scholarly journals Expression of Phytase gene in transgenic maize with seed-specific promoter 27-kDa γ Zein and constitutive promoter CaMV 35S

2021 ◽  
Vol 6 (3) ◽  
pp. 148
Author(s):  
Ririn Septina Arthasari ◽  
Rani Agustina Wulandari ◽  
Panjisakti Basunanda
Keyword(s):  
Enzyme Activity ◽  
Transgenic Plants ◽  
Animal Feed ◽  
Raw Materials ◽  
Transgenic Maize ◽  
35S Promoter ◽  
Constitutive Promoter ◽  
Phytase Gene ◽  
Camv 35S ◽  
Agrobacterium Tumefacien

Phytase enzymes are applied to animal feed to help animals absorb more nutrients. The use of feed raw materials containing phytase enzymes is expected to reduce the cost of animal feed production. Efforts to increase the phytase content in maize were carried out by improving genetics, in the way of assembling transgenic plants containing high phytase content. The 27-kDa γ Zein promoter is a specific promoter that expresses genes in caryopsis, and promoter CaMV 35S is a constitutive promoter that controls gene expression in all tissues and generally does not depend on the growth phase. Transgenic maize was transformed using Agrobacterium tumefacien infection method on maize B104. The reverse transcriptase polymerase chain reaction (RT-PCR) approach was used to examine the expression of phytase genes in leaves, roots, and caryopsis was done 10, 20, and 30 days after pollination (DAP). The phytase enzyme activity test was also carried out by using the colorimetric phosphomolybdate analysis method to see the phytase enzyme activity in unit µg-1. The results showed that the phytase gene in transgenic plants with the 27-kDa γ Zein promoter was highly expressed in maize caryopsis, but in line Z6.10 was also expressed in leaves, while in the CaMV 35S promoter the phytase gene was only expressed on the leaves. Phytase enzyme activity showed that transgenic maize was higher than non-transgenic maize.

scholarly journals Biolistic-mediated Transformation of Lilium longiflorum cv. Nellie White

HortScience ◽  
2008 ◽  
Vol 43 (6) ◽  
pp. 1864-1869 ◽  
Author(s):  
Kathryn Kamo ◽  
Bong Hee Han
Keyword(s):  
Transgenic Plants ◽  
Genomic Dna ◽  
Fusion Gene ◽  
Lilium Longiflorum ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Ms Medium ◽  
Camv 35S ◽  
Npt Ii

Slow-growing compact calluses were initiated from bulb scales of Lilium longiflorum cv. Nellie White that had been cultured for at least 6 months on Murashige and Skoog (MS) medium with 9 μm dicamba. To develop a reliable selection system, the sensitivity of nontransformed calluses and in vitro plants to different selective agents such as phosphinothricin, kanamycin, geneticin, paromomycin, and hygromycin was tested when grown on MS medium. Nontransformed calluses showed high sensitivity to 0.5 mg·L−1 phosphinothricin, 25 mg·L−1 geneticin, and 5 mg·L−1 hygromycin. Nontransformed plants grown in vitro died on either 2 mg·L−1 phosphinothricin or 75 mg·L−1 hygromycin. Plants did not die when grown on either 200 mg·L−1 kanamycin or 100 mg·L−1 geneticin, and 100 mg·L−1 paromomycin stimulated plant growth. Transformation was achieved using biolistics on callus bombarded with either the bar-uidA fusion gene under control of the CaMV 35S promoter or npt II and uidA under control of the CaMV 35S promoter. One week after biolistic bombardment, callus bombarded with the bar-uidA fusion gene was cultured for 1 month on MS medium supplemented with 9 μm dicamba and 0.1 mg·L−1 phosphinothricin and then transferred to 0.2 mg·L−1 phosphinothricin for 1 month followed by 1.0 mg·L−1 for the next 4 months. Regenerating shoots and well-established plants were cultured on MS medium lacking hormones and with either 0.2 mg·L−1 or 2.0 mg·L−1 phosphinothricin, respectively. Callus bombarded with the npt II gene was cultured on MS medium with 50 mg·L−1 geneticin until shoots regenerated. Regenerated shoots were cultured on MS medium lacking hormones. Under optimal conditions, 10 transgenic plants were selected from seven plates of callus bombarded with the bar-uidA fusion gene using phosphinothricin for selection. Both Southern hybridization of genomic DNA and polymerase chain reaction analysis verified the presence of the transgene in transformed ‘Nellie White’ plants. Transgenic plants were phenotypically normal, and they were crossed with nontransformed plants of L. longiflorum cvs. Sakai, Yin tung, Sakai, and Flavo. The presence of the bar gene in 41% of the T1 progeny plants confirmed stable integration of the transgene into the genomic DNA of transgenic lily plants. β-glucuronidase expression resulting from the uidA gene was demonstrated in leaves and roots of some of the transgenic lily plants by histochemical staining, determination of the specific activity of the β-glucuronidase enzyme, and Northern hybridization.

scholarly journals Delay of Disease Development in Transgenic Petunia Plants Expressing Cucumber Mosaic Virus I17N-Satellite RNA

1995 ◽  
Vol 120 (2) ◽  
pp. 353-359 ◽  
Author(s):  
Shin Je Kim ◽  
Kyung-Hee Paek ◽  
Byung-Dong Kim
Keyword(s):  
Transgenic Plants ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Unit Length ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Disease Development ◽  
Satellite Rna ◽  
Camv 35S ◽  
Transgenic Petunia

A cDNA clone of cucumber mosaic virus (CMV) 117 N-satellite RNA driven by the cauliflower mosaic virus (CaMV) 35S transcript promoter, was stably integrated into the genome of Petunia hybrida `Bluepicoti' tissues by Agrobacterium tumefaciens Ti plasmid-mediated transformation. Transgenic plants producing CMV satellite RNA showed delayed disease development when inoculated with CMV-Y, a helper virus for the I17N-satellite RNA. Furthermore, transgenic petunia plants showed delayed disease development against tobacco mosaic virus (TMV), a tobamovirus not related to CMV. Northern blot analysis revealed that large amounts of unit length satellite RNA (335 bp) were produced in CMV-infected transgenic petunia plants; whereas, mainly transcripts driven by the CaMV 35S promoter (approximately 1 kb) were produced in TMV-infected transgenic plants. SDS-PAGE and Western blotting showed that symptom reduction was correlated with a reduction in the amount of viral coat protein in transgenic plants.

Mining of Ruminant Microbial Phytase (RPHY1) from Metagenomic Data of Mehsani Buffalo Breed: Identification, Gene Cloning, and Characterization

2016 ◽  
Vol 26 (4) ◽  
pp. 252-260 ◽  
Author(s):  
Chandra Shekar Mootapally ◽  
Neelam M. Nathani ◽  
Amrutlal K. Patel ◽  
Subhash J. Jakhesara ◽  
Chaitanya G. Joshi
Keyword(s):  
Enzyme Activity ◽  
Animal Feed ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Significant Inhibition ◽  
Homology Search ◽  
Metagenomic Data ◽  
Protein Database ◽  
Phosphatase Domain ◽  
Phytase Gene

Phytases have been widely used as animal feed supplements to increase the availability of digestible phosphorus, especially in monogastric animals fed cereal grains. The present study describes the identification of a full-length phytase gene of <i>Prevotella</i> species present in Mehsani buffalo rumen. The gene, designated as RPHY1, consists of 1,251 bp and is expressed into protein with 417 amino acids. A homology search of the deduced amino acid sequence of the RPHY1 phytase gene in a nonredundant protein database showed that it shares 92% similarity with the histidine acid phosphatase domain. Subsequently, the RPHY1 gene was expressed using a pET32a expression vector in <i>Escherichia coli </i>BL21 and purified using a His60 Ni-NTA gravity column. The mass of the purified RPHY1 was estimated to be approximately 63 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal RPHY1 enzyme activity was observed at 55°C (pH 5) and exhibited good stability at 5°C and within the acidic pH range. Significant inhibition of RPHY1 activity was observed for Mg<sup>2+</sup> and K<sup>+</sup> metal ions, while Ca<sup>2+</sup>, Mn<sup>2+</sup>, and Na<sup>+</sup> slightly inhibited enzyme activity. The RPHY1 phytase was susceptible to SDS, and it was highly stimulated in the presence of EDTA. Overall, the observed comparatively high enzyme activity levels and characteristics of the RPHY1 gene mined from rumen prove its promising candidature as a feed supplement enzyme in animal farming.

Problems of ensuring metrological traceability of measurement results of indicators of quality for food products and food raw materials

2020 ◽  
pp. 64-70
Author(s):  
Mariya Y. Medvedevskikh ◽  
Anna S. Sergeeva
Keyword(s):  
Reference Materials ◽  
Mass Fraction ◽  
Nutritional Value ◽  
Animal Feed ◽  
Raw Materials ◽  
Meat Products ◽  
Metrological Traceability ◽  
Food Products ◽  
Freeze Dried ◽  
Measurement Results

The article raises the problem of ensuring metrological traceability of the measurement results of indicators of quality and nutritional value for food products and food raw materials: water (moisture), nitrogen (protein, crude protein), fat, ash and carbohydrates. The problem under consideration can be solved by applying reference materials of food composition, traceable to state primary measurement standards GET 173-2017 and GET 176-2019 and primary reference measurement procedures (PRMP), for attestation of measurement procedures and accuracy checking of measurement results. The article discusses the results of the PRMP development of mass fraction of fat, ash and carbohydrates in food products and food raw materials, as well as mass fraction of crude fat (oil content) in oil crops seeds and products based on them. The paper also presents metrological characteristics of reference materials of composition of dry dairy products, grain-milk dry porridges for nutrition of babies, grain dry porridges for nutrition of babies, egg powder, freeze-dried meat products, animal feed. The results of the work allow for building a chain of metrological traceability from GET 173-2017, GET 176-2019 and PRMP to routine measurement procedures, thereby ensuring the uniformity of measurements of nutritional value of food products.

The sugar and alcohol industry in the biofuels and cogeneration era: a paradigm change (part II)

2014 ◽  
pp. 97-104 ◽  
Author(s):  
Electo Eduardo Silv Lora ◽  
Mateus Henrique Rocha ◽  
José Carlos Escobar Palacio ◽  
Osvaldo José Venturini ◽  
Maria Luiza Grillo Renó ◽  
...  
Keyword(s):  
Large Scale ◽  
New Technologies ◽  
Process Integration ◽  
Animal Feed ◽  
Raw Materials ◽  
Biofuel Production ◽  
New Paradigm ◽  
Alcohol Industry ◽  
Feed Production ◽  
Steam Parameters

The aim of this paper is to discuss the major technological changes related to the implementation of large-scale cogeneration and biofuel production in the sugar and alcohol industry. The reduction of the process steam consumption, implementation of new alternatives in driving mills, the widespread practice of high steam parameters use in cogeneration facilities, the insertion of new technologies for biofuels production (hydrolysis and gasification), the energy conversion of sugarcane trash and vinasse, animal feed production, process integration and implementation of the biorefinery concept are considered. Another new paradigm consists in the wide spreading of sustainability studies of products and processes using the Life Cycle Assessment (LCA) and the implementation of sustainability indexes. Every approach to this issue has as an objective to increase the economic efficiency and the possibilities of the sugarcane as a main source of two basic raw materials: fibres and sugar. The paper briefly presents the concepts, indicators, state-of-the-art and perspectives of each of the referred issues.

scholarly journals Disintegration process in disc crushers

2013 ◽  
Vol 59 (No. 3) ◽  
pp. 98-104 ◽  
Author(s):  
P. Vaculík ◽  
J. Maloun ◽  
L. Chládek ◽  
M. Přikryl
Keyword(s):  
Food Industry ◽  
Fractional Composition ◽  
Animal Feed ◽  
Raw Materials ◽  
Specific Energy Consumption ◽  
Low Noise ◽  
Main Process ◽  
Industrial Sectors ◽  
The Subject

Grinding or crushing hard raw materials is usually a primary operation which precedes the follow-up technological processes in a number of industrial sectors. A great variety of machines using different principles of fragmentation are employed in the technology of pulverization. The food industry uses roller mills, in which the main process is the shear grinding. In the animal feed industry impact machines known as hammer mills are often used. In recent years, mills have been employed that use their frontal edges for grinding or crushing during the rotation of one of two adjacent discs. The modern design disc machines used for grinding grain have resulted from long development and their operation has a relatively low noise level with reduced dust. The separation process that occurs in the gap between the active edges of the discs can be described as shear grinding and is currently the subject of attention which is focused on the specific energy consumption and fractional composition of the product of grinding. &nbsp;

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