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     

村务管理专业学生的教育管理研究

随着国家乡村振兴战略的深入实施,村务管理专业学生招生规模迅速扩大,教育管理对乡村振兴发展、学院建设、个人成长具有重要意义.从分析学生的特点和教育管理现状入手,阐述了抓好村务管理专业学生教育管理的重要意义,提出把好招考入学、教书育人、考察使用关口,做好学院内部与外部管理结合、教育管理与精准服务结合、依法依规与沟通交流结合、教育引导与更新观念结合、宣传表彰与惩戒警示结合,加强学生的教育管理工作.     

<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Phenological and physiological evaluation of first and second cropping periods of sorghum and maize crops

Environmental conditions influence phenology and physiological processes of plants. It is common for maize and sorghum to be sown at two different periods: the first cropping (spring/summer) and the second cropping (autumn/winter). The phenological cycle of these crops varies greatly according to the planting season, and it is necessary to characterize the growth and development to facilitate the selection of the species best adapted to the environment. The aim of this study was to characterize phenological phases and physiological parameters in sorghum and maize plants as a function of environmental conditions from the first cropping and second cropping periods. Two parallel experiments were conducted with both crops. The phenological characterization was based on growth analyses (plant height, leaf area and photoassimilate partitioning) and gas exchange evaluations (net assimilation rate, stomatal conductance, transpiration and water-use efficiency). It was found that the vegetative stage (VS) for sorghum and maize plants was 7 and 21 days, respectively, longer when cultivated during the second cropping. In the first cropping, the plants were taller than in the second cropping, regardless of the crop. The stomatal conductance of sorghum plants fluctuated in the second cropping during the development period, while maize plants showed decreasing linear behaviour. Water-use efficiency in sorghum plants was higher during the second cropping compared with the first cropping. In maize plants, in the second cropping, the water-use efficiency showed a slight variation in relation to the first cropping. It was concluded that the environmental conditions as degree-days, temperature, photoperiod and pluvial precipitation influence the phenology and physiology of both crops during the first and the second cropping periods, specifically cycle duration, plant height, leaf area, net assimilation rate, stomatal conductance and water-use efficiency, indicating that both crops respond differentially to environmental changes during the growing season.     

Assessing seed desiccation responses of native trees in the Caribbean

New Forests - Native trees from the Caribbean were tested for seed desiccation responses, by adapting the “100-seed test” protocol. Ninety-seven seed lots of 91 species were collected...     

Prediction of Genetic Gain in Sweet Corn using Selection Indexes

Journal of Crop Science and Biotechnology - The cultivation of sweet corn is expanding in Brazil, but there are serious constraints about the availability of commercial cultivars. The selection of...     

Proteomic profiles during adventitious rooting of Eucalyptus species relevant to the cellulose industry

New Forests - Adventitious rooting (AR) is an obligatory step for vegetative propagation of commercial woody species. Paper industries have interest in Eucalyptus globulus Labill and its hybrids...     

Whitened kernel surface: A fast and reliable method for assessing Fusarium severity on cereal grains by digital picture analysis

Fusarium head blight (FHB) is a cereal disease of major importance responsible for yield losses and mycotoxin contaminations in grains. Here, we introduce a new measurement approach to quantify FHB severity on grains based on the evaluation of the whitened kernel surface (WKS) using digital image analysis. The applicability of WKS was assessed on two bread wheat and one triticale grain sample sets (265 samples). Pearson correlation coefficients between Fusarium‐damaged kernels (FDK) and WKS range from r = 0.77 to r = 0.81 and from r = 0.61 to r = 0.86 for the correlation between deoxynivalenol (DON) content and WKS. This new scoring method facilitates fast and reliable assessment of the resistance to kernel infection and shows significant correlation with mycotoxin content. WKS can be automated and does not suffer from the “human factor” inherent to visual scorings. As a low‐cost and fast approach, this method appears particularly attractive for breeding and genetic analysis of FHB resistance where typically large numbers of experimental lines need to be evaluated, and for which WKS is suggested as an alternative to visual FDK scorings.     

Effect of Kechuanning on airway remodeling and protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus

AIM:To investigate the effect of Kechuanning on airway remodeling and the protein level of p-ERK1/2 in lung tissues of asthmatic rats induced by virus. METHODS:The asthmatic rat model induced by respiratory syncytial virus was established. The experimental rats were divided into normal group, asthma model group, low dose (0.33 mL/kg), middle dose (3.0 mL/kg) and high dose (10 mL/kg) of Kechuanning groups, and PD98059 (3 mg/kg) group. The airway responsiveness of the rats was measured by animal ventilator. The pathological changes of the lung tissues were observed by HE staining. PAS staining and Masson staining were used to observe goblet epithelial cells metaplasia and airway collagen deposition. The expression of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in the lung tissues of the rats was detected by immunohistochemical staining. The protein levels of ERK1/2 and p-ERK1/2 were determined by Western blot. RESULTS:Compared with model group, the airway responsiveness of the rats in middle dose and high dose of Kechuanning groups was significantly decreased (P<0.01), the injury of lung tissues was significantly decreased, the goblet epithelial cells metaplasia and airway collagen deposition were significantly reduced (P<0.01), and the expression of MMP-9 and TIMP-1 in the lung tissues was also significantly decreased (P<0.01). In addition, the protein level of p-ERK1/2 in high dose of Kechuanning group was significantly decreased compared with model group (P<0.01). CONCLUSION:Kechuanning may treat asthma by regulating the expression of p-ERK1/2 in the lung tissues and improving the airway remodeling symptoms of asthmatic rats induced by virus.     

Evaluation of Particle Size Influence on Proximate Composition,Physicochemical, Techno-Functional and Physio-Functional Properties of Flours Obtained from Persimmon (<Emphasis Type="Italic">Diospyros kaki</Emphasis> Trumb.) Coproducts

The aim of the work was to study the influence of particle size in the composition, physicochemical, techno-functional and physio-functional properties of two flours obtained from persimmon (Diospyros kaki Trumb. cvs. ‘Rojo Brillante’ (RBF) and ‘Triump’ (THF) coproducts. The cultivar (RBF and THF) and particle size significantly affected all parameters under study, although depending on the evaluated property, only one of these effects predominated. Carbohydrates (38.07–46.98 g/100 g) and total dietary fiber (32.07–43.57 g/100 g) were the main components in both flours (RBF and THF). Furthermore, insoluble dietary fiber represented more than 68% of total dietary fiber content. All color properties studied were influenced by cultivar and particle size. For both cultivars, the lower particle size, the higher lightness and hue values. RBF flours showed high values for emulsifying activity (69.33–74.00 mL/mL), while THF presented high values for water holding capacity (WHC: 9.47–12.19 g water/g sample). The bile holding capacity (BHC) and fat/oil binding values were, in general, higher in RBF (19.61–12.19 g bile/g sample and 11.98–9.07, respectively) than THF (16.12–12.40 g bile/g sample and 9.78–7.96, respectively). The effect of particle size was really evident in both WHC and BHC. Due to their dietary fiber content, techno-functional and physio-functional properties, persimmon flours seem to have a good profile to be used as potential functional ingredient.