Construction of an Agrobacterium Mediated RNAI Genetic Transformation Vector Targeting the Replicase Gene of Indian cassava mosaic virus and Evaluation of Their Transformation Ability in Cassava Immature Leaf Lobes

Cassava mosaic virus is one of the major problems affecting cassava industry in India. Currently there are no effective strategy to completely protect cassava from cassava mosaic viruses. In order to attain cassava mosaic virus resistance RNAi vectors targeting the replicase gene of Indian cassava mosaic virus is constructed in this study. Their efficiency to transform cassava immature leaf lobes were also studied here. Replicase gene of Indian cassava mosaic virus in Tamilnadu are cloned and sequenced. Conserved domains are identified and sub cloned to CSIRO RNAi vector system and transformation studies are done in immature cassava leaves. Two different RNAi vectors were constructed, utilizing a conserved 440bp of 5’ end of ICMV Rep (AC1) gene which also corresponds to a part of AC4 gene, and functions as a viral RNAi suppressor protein. The partial Rep gene of ICMV was cloned in sense and anti-sense orientations in the RNAi intermediate vector, pHANNIBAL. After cloning into pHANNIBAL, the cloned RNAi gene cassettes of ICMV is released and cloned into the binary vector, pART27, which contains the kanamycin-resistance gene as a plant selectable marker. In order to use hygromycin as a selection agent in cassava genetic transformation, RNAi–Rep gene cassettes of ICMV were cloned into pCAMBIA1305.2. These constructs were named pICR1 and pICR2 respectively. The Genetic transformation studies in cassava leaves done using pICR2 vector could generate PCR positive plants. An agrobacterium mediated replicase RNAi vector is developed and that can be transformed into cassava immature leaf lobes. Their efficiency to silence the Indian cassava mosaic virus should be studied further.

The Characterization and Quality of 14 Accessions of Robusta Coffee

To produce superior varieties, gene sources of high diversity are needed. The study was intended to reveal the diversity of qualitative and quantitative characters and the quality of Robusta coffee germplasm. The study conducted at Pakuwon Experimental Garden, Sukabumi Indonesia, used 14 accessions of Robusta coffee grown by a single stem system. The observed characteristics were qualitative and quantitative characteristics, caffeine content, and cup quality. The data were analyzed descriptively and grouped by cluster analysis using the PBSTAT program. The results revealed variations in the angle of insertion of primary branches, stipule shape, immature leaf color, leaf shape, leaf apex shape, young shoot color, mature leaf color, ripe fruit color, and fruit shape. The number of bunches/branch is the characteristic of the highest variation coefficient. Eleven accessions had cupping scores between 80 and 86 with a caffeine content of 1.39 - 2.06%. The accession B4-17-1 had the lowest yield, but its final cupping test score was the highest. At the dissimilarity coefficient of 0.25, 14 accessions of Robusta coffee are divided into two large groups, while at the dissimilarity coefficient of 0.15 they consist of four groups.

Morphoagronomic characterization of sweet potato accessions from the germplasm bank of Embrapa hortaliças

Studies on the determination of genetic divergence among genotypes are important tools in breeding programs, contributing to the identification of parents with considerable productive potential. However, little is known about the combinatorial capacity of sweet potato (Ipomoea batatas) accessions and its adaptation to the different regions of Brazil. The objective of this study was to evaluate the morpho-agronomic traits from 102 sweet potato accessions from the Germplasm Bank of Embrapa Hortaliças. The experiment was laid out as an augmented block design comprised of 102 treatments. Nineteen above ground traits were measured using descriptors for the respective parts. Estimated values of broad sense heritability were high for the traits mean branch length (95.75%), immature leaf color (85.06%), and predominant branch color (90.57%). Coefficients of environmental variation were below 30.00% for all variables, except for branch weight (51.62%). The 102 clones analyzed presented broad genetic variability for the different traits evaluated, especially for branch weight, and branch length, and mature leaf color.

Antioxidant and Glycemic Regulatory Properties Potential of Different Maturity Stages of Leaf of Ceylon Cinnamon (Cinnamomum zeylanicum Blume) In Vitro

Dichloromethane:methanol (1:1, v/v) extracts of different maturity stages (immature, partly mature, and mature) of authenticated leaves of Ceylon cinnamon (CC) were used in this study. Antioxidant properties [total polyphenolic content (TPC) and total flavonoid content (TFC), 1, 1-diphenyl-2-picryl-hydrazyl (DPPH), 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS)), oxygen radical absorbance capacity (ORAC), and ferric reducing antioxidant power (FRAP)] and glycemic regulatory properties [antiamylase (AA); antiglucosidase (AG)] were evaluated using 96-well microplate based bio assays in vitro (TPC, TFC, DPPH, ABTS, ORAC n=4 each; FRAP, AA, AG n=3 each). Results clearly revealed significant differences (p<0.05) among different maturity stages of leaf of CC for both antioxidant and glycemic regulatory properties (except AG activity). The mean antioxidant and glycemic regulatory activities of immature, partly mature, and mature leaves ranged from TPC: 0.68 ± 0.02–22.35 ± 0.21 mg gallic acid equivalents/g of sample (GS); TFC: 0.85 ± 0.01–4.68 ± 0.06 mg quercetin equivalents/GS; DPPH: 0.42 ± 0.01–27.09 ± 0.65 mg Trolox equivalents (TE)/GS; ABTS: 3.57 ± 0.10–43.91 ± 1.46 TE/GS; ORAC: 0.71 ± 0.01–18.70 ± 0.26 TE/G, FRAP: 0.31 ± 0.02–69.16 ± 0.52 TE/GS; and AA: 18.05 ± 0.24–36.62 ± 4.00% inhibition at 2.5 mg/mL. Mature leaf had the highest antioxidant and AA activities for all the assays investigated. In contrast, immature leaf had the lowest. The order of potency for antioxidant and AA activities was mature leaf > partly mature leaf > immature leaf. This is the first study to report on antioxidant and glycemic regulatory properties of different maturity stages of leaf of Ceylon cinnamon and highlights its potential use in management of oxidative stress-associated chronic diseases including diabetes mellitus.

Extraction of Condensed Tannins from Tropical Plants as Affected by Leaves Maturity, Maceration Time, and Centrifugal Force

Condensed Tannin (CT) are secondary metabolites of the plant that synthesized along the phenylpropanoid pathway. It is known to suppress CH4emission in the rumen through protozoal defaunation as well as direct effects on methanogen bacteria. Recent studies have been reported to have anthelmintics activity to overcome gastrointestinal nematodes, appertain to theHaemonchus contortus. To obtain merits of CT, it can be applied in flour or infusion form. Infusion considered as a convenient alternative of CT application. Evaluation method to produce optimum levels of CT needs to be done for the merits of CT obtained optimally. This study aimed to evaluate the effect of leaf maturity, maceration time, and centrifuge force on CT levels produced in infusion leaf as a feed additive. Mature and immature leaves selected from Morindacitrifolia(CF), Muntingiacalabura(CA), Azadirachta indica(AZ), Hibiscus rosa sinensis (RS) and Hibiscus tiliaceus (HT).The results of this research showed that leaf age had a significant effect on CT, except on AZ and RS. Normally, CT content of immature leaf is higher than the mature leaf. CA has highest levels of CT, can be predicted because of lots of glandular trichomes in their mature leaves as a place to store secondary metabolite compounds. Optimum levels of CT from maceration occurring at 2h of immersion except on HTthat requiring 4h of immersion. Meanwhile, 372 g is the most effective centrifuge force in producing optimum CT, except in CA which requires 2.318 g. Based on this study, it can be concluded that leaf age, maceration time, and centrifuge force have a significant influence in producing an infusion with maximum CTat each stage of extraction. It is important for infusion production to contain CT in optimal amounts. This research is expected to be the first step in providing multifunctional feed additive for livestock.

Siliplant1 (Slp1) protein precipitates silica in sorghum silica cells

SummarySilicon is absorbed by plant roots as silicic acid. The acid moves with the transpiration stream to the shoot, and mineralizes as silica. In grasses, leaf epidermal cells called silica cells deposit silica in most of their volume by unknown mechanism.Using bioinformatics tools, we identified a previously uncharacterized protein in sorghum (Sorghum bicolor), which we named Siliplant1 (Slp1). Silica precipitation activity in vitro, expression profile, and activity in precipitating biosilica in vivo were characterized.Slp1 is a basic protein with seven repeat units rich in proline, lysine, and glutamic acid. A short peptide, repeating five times in the protein precipitated silica in vitro at a biologically relevant silicic acid concentration. Raman and NMR spectroscopies showed that the peptide attached the silica through lysine amine groups, forming a mineral-peptide open structure. We found Slp1 expression in immature leaf and inflorescence tissues. In the immature leaf active silicification zone, Slp1 was localized to the cytoplasm or near cell boundaries of silica cells. It was packed in vesicles and secreted to the paramural space. Transient overexpression of Slp1 in sorghum resulted in ectopic silica deposition in all leaf epidermal cell types.Our results show that Slp1 precipitates silica in sorghum silica cells.

Variasi Morfologi Ubi Jalar Lokal Asal Dataran Tinggi Minyambouw dan Anggi

Sweet potato is superior commodity of Papua because consumed as special food by society in the middle mountain of Papua. Therefore, sweet potato has vital value in economical, social and culture. In the highland Minyambouw and Anggi have been done exploration and characteristic morphology which is observed among other storage root, leaf and petiole. Result of research exploration and characterization are found around 18 local clons which consist of 8 clons in highland of Minyambouw and 10 clons in highland of Anggi. Clon of Simpengey has special characteristic of the primary colour of its storage root skin is brownish orange, the primary colour of its storage root flesh in crem and secondary colour its storage root is white. Clon of syeb Nntung Ninggos has characteristic among other storage root shape is elliptic, storage root skin primary colour is yellow and the most of the immature leaf colour is purple. Morphology can be proved that only 11 clone groups of sweet potato in the both area.