Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Marker‐assisted breeding for improvement of anaerobic germination in japonica rice (Oryza sativa)

Direct seeding is increasingly being practiced in rice cultivation areas because it saves labour and reduces cost of production and management. In this study, four japonica‐type breeding lines were developed from a cross of a japonica cultivar and an AG‐tolerant donor, KHO, using marker‐assisted selection. Validation of the target genes AG1 and AG2 in the selected lines was examined by PCR analysis and the background recovery rate by the recurrent parent was confirmed by genotyping using a 6K SNP chip. Compared to the recurrent, the survival rates of the selected lines were improved in a range from 33% to 115% in three conditions; in particular, the survival rate was increased in more than twice that of the recurrent parent under the hypoxic condition. The main agronomic trait, yield and quality‐related traits were similar to those of the recurrent parent. The use of these lines will contribute to the expansion of the cultivation area due to the proven economic benefits of direct seeding in Korea.     

Comparative energy and economic analyses of conventional and System of Rice Intensification (SRI) methods of rice production in Thai Nguyen Province,Vietnam

The consumption of energy inputs in agricultural production has been increasing rapidly during the past decades. However, given the limitations and costs of non-renewable energy, increasing production while using the least energy possible has become a major concern of most nations. Prompted by this concern, we conducted a face-to-face survey of 90 farming households in Thai Nguyen Province, Vietnam, to find out how energy is being used in agriculture and, specifically, in their rice production. Through analysis of energy input–output balances, combined with economic efficiency analysis, a comparison was made of conventional and SRI methods of rice production. The study found that applying the SRI method can save around 23% of energy inputs, while increasing energy outputs by 11%. Economic benefits per hectare also rise by more than 8 million dong (USD 364) compared to those under the conventional cultivation system. The study also showed conflicts between the energy and economic balances for manual compared with machine ploughing operations. This study contributes to providing an overview of energy consumption in rice cultivation at the household level. Its findings can help stakeholders to assess current policies and make better decisions on the uses of energy in agricultural production. In addition, the comprehensive approach taken here to analysing energy use and efficiency could expand the analysis and comparison of energy uses at sectoral or activity level—still a new field in Vietnam and many other countries.     

A study of experimental autoimmune encephalomyelitis in dogs as a disease model for canine necrotizing encephalitis

In the present study, the use of dogs with experimental autoimmune encephalomyelitis (EAE) as a disease model for necrotizing encephalitis (NE) was assessed. Twelve healthy dogs were included in this study. Canine forebrain tissues (8 g), including white and grey matter, were homogenized with 4 mL of phosphate-buffered saline for 5 min in an ice bath. The suspension was emulsified with the same volume of Freund''s complete adjuvant containing 1 mg/mL of killed Mycobacterium tuberculosis H37Ra. Under sedation, each dog was injected subcutaneously with canine brain homogenate at four sites: two in the inguinal and two in the axillary regions. A second injection (booster) was administered to all the dogs using the same procedure 7 days after the first injection. Clinical assessment, magnetic resonance imaging, cerebrospinal fluid analyses, necropsies, and histopathological and immunohistochemical examinations were performed for the dogs with EAE. Out of the 12 animals, seven (58%) developed clinically manifest EAE at various times after immunization. Characteristics of canine EAE models were very similar to canine NE, suggesting that canine EAE can be a disease model for NE in dogs.     

Fabrication of electropsun PLGA and small intestine submucosa-blended nanofibrous membranes and their biocompatibility for wound healing

In recent decades, tremendous research has focused on the production of nanoscale fibers using synthetic polymers, with the goal of fabricating nanofibrous scaffolds for wound healing. However, the hydrophobicity of such polymers typically hinders attachment and proliferation of the cells. In this study, we combined poly-d,l-lactide-co-glycolide (PLGA) and small intestine submucosa (SIS) to fabricate blended nanofibers for wound healing by electrospinning. PLGA and SIS were dissolved in 1,1,1,3,3,3-hexafluoro isopropanol to produce different weight ratios of PLGA/SIS-blended nanofibrous membranes (NFM). Physicochemical characterization of the electrospun NFM was performed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, water contact angle analysis, degradation test and tensile testing. The PLGA/SIS-blended NFM showed improved hydrophilicity and tensile strength. Better infiltration, attachment and proliferation of rat granulation fibroblasts of PLGA/SIS-blended NFMs compared to PLGA NFMs were identified by morphological differences determined by SEM and a water-soluble tetrazolium salt assay kit. Based on our results, the PLGA/SIS blended NFMs were found to be suitable for use as a potential material for wound dressing.     

Productivity of Sargassum linearifolium in potassium fortified inland saline water under laboratory conditions

Growing aquatic species in inland saline water (ISW) is one way to reduce the adverse impact of ISW to agriculture farms. This 84‐day laboratory‐trial was conducted to study the growth of Sargassum linearifolium cultured in ocean water (OW), ISW, ISW fortified with potassium equivalent to 100% (ISW100), 66% (ISW66) and 33% (ISW33) of potassium in OW at 35 g/L. The biomass and cumulative specific growth rate (SGR) of S. linearifolium increased significantly (p < .05) with increased potassium in ISW until 56 days and then declined. The ISW100 and OW resulted in similar growth patterns and yielded peak biomass at day 42, proving static biomass for the next 28 days before declining. The biomass of S. linearifolium cultured in ISW and ISW33 significantly (p < .05) decreased and was lower than in ISW100 and died after day 56. The SGR of S. linearifolium in OW, ISW100 and ISW66 levelled off and showed no difference during the first 56 days. The S. linearifolium biomass and SGR negatively and significantly (p < .05) correlated with the concentrations of nitrate, phosphate in all waters. The increased potassium concentration in ISW similar to its concentration in SW brought the growth of S. linearifolium cultured to a level that was similar in OW.     

The grafting of recycled polyol from waste polyurethane foam onto new polyurethane and its impact on shape recovery and water vapor permeation

Recycled polyols from waste polyurethane (PU) foams were grafted onto PU to improve the properties such as tensile strength, shape recovery, low-temperature flexibility, and water compatibility. The recycled polyol was either purified by column chromatography before grafting or was used directly for grafting. The soft segment melting temperature of PU did not notably increase with the addition of polyol, whereas the glass transition temperature increased with increased polyol content. The tensile strength sharply increased at low polyol content and decreased at high polyol content, while the strain at break did not significantly change with an increase in polyol content. The shape recovery at 10 oC notably improved compared with unmodified PU and remained high after four cyclic tests. Polyol-grafted PU demonstrated better lowtemperature flexibility and reduced the water vapor permeability of PU membranes. Overall, grafting recycled polyol onto PU significantly improved the tensile stress, shape recovery, and low-temperature flexibility of PU.     

Two reported cytotypes of the emergent orchid model species <Emphasis Type="Italic">Erycina pusilla</Emphasis> are two different species

Each species is characterized by a specific set of chromosomes, which is described as the chromosome portrait or karyotype. In general, such a karyotype is the same for all individuals in the population. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Here, we examined the karyotypes of the two cytotypes and found differences in arm ratios and heterochromatin patterns as well as in the presence of satellite chromosomes and in the number and location of rDNA and telomeric repeat sites. These differences are extensive and would have required multiple chromosome rearrangements to generate the differences between the two karyotypes. We also found that F1 hybrids between the parents with the two different chromosome numbers resulted in sterile offspring, in accordance with our previous findings. The combination of hybrid sterility and extensively rearranged chromosomes supports the hypothesis that these two reported cytotypes are, in fact, two different species.     

Effects of Fermentation Conditions Using Lactobacillus plantarum on the Charantin,Stigmasterol Glucoside and β-sitosterol Glucoside Contents of Bitter Gourd (Momordica charantia L.) Juice

Plant Foods for Human Nutrition - Bitter gourd fruits contain high amounts of charantin, stigmasterol glucoside and β-sitosterol glucoside, which have been shown to provide health benefits for...     

Transmission of resistance to common scab from the diploid to the tetraploid level via 4x-2x crosses in potatoes

Summary Diploid potato clones selected for their reaction to common scab and their ability to produce 2n male gametes were used in a series of crosses to a susceptible tetraploid female parent (cv. Shepody). In addition, two tetraploid clones were also selected for their reaction to common scab and crossed with Shepody as a female parent. Results indicated that resistance to common scab can be effectively transmitted from the diploid to the tetraploid level via 4x-2x crosses. Diploid parents producing 2n pollen via either first division or second division restitution can be used to transmit scab resistance. A relatively small proportion of resistant individuals could be recovered from susceptible x susceptible crosses in both 4x-2x and 4x-4x combinations. The data support a previously developed hypothesis that scab resistance is relatively simply inherited.