per plate with three replications were transferred to MS suplemented medium. The parameters observed were the color of callus before and after they were transfered to regeneration medium, the day of formation of globular, heart-shaped, tubular and cotyledonary phase, and the number of cotyledons formed. The results obtained showed the somatic embryogenic color of the 032 genotype was white with friable structure before being transferred to regeneration medium and was turned to yellowish white after being transferred to the regeneration medium. On the day sixth, friable embryogenic somatic of eggplant was developed into nodule on medium MS + NAA 4 mg / L, MS + CuSO4 2mM + BAP medium 1 mg / L, and MS + CuSO4 2mM + BAP 2 mg / L. Somatic embryogenic callus of accession 032 were able to pass complete globular, heart-shaped, tubular and cotyledonary phase. The most responsive medium for somatic embryogenic callus regeneration, based on the days of the callus phases formation and the number of early-phase cotyledons obtained, were MS medium suplemented with CuSO4 2mM + BAP,and CuSO4 2 mM + BAP 2 mg / L.Keywords: eggplant, Ralstonia solanacearum, regeneration, cotyledonary, clump, BA