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.     

Calibration and validation of optical chlorophyll‐measuring devices for use in predicting crude protein concentration in tropical grass herbage

The objective of this experiment was to evaluate the Fieldscout CM 1000 NDVI and Yara N‐Tester as easy‐to‐use and cost‐effective tools for predicting foliar chlorophylls (a, b and total) and crude protein (CP) concentrations in herbage from three tropical grass species. Optical chlorophyll measurements were taken at three stages (4, 8 and 12 weeks) of regrowth maturity in Guinea grass (Panicum maximum) and Mulato II (Brachiaria hybrid) and at 6 and 12 weeks maturity in Paspalum spp (Paspalum atratum). Grass samples were harvested subsequent to optical measurements for laboratory analysis to determine CP and solvent‐extractable chlorophylls (a, b and total) concentrations. Optical chlorophyll measurements and CP concentrations were highly correlated (Yara N‐Tester: r² = 0·77–0·89; Fieldscout CM 1000 NDVI: r² = 0·52–0·84). Crude protein prediction models from the Yara N‐Tester and Fieldscout CM 1000 NDVI accounted for 70–89% and 44–73% CP variability, respectively, in Mulato II and Guinea grass. The Yara N‐tester produced more accurate and reliable CP estimates based on very high concordance correlation coefficient [CCC (0·73–0·91)] and low rMSPE, mean and regression bias. It is concluded that the Yara N‐Tester produces more accurate and reliable CP estimates of tropical pastures.     

Carbohydrate and crude protein fractions in perennial ryegrass as affected by defoliation frequency and nitrogen application rate

The objective of this study was to evaluate the effects of defoliation frequency (either at two‐ or three‐leaf stage) and nitrogen (N) application rate (0, 75, 150, 300, 450 kg N ha^?1 year^?1) on herbage carbohydrate and crude protein (CP) fractions, and the water‐soluble carbohydrate‐to‐protein ratio (WSC:CP) in perennial ryegrass swards. Crude protein fractions were analysed according to the Cornell carbohydrate and protein system. Carbohydrate fractions were analysed by ultra‐high‐performance liquid chromatography. Sward defoliation at two‐leaf stage increased the total CP, reduced the buffer‐soluble CP fractions and decreased carbohydrate fractions of herbage (P < 0·001). The effect of defoliation frequency was less marked during early spring and autumn (P < 0·001) than for the rest of the seasons. An increase in N application rate was negatively associated with WSC, fructans and neutral detergent fibre (P < 0·001), and positively associated with CP and nitrate (N‐NO₃) contents of herbage. Nitrogen application rate did not affect CP fractions of herbage (P > 0·05). The fluctuations in CP and WSC contents of herbage resulted in lower WSC:CP ratios during early spring and autumn (0·45:1 and 0·75:1 respectively) than in late spring (1·11:1). The herbage WSC:CP ratio was greater (P < 0·001) at the three‐leaf than the two‐leaf defoliation stage and declined as the N application increased in all seasons (P < 0·001). The results of this study indicate that CP and carbohydrate fractions of herbage can be manipulated by sward defoliation frequency and N application rate. The magnitude of these effects, however, may vary with the season.     

Ray blight of pyrethrum in Australia: a review of the current status and future opportunities

Ray blight caused by Stagonosporopsis tanaceti is one of the most important diseases of pyrethrum (Tanacetum cinerariifolium), a perennial herbaceous plant cultivated for the extraction of insecticidal pyrethrins in Australia. The disease is responsible for complete yield loss in severe outbreaks. Infected seed is considered as the principal source of S. tanaceti. Infection hyphae remain only in the seed coat and not in the embryo, resulting in pre- and post-emergence death of seedlings and latent infection. Therefore, quantification of the level of infection by S. tanaceti within seed using a qPCR assay is important for efficient management of the disease. Stagonosporopsis tanaceti completes its life cycle within 12 days after leaf infection through production of pycnidia and can infect every tissue of the pyrethrum plant except the vascular and root tissues. Ray blight epidemics occur in pyrethrum fields through splash dispersal of pycnidiospores between adjacent plants. Besides steam sterilization, thiabendazole/thiram and fludioxonil are effective seed-treating chemicals in controlling S. tanaceti before planting begins. Ray blight is currently managed in the field through the foliar application of strobilurin fungicides in the first 1–2 years of crop establishment. Later on, difenoconazole and multisite specific fungicides in the next 2–3 years during early spring successfully reduce ray blight infestation. Avoiding development of resistance to fungicides will require more sustainable management of ray blight including the development and deployment of resistant cultivars.     

Replacement of fish oil in plant‐based diets for Pacific white shrimp,Litopenaeus vannamei,by stearine fish oil and palm oil

Stearine fish oil (SFO) and palm oil (PO) have emerged as promising alternatives for the replacement of fish oil (FO) in aquafeeds. This study evaluated the replacement of FO with alternative oils in practical diets for Litopenaeus vannamei. In a clear brackish water study (14.1 g/L) utilizing shrimp (0.29 ± 0.02 g, initial weight), FO was replaced by SFO at inclusion ratios of 100:0, 75:25, 50:50, 25:75, and 0:100 (FO:SFO) and PO as 90% of FO. After 55 days, no significant differences (p < 0.05) in final weight, growth, or survival of shrimp were observed. A second trial (8 weeks) in low‐salinity water (2.1 g/L) with shrimp (0.92 ± 0.02 g, initial weight) evaluated diets with 100% FO, 100% SFO, 90% PO, 90% soybean oil (SO), or 90% flaxseed oil (FXO) as a replacement for FO and four commercially produced diets with 2% of FO, SO, PO, or FXO. One treatment received half rations of the commercial FO diet, and one treatment was based entirely on natural productivity. Results show that the fatty acid profiles of the tail muscle conformed to the lipids of the feed, and highly unsaturated fatty acids (HUFAs) were preserved. Following 8 weeks of culture, there were no significant differences in production performance.     

Mapping QTLs using a novel source of salinity tolerance from Hasawi and their interaction with environments in rice

Background

Salinity is one of the most severe and widespread abiotic stresses that affect rice production. The identification of major-effect quantitative trait loci (QTLs) for traits related to salinity tolerance and understanding of QTL × environment interactions (QEIs) can help in more precise and faster development of salinity-tolerant rice varieties through marker-assisted breeding. Recombinant inbred lines (RILs) derived from IR29/Hasawi (a novel source of salinity) were screened for salinity tolerance in the IRRI phytotron in the Philippines (E1) and in two other diverse environments in Senegal (E2) and Tanzania (E3). QTLs were mapped for traits related to salinity tolerance at the seedling stage.

Results

The RILs were genotyped using 194 polymorphic SNPs (single nucleotide polymorphisms). After removing segregation distortion markers (SDM), a total of 145 and 135 SNPs were used to construct a genetic linkage map with a length of 1655 and 1662 cM, with an average marker density of 11.4 cM in E1 and 12.3 cM in E2 and E3, respectively. A total of 34 QTLs were identified on 10 chromosomes for five traits using ICIM-ADD and segregation distortion locus (SDL) mapping (IM-ADD) under salinity stress across environments. Eight major genomic regions on chromosome 1 between 170 and 175 cM (qSES1.3, qSES1.4, qSL1.2, qSL1.3, qRL1.1, qRL1.2, qFWsht1.2, qDWsht1.2), chromosome 4 at 32 cM (qSES4.1, qFWsht4.2, qDWsht4.2), chromosome 6 at 115 cM (qFWsht6.1, qDWsht6.1), chromosome 8 at 105 cM (qFWsht8.1, qDWsht8.1), and chromosome 12 at 78 cM (qFWsht12.1, qDWsht12.1) have co-localized QTLs for the multiple traits that might be governing seedling stage salinity tolerance through multiple traits in different phenotyping environments, thus suggesting these as hot spots for tolerance of salinity. Forty-nine and 30 significant pair-wise epistatic interactions were detected between QTL-linked and QTL-unlinked regions using single-environment and multi-environment analyses.

Conclusions

The identification of genomic regions for salinity tolerance in the RILs showed that Hasawi possesses alleles that are novel for salinity tolerance. The common regions for the multiple QTLs across environments as co-localized regions on chromosomes 1, 4, 6, 8, and 12 could be due to linkage or pleiotropic effect, which might be helpful for multiple QTL introgression for marker-assisted breeding programs to improve the salinity tolerance of adaptive and popular but otherwise salinity-sensitive rice varieties.

     

Herbage production,nutritive value and animal productivity within hardwood silvopasture,open and mixed pasture systems in Appalachia,United States

Demand for livestock food products is projected to increase dramatically through to 2050. Increased livestock production capacity on marginal lands will be critical to meeting this demand. A 5‐year research effort was undertaken to evaluate lamb and sward productivity within open and hardwood silvopasture (SP) systems in Appalachia, USA. Grazing began in mid to late April each year, with the grazing season averaging 141 d. Grazing system treatments during 2002 and 2003 grazing seasons were as follows: 100% open pasture (OP), 67% OP and 33% SP, and 67% OP and 33% SP with delayed SP grazing initiation (OSD). In 2004, a 100% SP (SP) system was added. Animals were rotationally stocked through either 6 (2002–2004) or 7 (2005–2006) paddocks. Open pasture produced greater (P < 0·001) grazing season herbage yield, while all systems generated similar animal performance. Based on summer solstice, herbage production in spring was greater (P < 0·001) than summer, except in 2003. Total non‐structural carbohydrate (TNC) content was greater (P < 0·05) in spring than in summer, except in 2004. Animal performance was superior in spring versus summer (P < 0·001). Animal plasma urea nitrogen (PUN) was lower (P < 0·05) for OP in 2003. When PUN was correlated with nutritive value indicators, the ratio of TNC to crude protein (CP) had the strongest correlation. The strong correlation indicates the need for synchronized ruminal energy and CP availability. Development of silvopasture from existing woodlots has potential to improve whole farm productivity on marginal lands.     

Synchytrium endobioticum – risk from biogas plants?

Synchytrium endobioticum is one of the most important pathogens of potato and is known for its persistent propagation structures. Under favourable conditions, infection of highly susceptible potato cultivars leads to clearly visible cauliflower‐like tissue warts, the typical symptom of potato wart disease. However, unfavourable infection conditions or low infection pressure may result in symptoms being overlooked. Thus, the introduction of pathogen structures into stages of the processing industry cannot be ruled out. As the amounts of processed potato products continue to rise, phytosanitary risks from processing discarded potatoes and potato waste in biogas plants have to be considered. Hence, the resilience of resting spores against mesophilic anaerobic digestion was analysed in stirred tank reactors. Laboratory‐scale results show that S. endobioticum not only withstands mesophilic anaerobic digestion but also subsequent storage of the digestate for at least 4 weeks. Large numbers of viable resting spores were detectable by microscopic assessment in all samples. Viability was proved and verified additionally by bioassay. Consequently, potatoes, potato waste and processing water from potato processing industries used in biogas plants pose a phytosanitary risk if the accruing digestates are returned as fertilizer to arable land.